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					                    Abstracts of Scientific Presentations
                                        2008 AALAS National Meeting
                                             Indianapolis, Indiana

                  Platform Sessions                                   10 times, led to seropositive recipients after embryo transfer.
                                                                      However, when oocytes in the cumulus-oocyte complex (COCs)
                                                                      were fertilized in vitro with cryopreserved spermatozoa (IVF)
PS1 Decrease in Helicobacter hepaticus Titers over Time in            in the presence of 104 TCID50/ml MMVp and the 2-cell embryos
BALB/c Mice                                                           were washed 10 times, recipients were seronegative. The lack of
CM Nagamine*1,2, Z Ge2, ME McBee3, KA Knox3, AB Rogers2,              transmission of infectious MMVp by IVF embryos may be due
JG Fox2, DB Schauer2,3                                                to the cumulus cells on the oocytes, which may have prevented
1Comparative Medicine, Stanford University, Stanford, CA; 2Com-       entry of MMVp to the zona pellucida or they may have absorbed
                                                                      the virus. To test this hypothesis, COCs or cumulus-free oocytes
parative Medicine, 3Biological Engineering, MIT, Cambridge, MA
                                                                      (CFOs) and spermatozoa were exposed to 104 TCID50 MMVp/
Helicobacter hepaticus induces clinical disease and confounds         ml in the fertilization medium (HTF). The resulting embryos
the interpretation of data in certain mouse strains. We recently      were washed 10 times in virus-free KSOM and kept in holding
reported that H. hepaticus appeared to be lost in BALB/cJ and         media (M2). Media and groups of 20 embryos were analysed
BALB-IL10 null mice starting at 13 wk postinoculation (wpi) based     using PCR and in vitro infectivity assay (IVia). Of 5 replications
on helicobacter-specific PCR on fecal pellets. We hypothesize that    with COCs, 1/5 holding drop and 2/5 groups of embryos were
the BALB/c genetic background inhibits long-term, high-titer          MMV-positive using PCR, while neither holding drops nor
colonization by H. hepaticus. To test this hypothesis, we gav-        embryos were positive using IVia. From IVF with CFOs, PCR
aged 35 BALB/cJ (BALB) females with H. hepaticus. Five 129S6/         revealed that 3/5 holding drops and 4/5 groups of embryos
SvEv-Rag2tm1Fwa (129RAG2 null) females were treated similarly         were MMV-positive, while 1/5 holding drops and no embryos
as positive controls. Negative control BALB females received          were positive for MMV using IVia. The present data show that
media only. Fecal pellets were collected from individual mice at      2-cell embryos from COCs did not transmit infectious virus to
2, 6, 10, 14, 18, 22, 26, and 30 wpi. Fecal DNAs were extracted and   the holding drops, most likely due to the presence of cumulus
used in an all-helicobacter, all-lactobacillus duplex PCR assay.      cells during fertilization.
All 129RAG2 null positive controls were helicobacter-positive,
and all BALB negative controls were helicobacter-negative.            PS3 Absence of Germline Transmission after Blastocyst Injec-
Inoculated BALB mice were helicobacter-positive at 2, 6, and 10       tion with Murine Embryonic Stem Cells Cultured with Mouse
wpi. However, at 14 wpi, 2 mice were helicobacter-negative. At        Hepatitis Virus and Mouse Minute Virus
18 wpi, 6 mice were helicobacter-negative and 1 of the 2 mice that
were initially negative reverted to being helicobacter-positive.      E Mahabir*1, K Reindl1, J Mysliwietz2, J Needham3, D Bulian1, K
Overall, 56% of inoculated BALB mice were helicobacter-negative       Markoullis1, J Schmidt1
at 1 or more time points. Quantitative PCR (qPCR) was per-            1Department of Comparative Medicine, 2Institute for Molecular
formed on the cecal DNA of 3 129RAG2 null positive controls,          Immunology, Helmholtz Center Munich—German Research
3 BALB mice that were helicobacter-positive at all time points,       Center for Environmental Health, Neuherberg, Germany; 3 The
and 7 BALB mice that were helicobacter-negative at 2 or more          Microbiology Laboratories, North Harrow, United Kingdom
time points. 129RAG2 null mice averaged 13 x 106 copies of H.
hepaticus in their ceca. BALB mice that were helicobacter-positive    The aim of this study was to determine the susceptibility of
at all time points had an average of 2.5 x 106 copies. BALB that      murine embryonic stem cells (mESCs) to infection with mouse
were helicobacter-negative at 2 or more time points had the least     hepatitis virus (MHV-A59) and mouse minute virus (MMVp)
number of H. hepaticus in their ceca (0.1 x 106 copies). This was     and the effect of these viruses on germline transmission (GLT),
24-fold fewer copies relative to BALB mice that were helicobacter-    and the serological status of the recipients and pups. The mESCs
positive at all time points. The data suggest that some BALB          were inoculated with 10-1 TCID50/cell MHV-A59 or MMVp and
mice develop low titers of H. hepaticus in their ceca, resulting in   cultured for 6 h or over 4 to 5 passages, with an interval of 2 d
H. hepaticus not being easily detected by fecal PCR. These data       between passages. mESCs were productively infected with MHV-
suggest caution in the interpretation of fecal PCR results used       A59 whereas infection was restricted in mESCs after exposure to
to screen for the presence of H. hepaticus.                           MMVp. Also, MHV-A59, but not MMVp, was cytolytic for the
                                                                      mESCs. Exposure of mESCs to the viruses over 4 to 5 passages,
                                                                      but not for 6 h prior to blastocyst injection and embryo transfer,
PS2 Lower Transmission of Mouse Minute Virus by In Vitro-             affected GLT to the extent that no chimeras were born. In contrast,
produced Embryos Due to Cumulus Cells                                 control mESCs led to 6.2 pups per germline chimera. Recipients
E Mahabir*, D Bulian, R Schmöller, J Schmidt                          and pups were seropositive for MHV-A59, but not for MMVp,
                                                                      when mESCs were cultured with the virus over 4 or 5 passages.
Department of Comparative Medicine, Helmholtz Zentrum
                                                                      The present data show that mESCs can transmit viruses during
München—German Research Center for Environmental Health,
                                                                      blastocyst injection and embryo transfer, depending on the virus
Neuherberg, Germany
                                                                      itself, and that germline transmission is dramatically affected by
In vivo-produced embryos, which had been exposed in vitro             virus-contaminated mESCs.
to 104 TCID50/ml of mouse minute virus (MMVp) and washed
Abstracts of scientific papers 2008 AALAS National Meeting

PS4 Mycoplasmas Affect Cell Parameters of Murine Embry-                detectable through 56 DPI. The mean peak concentration of MPV
onic Stem Cells, Germline Transmission, and the Resulting              DNA in BALB/c feces of approximately 1.5 million copies per mg
Chimeric Progeny                                                       of feces was 3 times higher than in B6 feces. The apex of viral DNA
                                                                       and mRNA transcript expression in tissues largely corresponded
K Markoullis1, D Bulian1, G Hölzlwimmer2, L Quintanilla-Fend2,
                                                                       to the pattern described for fecal shedding. In addition to being
H Zitzelsberger3, K Heiliger3, J Mysliwietz4, CC Uphoff5, HG
Drexler5, T Adler6, DH Busch6, J Schmidt1, E Mahabir*1                 delayed and at a lower level, MPV infection, as measured by qPCR,
                                                                       waned sooner in B6 than in BALB/c mice. Concentrations of viral
1Department     of Comparative Medicine, 2Institute of Pathology,      DNA and mRNA transcripts in tissue specimens were highest in
3Institute of Molecular Radiobiology, 4Institute of Molecular Im-
                                                                       mesenteric lymph nodes and lowest in spleens. At 56 DPI, viral
munology, Helmholtz Center Munich—German Research Center               DNA concentrations in tissues remained high, whereas viral mRNA
for Environmental Health, Neuherberg, Germany; 5Department             transcripts were no longer detectable. MPV-1 seroconversion was
of Human and Animal Cell Cultures, DSMZ—German Collection              first detected in a majority of BALB/c and B6 mice at 7 and 10 DPI,
of Microorganisms and Cell Cultures, Braunschweig, Germany;            respectively. In summary, the resistance of B6 mice to MPV infec-
6Institute of Medical Microbiology, Immunology and Hygiene,
                                                                       tion correlates with lower levels of virus replication and shedding,
Technical University Munich, Munich, Germany                           which can complicate detection and confirmation of MPV infection
Production of genetically modified mice implies frequent exchange      in mice that are B6 or on a B6 background.
of murine embryonic stem cells (mESCs) between laboratories,
which increases the risk of contaminations of cell cultures and        PS6 Comparison of Side Effects between Buprenorphine and
mice, thereby jeopardizing experimental protocols. To determine        Meloxicam Used Postoperatively in Dutch Belted Rabbits
the susceptibility of mESCs to mycoplasmas and the effects of          (Oryctolagus cuniculus)
mycoplasmas on cell parameters, the germline transmission
                                                                       CS Cooper*1, KA Pate1, C Barat2, L Swarthout1, J Anastasio1, D
(GLT), and the resulting chimeric progeny, 129/SvPas mESCs             Scorpio1
were inoculated at passage P13 with M. hominis, M. fermentans,
                                                                       1Molecular and Comparative Pathobiology, Johns Hopkins Uni-
and M. orale and cultured over 20 passages (P13 + 20). Com-
pared to controls, there was a 7-log reduction in the growth of        versity SOM, Baltimore, MD; 2Mathematics, Villa Julie College,
mycoplasma-contaminated mESCs. Karyotypic analysis revealed            Baltimore, MD
non-clonal chromosomal aberrations, which increased with the           One of the challenges faced by veterinarians and researchers who
duration of infection. At passage P13 + 6, where mycoplasma            use rabbits (Oryctolagus cuniculus) as a surgical model is choos-
infection was strongest, 46.3% of mycoplasma-infected mESCs            ing an appropriate postoperative analgesic that is efficacious at
expressed POU5F1 (OCT-4) and SSEA-1 markers, in contrast to            relieving pain without unwanted side effects. Buprenorphine
84.8% of control mESCs, indicating a decrease in the expression        and other opioid derivatives are common analgesics used to
of stem cell differentiation markers. The GLT of mycoplasma-           alleviate postsurgical pain in rabbits. However, this class of
infected mESCs at passage P13+5 decreased from 90%, as found           analgesics has been noted to cause anorexia and gut stasis in
in controls, to 50%. Similarly, the number of litters and the number   various mammalian species, including humans. The objective
of pups born decreased from 100% to 50%, and from 45% to 22%,          of this study was to evaluate the occurrence of buprenorphine
respectively. Some chimeras obtained from blastocyst injection         side effects in Dutch Belted rabbits when used postoperatively.
with mycoplasma-infected mES cells showed nasal discharge,             We evaluated meloxicam as an equal alternative to opioid use.
osteoarthropathia, and cachexia. The present data indicate that        We expected that meloxicam would provide the same quality
mycoplasmas affect cell parameters, germline transmission of           of pain relief for most surgical interventions in rabbits without
mESCs, and postnatal development, underlining the importance           unwanted side effects such as anorexia and gut stasis. Baseline
of screening mES cells for mycoplasmas prior to their use.             parameters including activity, pain assessment, food intake/fecal
                                                                       output (g), urine output, body weight (kg), physical examination,
PS5 Comparing Mouse Parvovirus Infection of Susceptible                abdominal palpation and auscultation, CBC/serum biochemis-
BALB/c and Resistant C57BL/6 Mice Using Quantitative Poly-             try, and fecal cultures were collected for 30 female Dutch Belted
merase Chain Reaction and Serologic Assays                             rabbits. A routine ovariohysterectomy was performed on each
                                                                       rabbit. Postsurgically, rabbits were randomly assigned to groups
KS Henderson, CL Perkins, LA Banu, SM Jennings, WR Shek*
                                                                       receiving either postoperative buprenorphine (n = 10), meloxi-
Research Animal Diagnostic Services, Charles River, Wilming-           cam (n = 10), or bupivicaine incision infiltration (n = 10). Rabbits
ton, MA                                                                in the meloxicam group demonstrated the fastest recovery to
In a previous study, we demonstrated that the dose of the MPV-1e       baseline parameters compared to rabbits treated with either
field isolate required to infect C57BL/6NCrl (B6) mice was 10 to       buprenorphine or bupivacaine. Normal eating and voiding be-
100 times higher than the infectious dose for BALB/cAnNCrl mice,       haviors of rabbits treated with buprenorphine gradually returned
whether the virus was inoculated by gavage or intraperitoneally.       to baseline; however, rabbits treated with bupivicaine needed
To investigate this susceptibility difference further, we inoculated   medical intervention to fully recover to baseline. Meloxicam is
B6 and BALB/c mice by gavage with approximately 1000 and 100           an appropriate alternative to buprenorphine for alleviation of
BALB/c infectious doses (ID50) of MPV-1e, respectively. Feces,         postoperative pain in the rabbit. Rabbits can be given meloxicam
mesenteric lymph nodes, ileum, spleen, and blood were collected        safely without onset of anorexia or gut stasis, which are docu-
at 11 time points, from 1 to 56 d postinoculation (DPI). Viral DNA     mented side effects associated with buprenorphine use. We also
and reverse-transcribed messenger (m) RNA copies were measured         recommend strong consideration and justification when using
in feces (DNA only) and tissue specimens by fluorogenic nuclease       bupivicaine infiltration as a sole method of pain relief.
qPCR assays. MPV seroconversion was assayed by several meth-
ods, including ELISA, IFA and MFIA. MPV DNA was detected               PS7 Identification of a Novel Low-pathogenicity Rabbit Calici-
in the feces of both B6 and BALB/c mice at 1 DPI; fecal shedding       virus in a Spontaneous Disease Outbreak in the United States
peaked in BALB/c mice at 5 DPI and 2 d later in B6 mice, and was
                                                                       I Bergin*, SR Bolin, M Kiupel, RK Maes, TP Mullaney, AG Wise
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

Michigan State University, Ypsilanti, MI                               with Taconic mice (0.850 ± 0.130; P < 0.01). These data indicate
                                                                       that the environment in which the mice are reared is more im-
We report a spontaneous disease outbreak in a privately owned
                                                                       portant than genetic relatedness. The notable difference between
Michigan rabbitry caused by a novel rabbit calicivirus related
                                                                       Taconic and Jax husbandry is that Taconic rederives their mice
to but distinct from rabbit hemorrhagic disease virus (RHDV)
                                                                       into the germ-free state with subsequent ASF colonization. In
and having low-level pathogenicity. At this facility, 35% of ap-
                                                                       contrast, Jax maintains their mice in SPF without rederivation and
proximately 200 New Zealand White rabbits died over a period
                                                                       colonization with ASF. This study suggests that the indigenous
of 1 wk. Pregnant does were predominantly affected; principal
                                                                       gastrointestinal microbiome plays a significant role in murine
lesions consisted of pulmonary hemorrhage and hepatocellular
necrosis. Calicivirus-like particles were detected in liver ho-        gallstone formation.
mogenate by electron microscopy. Initial RT-PCR amplification
of viral sequence using RHDV-specific primers was positive;            PS9 Single Nucleotide Polymorphisms That Differentiate
however, partial genomic sequencing demonstrated an overall            Common Inbred Strains and Substrains of Mice
79% nucleic acid identity with RHDV. Phylogenetic compari-             GW Bothe*1, JL Seymour1, JL Moran2, SM Festin1
sons of the partial viral genomic and amino acid sequences             1
were consistent with a unique calicivirus in the Lagovirus genus        Taconic, Rensselaer, NY; 2Brigham and Women’s Hospital,
that was related to but distinct from RHDV and other reported          Harvard Medical School, Boston, MA
lagomorph caliciviruses. We provisionally describe this virus          While microsatellite markers can be used to verify the identity of
as Michigan rabbit calicivirus (MRCV). To our knowledge, this          mice, single nucleotide polymorphisms (SNPs) are more easily
is the first report of a low-pathogenicity rabbit calicivirus in       available in current databases, and are more amenable to high-
the Lagovirus genus in the U.S., although low or nonpathogenic         throughput analytic technologies. SNP data on some mouse
rabbit caliciviruses have been reported in other countries. We         strains have been widely reported. However, the almost complete
performed an infection study using tissue homogenate from              absence of data on certain substrains made the development of
affected rabbits inoculated into specific pathogen-free rabbits.       a genetic quality control program difficult. Therefore, we first
This resulted in individual hepatocellular necrosis and positive       analyzed a panel of 768 mouse SNPs that contained a number
in situ hybridization within hepatocytes, but no overt clinical        of markers that differentiate C57BL strains. Markers were de-
disease or widespread histological changes. We propose that            tected using BeadArrays. Among major strains and substrains,
MRCV primarily causes subclinical infection of healthy rabbits         there were 19 differences between C57BL/6NTac (B6NTac) and
and may induce overt, RHD-like disease under certain field             C57BL/6J (B6J), 11 B6NTac and C57BL/6JBom (B6JBOM), and 8
conditions. In addition to the possibility of clinical disease, this   B6J and B6JBom, placing these strains into the expected positions
report calls attention to the potential diagnostic confusion of a      in the C57BL family tree. We found 459 differences between B6N-
low-pathogenicity calicivirus such as MRCV with the reportable         Tac and BALB/c, and no difference among BALB/c substrains
foreign animal disease agent RHDV.                                     AnNTac, ABomTac, J, and JBomTac. This result is unexpected
                                                                       given the long separation time of the substrains. However, it may
PS8 Role of the Gastrointestinal Microbiome in Cholesterol             be explained by a “marker identification bias” caused by nearly
Gallstone Pathogenesis                                                 all SNPs having been identified in screens of B6J compared with
                                                                       other strains. We used a 140,000 SNP DNA chip to search for
JJ Fremont*1, JG Fox1, MC Carey2, KJ Maurer1,2                         SNP markers differentiating BALB/c substrains. We were able
1Division of Comparative Medicine, MIT, Cambridge, MA; 2Divi-          to identify 8 candidate SNPs in the chip data, and have verified 3
sion of Gastroenterology, Department of Medicine, Brigham and          SNPs by PCR amplification and sequencing. We then developed a
Women’s Hospital, Harvard Medical School, Boston, MA                   genetic quality control program that would detect any mix-up of
                                                                       inbred mice after transport from one facility to another. For each
Cholesterol gallstone formation is a common costly disease with
                                                                       strain-specific panel, at least 2 markers were selected for each
no definitive nonsurgical treatment. Recently, we demonstrated
that BALB/c mice from different commercial sources exhibit             strain that could distinguish that strain from any other strain,
                                                                       and at least 2 additional markers were added to each panel to
markedly different prevalences in cholesterol gallstone forma-
tion. Specifically, BALB/c mice from The Jackson Laboratory            enable differentiation of substrains where necessary, resulting in
(BALB/cJ) demonstrated an increased prevalence compared                panels of 4 to 8 SNPs. This testing program combines maximum
to those from Taconic Farms. To determine if these differences         clarity of results with minimum cost. All SNP assays in the qual-
resulted from variations in the indigenous microbiota or genetic       ity control panels were verified by fluorogenic PCR.
differences between the substrains, we performed a prospective
study analyzing biliary phenotype in closely related BALB/c            PS10 Mouse Parvovirus Infection of Immunodeficient Mice
mice. Three groups (n = 10/group) of mice were analyzed for            J Macy*, PC Smith, J Wilson, SR Compton
gallstone phenotype: Group 1 was BALB/cJ mice; group 2 was
BALB/cByJ mice, which are from the same genetic substrain as           Comparative Medicine, Yale University, New Haven, CT
BALB/cAnNTac mice, but are housed at Jax; and group 3 was              The interplay of mouse parvovirus (MPV) and the host immune
BALB/cAnNTac. Eight-week-old mice were fed a lithogenic                system is not well understood. In order to determine how the
diet for 8 wk. At the conclusion of the study, gallbladder bile        immune system impacts MPV infection shedding, transmission,
was examined to determine biliary phenotype. BABL/cByJ                 and immune cell tropism 10 SCID (CB-17/Icr-Prkdcscid/Crl), 5
and BALB/cJ mice displayed similar prevalences of cholesterol          SCID-BG (CB-17/Icr.Cg-Prkdcscid Lystbg/Crl), 5 Igh (STOCK-
monohydrate crystals (70% and 90%) and cholesterol gallstones          Igh-Jtm1Dhu N?+N2), 5 CB17 (C.B-Igh1b/IcrTac) and 5 BALB
(50% and 70%). In contrast, BALB/cAnNTac mice developed                (BALB/cAnNTac) mice were infected with MPV. Fecal samples
cholesterol monohydrate crystals and cholesterol gallstones at         were collected weekly from each mouse for PCR. One SW sentinel
a significantly lower prevalence (10% for each; P ≤ 0.05). Mucin       was added to each cage at postinfection day (PID) 0 and replaced
gel score were increased in Jackson mice of either substrain           at PID 14 and PID 28. Sentinel mice were held for an additional 2
(BALB/cJ: 2.250 ± 0.367; BALB/cByJ: 2.150 ± 0.317) compared            wk and tested for seroconversion and fecal shedding. Index mice
Abstracts of scientific papers 2008 AALAS National Meeting

were euthanized on PID 14 (5 SCID) or PID 42 (all other mice)        tively allocate limited resources to multiple projects and requests
and mesenteric lymph nodes (MLN) were harvested. PCR and             from diverse end users. By using task and study time analysis
FACS were performed on MLNs to detect the presence of MPV in         methods during the scheduling process, we can track available
CD4, CD8, B220, and NK cells. The results showed that all BALB,      study support time compared to resources previously alloted.
CB17 and Igh, 8/10 SCID and 1/5 SCID-BG mice were shedding           Technicians supply actual time-in-motion data at the comple-
MPV in the feces on PID 7. At PID 14, feces from SCID, SCID-BG,      tion of each day, permitting the development of a database on
and Igh mice averaged 4.2 pg/µl of MPV DNA, whereas CB17             task performance. This practice has revealed a valuable “team
and BALB feces contained 1.6 and 2.8 pg/µl of MPV DNA, re-           effect”—the efficiency that occurs when 2 or more technicians
spectively. MLNs from SCID and CB17 mice contained 66.5 and          complete a specified task in less time than predicted by adding
7.4 pg/µl of MPV DNA, respectively. Seroconversion and fecal         time spent alone. We then assign daily schedules and study
shedding occurred in contact sentinels placed with SCID-BG or        work based on data collected from one technician performing
Igh mice on PID 14, but not with SCID, CB17 or BALB mice. By         each task, allowing for the team effect to provide flexibility in
PID 21, CB17 and BALB mice ceased shedding MPV, whereas              accommodating last-minute demands and increase the capac-
the immunodeficient mice were still shedding on PID 28, 35, and      ity of the team’s workload without strain to any individual. It
42. Sentinels placed on PID 28 with SCID-BG mice but not with        is driven by increasing participation from the technicians and
other mouse strains seroconverted and shed virus. At PID 42,         directly influences how the calendars are managed.
PCR detected 0.77 (Igh) to 6.7 (SCID-BG) pg/µl of MPV DNA
in feces and detected from 1.9 (CB17) to 86.4 (SCID) pg/µl of        PS13 Developing Meaningful Training in Laboratory Animal
MPV DNA in MLNs. FACS analysis indicated a low number of             Science: An Educational Methodology
B cells and NK cells contained MPV antigen on PID 42 (SCID
> SCID-BG > Igh). These results indicate that fecal shedding is      S Pack, P Clifford*
delayed in SCID and SCID-BG but virus levels in the feces and        ULAR, University of Pennsylvania, Philadelphia, PA
MLNs were high at PID 42. Although SCID and SCID-BG had
                                                                     A new concept to bring to the laboratory animal community is
high levels of virus in the feces and MLN, only the SCID-BG
                                                                     designing learning environments to promote understanding and
transmitted infection after PID28. This suggests a role for NK
                                                                     reason. There are a few basic instructional design techniques that
cells in controlling infection.
                                                                     can immediately be applied in training development to promote
                                                                     understanding. Applying a “backwards design” method can help
PS11 New Solutions for Management and Husbandry: RFID                in developing effective training by focusing on goals, evidence,
Census and Wash Cycle Tracking                                       and activities. These 3 simple words can make a difference for
B Scher*                                                             those needing to deliver training sessions to support or research
                                                                     staff. Before developing training, the goals of the session must
Dynasys, Clearwater, FL                                              first be clearly defined. Next, determine the acceptable evidence.
The process of arriving at an accurate census for large rodent       How are you going to assess the learners and be confident that
populations is time consuming, labor intensive, and prone to         they understand what is being taught? Lastly, develop the ac-
many errors. Principal investigators often challenge the ac-         tivities, which include presentations. Typically those delivering
curacy of their billed services and census reports. Tracking the     training sessions or presentations start with their activities first
current location of every cage housed within a large animal          (such as a PowerPoint presentation). This step in development
care facility requires continuous monitoring and extensive           should be last, hence the “backwards design” method. Assess-
recordkeeping. Radio Frequency Identification (RFID) tagging         ment and evidence are key and need to be clearly determined
of each cage allows for real-time, error-free electronic detection   prior to activity development. The 6 facets of understanding need
of hundreds of cages per second. The Electronic Product Code         to be considered to truly teach and train learners so that they gain
(EPC) is an open global standard designated for RFID labeling        an enduring understanding that is applied beyond the training
analogous to the Universal Product Code (UPC) barcode that has       session. These facets of understanding that are needed to effec-
been identifying all consumer goods for the past 30 y. EPC tags      tively assess knowledge building are as follows: can explain, can
can be directly attached to animal cages, racks, and cage card       interpret, can apply, have perspective, can empathize, and have
holders. These passive tags can survive autoclave sterilization      self-knowledge. Particularly, when working in our industry with
and can be directly attached to metal. EPC technology features       live animals, a deep and meaningful workplace understanding
anti-collision properties such that hundreds of tags can be read     is paramount. Therefore, by applying the above methodology
in seconds using a portable RFID reader, and the animal census       and building an awareness of the 6 facets of understanding,
of a room can be reconciled immediately. In addition to taking       trainers can gain an active awareness of their learners’ progress,
animal census, these tagged cages can be individually identified     while being assured that their learners are gaining practical and
as they pass through cage washers. RFID readers attached to the      meaningful knowledge.
washers identify each cage and integrate the process control data
with the cage tag identifiers, providing an accurate data record     PS14 Refining Rodent Paralysis Models through Animal Care
of each wash cycle. RFID technology will find many new ap-           Staff Involvement
plications within the laboratory facility.
                                                                     A Heiser*, E Westhoven, RJ Hurley, DM Jarrell
PS12 The Team Effect and Technician Efficiency at Genzyme            CCM, Massachusetts General Hospital, Charlestown, MA
M   Scribner*                                                        A recent advance in laboratory animal care is to leverage the
                                                                     daily interaction that front-line animal care staff already has
Department of Comparative Medicine, Genzyme Corporation,             with their assigned animals to notify appropriate parties when
Framingham, MA                                                       an animal has health or welfare problems. Delegation of such
When effectively scheduling animal work for a technical support      tasks can result in a faster and equally effective response rather
team, it is necessary to monitor the team’s workload to effec-       than waiting for higher-level employees (veterinary technicians,
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

veterinarians, investigators) to discover or confirm the same           PS16 Retaining Your Top Animal Care and Cage Wash Per-
problem. But IACUC-approved endpoints in potentially painful or         formers
distressful protocols may be unclearly defined or not adequately        C Kilcullen-Steiner*
understood by animal care staff to permit appropriate and timely
intervention. Murine models of amyotrophic lateral sclerosis (Lou       University Animal Care, University of Arizona, Tucson, AZ
Gehrig’s Disease) and Huntington’s Disease have helped identify         At academic institutions, keeping top performers in cage wash
new drug strategies, an important advance for these and other           and direct animal care positions is becoming more difficult.
neurodegenerative conditions for which there are no effective           These employees usually have the lowest remuneration, are
treatments. Such models routinely involve severe clinical signs         required to be on their feet during their whole shift, experience
that should require euthanasia rather than allow the animal’s           little variation in routine, and have no loyalty to the depart-
condition to decline further. IACUC-approved protocols at our           ment/institution. As research funding and other sources of
institution mandate euthanasia following hind limb paralysis,           revenue become harder to acquire, more principal investigators
the most common severe clinical sign in these models, if the            are recruiting for new employees from the animal care staff.
animal cannot right itself within 15 s. Our animal care staff used      The PI knows he will obtain a highly trained animal technician
multiple variations of the “15-s” rule, resulting in recommended        familiar with the laws, regulations, policies, and procedures
euthanasia at varying stages of disease. To standardize monitor-        that govern the care and use of research animals. He just trains
ing of an animal’s condition, we developed a precise and simple         them on his actual research. Usually the PI can offer the animal
decision tree for more consistent reporting and intervention. A         technician more money per hour and give them the opportunity
combination of pictorial guides, clearly defined clinical signs,        to conduct actual research in addition to direct animal care. In
and possible outcomes provided in an SOP format resulted in             order to slow or stop this outflow of trained staff, the animal
more effective training, simple interpretation, and reduction in        care facility had to move toward incentive methodologies, as
variability. Benefits of this approach include improved animal          well as good business practices to increase employee retention.
welfare without compromising research and increased awareness           As a start, new positions were created to provide more career
and involvement by a caring workforce.                                  paths. One job was created for an environmental enrichment
                                                                        specialist, two instructional specialist positions were created for
PS15 Development of Team Leaders in a Team-oriented Or-                 upward mobility in cage wash, back-ups were designated for the
ganization                                                              supervisors, and technician senior leads were created to give top
                                                                        employees experience in team leadership. In addition, monetary
C Gibbons*, J Fuller, DM Jarrell
                                                                        incentives were established for achieving certification, excelling
CCM, Massachusetts General Hospital, Charlestown, MA                    in job performance, etc. The outflow of staff has decreased, but
Today’s laboratory animal care workspace mirrors recent evolu-          we know not every employee can be promoted to a new position
tion in biomedical research by being more complex, expensive,           and money is limited. We continue to work on more novel ways
and potentially hazardous. However, the standard approach to            to retain valuable staff.
organizational structure and workforce development remains
unchanged. Conventional responses are to either increase su-            PS17 Developing and Implementing a Program for Standard-
pervisory oversight (greater control) or narrow the scope of each       izing and Harmonizing Training Requirements and Records
worker stratum (less risk) to avoid mistakes of greater impact. Both    for a Global Organization
approaches increase total labor expense while research funding is
                                                                        M Butler*1, I Garrod3, G Ma4, M Defeo1, SA Drinen2, SE Deboer1,
tighter and other costs (fuel, raw materials, supplies, equipment)
                                                                        R Burke1, KJ Rogers3, CF Fox3
are growing. Furthermore, employees are forced to wait for pro-
                                                                        1Worldwide   Comparative Medicine, Groton, CT; 2Worldwide
motions or lateral transfers before they attain additional skills and
enjoy career growth without leaving. By contrast, in line with using    Comparative Medicine, St. Louis, MO; 3Worldwide Compara-
Toyota Production Systems (TPS) as our main operational and             tive Medicine, Sandwich, Kent, UK; 4Worldwide Comparative
management strategy for improving work quality, we established          Medicine, Pfizer, La Jolla, CA
18 multi-functional teams of research animal specialists divided        Developing and implementing an effective training program to
amongst 6 facilities. Team responsibilities include all husbandry,      support quality animal care and use is critically important. Our
initial health assessment and basic veterinary care, operational        global organization, consisting of three sites in the United States
maintenance, and research services within their designated area.        and 1 site in the United Kingdom, was faced with the challenge
Strong team leadership is paramount in sustaining a highly func-        of developing such a program. To address this challenge, our
tional team. Team leaders are responsible for coordination of daily     organization formed a team consisting of select personnel from
tasks, team-directed evaluation, and improvement of work flow           each site. This team was given the task of developing a train-
processes, and assurance that team members work in a productive         ing program that was aligned across sites, creating a learning
and collaborative manner. An intensive team leadership training         environment with shared and measured accountability, and
program developed in-house includes a comprehensive review              developing and implementing an innovative learning approach
of problem-solving tools and principles of TPS, an emphasis on          that would save the organization both time and money. The
mature and clear communication skills for a strong researcher/          approach developed by this team, modeled after a successful
customer service focus, and building work style-focused interper-       approach used at our U.K. site, consists of four components.
sonal skills to support a highly functional team-environment. One       A master training index is used to identify all training require-
significant impact seen at MGH of improved team leaders and             ments specific to our global training program. Each item in this
highly functional teams has been our ability to increase staffing       index is listed under an appropriate training category and is
levels only 30%, while increasing our overall rodent cage census        supported by a corresponding training module. Each training
by 60% over the last 3 y. Financial impacts in less overtime and        module contains harmonized learning objectives agreed to by
increases in non per-diem revenues (such as research services           all sites and identifies the minimum requirements that an indi-
support) were also observed.                                            vidual must meet before they are considered proficient for that
Abstracts of scientific papers 2008 AALAS National Meeting

module. Qualified trainers use these training modules to ensure      hands-on classes were provided to learn postural modifications
that training is provided at an appropriate level of detail and      and exercises used to prevent injury when performing tasks such
that all required information is addressed during the training.      as rodent cage changing, dog cage cleaning, and moving heavy
Training schedules are used by managers at each site as a check-     equipment in feed and bedding storage areas and washrooms. A
list and guide to ensure that new personnel complete assigned        review of job specific exercises is presented here, as well as staff
training requirements within established timelines. Standardized     members’ comments by survey of the success of the program
training records mirror the contents of the master training index    in decreasing areas of discomfort and fatigue in the workplace.
and require that individuals are assessed while working under        Our departmental goal of decreasing injury rates by at least 10%
supervision, assessed as competent, and assessed as a competent      was met after implementing these changes and our future goals
trainer, if appropriate, for each assigned training module. This     related to safety will be addressed.
session details the approach taken to develop our global training
program and describes the tools developed to support it.             PS20 Development and Implementation of a Quality Control
                                                                     Program for Animal Care Activities
PS18 Ergonomic Improvements through Equipment Updates
                                                                     N O’Callaghan*, R Palmisano
in a Laboratory Animal Facility
                                                                     Yale University, New Haven, CT
R Starr*, N Montero, C Fuzzi, J Nolan, A Kopec, M Hargaden,
L Singer                                                             Monitoring the care and use of laboratory animals is an integral
                                                                     component of humane biomedical research and a responsibility
Department of Laboratory Animal Resources, Roche, Nutley,
                                                                     of institutional oversight and evaluation. Ongoing monitoring
NJ
                                                                     must be performed and documented to ensure humane and
The study of body movement and posture in relation to work           compliant direct care activities between federally mandated
demands, or ergonomics, has become a major point of interest in      inspections and accrediting site visits. This provides early detec-
places of work, including laboratory animal facilities. In 2007, a   tion and identification of noncompliance for corrective action and
global initiative to decrease work-related injuries and associated   quality control. To accomplish this, a position was created within
costs prompted the Department of Laboratory Animal Resources         the Regulatory and Safety Services (RSS) section with dedicated
(LAR) to invite an ergonomist to evaluate specific job functions     effort to monitoring animal care and related operations. Due to
in the animal facility. An internal safety committee comprised of    Yale University’s large, diverse, and geographically decentralized
an operations manager, building supervisors, laboratory animal       animal care and use program, a streamlined process and self-
technicians, clinical veterinary associates, and veterinarians       assessment tool was created to ensure that animal care activities
was responsible for working with the ergonomist to alleviate         are in accordance with institutional, federal, and state regulations
sources of repetitive motion or overuse injury and fatigue in        and policies. This assessment is conducted in every animal room
our employees. The committee evaluated new equipment and             and support space twice a year. The findings from each space
variations on the use of space to enhance safety in work areas,      monitored are immediately reported to the facility manager so
including cagewash rooms, large and small animal housing             that corrective action can be taken. Monthly reports with global
rooms, and food and bedding storage areas. Our evaluation            findings are given to all facility managers, and quarterly global
of several pieces of equipment, including hydraulic lifts, tote      reports are presented to all managers at a convened meeting.
tippers, table lifts, scooters, and ergonomic brushes and mop        Corrective action plans are then identified and implemented.
handles, will be discussed, as well as an overview of related        This quality control monitoring program promotes timely cor-
costs. Our departmental goal of decreasing injury rates by at        rective action and provides a global view of the entire program.
least 10% was met after implementing these changes. Our future       Additionally, this internal assessment provides a link to both
goals related to safety will be addressed.                           the refinement of the training program and the creation of and
                                                                     revision to the internal standard operating procedures (SOPs).
PS19 No-cost Solutions for Ergonomic Improvements in a               The institution’s investment in quality control monitoring affirms
Laboratory Animal Facility                                           its commitment to improving the quality of humane animal care
                                                                     while heightening the regulatory awareness of all levels of animal
C Fuzzi*, M Hargaden, L Singer                                       care staff and ultimately reducing noncompliance.
Department of Laboratory Animal Resources, Roche, Nutley,
NJ                                                                   PS31 Spontaneous Acute Tumor Lysis Syndrome in Swiss Webster
The study of body movement and posture in relation to work           Mice with Spontaneous Leukemias and Lymphomas
demands, or ergonomics, has become a major point of interest in      S Rasmussen*1,3, K LaPerle1,3, S Couto1,3, J White1,3, V Gillespie1,2,
places of work, including laboratory animal facilities. In 2007, a   FR Wolf1,4
global initiative to decrease work-related injuries and associated   1Postdoctoral Research, Tri-Institutional Postdoctoral Training
costs prompted the Department of Laboratory Animal Resources
(LAR) to invite an ergonomist to evaluate specific job functions     Program in Laboratory Animal Medicine and Science, New York,
                                                                     NY; 2Pathology, Animal Medical Center, New York, NY; 3RARC/
in the animal facility. An internal safety committee comprised
                                                                     CBC, Memorial Sloan-Kettering Cancer Center, The Rockefeller
of an operations manager, building supervisors, laboratory
                                                                     University, and The Weill Medical College of Cornell Univer-
animal technicians, clinical veterinary associates, and veterinar-
                                                                     sity, New York, NY; 4RARC, Memorial Sloan-Kettering Cancer
ians was responsible for working along with the ergonomist to
locate potential sources of repetitive motion injury or fatigue      Center and The Weill Medical College of Cornell University,
                                                                     New York, NY
and discomfort in our employees. In addition to providing our
employees with training in ergonomics, we consulted an exercise      We have observed a high incidence of spontaneous acute tumor
physiologist to address staff members’ individual concerns with      lysis syndrome (ATLS) in our colony of Swiss Webster mice used
their specific daily tasks. An exercise program was implemented      as dirty bedding sentinels. ATLS is thought to be rare in mice,
to increase awareness in posture and stretching, and small group     with a single reported case in a DBA/1J mouse. ATLS results
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

from a rapid destruction of neoplastic cells. When large volumes       findings. Vascular lesions, considered the cause of death in the
of byproducts from neoplastic cells are released, the body’s           published reports of GVHD in mice, were not observed in our
excretory mechanisms are overwhelmed, resulting in metabolic           cohort. X-GVHD has not been previously described in the NOD-
derangements, renal failure, and disseminated intravascular            scid IL2Rγnull strain, nor has the disease been fully characterized
coagulation and thrombosis. ATLS can occur spontaneously,              clinically and pathologically in mice. This comprehensive study
or more commonly following chemotherapy, radiation, or                 will further the understanding of X-GVHD in immunodeficient
corticosteroid administration. In humans, ATLS has a strong            mice and provide useful information for those using NOD-scid
association with high-grade lymphoproliferative malignancies,          IL2Rγnull mice in cancer immunobiology.
although also reported to occur with other tumor types. Com-
plete anatomic pathologic evaluations were performed on 146            PS33 Incidence, Evaluation of Sampling Techniques, Isolate
Swiss Webster mice that were either found dead without a history       Diversity, and Antibiotic Susceptibility of Corynebacterium
of clinical signs or were sacrificed due to lymphadenopathy or         bovis in Immunodeficient Mice
varying signs of morbidity. The mice were not subject to prior
experimental manipulation. Results from mice necropsied over           HN Burr*1,2, NS Lipman1,3, A Toma3, FR Wolf1,3
a 12-mo period revealed that 94 of 146 (64%) mice evaluated            1Tri-Institutional Training Program in Laboratory Animal
were found to have either lymphoma or leukemia. These find-            Medicine and Science, New York, NY; 2 Memorial Sloan-Kettering
ings are not surprising given that a high percentage of Swiss          Cancer Center, The Rockefeller University, and the Weill Medical
Webster mice are known to carry and express ecotropic murine           College of Cornell University, New York, NY; 3 Memorial Sloan-
leukemia viruses. Seven of 146 (4%) mice evaluated also had            Kettering Cancer Center and the Weill Medical College of Cornell
histologic lesions consistent with ATLS. Five of the 7 ATLS cases      University, New York, NY
died spontaneously. Of the remaining 2, 1 mouse was euthanized
                                                                       Corynebacterium bovis infection remains an endemic problem in
due to gross abdominal distension as a result of splenomegaly
                                                                       athymic nude mice at academic institutions. Infected athymic
and lymphadenopathy; the other due to neurologic signs. Four
                                                                       nude mice used in cancer research frequently exhibit signs of
of the 7 mice with ATLS had pathologic findings consistent with
                                                                       clinical disease, including hyperkeratotic dermatitis, septicemia,
both leukemia and lymphoma. The remaining 3 mice had find-
                                                                       and death, when experimentally manipulated. We sought to
ings consistent with disseminated lymphoma. All mice were
                                                                       evaluate C. bovis sampling techniques in immunodeficient hirsute
found to have had intravascular multifocal thrombi composed
                                                                       and glabrous rodents, incidence of infection, isolate diversity, and
of basophilic nuclear and eosinophilic cytoplasmic debris con-
                                                                       antibiotic susceptibility. Rodents in 2 animal holding rooms, each
sistent with ATLS.
                                                                       housing immunodeficient rodents including athymic nude mice
                                                                       and rats, were sampled and found to have a C. bovis infection
PS32 Clinical and Pathological Characterization of Xenogeneic          prevalence of 30% and 67%, respectively, with 95% of infections
Graft-versus-host Disease in NOD-scid IL2Rγnull Mice                   subclinical. Skin swab and buccal swab techniques were found
HN Burr*1,2, JC Markley3,4, JR White1,2, SS Couto1,2                   to be comparable for isolation of C. bovis from both hirsute and
1
                                                                       glabrous mice, but in rats, the buccal swab was more sensitive. In
  Tri-Institutional Training Program in Laboratory Animal
                                                                       hirsute rodents, the skin scrape technique was also evaluated and
Medicine and Science, New York, NY; 2 Memorial Sloan-Kettering
                                                                       found to be less sensitive. C. bovis isolates demonstrated 8 differ-
Cancer Center, The Rockefeller University, and the Weill Medi-         ent biochemical profiles with colorimetric biochemical tests; each
cal College of Cornell University, New York, NY; 3 Department          profile was confirmed by colorimetric carbon source metabolic
of Molecular Pharmacology, New York, NY; 4 Memorial Sloan-             fingerprinting, PCR and 16S rRNA sequencing to be C. bovis. C.
Kettering Cancer Center, New York, NY                                  bovis colony size also varied. Both a large and small colony-type
Immunodeficient mouse strains are commonly used to study               grew on Columbia colistin-nalidixic acid agar with 5% sheep blood
human tumor biology and adoptive immunotherapy. Conse-                 media. The Kirby-Bauer disk diffusion antibiotic susceptibility test
quently, they can develop xenogeneic graft-versus-host disease         revealed different sensitivities to penicillin between the large and
(X-GVHD) which may confound interpretation of experimental             small colony-type. Our findings demonstrate the skin and buccal
results. We characterized X-GVHD in NOD-scid IL2Rγnull mice to         swab sampling techniques in mice to be equally sensitive at C. bovis
aid in the differentiation between the disease and cancer-related      detection, contrary to published reports. Diversity within C. bovis
death. Eight-week-old male mice received intravenous human             isolates is newly reported and may lead to further understanding
Burkitt lymphoma cells (Raji cell line) alone (n = 6) or followed      of virulence and disease manifestation. Additional research that
by intravenous tumor-specific human T cells 6 d later (n = 19).        evaluates the efficacy of amoxicillin diet prophylaxis and treat-
After approximately 2 mo, mice receiving human T cells pre-            ment will be presented.
sented with nonspecific signs of illness including weight loss,
scruffy hair coat, and hunched posture. Histologically, a hetero-      PS34 Abnormally High Serum Potassium Levels in Laboratory
geneous infiltration of CD3 positive lymphocytes was observed          Guinea Pigs and Rodents Caused by CO2 Euthanasia–Induced
in several organs. In addition to dermatitis, colitis and hepatitis    Respiratory Acidosis
commonly seen in allogeneic GVHD, the mice had significant
lesions in the lungs, liver, salivary glands, prostate, thyroid        P Cowley, B Hayes, A Greene, WR Shek*
gland, and epididymis. Mild nephritis, arthritis, and meningoen-       R&D, Charles River, Wilmington, MA
cephalitis were also observed. Raji tumor cells were completely
                                                                       The carbonate/bicarbonate buffer system is essential in main-
eliminated in mice receiving human T cells. Mice receiving Raji
                                                                       taining pH homeostasis in animal physiology. CO2 euthanasia
cells alone presented with hindlimb paralysis and histological
                                                                       overwhelms the buffering capacity of the system, causing a
evidence of spinal cord and liver infiltration by Raji cells, but no
                                                                       respiratory acidosis that may lead to abnormal serum chem-
lesions compatible with X-GVHD. Mice suffering from X-GVHD
                                                                       istry values. We performed serum chemistry on specimens
likely die of multiorgan dysfunction with thyroid destruction
                                                                       collected from 16 specific pathogen-free Hartley guinea pigs
and pulmonary impairment being the most clinically relevant
                                                                       (Crl:HA ORIGIN), following ketamine/xylazine (40 to 80 mg/
Abstracts of scientific papers 2008 AALAS National Meeting

kg; 2 to 5mg/kg) anesthesia and after CO2 euthanasia. Serum          progression of disease have not been clearly identified. Glutamic
potassium (K+) levels were normal in specimens collected from        acid decarboxylase (GAD), found in mouse and human pancreatic
anesthetized animals (4.41 mmol/l). Following CO2 euthanasia,        beta cells, is one potential candidate autoantigen. We constructed
however, the serum K+ concentration increased 3-fold on average      mice with a human:mouse chimeric TCR recognizing GAD65
(12.95 mmol/l); moreover, less marked increases in glucose and       (555-567) which were crossed with murine MHC Class II knockout
inorganic phosphate levels were noted. Increases in K+ above         mice expressing human diabetes-associated MHC Class II alleles
normal concentrations (3.5 to 5.1 mmol/L) were also found in         (C57BL/6 background), with and without a recombination-
sera from CO2-euthanized mice (n = 182; K+ = 12.81 mmol/l)           activating gene deletion. Unexpectedly, we observed spontaneous
and rats (n = 572; K+ = 8.88 mmol/l).                                lymphoblastic lymphoma in 2 lines of GAD TCR transgenic
                                                                     mice (4.13αβ and 164αβ). Health records were examined over a
PS35 Poliomyelitis in SCID Mice Following Injection of               2-y period; mice presenting with clinical signs compatible with
Basement Membrane Matrix Contaminated with Lactate                   lymphoma such as combinations of abdominal or subcutaneous
Dehydrogenase-elevating Virus                                        masses, anemia, hunched, and ruffled fur plus cases confirmed
                                                                     by necropsy or histopathology were counted. Forty-four mice met
J Carlson*1, R Garg2, SR Compton1, C Zeiss1, E Uchio2                these criteria. Age ranges of affected 4.13αβ and 164αβ mice were
1Section of Comparative Medicine, 2Department of Surgery,            17.1 to 54.6 wk (average = 28 wk) and 18.7 to 53 wk (average =
Section of Urology, Yale University School of Medicine, New          34.5 wk), respectively, with no sex bias. The incidence of disease
Haven, CT                                                            in 4.13αβ mice was 19.7% and 13.9% in 164αβmice. Interestingly,
                                                                     mice transgenic only for the 164β chain of the TCR had a disease
Five 30- to 40-wk-old female ICR/SCID mice presented for
                                                                     incidence of 0.03% based on the above criteria. A large thymus,
para- or quadriplegia. The mice were part of a 50-mouse study
                                                                     splenomegaly, and lymphadenopathy were the most common
involving subcutaneous injection of carcinoma cells suspended
                                                                     gross findings. Histopathology performed on 6 animals revealed
in basement membrane matrix. Ruleouts included mouse
                                                                     extensive lymphoma in multiple organs with bone marrow in-
hepatitis virus, mouse encephalomyelitis virus, lactate dehy-
                                                                     volvement where examined. The necropsy findings combined with
drogenase-elevating virus (LDV), and CNS metastasis. Upon
                                                                     characteristic lymphoblastic cells strongly support a T cell origin;
initial investigation, it was discovered that the matrix was con-
                                                                     however, immunophenotyping will aid in further characterization.
taminated with LDV. Twenty-one mice were analyzed to confirm
                                                                     Clinicians and investigators are reminded that lymphoma may be
LDV as a cause of clinical findings; tissues from 14/21 mice were
                                                                     an unintended consequence in genetically engineered mice.
positive for LDV by RT-PCR. Histology was performed on 18
mice, of which 4 presented with paralysis; 12/18 mice had CNS
lesions consisting of neuronal degeneration, marked astrocytic       PS37 Development and Implementation of Multimedia Con-
hypertrophy and gliosis and myelin loss. The spinal cord was         tent for Web-based Rodent Microsurgery Courses
most commonly affected, with less consistent involvement of the      S Baran*1, J Kehler2, E Johnson3, F Hankenson4
brainstem and caudal midbrain. Cord lesions co-localized with        1Department   of Surgery, Veterinary Bioscience Institute, Har-
focal marked astrogliosis on GFAP immunohistochemistry. Wal-
                                                                     leysville, PA; 2Department of Clinical Studies, University of
lerian degeneration of sciatic nerves was detected in 4/18 mice.
                                                                     Pennsylvania, Kennet Square, PA; 3 Harleysville Veterinary
Lymphoma was present in 5/18 mice. Additionally, 21/21 mice
                                                                     Hospital, Harleysville, PA; 4ULAR, University of Pennsylvania,
were positive for murine leukemia virus (MuLV) by RT-PCR.
Cell lines were negative by PCR for viral contaminants, while        Philadelphia, PA
matrix was positive for LDV. These results strongly implicated       Microsurgery is commonly used in the human, veterinary and
LDV-contaminated matrix as a cause for clinical findings. LDV is     biomedical fields. Advances in microsurgical equipment and
a mouse arterivirus that is generally asymptomatic. MuLVs are        rodent technologies have increased the demand for biomedical
endogenous retroviruses found in all laboratory mice as provi-       microsurgeons. While a limited number of microsurgical training
ruses integrated in the chromosome, although only strains that       courses are available, ultimately many rodent biomedical proce-
carry the Fv-1n/n allele express infectious virus. Co-infection      dures are self-taught, introducing a bias of technical proficiency
with MuLV and LDV in immunosuppressed Fv-1n/n mice results           into experimental outcomes. Electronic-learning (e-learning) has
in the development of age-dependent poliomyelitis (ADPM), a          been successfully incorporated into the human surgical curricula.
model of human amyotrophic lateral sclerosis. In this case, clini-   E-learning allows practitioners to study and improve their skills
cal disease consistent with ADPM developed due to incidental         at their convenience. Translating this model to the biomedical
inoculation of LDV into immunocompromised mice. This report          research field provides users with an opportunity to learn rodent
highlights the importance of screening biological materials before   surgical techniques through electronic resources like interac-
inoculation into laboratory animals.                                 tive and non-interactive videos, animations, presentations, and
                                                                     standard operating procedures. This training can be completed
PS36 Spontaneous Lymphoma in Glutamic Acid Decarboxylase             prior to using live animals, which promotes the principle of re-
T-cell Receptor Transgenic Mice                                      duction since animals are not used and enhances the experience
                                                                     of hands-on courses. The goals of the project were to develop
AJ Burich*1,2, PM Treuting2, JA Gebe1, TL Brabb2                     web-based interactive learning for rodent surgical techniques.
1Benaroya   Research Institute at Virginia Mason, Seattle, WA;       Unique e-learning microsurgical modules were incorporated into
2Department    of Comparative Medicine, University of Washing-       a training class of undergraduate laboratory animal students
ton, Seattle, WA                                                     (n = 60). The tested modules included laws, regulations, and
                                                                     guidelines; surgical patient preparation; and proper tissue han-
T-cell receptor (TCR) transgenic mice are used to understand
                                                                     dling and dissection. The project was evaluated using postcourse
thymocyte development, peripheral tolerance, and T cell:antigen
                                                                     survey data. Within the training class, all students responded
interactions in infectious and autoimmune diseases. Type 1
                                                                     favorably to videos, step-by-step photographs of procedures, and
diabetes is thought to be a T-cell mediated autoimmune process,
                                                                     24-h availability of the course. Voiceovers with the videos were
although the autoantigens responsible for the initiation and
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

preferred by 65% of students, and short video clips embedded          been previously described in these animals; however, there are
in the text over compilation videos were preferred by 87% of          currently no reports pertaining to other clinical findings when
students. Also, 95% responded positively to review questions          these rabbits are placed on a high-cholesterol diet. In the current
provided after each section of the course. Critiques included         study, Watanabe rabbits (n = 14) were placed on a diet contain-
improving video resolution and the quality of the voiceover           ing 0.5% cholesterol. In addition to the high-cholesterol diet,
performance. These preliminary results support the continued          2 groups of these rabbits also received atherosclerotic plaque-
development and implementation of electronic and interactive          reducing drugs. Bloodwork, serial cholesterol, and triglyceride
rodent surgical technique courses to the growing laboratory           levels were monitored throughout the study. After 2 mo, the
animal and biomedical research communities.                           rabbits began to develop some unusual lesions. Complete oph-
                                                                      thalmic examinations revealed that all rabbits had a significant
PS38 Cesarean-derived SPF Sheep for Biomedical Research               lipid arcus and uveitis, and 2 out of the 14 had scleral and uveal
                                                                      eruptive xanthomas. Six rabbits developed complete bullous
RJ Hurley*1, JJ Hurley2                                               retinal detachments and glaucoma secondary to the uveitis. The
1CCM, Massachusetts General Hospital, Charlestown, MA;            2   retinal detachments and secondary glaucoma led to vision loss
New England Ovis, LLC, Rollinsford, NH                                in this population. In addition, integumentary xanthomas and
                                                                      weight loss were seen clinically. At necropsy, affected animals
Great strides have been made in recent decades to minimize or
                                                                      had granulomatous inflammation in the pyloric region lead-
eliminate unintended environmental and biological variables in
                                                                      ing to partial gastric obstruction. Histological analysis of the
research animals. Pathogens have been eliminated from many
                                                                      ocular lesions revealed that the sub-retinal space was markedly
commonly used species. This has resulted in healthier animals,
                                                                      expanded by lipid-filled macrophages and cholesterol clefts.
of which fewer are needed to generate reproducible and sta-
                                                                      Additional histopathological findings included severe diffuse
tistically significant data. Many of these advantages, however,
                                                                      hepatic lipidosis and lipogranulomatous inflammation of the
have not been realized in research using sheep because their
                                                                      meninges, ependymis, renal papilla and choroid. Although clini-
commercial production does not employ the same “barrier” ap-
                                                                      cal complications are expected in these rabbits, especially when
proach applied to smaller mammals. In particular, it has been
                                                                      they are fed a high-cholesterol diet, the severity and extent of
the experience at our facility that pulmonary and cardiovascular
                                                                      systemic involvement seen in these animals was unprecedented
studies using sheep are very dependent on the health status of
                                                                      and warrants disclosure to the scientific community.
the animal subject. Sheep from commercial flocks would ar-
rive clinically normal, but, following shipment-related stress,
changes in housing and diet, and experimental manipulation,           PS40 Use of Tricaine Methanesulfonate for Euthanasia of
they often developed respiratory signs. In addition, some were        Reptiles
discovered during thoracotomy to have bacterial or mycoplasmal        NE Hahn*
pneumonia that excluded them from sensitive physiological
studies. Other occasional infections such as contagious ecthyma       Office of Laboratory Animal Care, University of California
and clinical coccidiosis imposed additional health management         Berkeley, Berkeley, CA
burdens in a vivarium setting. In response, a flock of cesarean-      Tricaine methanesulfonate (MS-222) injected into the intrac-
derived Polypay sheep was established. Lambs were aseptically         oelomic (ICm) cavity of reptiles was evaluated as a chemical
and immediately removed from their ewes and hand-raised at a          euthanasia method. This euthanasia method was developed
separate facility isolated from other ruminants. Extensive test-      as an alternative to sodium pentobarbital, which due to Drug
ing (serology, PCR, culture) has shown these animals to be free       Enforcement Agency control is problematic to use, particularly
of bacterial, viral, and parasitic organisms commonly found in        for international fieldwork. Three western fence lizards, 2 des-
conventional flocks. Monitored and excluded agents include all        ert iguanas, 4 garter snakes and 6 geckos were successfully
intestinal parasites (helminthes, cryptosporidia, giardia), lung      euthanized using this protocol. The reptiles were given an ICm
worms, ectoparasites, bluetongue virus, bovine viral diarrhea         injection of 250 to 500 mg/kg of 0.7 to 1% sodium bicarbonate-
virus, contagious ecthyma, ovine progressive pleuropneumonia,         buffered MS-222. Loss of response to toe or skin pinch occurred
Toxoplasma, Brucella, chlamydia, Q-fever, Mycobacterium paratu-       for most reptiles within 2 to 3 min, at which point a second ICm
berculosis, Leptospira, and all respiratory bacterial pathogens       injection of 0.1 to 1.0 ml unbuffered 50% (v/v) MS-222 solu-
(culture negative). Coccidia were detected 1 y into the project,      tion was administered. Respiratory and cardiac arrest occurred
but no clinical cases have been seen. When used in research pro-      within seconds in all animals, and was confirmed visually after
tocols, these animals have been free of clinical signs and lesions,   opening the coelom. This 2-stage protocol for the euthanasia of
providing investigators with consistently reliable quality along      reptiles MS-222 can be easily followed in the laboratory or field.
with an improvement in animal welfare.                                In conclusion, MS-222 offers an acceptable alternative to sodium
                                                                      pentobarbital for euthanasia of reptiles.
PS39 Clinical Complications in Watanabe Heritable Hyper-
lipidemic Rabbits on a High-cholesterol Diet                          PS41 Does Sex Matter in a Rhesus Macaque (Macaca mulatta)
                                                                      Model of Focal Stroke?
LGallaugher*1,  C Forrider1, S Stone2, L Wancket2,   D   DaCunha1,
S Lewis1, B Farrar3, M Kariisa3, S Rajagopalan3                       SJ Murphy*1, JR Kirsch1, W Zhang1, M Grafe1,2, PD Hurn1
1UniversityLaboratory Animal Resources, 2College of Veteri-           1Anesthesiology and Peri-operative Medicine, 2Pathology, Or-
nary Medicine, 3College of Medicine, The Ohio State University,       egon Health and Science University, Portland, OR
Columbus, OH                                                          Gender differences, sex steroid effects, and sex-specific candi-
The Watanabe heritable hyperlipidemic (WHHL) rabbits are the          date therapeutics in experimental ischemic stroke have been
most commonly used genetically altered strain of rabbits used         studied primarily in rodents. Many preclinical stroke studies
today for atherosclerosis research. Clinical complications such as    involving nonhuman primates use only males, do not indicate
lipid keratopathy, eruptive xanthomas and hepatic lipidosis have      sex, or do not stratify outcomes relative to sex. The lack of data
Abstracts of scientific papers 2008 AALAS National Meeting

in female higher order species severely limits the translation           cord, spinal nerve roots, and peripheral nerves were submitted
of experimental data to women requiring treatment for stroke             for pathologic evaluation. Light and electron microscopy demon-
or neuroprotection during stroke. In this feasibility study (n =         strated severe, diffuse spongiform changes in the central nervous
3 per group), we developed a model of transient focal cerebral           system only. Widespread intracellular vacuolation of the astro-
ischemia in adult male and female rhesus macaques that consis-           cytes was seen predominantly in the gray matter of the brain and
tently includes white matter injury. We also questioned whether          spinal cord. The brain stem (medulla oblongata), cerebellum, and
gender-linked differences in histopathological outcomes could be         midbrain were more severely affected than the cerebral cortex.
evaluated in this model for future, larger preclinical trials. Male      Future characterization will focus on differentiating a primary
and female rhesus macaques aged 7 to 11 y were subjected to 90           astrocyte membrane dysfunction from vacuolation secondary to
min of reversible middle cerebral artery occlusion (MCAO) using          a metabolic disorder. These ataxic OFA rats constitute a novel
the transorbital approach with temporary aneurysm clip place-            model for studying the etiology, pathology, and treatment of
ment. Histological evaluations of the caudate nucleus, putamen,          astrocyte associated neurodegenerative diseases.
globus pallidus, claustrum, internal capsule, external capsule and
hemisphere were done 4 d following MCAO using cresyl violet              PS43 Normal Feeding and Body Weight in Fischer 344 Rats
staining. Intra-ischemic and reperfusion blood glucose, blood            Lacking the Cholecystokinin-1 Receptor Gene
pressure, heart rate, oxygen saturation, and rectal temperatures
were determined and found equivalent between females and                 J Blevins3, J Overduin3, J Fuller2, D Cummings3, K Matsumoto4,
males. Although there were no differences in regional infarction         DH Moralejo*1
volumes between male and female subjects in this small sample,           1Comparative   Medicine, 2Medicine, University of Washington,
females showed greater variability in infarction volume (mm3;            Seattle, WA; 3Division of Endocrinology/Metabolism, VA Puget
% contralateral structure) than did males in the caudate nucleus         Sound Health Care System, Seattle, WA; 4Division for Animal
(125 ± 109 mm3; 45 ± 38% compared with 217 ± 63 mm3; 97 ±                Research Resources, The University of Tokushima, Tokushima,
16%), putamen (283 ± 184 mm3; 62 ± 42% compared with 499 ±               Japan
58 mm3; 106 ± 11%), and hemisphere (781 ± 437 mm3; 10 ± 8%
                                                                         Cholecystokinin (CCK) is hypothesized to inhibit food intake by
compared with 1261 ± 383 mm3; 18 ± 6%). This is the first report
                                                                         acting at CCK1 receptors (CCK1R) on vagal afferent neurons that
to compare cerebral ischemic outcomes in female nonhuman
                                                                         innervate the gastrointestinal tract and project to the hindbrain.
primates to that of their male counterparts. Infarction volume
                                                                         Recent studies have shown that OLETF rats, which carry a spon-
variability in male compared with female macaques was com-
                                                                         taneous null mutation of the CCK1R, are hyperphagic, obese, and
parable to what is observed in human focal cerebral ischemia
                                                                         predispositioned to type 2 diabetes. In contrast, we have gener-
and in other nonhuman primate proximal MCAO models. The
                                                                         ated a congenic strain in which the OLETF-derived CCK1R gene
female macaque ischemic stroke model could be used follow-
                                                                         identified by the QTL (Niddm10/of) on chromosome 14 was in-
ing rodent studies in providing vital preclinical data for clinical
                                                                         trogressed into the normoglycemic and nonobese Fischer 344 rat
trials in women.
                                                                         genetic background, producing a CCK1R-deficient Fischer 344 rat
                                                                         (F344.Cck1r-/-) with a lean and normoglycemic phenotype. Here
PS42 Characterizing the Neurological Phenotype of the Spon-              we characterized the behavioral and neurobiological phenotype
taneous Mutant Ataxic OFA Rats                                           of this rat strain more fully. F344.Cck1r-/- and F344.Cck1r+/+
ST Chou1, KL Walter*2, E Ahearn1, A Harper1, R Driscoll1, LA             rats did not differ in body weight and showed comparable weight
Conour1                                                                  gain when maintained on Ensure Plus for 3 wk. No difference
1 Charles River, Wilmington, MA;           2
                                                                         was found in 24-h food intake and dark-phase meal frequency
                                               Harleysville Veterinary   or meal size between F344.Cck1r+/+ and F344.Cck1r-/- rats.
Hospital, Harleysville, PA                                               As expected, intraperitoneal injections CCK-8 inhibited intake
A spontaneous autosomal recessive mutation in a colony of out-           of chow and Ensure Plus in F344.Cck1r+/+ rats but not in F344.
bred Oncins France Strain A rats (Crl:OFA[SD] resulted in a novel        Cck1r-/- rats. CCK-8 failed to induce Fos in the area postrema
neurological phenotype. A clinical scoring system was used to            and the gelatinosus, commissural and medial nucleus tractus
establish a timeline for onset and progression of phenotype. The         solitarius of F344.Cck1r-/- rats, whereas robust Fos responses
average body weight of the mutants is higher at weaning, but             were seen in these areas in the F344.Cck1r+/+ rats. Together,
falls behind in growth curve by 6 to 7 wk of age. There is no gross      these results confirm that CCK-1R mediates CCK-8-induced
difference in clinical disease progression by sex. Whole body            inhibition of food intake and Fos activation in the hindbrain, but
tremors are noted at weaning and progress into an exaggerated            we find that in rats with a F344 background, genetic ablation of
“bouncing” of hindlimbs during ambulation. This is evident in            CCK1R is not associated with obesity or hyperphagia.
>90% of the mutants by 7 wk of age. Gait analysis at 4 wk of age
shows the affected rats to have a shorter stride length and wider        PS44 A Novel Neonatal Non-invasive Motor Function Test with
base stance than the unaffected rats (P ≤ 0.01). By 8 wk of age,         Potential Application for Neuromuscular Diseases
100% of the affected rats exhibit ataxia and incoordination. Print
interpretation from gait analysis becomes increasingly difficult         C Pater*1, S Ramboz1, M Winberg2, A Chen1, N Edgar1, B El-
as their gait leaves behind multiple sets of hindlimb prints with        Khodor1
each step taken. Despite abnormal gait, open field assessment            1PsychoGenics,   Inc., Tarrytown, NY; 2SMA Foundation, New
showed a significantly higher number of rearing events (P ≤ 0.01)        York, NY
and no difference in distance traveled between rat groups within
                                                                         Comprehensive motor and sensorimotor studies have been per-
10-min test sessions. The neurological phenotype fluctuates in
                                                                         formed on a variety of neonatal neuromuscular disease models.
progression rate and severity as animals age. By 6 mo of age,
                                                                         We realized that the existing neonatal neuromuscular tests do not
clinical signs include loss of body condition and righting reflex,
                                                                         adequately measure muscle strength and possible subtle drug ef-
recumbency, and absence seizures. The onsets of these clinical
                                                                         fects. Such a test would ideally be a scored, reflex-based motor test
signs are used as humane endpoint criterions. The brain, spinal
                                                                         that could be easily and reliably administered to rodent infants as
                                                                                          Abstracts of scientific papers 2008 AALAS National Meeting

early as the day of birth and would remain valid for at least the first   Most experimental approaches in stroke research are carried out
2 wk of life. The hind-limb suspension test (the tube test) evaluates     in rodents, but given differences between rodents and primates,
the proximal hindlimb muscle strength, weakness, and fatigue in           nonhuman primate stroke models may provide a valuable tool
mouse or rat neonates. It uses simple equipment found around the          to study therapeutic approaches in stroke. We have developed
lab, such as 50 ml conical tubes or beakers for larger mice. There        a reproducible surgical method for cerebral infarction of the
are 2 consecutive trials. The mouse is placed head down, hanging          M1 branch of the middle cerebral artery (MCA) using 7 African
by its hind limbs in a 50 ml conical tube with a cotton ball at the       green monkeys (Chlorocebus aethiops). A left fronto-temporal
bottom to cushion its fall. Three parameters are evaluated: latency       craniotomy was performed and after dural exposure, the M1
to fall from edge of tubes; number of pulls that animal performs          branch was surgically exposed. A vascular clip was applied to the
during its attempt to escape the tube; and hindlimb score (HLS),          distal portion of M1 for 2 h. After releasing the clip, cranioplasty
which assesses the positioning of the legs and tail. Observations         and repair of the temporalis muscle were carried out. Neuro-
with neonates exposed to the tube test showed that WT mice readily        logical examinations and behavioral testing were developed
held on with their hind limbs from postnatal day (P) 2 until P12. The     for functional assessments. Postsurgically, 4 animals showed
effects of various factors known to influence motor performance           typical right hemiparesis, similar to human patients with M1
in neonates such as hypothermia have been evaluated in the tube           infarction. Magnetic resonance imaging (MRI) demonstrated
test. In addition, using a neonatal mouse model of spinal muscular        extensive infarction throughout the left MCA territory (complete
atrophy (SMA), we showed that the performance of the KO was               occlusion group). Three animals showed mild hemiparesis, and
significantly different from WT in the 3 parameters of the tube test      MRI demonstrated smaller areas of infarction (partial occlusion
as early as P4. This test would enable anyone to obtain a reliable        group). During M1 clipping, we observed darkening of cerebral
quantitative assessment of motor function in mice as early as the         venous blood, small vessel collapse, and blood pooling of corti-
day of birth. We would like to acknowledge the SMA foundation             cal veins, evidenced by back-and-forth movement of stagnant
for their help and support in this project.                               blood during brain pulsation. In animals with partial occlusion,
                                                                          we did not observe blood pooling of cortical veins. These data
PS45 Validation of Chronic Mild Stress in Rodents as a Trans-             indicate that a highly reproducible MCA occlusion model with
lational Model of Clinical Depression                                     consistent functional deficits in the nonhuman primate can be
                                                                          obtained with the clipping of the distal portion of M1 for 2 h,
TA Swanson*1, G Lessard1, K Muravnick2, RP Giovanelli2, L                 if the above visual criteria of the infarcted cortex are met. The
Badura3, PA Seymour4, S McLean4, M Moalli1                                reproducibility of the model is important for future experimental
1Comparative     Medicine, 2Pathology, 3Translational Medicine,           therapeutic strategies.
4Neuroscience,   Pfizer Global Research and Development, Gro-
ton, CT                                                                   PS47 The Role of Positive Reinforcement Training in Creating
Chronic mild stress (CMS) is considered to be a predictive ro-            a Reproducible Surgical Stroke Model
dent model that recreates some of the pathophysiology present             S Rubino*1,2, M Sasaki2, J Kocsis2
in human depression. Anhedonia, a core symptom of clinical                1Comparative  Medicine, Pfizer, Inc., Griswold, CT; 2Neurosci-
depression, is characterized by the loss of interest in normally
                                                                          ence, Yale University, New Haven, CT
rewarding stimuli. This can be quantitatively assessed in the
rodent by measuring decreases in consumption of sucrose                   During the development of a reproducible surgical stroke
solution. Clinical depression has also been associated with a             model, it was necessary to develop neurological examinations
disruption in circadian rhythm. These two outcome measures                and behavioral tests for functional assessments. As part of the
were used to validate chronic mild stress as a translational model        neurological exam, African green monkeys (Chlorocebus aethiops)
of clinical depression. Experiments focused on optimization of            were trained to touch a target stick with their left hand or right
the stress paradigm, adding the use of telemetry to monitor ac-           hand and to track an object through horizontal and vertical
tivity rhythms, and validation of biomarkers. Approved by the             movements. Animals were also exposed to a puzzle feeder that
Pfizer IACUC, this model consists of exposure to a series of mild,        required the use of 2 hands to remove food rewards. For func-
random, unpredictable stressors which result in a state of anhe-          tional assessments, animals were trained to perform fine motor
donia, disruption of normal circadian rhythms, and weight loss.           grasping tasks including a well/reach task and a staircase task.
Stressors include periods of cage tilt, exposure to stroboscopic          These tests were timed and assessed prior to surgery as a baseline
light, white noise, and wet bedding. Strain and sex differences           for postoperative data collection. Behaviors were taught using
in response to CMS were examined in both Sprague-Dawley                   positive reinforcement. The training plans developed for these
and Wistar rats. Biomarkers such as serum and urinary corti-              tasks allowed us to successfully train all animals selected for the
costerone and lymphocyte counts were assessed. As predicted,              preliminary project.
sucrose consumption was decreased in response to the chronic
stress paradigm. However, reversal by antidepressant therapy              PS48 Establishment of a Nonhuman Primate Cognition Labora-
was inconsistent with published data and warrants further in-             tory
vestigation of this model.
                                                                          JA Vivian*1, JL Basile1, MR Tinsley1, R Morris1, N Poy2, D Brandt2,
                                                                          C Waugh2, L Santarelli1
PS46 Establishment of a Reproducible Surgical Stroke Model
                                                                          1Neuroscience, 2Comparative Medicine, Roche Pharmaceuticals,
in the Nonhuman Primate
                                                                          Palo Alto, CA
M Sasaki1, J Kocsis1, S Rubino*2,1
1Neuroscience,
                                                                          Due to their expanded social and behavioral repertoire, and exten-
               Yale University, New Haven, CT; 2Comparative               sive frontal cortical development, the establishment of a cognition
Medicine, Pfizer, Inc., Groton, CT                                        facility using nonhuman primates would appear relevant and an
Stroke is a leading cause of death in adult humans, and extensive         important endeavor for basic, pharmaceutical, and translational
research is needed to develop new therapies for stroke patients.          (preclinical to clinical) science. However, issues including the 3 Rs,
Abstracts of scientific papers 2008 AALAS National Meeting

and the special requirements for monkey research, have proven             (9 mitral, 2 aortic). We originally assumed that the main cause of
challenging. In this presentation, an overview will be provided for       mortality was due to surgeon error and immediately instituted more
the development of a research effort in neuroscience (specifically,       conservative measures for de-airing the heart and assuring hemo-
cognition) using nonhuman primates, explaining why monkeys are            stasis. After analysis of the data, we saw that while this was true
used, and providing some detail for key decision points, includ-          for aortic valves, for mitral implants the key cause of mortality was
ing the choice of monkey strain and gender; handling, housing             sheep-related. Since introducing the procedure changes we have
and enrichment concerns; veterinary and staff oversight; and              reduced surgeon-related mortality, and surprisingly also reduced
test model development (training and testing strategies). Briefly,        the incidence of fatal GI dysfunction and pulmonary edema. The
socially housed adult male cynomolgus monkeys were chair- and             exact mechanism for this improvement is unclear. This study shows
test system-trained using a modified Wisconsin General Test               that it is important to review records to determine the actual causes
Apparatus, operant chambers (lever boxes) and computer touch-             of mortality and that solving one problem can actually improve
sensitive screens (CANTAB). Subsequently, they were tested with           outcomes in other areas.
prototypic drugs effective in the treatment of Alzheimer disease
and schizophrenia. Monkeys readily learned the various tasks, and         PS51 Primate Tuberculosis Testing Comparison: Gamma In-
demonstrated cognition-enhancing effects after drug treatment.            terferon Assay and Mammalian Tuberculin
Taken together, these results marshal support for the unique value
of monkeys in the assessment of complex cognitive behavior, and           ST Shipley*1, T Coksaygan1, C McLeod1, DK Johnson2, LJ De-
hopefully, their predictive and translational value for the clinic.       Tolla1
                                                                          1Comparative   Medicine/Veterinary Resources, University of
PS49 Nasal Intubation in the Rabbit: A New Twist on a Dif-                Maryland School of Medicine, Baltimore, MD; 2Scientific Advi-
ficult Procedure                                                          sor, Harlan, Chester Springs, PA
J Stephens-DeValle*                                                       Tuberculosis (TB) is a highly infectious disease in nonhuman pri-
                                                                          mates (NHPs) causing high morbidity and mortality that is also
Department of Clinical Investigations, DD Eisenhower Army
                                                                          important for its zoonotic potential. A comprehensive diagnostic
Medical Center, Fort Gordon, GA
                                                                          program is essential for primate colony health management. The
Although nasal intubation has been described in the rabbit, there         gold-standard assay for TB screening in NHP colonies is the delayed
is limited literature concerning this technique. At our facility we       hypersensitivity test using mammalian old tuberculin (MOT) to
needed to intubate rabbits for a protocol that required the animals       marshal a delayed dermal hypersensitivity response over a 72-h
to be placed under general anesthesia twice, with a 2-min time span       period. A new primate diagnostic test is available measuring gamma
between the initial intubation and the final intubation. Initially, the   interferon release from sensitized lymphocytes in response to ex-
animals were orally intubated; however, after encountering difficul-      posure to TB antigens. This interferon release is then measured by
ties with oral intubation, nasal intubation was performed. During         ELISA assay. This gamma interferon assay patterns itself after an
the development of this technique, we learned several essential key       analogous test of human tuberculosis testing. The antigens used
points: total relaxation, modification of the classic blind position,     for the primate gamma interferon assay are 2 strains of myco-
the proper approach, and securing the tube. Contraindications for         bacterium (M. bovis and M. avium complex), whereas the human
nasal intubation include the possibility of introducing pathogens         assay uses purified protein derivative of human mycobacterium
into the lungs and the need for high oxygen flow rates. However,          (M. tuberculosis). A 2-y comparative evaluation of tuberculosis test-
clinically, neither item was an issue. At our facility, after a few       ing in primates was performed by alternating between MOT and
modifications and practice, nasal intubation was preferred as an          primate gamma interferon assays during both primate quarantine
easy, less traumatic method of rabbit intubation.                         and semi-annual facility TB testing postquarantine. Results from
                                                                          both types of testing were found to be equivalent, and both tests
PS50 Analysis of Early Postoperative Mortality in a Sheep (Ovis           correctly identified animals positive for naturally acquired TB
aries) Model of Mitral and Aortic Valve Replacement                       (culture-confirmed infections). The advantage of the interferon as-
                                                                          say is the ability to have results within 24 h of blood draw, which
JM Olin*, A Okos, S Meyer                                                 is 48 h faster than the MOT assay. However, the initial steps of the
Biological Resource Center, Edwards Lifesciences, Irvine, CA              interferon assay must be performed in-house within 6 to 12 h of
                                                                          blood collection using validated lymphocyte culturing procedures
Higher than expected mortality in early 2008 prompted us to de-
                                                                          by highly trained technicians. The interferon assay has limits based
velop a plan to improve early postoperative outcomes for surgical
                                                                          on cost of reagents, technician labor time, and intermittent delays of
valve replacement in sheep. The records of 99 consecutive animals
                                                                          test kit availability. Although the USDA has licensed this interferon
from Jan 2007 through Apr 2008 were reviewed to analyze the cause
                                                                          test in several species of NHPs, the MOT (delayed dermal hyper-
of death. Early mortality was considered to be any death or illness
                                                                          sensitivity) assay remains the principal recognized diagnostic test
requiring euthanasia within the first 7 d postimplant. Mortality was
                                                                          by regulatory agencies such as CDC and USDA. For these reasons
categorized as surgeon error, device complication, or spontaneous or
                                                                          the gamma interferon assay cannot be used alone in absence of the
sheep-related. Surgeon error included hemorrhage, emboli, or injury
                                                                          gold standard mammalian intradermal test.
to other cardiac structures. Some of the devices were prototypes or
atypical configurations; any mortality that was linked to a unique
device feature was categorized as a device complication. Spontane-        PS52 Methicillin-resistant Staphylococcus aureus Infection
ous or sheep-related mortality included untreatable GI dysfunction        and Cranial Abscessation in a Rhesus Macaque (Macaca
and pulmonary edema, and deaths for which no other cause could            mulatta)
be assigned. All implants were performed by the same surgical             BH Jasmin*1, JI Gold2, CH Vite3, SC Rankin4, J Baxter1, F Han-
team using sheep from the same source. In the 16-mo period, 57            kenson1,4
mitral and 42 aortic valve replacements were performed, with early        1University Laboratory Animal Resources, 2Department of Neu-
mortality distributed across surgeon error (4 mitral, 6 aortic), device
complication (1 mitral, 3 aortic) and spontaneous or sheep-related        roscience, 3Department of Clinical Studies-Philadelphia, Section
                                                                                          Abstracts of scientific papers 2008 AALAS National Meeting

Chief of Neurology,   4Department
                               of Pathobiology, University of             that the quinolone resistance for this isolate was plasmid-mediated
Pennsylvania, Philadelphia, PA                                            and transferable to a quinolone-susceptible E. coli. PMQR is clini-
                                                                          cally important when treating macaques prophylactically and for
A 7-y-old female rhesus macaque (Macaca mulatta) presented for
                                                                          bacterial infections, especially given the common clinical usage
clinically abnormal behavior within 24 h of surgery to relocate a
                                                                          of fluoroquinolones. Of the antibiotics tested, the shigella isolates
cylinder in a pre-existing head stage implant. This animal was used
                                                                          were only susceptible to trimethoprim/sulfamethoxazole and that
in an IACUC-approved behavioral study exploring mechanisms
                                                                          antibiotic was used to eradicate S. flexneri from the macaques. To
of learning and visual discrimination. Clinically, the animal was
                                                                          our knowledge, this is the first report of PMQR in S. flexneri isolated
intermittently depressed over the first 3 postsurgical weeks despite
                                                                          from monkeys.
antibiotic therapy and pain management. Serum chemistry and
complete blood count were within normal limits. During routine
cleaning and maintenance of the implant cylinders, abnormal tissue        PS54 Corynebacterium spp in Ferrets (Mustela putorius furo)
growth within the caudal cyclinder was noted. Cultures from the           and Macaques (Macaca mulatta)
cylinders grew methicillin-resistant Staphylococcus aureus (MRSA).        RP Marini*1, EM Buckley1, Y Peters1, P Cassiday2, JG Fox1
Paired samples were collected and submitted to a contract diag-           1Comparative
nostic laboratory specializing in culturing MRSA for confirmation                       Medicine, Massachusetts Institute of Technology,
and bacterial typing. Topical flushing treatments of the dura around      Cambridge, MA; 2pertussis and Diphtheria Laboratory, Centers
the implant were attempted with sterile saline, 1% chlorhexidine,         for Disease Control and Prevention, Atlanta, GA
and appropriate antibiotics without successful MRSA eradication.          The genus Corynebacterium contains organisms responsible for
A routine MRI for assessment of cylinder placement unexpectedly           diphtheria of humans (C diphtheriae and, sporadically, C ulcerans
identified a 1.7-cm space-occupying lesion in the left occipital lobe.    and C pseudotuberculosis) and a number of diseases of veterinary
A repeat MRI with contrast was then performed and the lesion              importance. Recent publications reporting the zoonootic trans-
was classified as an abscess with evidence of hemorrhage and              mission of the diphtheroid bacterium Corynebacterium ulcerans
meningitis. Approximately 5 mo after initial MRSA diagnosis, the          from cats and dogs to people motivated a survey of bacteriologi-
animal’s cranial implant was removed. During removal, purulent            cal isolates from cephalic implants (n = 73) and oropharynges (n =
material found at the implant site yielded MRSA. MRSA was also            78) in a colony of macaques (Macaca mulatta) and ferrets (Mustela
isolated from a sample of the actual hardware pieces. Following           putorius furo; n = 3 and n = 1, respectively). Bacteria identified
implant removal, the skull and dural surface were thoroughly              from cephalic implants by the apiCoryne system included C
flushed with alternating 1% chlorhexidine and saline. Cultures of         ulcerans (n = 40), C renale (n = 3), C pseudotuberculosis (n = 6),
these disinfected tissues were negative for MRSA. Due to its loca-        and C striatum (n = 1) in macaques and C ulcerans (n = 3) and C
tion, the cranial abscess could not be readily accessed for culture       pseudotuberculosis (n = 1)in ferrets. Oropharyngeal recovery of
but was presumed to be related to the chronic bacterial infection         coryneforms from macaques included C ulcerans (n = 7), C propin-
in this animal; no neurological abnormalities have been observed          quum (n = 2) and C auris (n = 1). C pseudotuberculosis was cultured
in this animal. This case exemplifies the difficulty in eradication of    from the oropharynx from the ferret harboring that organism in
MRSA, an emerging infectious disease in laboratory animal species,        its implant. Mixed infections of Corynebacterium spp and other
from nonhuman primates with chronic cranial implants.                     microorganisms were common in both macaques and ferrets.
                                                                          Human contacts of these animals (n = 34) were subjected to
PS53 Plasmid-mediated Quinolone Resistance in Shigella                    oropharyngeal culture to evaluate carriage and zoonotic spread
flexneri Isolated from Macaques (Macaca mulatta)                          of Corynebacterium spp. No humans shared organisms with ani-
                                                                          mals with which they had direct contact, although carriage of C
HR Martin*, Z Shen, EM Buckley, JG Fox                                    pseudotuberculosis (n = 1), C jejeukium (n = 3) and C striatum (n = 1)
Division of Comparative Medicine, Massachusetts Institute of              was demonstrated. Diphtheria toxin activity of select C ulcerans,
Technology, Cambridge, MA                                                 C renale, and C pseudotuberculosis isolates were negative in Elek
                                                                          tests. Sequence analysis of 16s ribosomal RNA genes of select
Plasmid-mediated quinolone resistance (PMQR) is an emerging is-
                                                                          macaque isolates revealed over 99% homology with reference
sue with bacterial species in the family Enterobacteriaceae. Typically,
                                                                          sequences for C ulcerans (n = 5) and 98.9% homology for C renale
quinolone resistance is chromosomally mediated; however, PMQR
was first reported in 1998 in Klebsiella pneumoniae. The first PMQR       (n = 1). To our knowledge, this is the first demonstration of puta-
determinant, QnrA, was a 218 amino acid protein protecting DNA            tive C ulcerans and C pseudotuberculosis infection of ferrets and
from quinolone binding. Two other Qnr-type determinants, QnrB             the first demonstration of cephalic implant infection of macaques
and QnrS, confer resistance to the quinolone, nalidixic acid, and         with C renale and C pseudotuberculosis. Molecular epizootiology,
increase the mean inhibitory concentrations of fluoroquinolones.          evaluation for virulence factors, and confirmation of isolate
Five different isolates of multi-drug resistant Shigella flexneri were    identity with 16s ribosomal RNA gene sequencing is ongoing.
isolated from the feces of 3 macaques. Antibiotic susceptibility test-    Personal protective practices used in the vivarium appear to have
ing was performed using disc diffusion and resistance to ampicillin,      prevented zoonotic transmission of these organisms.
amoxicillin + clavulanic acid, cephalothin, gentamicin, tetracycline,
enrofloxacin and nalidixic acid was noted. PCR was performed on           PS55 Diabetic Ketoacidosis in a Pregnant Rhesus Macaque
the shigella isolates for the presence of the qnrA, qnrB, and qnrS        (Macaca mulatta)
genes. Genomic DNA from all isolates was positive for qnrS. Plas-         E Paster*, L Colgin, J Wilk, G Maginnis
mid DNA was extracted and 4/5 isolates tested positive for qnrS.
A standard transformation protocol was followed using plasmid             Portland VA Medical Center, Portland, OR
DNA and E. coli competent cells. E. coli competent cells were used        A 13-y-old pregnant female rhesus macaque presented to the
as a control and were susceptible to nalidixic acid by disc diffusion.    clinic for partial anorexia. A fetal ultrasound had been performed
A transformed E. coli colony was isolated in the zone of susceptibil-     under sedation earlier that week. She had previously delivered
ity for nalidixic acid and confirmed to be resistant to nalidixic acid.   live infants vaginally in February of 2005 and 2006 and the infants’
The zone of enrofloxacin sensitivity was decreased, demonstrating         birth weights were 0.54 kg and 0.60 kg, respectively. Ultrasound
Abstracts of scientific papers 2008 AALAS National Meeting

revealed that the placental attachments were normal, there was no          R Duran-Struuck*1, PS Cho1, SG Moran1, A Teague1, K Fergu-
evidence of fetal distress, and the estimated gestational age was          son1, BV Lutton1, I Hanekamp1, KJ Wikiel1, B Horner1, DP Lo1,
118 d, based on biparietal diameter measurements. Two days later,          M Duggan1, SJ Arn1, B Billiter1, S Houser1, SV Westmoreland2,
there was suspected vomitus in the cage and the patient developed          DH Sachs1, CA Huang1
diarrhea. A follow-up ultrasound was performed and no fetal                1Surgery, Transplantation Biology Research Center, Massachu-
heartbeat was detected. A cesarean section was performed and               setts General Hospital—Harvard Medical School, Boston, MA;
a nonviable 0.273 kg fetus was removed. An in-house chemistry              2Pathology, New England Primate Center, Harvard Medical
analysis performed during surgery revealed metabolic acidosis.             School, Southborough, MA
Blood chemistry results revealed hyperglycemia, azotemia,
hypertriglyceridemia, hyperphosphatemia, hyperkalemia, and                 The Massachusetts General Hospital (MGH) miniature swine
panhypoproteinemia. Hematologic abnormalities demonstrated                 characterized at the major histocompatibility complex (MHC)
a mild leukocytosis with a minimal left shift and a glycosylated           serve as a model system for human clinical transplantation.
hemoglobin of >13%. Urinalysis showed a specific gravity of 1.011,         Sublines of these animals have been successfully inbred and now
mild glucosuria, and significant ketonuria. Gestational diabetic ke-       permit adoptive transfer studies in a large animal preclinical
toacidosis was suspected; however, as previous blood profiles were         model. We report here 5 cases of spontaneous chronic myeloid
lacking in this patient, pre-existing diabetes mellitus could not be       leukemia with similarities to human disease that have occurred
ruled out. Fetal macrosomia, a consequence of gestational diabetes         in our inbred miniature swine herd. Four of these cases occurred
in humans, was not present in this case. Despite aggressive fluid          within the highly inbred histocompatible sublines. All animals
therapy, the patient continued to decline and was euthanized due           presented with symptoms of weight loss, lethargy, and marked
to poor prognosis. Necropsy findings of the mother included acute          leukocytosis. Samples were obtained for complete blood count,
tubular necrosis, hepatic lipidosis, lymphatic ectasia and fibrin          peripheral blood smear, and flow cytometry analysis. Animals
thrombi in the heart and liver, along with placental infarcts. A           that were confirmed to have neoplasms were then euthanized. At
review of the current literature reveals that this is the first reported   necropsy, all animals had diffuse disease involvement of vascu-
case of gestational DKA in a rhesus macaque. To improve maternal           larized organs and severe hepatosplenomegaly. Peripheral blood
and fetal outcome, screening for diabetes should be considered             mononuclear cells and tissues were phenotyped by fluorescence
in obstetric cases that have a history of large fetuses or in those        activated cell sorting (FACS). In addition, all affected tissues were
patients that develop signs of morbidity.                                  assessed histopathologically. Taking under consideration the clini-
                                                                           cal presentation, FACS, gross pathology and histopathology we
                                                                           diagnosed these cases as chronic myelogenous leukemias. Some
PS56 Acute Respiratory Distress Syndrome in Two Rhesus
                                                                           of these tumors have been successfully established as in vitro cell
Macaques (Macaca mulatta)
                                                                           lines and are candidates to become the first leukemias to be used as
JJ Fremont*, RP Marini, JG Fox, AB Rogers                                  transplantable tumors in inbred and MHC-defined large animals.
Department of Comparative Medicine, MIT, Cambridge, MA                     Thus, MGH MHC-inbred miniature swine and established porcine
                                                                           tumors may provide useful preclinical translational models to
Acute respiratory distress syndrome (ARDS) is an important                 study immunotherapeutic approaches to treat human leukemia.
and potentially life-threatening complication in humans subse-
quent to a variety of primary insults, including noxious fume
                                                                           PS58 Epidemiologic Investigation of a Multidrug-resistant
inhalation, infection, and trauma. Clinically, animals with ARDS
                                                                           Staphylococcus aureus Outbreak in Two Macaque Colonies
present with progressive dyspnea and pulmonary edema. Physi-
ologically, ARDS is characterized by severe hypoxemia (partial             JR Baxter*1, BH Jasmin1, F Hankenson1, A Behrman2, S Rankin3
pressure of oxygen [PaO2]/fractional concentration of oxygen               1University Laboratory Animal Resources, University of Penn-
in inspired air [FIO2]<200). Histopathologically, the disease is           sylvania, Philadelphia, PA; 2Occupational Medicine, Hospital of
characterized by 2 phases. In the initial phase, the lungs are             University of Pennsylvania, Philadelphia, PA; 3Clinical Microbi-
edematous admixed with variable numbers of neutrophils,                    ology, Ryan Veterinary Hospital, Philadelphia, PA
alveolar hemorrhage, and atelectasis. The disease progresses
to the fibrinoproliferative phase 7 to 10 d postinsult. Here we            Methicillin-resistant Staphylococcus aureus (MRSA) is a multi-drug
describe for the first time ARDS in 2 rhesus macaques associated           resistant bacterium that has become a prevalent pathogen among
with complications following experimental surgical craniotomy.             people and is now emerging among the veterinary patient popula-
ARDS in 1 monkey was attributed to sepsis resulting from                   tion, underscoring its potential as a zoonotic. This microorganism
meningeal infection with Staphylococcus aureus, group A Strepto-           has not been reported in nonhuman primates (NHP). MRSA was
coccus pyogenes, and Corynebacterium ulcerans. ARDS in the other           diagnosed in 3 clinically compromised animals within 2 macaque
monkey was ascribed to direct neurogenic injury. Despite the               colonies, Colony A and Colony B, in one of our facilities. The 2
different etiologies, both monkeys developed clinical features             rhesus macaques in Colony-A were used for IACUC-approved
of ARDS and syndrome-defining histopathologic criteria includ-             neurobehavioral studies requiring invasive cranial implants, while
ing edema with intra-alveolar neutrophils, fibrinohemorrhagic              the cynomologus macaque in Colony B was used for IACUC-
effusions with crescentic membranes, and interstitial vascular             approved transplant research requiring immunosuppression.
degeneration. ARDS may be underreported in macaques and                    Containment and eradication planning was critical because these
other nonhuman primates. Recognition and aggressive treatment              chronic studies involve numerous animals. Additional samples
of ARDS at an early stage may improve survival rates in these              were analyzed from colony animals using chrome agar enrichment
highly valuable animals.                                                   techniques specific for MRSA. Colony B had no additional positive
                                                                           animals, while Colony A had 2 additional implants culture posi-
                                                                           tive from a pool of 5 clinically healthy animals, yielding an overall
PS57 Immunophenotypic Characterization of Spontaneous
                                                                           colonization rate of 57% in Colony A. Body swabs were obtained
Myeloid Leukemias in Adult MHC-defined and Histocompat-
                                                                           from the individuals with colonized implants to determine if
ible Miniature Swine: Towards the Development of a Large
Animal Transplantable Tumor Model                                          NHP can be colonized in a pattern similar to humans colonized
                                                                           with MRSA. Nasal swabbing is the gold standard test for human
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

colonization; however, NHP with colonized axillary and perineal        structurally complex housing configuration with an exercise
areas did not consistently have positive nasal cultures. Our vet-      program. In the initial housing system, cats were housed either
erinary and husbandry team reviewed personnel and equipment            singly (males) or in isosexual (females) dyads or triads in pens
trafficking and animal practices that may have contributed to the      with minimal PVC furniture for climbing. Two new housing rooms
outbreak. An interdepartmental collaborative effort was made to        with 6 pens in each room were designed from recycled dog pens
screen all personnel with NHP contact (n = 44) for MRSA. DNA           so the cats could be divided into 2 groups. In each housing room
fingerprint analysis of the single human isolate and all NHP           the 6 pens were contiguous by wall-mounted tunnels and side
isolates were of the healthcare-associated (HA) strain of MRSA.        doors to allow the cats continuous free access among all of the
Our investigation led to improved hand washing, sanitization,          pens. Vertical complexity was achieved through placement of wall-
aseptic technique, and heightened institutional awareness of the       mounted staggered perches, varying height tunnels between pens,
importance of consistent use of personal protective equipment to       and commercially available climbing furniture with hammocks for
prevent the potential transmission of zoonotic agents.                 resting and sleeping. Eventually all 16 cats were combined into 1
                                                                       of the 2 rooms. The other room was converted into an exercise/
PS59 Animal Health and History: Electronic System for Captur-          play room with an area rug, floor toys, and suspended toys. The
ing Medical and Husbandry-related Data                                 housing room and exercise/play room were connected to one
                                                                       another through a walled off ante-room to allow cats direct access
J Sims*2, SA Mischler1, J Ashline1                                     to the exercise/play room 4 d per wk for 4 to 6 h per d. The incor-
1Drug   Safety, 2Information Services, Wyeth, Chazy, NY                poration of the exercise/play room into the enrichment program
                                                                       resulted in the desired weight loss to a healthy weight range and
Our facility’s husbandry and veterinary activities in drug safety
                                                                       body condition score. Additionally, the response of care staff and
were originally built around a paper-based system. All informa-
                                                                       regulatory agencies to this program was positive.
tion for animals was recorded and maintained on paper, including
medical records, scheduled tasks (physical exams, body weights,
vaccinations, and so forth), and training activities. The paper        PS61 Cage-level Barrier System for Primates and Other Non-
system was an effective way to maintain compliance with the            rodent Species Application for Biodefense Select Agent and
Food and Drug Administration’s (FDA) 21 CFR Part 58 (Good              Infectious Disease Studies (ABSL-3)
Laboratory Practice for Non-clinical Laboratory Studies) and 21        DK Johnson*1, D Allen1, LJ DeTolla1, Y Gal1, J Huss1, SD Reyn-
CFR Part 11 (Electronic Records and Electronic Signature) guide-       olds2
lines. The paper system, however, was time consuming and made          1Harlan
                                                                             Sprague Dawley Inc, Indianapolis, IN; 2CAES Group,
it difficult to perform certain tasks, including analyzing trends,
                                                                       M/E Engineering P.C., Syracuse, NY
inspecting data for internal quality control, reviewing data for
poststudy activities, and analyzing animal care staffing require-      A cage-level barrier system for primates that is also suitable
ments. Data for animals was also stored in disconnected systems        for other large laboratory animals was engineered with reason-
that required duplicate data entry and complicated data searches.      able adaptations of flexible film isolators, along with minor
Wyeth formed a team of veterinary, husbandry, and information          modifications of conventional cages. The rapid growth of funded
services staff to gather requirements and develop a custom ap-         laboratory animal research for biodefense studies, as well as in-
plication to replace the paper system. The project had a broad         fectious disease investigations, require the use of special agents,
scope so the agile software development framework was used to          highly contagious infectious organisms, aerosols, toxins, and new
incrementally deliver functionality. The result was a web-based        vaccine testing in primates and non-rodent species. This com-
application called Animal Health and History (AHH), which is           bined flexible film isolator and cage system establishes a barrier
currently at version 2.1. Wireless tablet PCs are used to record       at the cage level for these larger laboratory animals. It allows the
data. Other validated systems were also integrated with AHH to         safe conduct of infectious disease studies and meets occupational
minimize duplicate data entry and facilitate searches. AHH has         health requirements for containment of infected animals. The
been in production in a validated state since January 31, 2005. Data   safety protection to personnel and the environment meet insti-
and reports from AHH have been used to meet both USDA and              tutional and governmental guidelines. A validation engineering
FDA requirements. Quality control activities have been facilitated     study was conducted to verify the operational effectiveness by
by making the data available electronically. Retrospective pain and    using the technology of Computational Fluid Dynamics (CFD).
census information is easily recorded and reported to the USDA.        This credible engineering program accurately determines airflow,
Husbandry and veterinary activities required in FDA submissions        velocity, distribution, thermal loads, and related aerodynamic
are easily analyzed and retrieved. AHH continues to evolve as new      parameters. Multiple examples of leak scenarios, including van-
functionality is added. The ultimate goal is to provide a completely   dalism, were evaluated. The CFD data provides documentation
paperless system to capture husbandry and veterinary data, while       for the successful approval of this cage-level barrier system. In
leveraging related data in other systems and maintaining compli-       addition, the system has application for large laboratory animals
ance with applicable FDA and USDA guidelines.                          assigned to immunodeficiency studies.

PS60 Development of a Complex Housing Arrangement with                 PS62 Working with State Regulating Agencies: The Value of
Integration of an Exercise Program for Laboratory Cats                 State Biomedical Research Organizations
M Wilkinson*3, L Wood1, A Boyd1, S Watson2, A Wilkinson1               JS Ellis*
1 Animal Resources Unit, Alcon Labs, Fort Worth, TX; 2De-              PA Society for Biomedical Research, Camp Hill, PA
partment of Psychology, University of Southern Mississippi,            In addition to federal laws and regulations, in many states
Hattiesburg, MS; 3Veterinary Sciences, Tulane National Primate
                                                                       research facilities must also comply with state laws and regula-
Research Center, Covington, LA                                         tions. Most often research facilities focus on the federal laws and
Over the course of 36 mo, 16 domestic shorthair cats were transi-      regulations, which are well known and discussed at national
tioned from a standard feline housing system to a group-housed,        meetings. State laws and regulations may not be as well known
Abstracts of scientific papers 2008 AALAS National Meeting

throughout the research community, and state agencies may not          Animal Resources Center, University of Texas Medical Branch,
fully understand the research process. Research facilities may         Galveston, TX
not even be aware that certain state agencies have regulations         The Galveston National Laboratory (GNL) at the University of
that apply to them. The state biomedical research associations         Texas Medical Branch in Galveston Texas is 1 of 2 state-of-the-art
can play a key role in educating both its membership and state         national biocontainment laboratories constructed under grants
agencies. This is only one of the many benefits of membership          awarded by the National Institute of Allergy and Infectious
in the state biomedical research organizations. This presenta-
                                                                       Diseases/National Institutes of Health (NIAID/NIH) that will
tion will discuss 2 such examples that have recently taken place       support Biosafety Level 2 through Biosafety Level 4 research.
in Pennsylvania. The first example is the education of the PA
                                                                       While the GNL will provide much-needed research space for
Department of Agriculture on why federally registered research
                                                                       naturally occurring emerging diseases, as well as for viral and
facilities should be exempt from legislation designed to improve
                                                                       bacterial agents, attracting an exceptionally talented and mul-
the conditions of dogs living in commercial kennels. The second
                                                                       tidisciplinary team of techs for high-biocontainment work is a
case involves efforts to educate the PA Game Commission on
                                                                       daunting endeavor. Our initial criteria for a Animal Resources
research needs, as well as to educate research facilities on the
                                                                       Biohazard Technologist required AALAS certification at the LAT
Game Commission’s regulations. Since many voices working
                                                                       or LATG level, a minimum of 3 y experience in research, and 1
together are stronger than a single voice, working collectively as
                                                                       y experience in biocontainment. Finding ideal candidates that
a research community makes sense. The value of state biomedi-
                                                                       possessed our requirements was found to be nearly impossible.
cal research organizations is exemplified by the coordination of
                                                                       After seeing many applications that had met 1 or 2, but not all,
this research community.
                                                                       of the requirements, we modified our criteria to fit the applicants
                                                                       we were seeing. These modifications opened a larger pool of
PS63 A Novel Web-based Approach to Ethical Committee                   applicants for us, and we used novel recruiting strategies. We
(IACUC) Functions: Managing Protocols, Meeting Minutes,                took our show on the road, using platform sessions at AALAS
and Inspections                                                        branch meetings and attending job fairs at regional universi-
CM Pate*1, B Bell1, R Fosse2, A Staubi3                                ties. Our plan was to hire the traditional applicants, give them
1Laboratory
                                                                       intensive biocontainment training in animal husbandry and
            Animal Science & Welfare, US, Sanofi-Aventis,              procedures, provide AALAS certification “boot camp,” and pro-
Bridgewater, NJ; 2 Laboratory Animal Science & Welfare,                vide concurrent training in quality system and GLP standards.
Northern Europe, Sanofi-Aventis, Montpellier, France; 3 a-Tune         AALAS certification at the LAT level within 1 y is a condition
Software AG, Darmstadt, Germany                                        of employment. The proliferation of Animal Biosafety Level 3
The Consolidated Laboratory Animal Database (COLADA) is a              and 4 labs throughout the country has left an increased gap of
novel web-based solution we developed to provide our organiza-         demand outweighing the supply of biocontainment technicians.
tion with a flexible system that automates the office workflow in      The most realistic solution is to hire traditional animal techni-
a completely transparent manner. COLADA provides automated             cians and train internally while time is not on your side. Doing
reminders to researchers, management, and the ethical committee        this entails an entirely new problem with retention.
on required actions such as notices of annual reviews or inspec-
tion findings that may require action. The system creates protocol     PS65 Handling during Routine Care Affects Responses of
documents using a “dictionary” system populated with techniques        Laboratory Rats toward Humans
and methods that are approved by the ethical committee. The
researcher is able to create new protocols using a plug-and-play       S Cloutier*, J Panksepp, RC Newberry
system of adding items listed within the dictionary. Because the       VCAPP, Washington State University, Pullman, WA
system is web based, each protocol is easily tracked through the       We hypothesised that the type of handling used by caretakers
workflow by researchers, management, and the ethical committee.        during routine care of laboratory rats affects fear and anxiety
The program checks training records of individuals by technique        responses in behavior tests used in biomedical research. We as-
and immediately shows if an individual has the training required       sessed caretaker handling effects on male Sprague-Dawley rats
to perform the task. This feature provides the principal investi-      (Rattus norvegicus; Sim:[SD]fBR Albino; found free of internal
gator immediate feedback regarding the training qualifications         and external parasites and disease by quarterly serology and
without having to cross-reference outside paper or electronic files.   parasitology, and yearly necropsy, evaluations) in 4 treatments
COLADA is an improvement on the current internally written pro-        (n = 8 rats/treatment) between 57 and 74 d of age. The standard
tocol review software, which only provides a platform for protocol
                                                                       (control) treatment involved rats being handled on cage cleaning
document generation and review but does not manage workflow            days (2 times/wk) only (picked up by tail and placed in clean
or any other administrative aspects of the ethical committee. CO-      cage). The passive hand treatment involved rats being exposed
LADA is an integrated system that also provides a platform for         to a motionless hand for 2 min daily. Pinned rats were restrained
managing meeting minutes and inspections, generating reports,
                                                                       on their backs for 2 min daily. Tickled rats were tickled for 2 min
and documenting workflow of review and approval of the re-             daily. Behavior was assessed between 78 and 80 d of age in a hu-
ports. Our company continually seeks to improve animal welfare         man approach test (2 min without hand [HA], followed by 3 min
compliance and ethical committee review. A global initiative of        with a familiar or an unfamiliar hand); an elevated plus maze
harmonizing ethical review of research protocols across multiple       (EPM), and a cat odor (CAT) test. Ultrasonic vocalizations (50
countries has been initiated. COLADA was developed as a web-           kHz, interpreted as evidence for positive affective states) were
based tool to manage compliance for the German, American, and          recorded during each test. Tickled and pinned rats emitted more
French sites using animals for research.                               50-kHz calls with a familiar hand than rats from the passive hand
                                                                       and standard treatments (P = 0.01). Tickled rats reared more than
PS64 Recruiting Qualified Animal Technicians for a High-               rats from the pinned and standard treatments in the HA (P =
biocontainment Animal Facility                                         0.05) and with a familiar hand (P = 0.01). Tickled rats spent more
NS Linde*, MK Tate, M Maccallum                                        time near, and made more contact with, the familiar hand than
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

rats from the standard and passive hand treatments (P ≤ 0.01).        contract. The members could explain findings and corrective
Rats in the standard treatment made the fewest entries into the       action in terms that coworkers would relate to and would be
area close to the unfamiliar hand (P = 0.02). Treatment did not       more likely to institute since they were not being directed from
affect behavior in the EPM and CAT. In conclusion, incorporat-        a supervisor or management personnel. The goal was to try to
ing playful handling (such as tickling) into the routine care of      bridge the gap between staff and management by including
laboratory rats reduces fear of familiar and unfamiliar humans        staff in the resolution process, teach staff to identify and correct
but not general anxiety.                                              problem areas, improve communication between management
                                                                      and staff regarding improvements to processes, and ultimately
PS66 Benefits of Postapproval Monitoring                              reduce the number of findings for each department.
CM Malinowski*
                                                                      PS68 Extracellular Matrix Adjuvant for Cancer Vaccines
Center for Comparative Medicine, Northwestern University,
Chicago, IL                                                           MA Suckow*1, P Hall2, W Wolter1, V Sailes1, M Hiles2
                                                                      1Biological Sciences, University of Notre Dame, Notre Dame, IN;
Monitoring of animal use protocols for compliance is a require-       2Cook
ment of the Animal Welfare Act, the Guide for the Care and Use of             Biotech, Inc., West Lafayette, IN
Laboratory Animals, the Public Health Service Policy on Humane        Most vaccines include an adjuvant of some type, a compound
Care and Use of Laboratory Animals, and the United States             which enhances the immune response to the immunogens.
Government Principles for the Utilization and Care of Vertebrate      Currently, most commercial vaccines use adjuvants based on
Animals Used in Testing, Research and Training. However, the          aluminum salts or combinations of such salts, such as alum.
methods used to accomplish this monitoring are not specified.         Some studies have used specific cytokines as adjuvants, though
Postapproval monitoring can therefore be a major hurdle for           cytokines typically enhance specific features of the immune re-
institutions to face. In order to accomplish this task, a postap-     sponse and may be unstable outside of very controlled storage
proval monitoring program was developed and implemented at            conditions. We have noted that an extracellular matrix material,
our institution. Postapproval monitoring is a program through         small intestinal submucosa (SIS), attracts large numbers of an-
which an IACUC and the institution itself can ensure and              tigen processing cells (macrophages) as it is incorporated into
document program integrity, compliance with the regulations           surrounding tissue; this feature suggests that SIS could serve
and guidelines, and adherence to protocols. At our institution,       as an effective vaccine adjuvant. To test the hypothesis that SIS
postapproval monitoring is conducted on individual protocols.         can enhance the immune response to cancer antigens, groups of
Procedures are observed with focus on the following areas of          5 Lobund-Wistar rats were administered 1 × 106 PAIII prostate
the protocol: personnel, study procedures, anesthesia, surgery,       cancer cells subcutaneously over the flank. The animals under-
postsurgical care, euthanasia, recordkeeping, and the labora-         went aseptic surgical debulking of the tumor mass 14 d later. At
tory environment. The benefits of postapproval monitoring are         the time of tumor resection, animals were either left untreated,
numerous and include the ability to identify any problems that        vaccinated with 5 × 106 glutaraldehyde-fixed tumor (GFT) cells
may occur and deal with them before they get to a point where         harvested from a subcutaneous PAIII tumor; vaccinated with
they need to be reported to regulatory agencies. The program          GFT cells that had been grown on SIS for 3 d prior to fixation,
also helps foster an environment of openness, encourages the          or vaccinated with fixed SIS without GFT cells. In cases where
investigative staff to engage the animal facility and IACUC per-      animals were vaccinated, vaccines were administered directly
sonnel when they need assistance, and provides an educational         onto the tumor bed. Animals were euthanized 21 d later, and
and training forum for the research staff. This program has met       regrown tumors weighed. While there were no significant dif-
with much success at our institution, and to date 39 visits have      ferences in rats left untreated or vaccinated with GFT cells or SIS
been conducted.                                                       alone, tumor weight was reduced by greater than 50% in rats
                                                                      vaccinated with GFT cells on SIS, significantly less (P < 0.005)
PS67 New Approach to Quality Assurance                                than controls. Further, only 40% of rats vaccinated with GFT cells
                                                                      on SIS had pulmonary metastases, compared to 100% of controls.
K DeMuro*                                                             These data demonstrate that cells and immunogens affixed to
1   SoBran, Bethesda, MD; 2 NIH/DVR, Bethesda, MD                     extracellular matrices such as SIS offer an outstanding platform
                                                                      for vaccine adjuvant development.
In an animal facility, quality assurance (QA) is a part of each
individual’s responsibility and can be demonstrated as review-
ing one’s own work, supervisory checks at the end of the day,         PS69 Clinical Monitoring Study of Rhesus Macaques (Macaca
management walk-throughs, and a formal QA program. It can             mulatta) Given Humanized Monoclonal Antibody
be difficult to incorporate a QA mentality into staffs that are       W Porter*1, A Wickham1, C Johnson1, A Stadheim2, J Drummond3,
not familiar with the QA process. In order to improve upon our        B Doerning1, W Strohl3
performance and decrease the number of QA findings, we imple-         1Laboratory
mented QA committees that were made up of the QA manager,                         Animal Rescources, Merck Research Laboratories,
                                                                      Rahway, NJ; 2GlycoFi, Lebanon, NH; 3Biologics Research, Merck
program manager, training staff, and a representative from each
                                                                      Research Laboratories, West Point, PA
building within the contract. There was a separate committee for
the different areas of the facilities (cage wash, animal caretaker,   When humans are intravenously dosed with humanized
and veterinary technician). During these meetings, the repre-         monoclonal antibody (huMab), any resulting infusion reactions
sentatives discussed routine findings from the buildings and          are usually minor; however, on rare occasions fatalities have
the corrective actions and recommendations to prevent their           occurred. Due to this phenomenon, the short- and long-term
occurrences. The representatives were then asked to present the       health status of nonhuman primates on huMab research studies
information from the meeting to their peers at their building’s       could be at risk and make their use on future studies prob-
next weekly training session. This provided a more consistent         lematic. These considerations led us to closely monitor a very
corrective action plan amongst the 6 facilities that are under this   comprehensive range of clinical parameters in unanesthetized
Abstracts of scientific papers 2008 AALAS National Meeting

adult, male rhesus macaques (Macaca mulatta) during and after          of tissues prior to spontaneous death without significantly im-
dosing with huMab. Twelve animals were intravenously dosed             pacting longevity outcomes.
with huMab; repeat dosing occurred 21 d later. Four animals
were dosed once with vehicle control. Animals were monitored           PS71 Refinement of Mouse Telemetry Transmitter Implantation
extensively for 24 h after each dosing, and were followed for          using 3D Microultrasound-guided Surgery
up to 135 d for changes in general health and for potential on-
set of immune-mediated complications such as anaphylaxis or            X Shen*1, C Ahn2, B Campbell3, P Zhou2, M Bunzel3, J Mane1, C
immune-complex disease. We monitored and tracked the follow-           Zhang3, P Cheng2, R Sophie2, D Cully2, B Doerning1, M Walker
ing; skin/mucous membrane coloration, physical examination             III3, T Cai2, C Johnson1
including cardio-pulmonary auscultation and heart/respiration          1Laboratory Animal Resources, 2Cardiovascular Diseases, 3Im-
rate, body weight/temperature, behavior, blood cell count and          age, Merck & Co., Inc., Rahway, NJ
chemistries, total serum complement activity (CH 50 assay),
                                                                       Previously, we reported implementation and characterization of
erythrocyte sedimentation rate, urinalysis, and other aspects
                                                                       a radiotelemetry system measuring blood pressure with C57BL/6
of health status. Only 2 parameters were altered with statistical
                                                                       mice. In the process of expanding the system to other strains of
significance (decreased lymphocyte counts and increased CH
                                                                       mice, we found that the length of the carotid artery varies from
50 serum activity) in the huMab group compared with the con-
                                                                       strain to strain of mice, or even animal to animal within the
trol group, and these transient physiological changes occurred
                                                                       same strain, making consistent catheter placement a challenge.
only during the first 24 h after the initial dosing. From this we
                                                                       Surgical placement of the PA-C10 transmitter catheter resulted
conclude that huMab administration did not adversely affect
                                                                       in a high variability in the blood pressure readout, and a sub-
the health of the animals in the study, through at least 135 d. We
                                                                       sequent lower success rate of the telemetry modeling system.
consider these animals to be clinically normal and suitable for
                                                                       Ultrasound-guided surgical techniques have been successfully
use in other types of studies. To our knowledge, this is the first
                                                                       used to minimize invasiveness and to increase effectiveness, lead-
report of a comprehensive monitoring of rhesus macaques for
                                                                       ing to a reduced procedure and recovery time in animals. Our
acute and chronic health effects after dosing with therapeutic
                                                                       group introduced a quantitative 3D-microultrasound (Q3DMU)
levels of huMab.
                                                                       to improve the surgical catheter placement accuracy and blood
                                                                       pressure recording stability in several strains of mice. Standard
PS70 Endpoints Related to Imminent Death in Mice Used for              surgical procedure was followed in implanting a PA-C10 trans-
Longevity Research                                                     mitter to the branch level of the aorta by way of the left carotid
MA Ray, N Johnston, S Verhulst, LA Toth*                               artery. Real-time adjustments were then applied while imaging
                                                                       with the Q3DMU such that the probe tip is precisely placed at
Southern Illinois University School of Medicine, Springfield,
                                                                       the aortic arch region of the left carotid. Following a standard
IL                                                                     postsurgical recovery process (2 wk), the blood pressures were
Despite anticipated health problems, aged mice can be found            recorded to document successful placement. We have applied
dead without preceding signs of illness. The goal of this study        this method with 3 strains of mice (C57, BPH and BALB/c), and
was to identify objective criteria for prediction of imminent death    a success rate of over 90% was achieved with ultrasound-guided
in mice used in longevity research. ICR mice (retired breeders;        technique, based on quality of blood pressure measurements.
58 females and 47 males) were subcutaneously implanted with            This modeling system addresses the 3Rs by refining the radiote-
an identification chip that also allowed remote measurement            lemetry placement procedure and reducing the experiment time
of body temperature. Male mice were housed individually                and number of animals required.
to avoid injury due to fighting; female mice were assigned to
either group (10 mice per cage) or individual housing. Mice            PS72 Sustained Delivery of GLP-1 to Treat a Rat Model of Type
were evaluated weekly for body temperature, weight, body               2 Diabetes and Obesity
condition, respiratory rate and character, dehydration, posture,
hair coat condition, spontaneous ambulation, and response to           DH Moralejo*1, O Yanay3, J Fuller2, R Tatake4, R Barton4, A Lern-
manipulation. Twenty-four mice underwent euthanasia based              mark2, W Osborne3
on clinical signs (inability or unwillingness to walk, lack of overt   1Comparative    Medicine, 2Medicine, 3Pediatrics, University of
response to manipulation, large or ulcerated tumors, or palpable       Washington, Seattle, WA; 4Boehringer Ingelheim Pharmaceuti-
hypothermia), and 81 were monitored until spontaneous death            cals, Inc., Ridgefield, CT
occurred. Mice that underwent euthanasia were killed at a mean
                                                                       Type 2 diabetes (T2D), a common disorder with a prevalence
age of 25.3 ± 1.3 mo, whereas the mean age at death among mice
                                                                       of 4% to 5% that is increasing worldwide, is associated with
that died spontaneously was 24.5 ± 0.7 mo. Housing and gender
                                                                       shortened life expectancy due to late diabetes complications;
did not significantly influence outcome measures. Weight loss
                                                                       thus, novel gene therapy treatments are warranted. We treated
was the most common and earliest sign of imminent death. Mice
                                                                       a leptin receptor-deficient rat model (DR.lepr/lepr) of T2D and
lost about 20% of their weight during the 3-mo period that pre-
                                                                       obesity with encapsulated allogeneic vascular smooth muscle cells
ceded death, with about half of that loss occurring during the last
                                                                       (VSMC) genetically modified to secrete glucagon like-peptide-1
week of life. Body temperatures fell by over 1 °C during the last
                                                                       (GLP-1). GLP-1 exhibits biological and clinical effects that include
week of life. Within 24 h of death, 43% showed slow or labored
                                                                       stimulation of glucose-dependent insulin secretion, insulin biosyn-
respiration. Thus, monitoring weight as a preliminary marker,
                                                                       thesis, beta cell proliferation, neogenesis and induction of a satiety
with hypothermia, respiratory difficulty, and/or precipitous
                                                                       signal. Rat VSMC transduced with a retrovirus encoding GLP-1
weight loss as penultimate markers, appears to allow prediction
                                                                       and selected in G418 secreted 294.4 pg GLP-1 per 106 cells per 24
of death to within 1 wk. Among the mice used in this study, 1 wk
                                                                       h of GLP-1. We seeded encapsulating devices with 107 transduced
is about 1% of the median life span. Closer evaluation of high-
                                                                       cells and implanted a single device SQ into random fed male
risk mice could potentially allow even more accurate prediction.
                                                                       DR.lepr/lepr rats and monitored blood glucose, weight, and islets.
Euthanasia of mice based on these criteria may allow collection
                                                                       Blood glucose levels decreased after surgery and fell within the
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

normal range (90 to 120 mg/dl) 5 d after cell implantation. For the      macaques. Currently, there are 5 well-described serotypes of SRV
27-d period before devices were removed, the blood glucose level         that are all genetically related to the prototypic type D retrovi-
mean was 112.8 ± 41.7 mg/dl (n = 2) without hypoglycemia. After          rus, Mason-Pfizer monkey virus (MPMV or SRV3). Retroviruses
27 d we removed the encapsulation devices, and although blood            are enveloped, single-stranded RNA viruses that can reverse
glucose levels rose to a mean of 160.7 ± 46.1 mg/dl (n = 2), they        transcribe their RNA genome to DNA and then integrate into
did not return to the hyperglycemic levels recorded before surgery       the host’s genomic DNA, resulting in a persistent, lifelong virus
(>350 mg/dl). In rats treated with GLP-1 delivery, pancreatic sec-       infection. Depending upon various host, virus, and environmen-
tions stained for insulin and H&E showed enlarged islets without         tal factors, SRV infection can vary from nonpathogenic to an
multifocal vacuolization and with normal morphology, and an-             immunosuppressive AIDS-like disease that results in death from
giogenesis with islet proliferation and neogenesis, respectively.        opportunistic infections. Diagnosis of SRV infection is difficult,
These data indicate we achieved sustained delivery of GLP-1 to           since not all SRV-infected macaques mount a robust humoral im-
treat diabetes in our rat model of T2D and obesity.                      mune response to the virus and SRV-infected macaques are often
                                                                         negative by polymerase chain reaction (PCR). This presentation
PS73 Comparison of Two Euthanasia Methods for Zebrafish                  will describe the development of a truncated, recombinant SRV
(Danio rerio)                                                            glycoprotein 20 (rgp20) antigen for use in SRV serodiagnosis.
                                                                         The antigen was produced as a chimeric fusion protein with a
JM Wilson*, AJ Carty                                                     polyhistidine tag in a baculovirus expression system, and was
University Laboratory Animal Resources, University of Penn-              partially purified by metal affinity chromatography. The rgp20
sylvania, Philadelphia, PA                                               antigen was then used to develop a Luminex xMAP-based as-
                                                                         say for SRV1-5. Diagnostic results from a single accession of 45
The 2007 “AVMA Guidelines on Euthanasia” and the ILAR report
                                                                         cynomolgus macaques (Macaca fascicularis) from an SRV endemi-
Recognition and Alleviation of Pain and Distress in Laboratory Animals
                                                                         cally infected colony revealed that 42 of 45 animals tested were
state that hypothermia (also referred to as rapid cooling) is an
                                                                         positive for SRV by serology, PCR, or both. Of the 42 animals
unacceptable method of euthanasia in fish. Although no scientific
                                                                         that were positive for SRV, 36 were positive by serology using
justification is provided as to why rapid cooling is unacceptable,
                                                                         SRV2 whole virus lysate as antigen and 41 were positive by
there is speculation that ice crystal formation occurs in the tissue
                                                                         serology using rgp20 as antigen. All 5 samples that were serol-
during rapid cooling, which is presumably painful. In order to
                                                                         ogy positive using rgp20 but negative using SRV2 whole virus
assist Institutional Animal Care and Use Committees about the
                                                                         lysate were confirmed as SRV-infected by PCR. When compared
most humane method of euthanasia in zebrafish (Danio rerio), we
                                                                         to assays that use partially purified SRV2 whole virus lysates as
hypothesized that rapid cooling in zebrafish is a humane method
                                                                         antigen, the rgp20 assay has enhanced diagnostic performance
of euthanasia, due to a more rapid time between cold exposure and
                                                                         characteristics, including improved diagnostic sensitivity and
death, with minimal signs of distress. Zebrafish were individu-
                                                                         specificity.
ally euthanized by immersion in 1 of 2 solutions. Twenty-three
zebrafish were placed in an ice-water bath (<4 °C) and 23 were
placed in MS-222 solution (250 mg/l; 25 to 30 °C). The time from         PS75 Intracardiac Pacing Lead in Telemeterized Cynomolgus
exposure to the solution until the fish were unable to swim and          Monkey (Macaca fascicularis): Evaluation of Refinements
right themselves, as well as the cessation of gill movement (death),     from 2005 to 2008
were noted. Specific signs of distress were also recorded. Two           M Volberg*, N Poy, P Harris, M Engwall
minutes following the last opercular movement, animals were
placed in a “recovery tank” of system water to determine the             Pfizer Global Research & Development, Groton, CT
effectiveness of each euthanasia solution. Tissues were then col-        Telemetry is widely used as a noninvasive method of collecting
lected for histologic analysis. Time to cessation of gill movement in    continuous blood pressure and ECG data in canine and nonhu-
animals euthanized by rapid cooling ranged from 5 to 9 s (average        man primates particularly for safety pharmacology studies. The
= 7.13 s). Time to death with MS-222 was significantly longer (P =       use of an intracardiac bipolar pacing lead in conjunction with
0.0005) and ranged from 29 to 124 s (average = 53.52 s). Signs of        a telemetry transmitter has many advantages over traditional
distress occurred in 39% of animals euthanized by rapid cooling          subcutaneous lead placement. Advantages include improve-
as opposed to 100% of fish euthanized with MS-222 (P < 0.0001).          ment of signal quality, higher signal-to-noise ratio and minimal
Of the zebrafish euthanized with MS-222, 17.4% animals regained          movement artifacts, all resulting in more consistent waveforms
consciousness in the recovery tank after exposure to MS-222 solu-        to enhance data analysis. During the early development of the
tion for 2 min past the last observable opercular movement. This         model, the bipolar pacing lead with 3 cm distance between
phenomenon did not occur with any animals euthanized by the              electrodes was used. In the course of study use, inconsistency
rapid cooling method. Based upon the more rapid, less stressful,         in size and morphology of the P-wave was noted, and the inter-
more effective death of zebrafish euthanized by rapid cooling, we        val between electrodes became progressively longer or wider.
advocate this method as a humane practice.                               Data suggested that the most favorable placement consisted of
                                                                         the terminal/tip electrode in the right ventricular apex and the
PS74 Development of a Recombinant Serodiagnostic Assay                   proximal/ring electrode inside or just above the right atrium.
for Simian Type D Retrovirus Infection in Macaques (Macaca               Refinement for capturing optimal waveforms has resulted in 5-
fascicularis)                                                            or 6-cm–spaced electrodes for cynomolgus weighing between 4
                                                                         to 7 kg. As an additional refinement of the surgical procedure
JH Simmons*, J Cowley, ML Wunderlich, BA Bronson, RK Dha-                with no impact on data capture, the transmitter body is located
wan                                                                      in an intramuscular pocket placed by way of laparotomy. Eighty-
Research Animal Diagnostic Services, Charles River, Wilming-             five cynomolgus monkeys have been instrumented to date with
ton, MA                                                                  these methods and have experienced minimal or no postopera-
                                                                         tive complications. A review of the surgical procedure and ECG
Exogenous simian type D retroviruses (SRV) are a closely re-
                                                                         enhancements will be presented.
lated group of viruses that infect free-ranging and laboratory
Abstracts of scientific papers 2008 AALAS National Meeting

PS76 Circadian Disruption Induced by Dark-phase Light                for measurement of AFP, a biomarker that reflects presence and
Contamination in Laboratory Animal Facilities                        growth of human tumor cells. Tumors were collected at various
                                                                     times throughout the study for histological analysis of vessel
RT Dauchy*, EM Dauchy, RP Tirrell, LK Davidson, DT Lynch,
                                                                     density using Factor VIII staining. Following 7 d of treatment
PC Tirrell, DE Blask
                                                                     with an antiangiogenic drug, uptake of microbubble contrast
Bassett Research Institute, Cooperstown, NY                          was assessed and showed a 35% decrease (P = <0.05) when
Diurnal light cycles and circadian rhythms are important to          compared to vehicle-treated tumors. After 10 d there was a 30%
humans and laboratory animals alike, and have been investi-          difference (P = <0.1) between treatment and control groups.
gated for many years. Previous studies from this laboratory          Blood pressure showed no significant change between treatment
demonstrated that minimal light contamination (0.25 lux) during      groups throughout the study, indicating that the changes seen in
an otherwise normal dark phase resulted in suppression of the        tumor blood flow are independent of anesthetic effects. We have
circadian neurohormone melatonin. Here, 36 adult male Sprague        demonstrated a correlation between changes in tumor vascular-
Dawley were randomized into 6 designated groups (n = 6/              ity and response to compound treatment. While power Doppler
group), housed and maintained in light exposure chambers and         imaging is somewhat limited to large vessels with high flow, the
provided free access to food and water. Animals were maintained      use of microbubble contrast enhancement provides much greater
on either control (I) 12 L (345 lux; 141.5 µW/cm2; lights on 0600    detail about flow and microvessels within a tumor.
h):12 D or experimental light-at-night (LAN) lighting cycles as
follows: (II) 12 L:12 D–0.02 µW/cm2; (III) 12 L:12 D –0.05 µW/cm2;   PS78 Facility Design Requirements for Integrating Gaseous
(IV) 12 L:12 D–0.06 µW/cm2; (V) 12 L:12 D–0.08 µW/cm2; and           Chlorine Dioxide Decontamination Systems
constant light (24L). Following 2 wk on these lighting regimens
                                                                     P Lorcheim, M Czarneski*
animals were subjected to a series of 6 low-volume blood draws
by cardiac puncture at 4-h intervals (beginning at 0400 h). Plasma   Clordisys Solutions Inc, Lebanon, NJ
samples were measured for total fatty acids (TFA), triglycerides     This presentation provides detailed information on designing
(TGA), free fatty acids (FFA), phospholipids (PL), cholesterol       and integrating a gaseous chlorine dioxide decontamination
esters (CE), glucose, lactic acid, corticosterone, and melatonin     system into a new animal research facility. Topics to be covered
levels. Plasma TFA and LA, respectively, for groups I–V revealed     include reasons for choosing gaseous chlorine dioxide, facility
an identical diurnal rhythm, highest during the dark phase (4.66     HVAC design requirements, and room construction require-
and 1.61 mg/ml, 0400 h) and lowest during light phase (0.91 and      ments, as well as ways to interface the decontamination system
0.30 mg/ml, 1600 h), but remained at the same level throughout       to rooms, pass-through chambers, and entire suites such as that
the day in group VI (2.72 and 0.92 mg/ml, constant light; TGA >      of a BSL facility. A distributed installation configuration layout,
FFA > PL > CE). Diurnal plasma glucose levels, similar for groups    where the decontamination system could remotely decontami-
I–IV (peak, 2000 h; nadir, 0400 h), were significantly elevated (P   nate any room of a BSL-3 suite area, will be presented, and the
< 0.05) and phase-delayed in groups V (peak, 1200 h) and VI          use of decontamination chambers to decontaminate equipment
(peak, 2400 h), respectively; melatonin rhythms were normal for      and supplies entering a barrier facility or exiting a BSL facility
groups I–IV (130.8 pg/ml, dark phase; 6.1 pg/ml, light phase),       will be explored. The benefits of a complete suite or facility
but disrupted and low diurnally for groups V and VI (6.1 pg/         decontamination system will be compared to a room-by-room
ml, light phase). These findings argue that even minimal light       decontamination system.
contamination during an otherwise normal dark phase in the
laboratory animal facility causes disruptions in normal physi-
                                                                     PS79 Circadian Disruption Induced by Dark-Phase Light
ologic and metabolic circadian rhythms.
                                                                     Contamination in Animal Facilities Stimulates Human Tumor
                                                                     Growth and Metabolism in Nude Rats
PS77 Application of High-frequency Ultrasound to Assess
Tumor Neovascularization and Compound Response                       RT Dauchy*, EM Dauchy, LK Davidson, PC Tirrell, DT Lynch,
                                                                     RP Tirrell, MW Greene, LA Sauer, DE Blask
TA Swanson*, KJ Patterson, CJ Andresen, C Bagi, MR Moalli
                                                                     Bassett Research Institute, Cooperstown, NY
Comparative Medicine, Pfizer Global Research and Develop-
ment, Groton, CT                                                     Laboratory animal facility diurnal lighting protocols significantly
                                                                     impact the chronobiologic nature and outcome of scientific inves-
The use of high-frequency ultrasound (HFUS) in preclinical drug      tigations, and strict adherence to such protocols, as outlined in the
development has many applications. Ultrasound measures of            Guide, is often overlooked. Previously, our laboratory demonstrated
blood flow and vascularity are directly translatable to the clinic   that animal facility dark-phase “light contamination” with as little
and are comparable to clinical ultrasound. Our current studies       as 0.25 lux (0.08 μW/cm2) suppressed production of the circadian
involve the validation of high-frequency ultrasound power            neurohormone melatonin and stimulated rodent and human breast
Doppler and contrast-enhanced imaging on xenograft tumor             tumor growth and metabolism. This occurs by way of an MT1
models to assess growth kinetics and response to drugs that are      melatonin receptor-mediated suppression of signal transduction,
known to effect neovascularization. Our objective was to assess      resulting in inhibition of linoleic acid (LA) uptake and conversion to
neo-vascularization events in mouse and rat tumor xenografts,        the mitogen 13-hydroxyoctadecadienoic acid (13-HODE). Here we
quantify changes in vascularization detected by HFUS, and cor-       tested whether dark-phase light contamination disrupts temporal
relate those changes with biomarkers such as alpha-feto protein      coordination of human breast tumor–host metabolic mechanisms.
(AFP) and tumor histochemistry. Nude rats were implanted with        Female nude rats (170 g; n = 36/group) bearing “tissue-isolated”
Huh7.5 hepatocarcinoma cells injected subcutaneously into the        human breast cancer xenografts were maintained on either control
flank. Tumor size and vascularity were assessed using 3D gray-       (I) 12 L(345 lux; 141.5 μW/cm2):12 D or experimental (II) 12 L:12
scale volume imaging, power Doppler and microbubble contrast         D–0.08 μW/cm2 lighting regimen (lights on 0500 h). Tumor latency
bolus injections. Blood pressure and body temperature were           to onset (first palpable mass) and growth rates for groups I and II
monitored during these studies. Blood samples were collected         were, respectively, 16 and 11 d, and 0.26 ± 0.03 and 0.58 ± 0.04 g/d.
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

When tumors reached an estimated weight of 5 to 6 g, animals            animal care staff and facility managers overseeing time manage-
were prepared for arteriovenous measurements over 6 circadian           ment and per diem rates. Our objective was to determine the
timepoints in a 24-h period (n = 6/timepoint). Results showed           impact of microisolation cage bedding volume and cage change
plasma melatonin levels in group I were high in dark-phase, and         interval on microenvironmental conditions, mouse health, and
low in light-phase (I) or group II animals. Diurnal tumor glucose       behavior. Fifteen cages housing ICR mice female (n = 5/cage)
uptake, lactic acid release, cAMP levels, LA-uptake, 13-HODE            were monitored for 17 d. The mice were obtained at 12 wk of age
release, and DNA [3H]thymidine incorporation were elevated              and allowed a 2-wk acclimation period. Three different volumes
significantly (P < 0.001) in daytime (I) and group II by over 60%,      of autoclaved aspen bedding chips were used: minimal volume
400%, 700%, 1000%, and 900%, respectively, compared to dark-            (approximately 250 ml/40 g), large volume (approximately 550
phase (I). Tumor Akt, MEK, ERK 1/2 activation was elevated for          ml/100 g), or normal volume (approximately 400 ml/70 g). Mice
groups I (light-phase) and II. These findings are the first to show     were evaluated (physical exam, weight) on d 7, 10, 14, and 17,
in vivo that temporally coordinated and integrated metabolic and        and a qualitative assessment of cage soiling was photographed
signal transduction mechanisms underlying human breast cancer           at these timepoints. Pooled feces were collected for corticos-
growth can be disrupted by dark-phase light contamination in the        terone analysis, and bedding mass was also measured at these
animal facility.                                                        timepoints. On d 17, 1 mouse from each cage was euthanized and
                                                                        bronchoalveoloar lavage was performed for cytologic analysis.
PS80 A Comprehensive Rodent Quarantine System That Jump-                Mice were videotaped for approximately 10 min during the active
starts Research                                                         periods on d 1, 8, and 15. Atmospheric analysis was performed
                                                                        to determine intra-cage ammonia, particulate matter concen-
J Watson*                                                               tration, and cage humidity/temperature on a daily basis. The
MCP, Johns Hopkins, Baltimore, MD                                       atmosphere was measured using a photoionization detector and
                                                                        particulate size and number was recorded by an aerodynamic
This presentation outlines the logistics of setting up a full-service
                                                                        particle sizer. There were no indications that the animals were
rodent quarantine program geared for research support. Statistics
                                                                        stressed, experienced adverse health effects, or exposed to higher
from our first year of implementation will be presented. Rodent
                                                                        than recommended levels of particulates or adverse environ-
quarantine is a necessary component of most research facilities to
                                                                        mental conditions during the 14-d period. Significant findings
prevent introduction of unwanted pathogens. Interinstitutional
                                                                        included variations in humidity depending upon cage location
rodent shipments are increasingly common due to NIH sharing
                                                                        and ammonia concentration among cage bedding levels. Cages
agreements, research collaborations, faculty recruitment, and
                                                                        were visibly soiled beyond what would be considered accept-
international vendor sources. However, there is no universal
                                                                        able to most observers by d 14. This study demonstrates that a
standard regarding which organisms should be excluded from
                                                                        2-wk interval between cage changes did not impact measurable
rodent colonies, and the varied rodent housing arrangements
                                                                        animal welfare indices, despite the appearance of excessive soil-
and systems for colony health surveillance make it hard to assess
                                                                        ing by this timepoint.
infection risk. Thus, it is usually necessary to quarantine incom-
ing animals to evaluate their infection status, which can take 8 to
12 wk. In some cases, this process may impede or even prevent           PS82 Ammonia Concentrations at the Cage Level for Mice
research. Colony founders may be too old to breed by the time           and Rats: Effects of Strain, Housing Density, and Ongoing
they are released, and age-sensitive phenotypes may occur during        Breeding
the quarantine period. Some shipments may be rejected because of        A Ostdiek*2, A Frazier1, N Johnston1,4, T Liberati1,4, R Trammell4,
infections in the source colony or incur additional delays and costs    LA Toth3,5
for surgical rederivation. We developed a comprehensive quar-           1Division  of Laboratory Animal Medicine, Southern Illinois
antine program to address these problems. Research delays were
                                                                        University School of Medicine, Springfield, IL; 2College of
mitigated by providing technical services in quarantine, including
breeding, identification, sample collection, genotyping, and basic      Veterinary Medicine, University of Illinois, Urbana, IL; 3Phar-
                                                                        macology, 4Internal Medicine, 5Associate Dean for Research and
experimental manipulations. Shipments that would previously
                                                                        Faculty Affairs, Southern Illinois University School of Medicine,
have been rejected or delayed because of infection were instead
                                                                        Springfield, IL
placed in high-risk quarantine and tested individually. Infected
animals were rederived by neonatal transfer. Our first year, 59         High ammonia in the microenvironment can adversely impact
of 230 shipments were placed in high-risk quarantine, and 58 of         rodent health. Many factors influence ammonia concentrations
them tested free of infection. Thirty-four shipments used technical     in the cage, including housing density, air flow rate, room tem-
services, 118 shipments were bred, and 17 genetic lines were red-       perature and humidity, bedding type, type of caging system,
erived to eliminate Helicobacter sp. Our experience demonstrates        and the frequency of cage changes. The NIOSH-established
that rodent quarantine, far from impeding research, can actively        exposure limit for people is a time-weighted average of 25 ppm
promote an efficient start to research following interinstitutional     over an 8-h period. Levels above this are reported to have det-
rodent transfers.                                                       rimental effects in people and rodents. We performed this study
                                                                        to examine the relationship between ammonia levels and strain
PS81 Bedding Change Interval Influence on the Microenviron-             differences, housing density, and breeding animals. To verify
ment of Mice in Ventilated Microisolation Housing                       that our standard housing conditions did not generate damag-
                                                                        ing concentrations of ammonia in the cage, we used a hand-held
M Rosenbaum*1, S VandeWoude1, T Johnson2                                piston pump and ammonia indicator tubes to monitor ammonia
1Microbiology, Immunology, and Pathology, 2Environmental                of 2 strains of rats (GEPR-9 and Sprague Dawley) and 3 strains
and Radiation Health Sciences, Colorado State University, Fort          of mice (Maf/D2, Ames dwarf, and PEPCK-BGH1). These strains
Collins, CO                                                             were selected due to their frequent use in our facility or to their
                                                                        apparent tendencies to produce relatively high amounts of urine.
The frequency of mouse cage bedding changing has significant
                                                                        The animals were housed in static micro-isolator cages that were
implications not only for the animals themselves, but also for the
Abstracts of scientific papers 2008 AALAS National Meeting

changed weekly. Breeding cages contained 1 to 3 adults with as        The fast increase in the production of new lines of genetically
many as 17 pups. Single-sex cages contained 1 to 5 animals. For       modified mice has raised concern about how gene manipula-
each strain, 2 breeding cages and 2 non-breeding cages were           tion techniques affect the welfare and normal development of
evaluated over a 2-wk cycle, providing a total of 4 cages per         the resulting animals. The phenotype of the mice obtained by
strain that were sampled over 2 cage changes. For both rats and       those techniques is influenced by the background strains, the
mice, ammonia concentrations rose above 25 ppm between 4              environment, and possibly by the occurrence of compensatory
and 6 d after a cage change. To determine whether this degree         processes. Unexpected phenotypes may result in welfare prob-
of exposure caused a pulmonary inflammatory response, we              lems and excessive suffering, which justify the establishment
compared concentrations of 10 cytokines in the lungs of mice          of humane endpoints. While the need for the development of
maintained in a constant low ammonia environment by more              monitoring routines at an early age has been discussed, no pro-
frequent cage changes to those in lungs of mice that were exposed     tocols for preweaning characterization of genetically modified
to a high-ammonia environment for 2 d. Concentrations of 4 of         and mutant mice have so far been validated. This study com-
the 10 cytokines were below limits of assay detection. Significant    prises a first assessment of a characterization routine that has
differences between groups were not detected. We conclude that        been developed in our laboratory. A total of 56 mouse pups of 3
the practice of weekly cage changes does not cause inflammatory       inbred strains and 1 hybrid commonly used as backgrounds for
changes in the lungs of mice, despite their exposure to levels of     the construction of transgenic and knockout mice (C57BL/6Bkl,
ammonia over 25 ppm for periods of up to 2 d.                         BALB/cBkl, 129S6Bkl, and B6CBAF1Bkl) were monitored from
                                                                      birth to weaning on key days of morphological, physiological,
PS83 Radical New Approach to IVC Housing for Rats                     and behavioral development. The results presented show that
                                                                      the developed characterization routine is efficient in detect-
M Jackson*                                                            ing differences in physical and sensory development between
BSF, University of Manchester, Manchester, United Kingdom             strains, namely concerning weight gain, reaction to handling,
                                                                      physical condition at weaning, and emotional reactivity. A future
A rat focus group featuring experienced animal technologists
                                                                      study will address its efficiency in detecting differences in the
from academia, contract research, and the pharmaceutical sectors
                                                                      development of genetically modified pups and their wild-type
was formed to address the need for a new rat IVC cage. The key
                                                                      litter mates.
considerations were to produce a cage which is compliant with
UK and European legislation. It should hold up to 5 adult rats
or a breeding pair in an enriched environment that allows rats to     PS85 Predicting the Impact of Noise from Construction on
express a full range of behaviors including bi-pedal posture. The     Mouse Gestation and Nursing Behavior
cage must also have good manual handling characteristics and          S Rasmussen*1,3, GM Glickman2, FW Quimby1,3, R Tolwani1,3
be as light and well balanced as possible. Flexibility is important   1CBC,   The Rockefeller University, New York, NY; 2 Wilson,
so as to be acceptable to a range of research environments. The
cage should be operational at positive or negative pressure and       Ihrig & Associates, Inc., New York, NY; 3Postdoctoral Research,
allow use of various water delivery systems. As automation is         Tri-Institutional Postdoctoral Training Program in Laboratory
increasingly used for washing and bedding replacement in many         Animal Medicine and Science, New York, NY
institutes, the cage must withstand being handled by robotics.        A comprehensive study was undertaken to identify the impact
The cage design includes a base and a high, domed lid. A horizon-     of noise from a planned expansion of our animal facility on
tal U-shaped divider provides an extra story, effectively doubling    mouse gestation and nursing behavior. Ambient noise levels
floor space in the same footprint. Two test cages were fabricated     were analyzed first in order to gain an understanding of daily
using stereolithography and connected to an IVC rack system           noise exposure. We then compared the ambient noise levels to
for 4 wk. Three male S/D rats were allocated to each test cage.       that predicted from construction activities to see if noise from
Animals were weighed daily, and food and water consumption            construction would impact mouse gestation or maternal nursing
were noted. Growth rates and diet consumption were normal             behavior. We hypothesized that high noise levels would result in
for this strain. Video monitoring was used to study activity both     fewer live births and higher pup mortality as a result of disrupted
in light and dark phases of the daily cycle and after weighing or     nursing behavior. To study the impact of noise on mouse gesta-
cleaning of cages. Observations revealed rats using all areas of      tion, 70 8-wk-old female ICR mice underwent embryo transfer.
the cage. Sleeping was confined mainly to the lower half of the       Mice were divided into groups of 10, with each group receiving
cage, while play occurred on both levels. Exploratory behavior        exposure to a single intensity of simulated noise analogous to
and climbing are seen as positive welfare aspects of the design.      concrete saw cutting at 70 dBA, 80 dBA, or 90 dBA for 1 h daily.
Animal technologists of varying experience were encouraged to         The frequency of the simulated noise ranged from 2 to 4 kHz.
use the cage and complete a questionnaire to elicit their opinions.   Thirty mice received noise exposure d 1 through 7 following
The paper shows that the design brief was met and an enriched,        embryo transfer, 30 were exposed d 8 through 14, and 10 mice
radical new rat cage is ready to go to mass production.               received ambient noise only. The number of viable pups born was
                                                                      recorded. An additional 20 mice were used for a second study to
PS84 Characterization Routine for Improved Animal Welfare             examine the effects of construction noise on nursing behavior. A
in Preweaning Mice                                                    control group of 5 mice received normal ambient daily noise. The
                                                                      remaining 15 mice were divided into groups of 5. Each group was
JM Marques1,2, H Augustsson*3, S Ögren4, K Dahlborn1                  exposed to a single intensity of noise simulated from concrete
1Department of Clinical Sciences, Swedish University of Ag-           saw cutting at 70 dBA, 80 dBA, or 90 dBA for 1 h daily during
ricultural Sciences, Uppsala, Sweden; 2Laboratory Animal              the first 7 d following parturition. Female mice were monitored
Science, Institute for Molecular & Cell Biology, Porto, Portugal;     for cessation of nursing behavior. Pups were weighed daily
3Department for Animal Welfare and Health, Swedish Board of           and monitored for increased mortality. Based on these studies,
Agriculture, Jönköping, Sweden; 4Department of Neuroscience,          specific construction noise limits were established to minimize
Karolinska Institute, Stockholm, Sweden                               any negative impact on mouse reproduction.
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

PS86 Novel Means of Continuously Monitoring Laboratory                 packaging and transport, production and distribution of IVCs,
Rodents                                                                and use and disposal of IVCs throughout the life cycles of the
                                                                       alternative systems. The analysis was performed using a system
SM Niemi*1, M Dempsey2, N Sims2, M Wollowitz3
                                                                       perspective and the LCA methodology. The functional unit was
1Center for Comparative Medicine, 2Center for Integration of           set to the use of 1 IVC (disposable or multiuse) for a period of
Medicine and Innovative Technology, Massachusetts General              1 y. The yearly number of replacements and the disposal of the
Hospital, Boston, MA; 3 Tangible Design, Inc., Chatham, NY             single-use PET IVCs compared with the yearly number of wash-
The dramatic growth of genetically engineered mice in biomedi-         ing and sterilization cycles of the PSU IVC were considered. The
cal research has resulted in more complex breeding programs,           results of the LCA study are presented both in terms of direct
sophisticated endpoints, subtly different phenotypes, occasional       inventory data, including mJ of primary energy from nature, by
difficult breeders, and consequently greater financial losses in the   fuel type, and in terms of impact assessment, meaning tons of
event of husbandry failures. But because mice are nocturnal, they      each greenhouse gas emitted to the atmosphere by all life cycle
are more likely to display abnormal behavior or clinical signs of      processes, and estimates of human health consequences and
disease or injury at night. All of these considerations demand         ecological consequences of the pollutant releases to air, water,
better observation of cages than a perfunctory visual check once       and soils over the life cycle. In addition, a sensitivity analysis
daily. However, there may be difficulties or inconsistencies in        was performed considering particularly different IVC disposal
observing every mouse on a routine basis because the bottom            scenarios so as to understand how to reduce the environmental
and top rows of high-density cage racks are not easily viewed. To      burden at the end of an IVC’s life.
address these problems, a novel approach was devised in which
a series of gantry-mounted infrared cameras scan all cages on
a rack; video (at programmable intervals and of programmable
durations) images are recorded and stored for later viewing.
                                                                                           Poster Sessions
Each gantry communicates to an on-site or remote web server.
With appropriate authentication, live video feeds or stored data       P1 Model for Clinical Evaluation of Perioperative Analgesia
can be viewed through any web browser. Features include ob-            in the Rabbit
servation of mice 24/7, digital documentation of cage location         AZ Karas*, LA Weaver, KA Andrutis, CA Blaze
and animal signalment, storage of retrievable images, restricted
                                                                       Clinical Sciences, Tufts University Cummings School of Veteri-
access to images for each user, and remote override of scanning
schedules. Software enhancements will include automated alerts         nary Medicine, North Grafton, MA
in the event of excessive activity (such as fighting, stereotypy)      Assessment of pain and distress in rabbits is problematic, and
or absence of activity (moribundity or death), flooded cages,          studies of effective surgical analgesia are lacking for this species.
empty feed hoppers and so forth. Advantages of this solution           Seeking potential indicators of postoperative pain or distress,
with respect to discovery research and GLP testing include             we performed ovariohysterectomy and telemeter placement as
continuous observation for subtle changes in behavior or unan-         a form of moderate surgical injury in 20 female rabbits. Rabbits
ticipated phenotypes in neuroscience protocols, more precise           were randomly assigned to 1 of 4 treatment groups (n = 5 per
determination of impending morbidity for toxicology, and safe          group): buprenorphine (0.03 mg/kg SC, every 12 h for 3 d); fen-
(remote) observation for select agent protocols and other highly       tanyl (25 mcg patch placed 24 h prior to surgery); ketoprofen (1
hazardous studies. In addition, this solution ensures that every       mg/kg, SC, every 24 h for 3 d) and control (no treatment given).
animal is, indeed, observed daily per regulatory requirements          Various physiologic and behavioral parameters were recorded or
and requires fewer staff to do so.                                     scored by blinded observers, including heart rate and blood pres-
                                                                       sure, temperature, food and water consumption, fecal output,
PS87 A Comparison of Single-use and Multi-use Indi-                    and remotely recorded video-recorded behaviors during daily
vidually Ventilated Cages: CO 2 and Energy Environmental               exercise in 1.8 × 2.4 m floor pens. Compared to preoperative
Performance                                                            values, declines in behaviors occurred on postoperative day 1:
                                                                       mean travel distance (70%), vertical exploring (94%), decrease
GA Norris*1, V D’Incognito2                                            in food consumption (87.6%). Many values had not returned to
1School of Public Health, Harvard University, Boston, MA; 2Take        baseline levels after 7 d. Fecal output decreased significantly as
Care, Milano, Italy                                                    well. Although no single treatment proved significantly better
                                                                       than another, some analgesic treated rabbits did suffer a change
Individually ventilated cages (IVCs) have been under close
                                                                       in behavior values the day after cessation of therapy. Our results
scrutiny recently, especially the choice of plastic materials for
                                                                       demonstrate substantial inappetance and reduction of normal
single-use or multi-use cages. Many different requirements
                                                                       activity levels in rabbits after surgery. Although empirical doses
such as medical safety, rodent care, treatment functionality,
                                                                       (those typically recommended) of analgesics did not appear
costs and efficiency, and environmental performance have to be
                                                                       substantially better than placebo, comparison of other doses
met. For several years now, the most commonly used materials
                                                                       and multimodal analgesic techniques using these monitoring
for IVC manufacturing have been polysulfone (PSU) for multi-
                                                                       strategies may prove useful in future studies aimed at optimiz-
use cages and, more recently, polyethilene terphthalate (PET)
                                                                       ing postoperative analgesia in rabbits.
for single-use cages. A life cycle assessment (LCA) study was
used to compare the environmental performance of the 2 cage
types, with focus on the energy consumption and global warm-           P2 Evaluation of Supplemental Warming Methods for Preven-
ing contribution due to greenhouse gas (GHG) emissions. The            tion of Hypothermia in Rats
LCA study is based primarily on the use of publicly available          A Koch*2,1, DG Scorpio1, D Ruben1
life cycle inventory databases that quantify raw material input        1 Molecular and Comparative Pathobiology, Johns Hopkins
from nature and releases of pollution to air, water, and land as-
                                                                       University, Baltimore, MD; 2Cummings School of Veterinary
sociated with the input of primary and intermediate materials,
                                                                       Medicine, Tufts University, North Grafton, MA
Abstracts of scientific papers 2008 AALAS National Meeting

Prolonged anesthesia and hypothermia is a common problem in               mg/dl). The macaques that received Telazol for induction had an
rodents undergoing experimental procedures. Previous studies              average temperature decrease of 3.5°F (preoperative values = 100.5
conducted with Mylar-backed drapes in anesthetized rats revealed          ± 1.3 °F (x¯ ± SD); range 98.7 to 102.2 °F)(postoperative values = 99
reduced hypothermia compared to conventionally draped and                 ± 2.1 °F (x¯ ± SD); range 96.1 to 101.5 °F) and blood glucose levels
undraped rats. Mylar reduces radiant heat loss in rodents under-          increased an average of 30.7 mg/dl (preoperative values = 62.4
going procedures in ventilated hoods or atop laboratory benches.          mg/dl ± 6.9 mg/dl (x¯ ± SD); range 55 to 75 mg/dl; postoperative
This study was performed to compare Mylar-backed drapes to                values = 93 mg/dl ± 23.1 mg/dl (x¯ ± SD); range 69 to 127 mg/
other warming methods in anesthetized rats undergoing sham                dl). In conclusion, although the group premedicated with Telazol
laparotomy. Conventional surgical drapes and Mylar drapes were            showed increased postoperative glucose concentrations compared
compared, both with and without a heat-emitting gel pad. A control        to preoperative values relative to the ketamine group, the change
group included no drapes or gel pad. Rats were anesthetized with          was not statistically significant; thus the choice of one sedative
intraperitoneal ketamine/xylazine. A ventral midline incision was         over the other does not impact surgical recovery.
created through a drape fenestration covering each rat’s ventrum.
Temperature was monitored with digital rectal probes for 60 min.          P4 Suppurative Osteomyelitis, Meningoencephalitis, and
Rats were euthanized at conclusion of experiments. All groups             Multifocal Suppurative Bronchopneumonia in the Gambian
developed hypothermia except for the group with the isothermal            Rat (Cricetomys gambianus)
pad combined with the Mylar drape. These rats exceeded nor-
motherrmia with temperatures between 107 to 109 °F, due to gel            BP Herrod*, N Bryant, D Carroll, E Jackson, N Powell
pad isothermal release of 108 °F+ reflecting off the Mylar radiant        Animal Resources Branch, Pathology Laboratory, Centers for
drape. The standard drape with gel pad allowed an average 2.19%           Disease Control, Atlanta, GA
heat loss from baseline; conventional and mylar drapes used alone
                                                                          The African giant pouch or Gambian rat (Cricetomys gambianus)
allowed 8.44% and 9.65% heat loss from baseline, respectively,
                                                                          is a nocturnal rodent from central and southern Africa, where it
though far less than control rats (14.39%). Heat loss likely occurred
                                                                          is used as a meat source by natives. Gambian rats were popular
from the abdominal incision, thus both drape types yielded similar
                                                                          pets in the US until June of 2003, when they were suspected to
results. Rats completely wrapped in the Mylar drape without gel
                                                                          be a source of a monkeypox outbreak. Health officials suspected
pad sustained 4% to 5% heat loss from baseline. Mylar drapes and
                                                                          that they were the source of infection because tests have shown
heat-emitting pads serve to prevent hypothermia during anesthesia
                                                                          that these rats have antibodies to monkeypox. Presently there
in rodents. Used together, however, rat core body temperature rises
                                                                          are no licensed therapies for human monkeypox; therefore, the
quickly. Using both methods together is beneficial for hypothermic
                                                                          human population is at risk to this zoonotic agent. Due to this
rats. Other supplemental heat modalities such as warm water recir-
                                                                          susceptibility, there are fears that the monkeypox virus could be
culation or forced air should be ideal when used with Mylar-backed
                                                                          used as a bioterrorism agent. Gambian rats, in addition to other
drapes because they generate less heat than the gel pad; both will
                                                                          animal species, are being studied to investigate the pathogenesis
be further evaluated.
                                                                          and treatment of orthopoxvirus infections. We received wild-
                                                                          caught, female Gambian rat, which died suddenly, for necropsy.
P3 Perioperative Effect of Ketamine Compared with Telazol                 The animal was negative for the monkeypox virus and had not
on Body Temperature and Blood Glucose Concentration with                  been used in a study. At gross necropsy, a few abnormalities were
Rhesus Macaques (Macaca mulatta)                                          noted. An abscess was found within the right temporomandibu-
BC Davisson*, M Patterson, RP Marini                                      lar region which extended into the right temporomandibular
                                                                          joint and the base of the skull. We also found multifocal white
Division of Comparative Medicine, Massachusetts Institute for             nodules along the surface of the both lungs. Aerobic bacterial
Technology, Cambridge, MA                                                 cultures were taken in an effort to identify the microbial agents
Variables likely to impact intraoperative and postoperative man-          involved. The results from the mandibular region revealed a
agement of macaques include anesthetic induction regimen, body            heavy growth of Pasteurella multocida, light growth of Escherichia
temperature, and blood glucose concentration. A sample group of           coli, and scant growth of alpha-hemolytic streptococci. Bacterial
macaques (n = 30) undergoing extensive cranial surgeries (rang-           cultures taken from the base of the skull revealed moderate
ing from 2 to 11 h) was examined to compare these parameters              growth of Pasteurella multocida and scant growth of a Coryne-
and determine what impact induction agent had in maintaining              bacterium spp. Histopathology revealed evidence of glossitis,
normothermia and blood glucose concentrations. All macaques               pneumonia, meningoencephalitis, osteomyelitis, and myositis. In
underwent at least a 12-h fast and then received either intramus-         this presentation, we will further characterize the histopathologic
cular ketamine for induction (n = 23) or intramuscular Telazol            lesions through special stains and other molecular diagnostic
(n = 7). After sedation, rectal temperatures were recorded and            procedures. We will provide histopathologic evidence of an in-
blood glucose concentrations were measured using a handheld               fection that we encountered in a Gambian rat. After an extensive
glucometer. Animals were intubated and maintained on 1% to 3%             literature review, we have concluded that this is the first case
isoflurane in balance oxygen and were intravenously administered          report of this type of infection in a research Gambian rat.
a lactated Ringers solution containing 5% dextrose. Immediately
postoperatively, a second blood glucose sample was tested from            P5 Uterine Epithelioid Trophoblastic Tumor in a Cynomolgus
each animal and postoperative rectal temperatures were recorded.          Macaque (Macaca fascicularis)
In the group that received ketamine for induction, body tempera-
tures decreased an average of 3.7 °F (preoperative values = 101.2 ±       CL Emerson*1, J Hutt1, TK Cooper2
1.6 °F (x¯ ± SD); range 97.3 to 102.6 °F; postoperative values = 97.9     1Lovelace Respiratory Research Institute, Albuquerque, NM;
± 1.7 °F [x¯ ± SD]; range 94.3 to 100.8 °F), while blood glucose levels   2Comparative  Medicine, Penn State Milton S. Hershey Medical
increased an average of 22.9 mg/dl (preoperative values = 61.3            Center, Hershey, PA
mg/dl ± 12.7 mg/dl [x¯ ± SD]; range 41 to 97 mg/dl; postopera-
                                                                          An adult female cynomolgus macaque was evaluated for lethargy
tive values = 84.1 mg/dl ± 21.4 mg/dl [x¯ ± SD]; range 52 to 126
                                                                          and dehydration. Results of blood work were unremarkable,
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

and the animal was rehydrated. The animal’s clinical signs im-        1Microbiology,  Immunology and Pathology, 2LAR, Colorado
proved the day after admission, but declined dramatically by          State University, Fort Collins, CO
the following day and the animal was euthanized. At necropsy, a       A wildlife sanctuary presented a postmortem adult female cot-
sessile mass was identified in the wall of the uterine body, with a   tontail (Sylvilagus spp.) rabbit, age unknown, to the Colorado
portion of the mass extending into the lumen. The mass was ap-        State University Pathology service for necropsy. The available
proximately 3 cm in diameter, yellowish-white, and friable in the     history on this animal was limited, but it was reported that at
center. There were numerous multifocal to coalescing metastatic       least 1 other Sylvilagus spp. rabbit in the same area was reported
nodules of similar appearance present in the lungs, with efface-      to exhibit similar gross cutaneous lesions. On gross exam, the
ment of approximately 80% of the normal lung parenchyma.              animal had numerous pigmented wartlike lesions arising from
Scattered metastases were also present in the liver and spleen.       the skin of the head surrounding the ears, eyes, nares, mouth, and
Based upon the histological appearance of the uterine mass and        a few areas along the dorsum. Masses were firm, greasy, friable,
select immunohistochemical stains, the tumor was identified           and easily detached from the underlying skin. Affected cutaneous
as an epithelioid trophoblastic tumor (ETT). ETT is one type of       tissue was fixed in formalin, embedded in paraffin, and sectioned
gestational trophoblastic disease derived from the placental tro-     by microtome. The slides were stained with hematoxylin and
phoblast. Gestational trophoblastic diseases present with a wide      eosin for histopathology. Histologic evaluation revealed multiple
range of clinical behaviors and histological appearances, some of     papillary masses lined by stratified squamous epithelial cells
which are aggressive and malignant. These tumors are distinct         with central cores of fibrovascular connective tissue arising from
from all other tumors in that they are fetal tumors (products of      the epidermis. Cells of the stratum spinosum were frequently
conception) arising in the mother’s body. ETT was first described     swollen with abundant perinuclear cytoplasmic clearing and
in women in 1998 and has features that are similar to carcinoma.      occasional intranuclear amphophilic glassy spherical inclusions
ETT has been diagnosed in nonhuman primates and found in              up to 3 µm in diameter. The lesions were consistent with rabbit
the uteri of a red-tailed guenon (Cercopithecus ascanius) and an      Shope papilloma virus. Differential diagnoses included Shope
African green monkey (Chlorocebus aethiops). A germ cell tumor        fibroma virus and myxoma virus. To definitively confirm the
with features of ETT (nongestational ETT) has been described in       Shope papilloma virus diagnosis, histopathology slides were
the ovary of a cynomolgus macaque (Macaca fascicularis).              submitted for immunohistochemistry; results were positive.
                                                                      Affected tissue was fixed in glutaraldehyde and submitted for
P6 Comprehensive Dental Health Assessment and Treatment               electron microscopy, through which the virus was identified. This
Program                                                               case presents an opportunity for review of the numerous virally
CL Gai*1, T Trinh1, A Parlapiano2, GO Voronin1                        induced cutaneous lesions due to papillomavirus and poxvirus
1Laboratory Animal
                                                                      infection in rabbit species, and it illustrates an important model of
                    Resources, 2Lead Optimization, Merck and          viral carcinogenesis as well as a disease that represents different
Co., Inc., Rahway, NJ                                                 disease spectrums following cross-species transmission between
Dental health and hygiene are important components of the over-       Sylvilagus and Oryctolagus spp. A detailed explanation of this
all health and well-being of animals. Undiagnosed or overlooked       differential disease spectrum is included in the discussion.
dental disease can lead to poor patient health and discomfort,
disease in other body systems, and may be a confounding factor        P8 Use of Behavior Conditioning to Facilitate Venipuncture
in research. Providing adequate dental care to a large number of
animals presents a logistical challenge. Mandating dental clean-      C Lanman*
ings for all animals on a predetermined schedule could lead to        Pet Care, Proctor and Gamble, Lewisburg, OH
animals receiving unnecessary treatment and exposure to anes-         In every capacity of research and in every facility, it can be as-
thesia. It would lead to the expenditure of animal care resources     sumed that methodology improvements come to the minds of
and manpower that would be better used elsewhere. Tailoring           technicians quite often. In the case of feline restraint for blood
care to the individual needs of an animal would require a greater     collections, new ways to reduce the stressful nature of these
amount of effort for each animal, but would also ensure that each     procedures for both the felines and the technicians have been
animal receives the appropriate level of care in a timely manner.     difficult to uncover. This innovative venipuncture method not
In order to deliver animal-specific and -appropriate dental health    only decreases the stress sometimes associated with restraint and
care, a comprehensive dental health assessment and treatment          venipuncture, but also allows for blood collection by a single
program was developed, tested and implemented in our canine           technician. The method involves first conditioning felines at a
colony. Initially, a simplified dental health scoring system was
                                                                      younger age to be comfortable with venipuncture, toenail trims,
developed that was used to grade the current dental health status     and veterinary treatments. Training begins at 2 d of age. At 3 wk
of the colony. A spreadsheet was designed that could not only         of age, the kittens are acclimated to being placed dorsally in a
be used to keep track of dental health, but would also generate       technician’s lap with the sound of clippers running. From this
the appropriate dental care recommendation based on the data
                                                                      point, the technicians continue the dorsal placement training
entered. Trained observers examined100 dogs, and their recom-         while adding different collection techniques such as stretching
mendations were evaluated for interobserver reliability by use of     the neck, stroking the jugular furrow, and clipping the hair along
a kappa statistic. Veterinarians and veterinary associates shared     the furrow. This training is ongoing to ensure the safety of the
the spreadsheet to generate a plan to deliver dental health care      felines and technicians. These training protocols and venipunc-
to individual animals during the course of 6 mo. This poster          ture method have proven to be instrumental in improving feline
presents the development, implementation, and results of our          well-being and technician aptitude in this research facility.
comprehensive dental health plan during this time period.
                                                                      P9 Research Opossum (Monodelphis domestica) with Perior-
P7 Cutaneous Lesions in a Cottontail Rabbit (Sylvilagus               bital Swelling
spp.)
                                                                      D da Cunha*1,2, LM Wancket2, C Freed1, L Gallaugher1
C Weiner*1,2, M Rosenbaum1,2, K Fox1, S VandeWoude2,1
Abstracts of scientific papers 2008 AALAS National Meeting
1 University of Laboratory Animal Resources, 2 College of              tion program. Newly replaced lines developed the same level of
Veterinary Medicine—Veterinary Biosciences, The Ohio State             ATP as used lines after the first week of use. Although the ATP
University, Columbus, OH                                               levels in lines associated with clinical signs were lower than those
                                                                       without, the sanitation procedure reduced the overall incidence
There are around 1,500 publications in the PubMed database
                                                                       of clinical signs to 0.4%. This study demonstrates that use of
pertaining to the use of marsupials in experimental, biomedical-
                                                                       ABS systems in rats is associated with development of resident
oriented research. Monodelphis domestica are naturally found in
                                                                       bacteria in the lines that can result in clinical infections. The oc-
arboreal habitats in South America. Here we report a case of a
                                                                       currence of infection is not predicted by bacterial load alone, but
3-y-old breeding female, part of an endocrinology study, that
presented with right enophthalmia, moderate periorbital swell-         is likely related to other factors. However, controlling the overall
                                                                       level of contamination through weekly sanitation reduces clinical
ing, prolapsed and inflamed third eyelid, blepharospasm, and
                                                                       infections, but complete sterility cannot be achieved, even when
blepharitis with mild purulent discharge causing irritation and
medial canthus alopecia. Triple antibiotic ophthalmic ointment         new lines are used. A luminescent bacterial-ATP assay used in
(TAO) with hydrocortisone was initiated, and systemic treat-           conjunction with bacterial culture and identification is a useful
ment with ibuprofen (15 mg/kg) provided for 7 d, after which           tool for monitoring and managing ABS line infections.
no signs of inflammation remained. Two months later (January
11th) the same animal presented with ventral periorbital swell-        P11 Regimens and Results for Treating Myobia musculis
ing, mildly prolapsed third eyelid, and buphthalmia in the right       D Newsom*1, IM Washington1, K Chase2, TL Brabb1, L Maggio-
eye. Oral cavity examination under isoflurane anesthesia ruled         Price1, S Dowling1
out tooth root abscess. Systemic treatment with enrofloxacin (5        1 Comparative
mg/kg), ibuprofen (5 mg/kg) and topical TAO was initiated.                           Medicine, University of Washington, Seattle,
On re-evaluation 3 d later, the animal presented with severe           WA; 2Veterinary Services, California Institute of Technology,
buphthalmia, blepharoedema, third eyelid thickening, and hy-           Pasadena, CA
peremia. Self-trauma to the periorbital area resulted in alopecia      The University of Washington (UW) was notified that a mouse
and hyperemia of the surrounding tissue. Based on the clinical         sent to another university tested positive for fur mites. Upon
presentation with poor prognosis and the age of the female, the        notification, the UW colony from which the positive mouse origi-
investigator consented to euthanasia and postmortem evalua-            nated was tested by tape test and confirmed positive for Myobia
tion. Blood was collected for CBC analysis. Grossly, the surface       musculis. Additionally, over a 1000 tape tests were performed
of the spleen appeared irregular, but otherwise within normal          on mice from 7 different rooms, 5 investigators, and 13 racks; 15
limits. The liver appeared muddy-brown in color with a smooth          positive animals were identified. The positive animals were from
surface. CBC findings included neutrophilia without bands,             2 investigative groups, and were of 8 different strains, some im-
accompanied by slight lymphopenia resulting in normal total            munocompetent and some immunodeficient. All sentinels were
white cell count. These results were supportive of transient           negative for mites by pelt testing. All mice from the 2 positive
stress most likely due to disease. H&E-stained sections showed         investigative groups (approximately 3400) were treated with
severe unilateral suppurative maxillary cellulitis with granula-       a nestlet impregnated with fipronil and methoprene (1.0 mg/
tion tissue that displaced the right globe. The lesion contained       nestlet). The treated nestlet was replaced 2 wk later, for a total
both Gram-positive cocci (consistent with Staphylococcus or Strep-     exposure of 28 d. A single dose of topical selamectin (10 mg/kg)
tococcus sp.) and filamentous bacteria (consistent with Nocardia       was applied to the intrascapular region of mice in any cage with
or Actinomyces sp.).                                                   an incompletely torn nestlet and to all mice in cages containing
                                                                       the 15 positive animals. No side effects from treatment were
P10 Management of Line Infections in an Automated Blood                observed. Culled animals were submitted posttreatment for pelt
Sampling System for Rats                                               examination. The treated mice remained fur mite negative for 5
                                                                       mo. In January 2008, live Myobia musculis mites were noted on
K Carpenter3, DM Cooper*1, K Heinz-Taheney2, J Sullivan2               1 mouse that had been treated with impregnated nestlets alone.
1Veterinary  Resources, 2Discovery Pathology, 3Animal Studies,         Based on this finding, it was assumed that treatment with only
Eli Lilly and Company, Greenfield, IN                                  fipronil/methoprene neslets failed to eliminate all mites. A sec-
                                                                       ond treatment was carried out, in which all haired mice were
We observed a 6.8% incidence of rear limb paralysis in femoral
                                                                       treated topically with a combination of selamectin (10 mg/kg)
artery– and vein–cannulated rats within several days of being
                                                                       and fipronil/methoprene (8 mg/kg) applied to the intrascapular
placed on an automated blood sampling (ABS) system. This prob-
                                                                       region. Mice were treated twice, 30 d apart. Pelt examination
lem was not seen with a previously used system and was not seen
                                                                       continues. No mice have tested positive for fur mites at 5.5 mo
in animals from the same shipment not used on study. Necropsy
of affected animals showed catheter-associated phlebitis or cel-       posttreatment. All sentinels remained negative throughout this
lulitis of the lumbar epaxial muscles or spinal cord. Cultures of      outbreak.
lesions and the pump lines detected Stenotrophomonas maltophilia,
Escherichia coli, Staphylococcus aureus, Burkholderia cepacia group,   P12 Effects of Altitude on Red Blood Cell Count, Hemoglo-
Klebsiella oxytoca, and K pneumoniae. A luminescent assay for          bin, and Hematocrit in African Green Monkeys (Chlorocebus
bacterial ATP in the pump lines had average readings of 98,635         aethiops)
relative light units (RLUs) from all pumps tested, even ones that      DC O’Donnell*, CL Emerson
were not associated with clinical signs. As the lines for the ABS
were not designed for replacement between studies, we started a        Animal Resources, Lovelace Respiratory Research Institute,
weekly sanitation cycle in addition to that recommended by the         Albuquerque, NM
manufacturer. ATP levels were markedly reduced immediately             Lovelace Respiratory Research Institute is in Albuquerque, NM
following sanitation, and further decreased after the unit was idle    with an altitude of approximately 5,300 ft. African green monkeys
over a weekend (average of 6,135 RLUs.) Lines did not develop          (AGMs) arrived at the facility in November 2006 from St. Kitts,
an increased baseline in ATP after several months on the sanita-       West Indies, Eastern Caribbean (n = 16), and February 2008 from
                                                                                    Abstracts of scientific papers 2008 AALAS National Meeting

Barbados (n = 20). It was noted that results of hematologic tests    Portland VA Medical Center, Portland, OR
performed on AGMs indicated that a few red blood cell (RBC)          B6.129-Slc6a4tm1KplN10 strains of mice have a mutation resulting
indices were significantly increased, compared with published        in reduced serotonin-transporter function, which causes phe-
reference intervals obtained at sea level. Analysis of hematologic   notypic changes such as gut dysfunction, bone weakness, and
tests performed on AGMs received in February 2008 and sampled        stress-related behaviors. They are used at our facility for osteopo-
approximately 2 wk after arriving at the facility revealed RBC       rosis studies. We have received clinical reports in heterozygous
counts were high in 13 of 20 AGMs and hemoglobin and hema-           mice at about 100 d of age describing lethargy, dehydration, and
tocrit values were high in 17 of 20 AGMs. Additionally, analysis     hypersalivation. These clinical reports represented over 60% of
of hematologic tests performed on AGMs received in November          the heterozygous mice of this age. Bloodwork was consistent
2006 and sampled approximately 1.5 y after arriving at the facil-    with dehydration. Necropsy revealed a flaccid, dilated distal
ity revealed similar increases in RBC counts (12 of 16 AGMs),        esophagus impacted with ingesta, which are findings consistent
hemoglobin (14 of 16), and hematocrit (13 of 16). In most AGMs       with achalasia. Literature has suggested small intestinal motility
from both sources, values for mean corpuscular volume, mean          is decreased in these strains, but this is the first time alteration
corpuscular hemoglobin, and mean corpuscular hemoglobin              of esophageal motility or lower esophageal function has been
concentration were within published reference intervals. Simi-       described. Microscopic evaluation of the esophagus around the
lar changes in RBC indices have been observed in humans and          impaction in these mice has been largely unremarkable to date.
laboratory rodents at altitude. These results may be important       Esophageal achalasia is primary motility disorder in which peri-
to consider during acclimation of research animals, as well as in    stalsis is lacking in the esophageal body and there is abnormal
studies that would potentially be affected by RBC mass.              relaxation in the lower esophageal sphincter. In humans, the
                                                                     etiology of esophageal achalasia is ill-defined, although an au-
P13 Use of Dietary Fiber in the Management of Chronic and Re-        toimmune component is suspected. The diagnostic test of choice
occurring Diarrhea in Pigtail Macaques (Macaca nemestrina)           for human esophageal achalasia is manometry.
K McGee2, D Patel*2, J McKenna2, F Banks2, S Sekou2, K Prevost1,
D Barnard1, B Skopets1                                               P15 Sublingual Blood Collection in Rats: An Alternative to
1DVR, 2SoBran,                                                       Tail Vein and Orbital Sinus Collection
                 Inc., NIH, Bethesda, MD
                                                                     EM Wolak*, A Freeman
Chronic diarrhea is a common syndrome in cage-housed nonhu-
man primates. Six 3- to 12-y-old male pigtail macaques (Macaca       Toxicology, Novartis Pharmaceuticals, East Hanover, NJ
nemestrina) with chronic or reoccurring diarrhea tested negative     Nonterminal blood collection for rats has routinely been accom-
for common bacterial and parasitic intestinal agents. All animals’   plished using the tail vein and orbital sinus puncture (primary)
diarrhea had been successfully managed for several months            procedures. These techniques require more skill to master and
by various repeated treatments with exception of dietary fiber       may lead to tail lesions as well as lesions in the orbital plexus,
supplementation. Two animals were fed a commercial labora-           resulting in damage to the eyes. Ophthalmoscopic examinations
tory primate diet containing 5% crude fiber. Four animals were
                                                                     and obtaining quality blood samples for use in clinical pathology,
fed a commercial laboratory primate diet containing 8.5% crude       toxicokinetics, and genomics are important in all toxicity studies.
fiber. Studied animals were not involved in ongoing research.        Currently, we are collecting blood sublingually from rats on our
We hypothesized that higher fiber content may be curative for        toxicity studies. Sublingual tongue vein collection has proven
chronic diarrhea. To test this hypothesis, we arbitrarily supple-    to be a successful and suitable method for collection and yields
mented 6 animals’ diet with 1 tsp of Metamucil once daily and        sufficient blood volume for quality clinical pathology and toxi-
gradually discontinued other treatments. As a result, 4 out of 6     cokinetics analyses. In addition, the site can be drawn from more
monkeys still had diarrhea, which was addressed by increas-
                                                                     than once in a day and the animals recover more quickly. The use
ing the dose of Metamucil to 1 tsp twice daily. The increase of
                                                                     of this procedure has no impact on the animal’s health or ability
Metamucil resolved diarrhea in 3 out of 4 animals. Animals           to eat and the site of collection heals quickly. The technique is
were maintained on Metamucil for a period of several weeks,
                                                                     relatively easy learn and can be applied for all types of blood
after which the decision was made to switch them to a higher         collections on research and toxicity studies.
fiber (approximately 15.5%) commercial diet. Initially, the ani-
mals were unwilling to eat the high-fiber diet. The problem was
overcome by gradually transitioning the monkeys from the low-        P16 Development of a Species-specific Giardia PCR Assay
fiber diet to the high-fiber diet and moistening the biscuits with   EK O’Connor*, LK Riley, CL Franklin, R Livingston
orange juice. During the transition, 2 animals had 1 to 2 episodes
                                                                     Pathobiology, University of Missouri, Columbia, MO
of diarrhea, which resolved without treatment or was quickly
reversed by administering Pepto-Bismol. In conclusion, chronic       Giardia muris is a flagellated protozoan that colonizes the small
or reoccurring diarrhea of unclear etiology in pigtail macaques      intestine of rodents and is generally considered nonpathogenic.
can be successfully managed by daily supplementation of 1 to         However, immunodeficient mice or mice with large parasite bur-
2 tsp of Metamucil or by feeding a diet containing 15.5% crude       dens may develop clinical signs associated with malabsorption
fiber. However, the study showed that primate diets containing       and maldigestion, including a rough haircoat, weight loss, and
15.5% crude fiber are not highly palatable to macaques and re-       abdominal distention. Current methods used to identify Giardia
quire a very gradual transition period and possibly a temporary      infections in rodents include histopathologic evaluation of the
flavoring of the new diet (such as orange juice).                    small intestine or visualization of Giardia trophozoites on direct
                                                                     smears of small intestinal contents. Drawbacks to these methods
P14 Dehydration and Excessive Salivation in B6.129-Slc6a4t-          include their inability to accurately speciate Giardia organisms
m1KplN10 Mice                                                        and the requirement for postmortem sample collection. Our
                                                                     hypothesis is Giardia PCR assays performed on feces will allow
E Paster*, D Hickman, R Baldwin                                      antemortem detection of infected mice and provide more sensi-
Abstracts of scientific papers 2008 AALAS National Meeting

tive and specific identification of Giardia infections compared      Tissues from a 5-y-old privately owned, male guinea pig that
to current detection methods. In this study, we developed PCR        died suddenly were evaluated histologically. The stomach con-
assays that target rRNA gene sequences to detect all Giardia spp.    tained a spherical (approximately 7 mm diameter) expansile
or to specifically identify G. muris. Feces and duodenal tissue      mass originating within the muscularis externa and elevating
from 10 mice from a colony with a history of Giardia infection       the submucosa and mucosa. The mass was densely cellular and
were tested using the developed PCR assays. All 10 mice tested       consisted of spindle-shaped cells with ovoid to flattened nuclei
positive on both PCR assays, while trophozoites were only            arranged in tightly interwoven bundles and whorls within a scant
visualized in 5/10 mice on direct smear. Additional testing of       collagenous stroma. There were foci of central and superficial
samples from pet store hamsters and mice with the G. muris           necrosis and a low mitotic index. The initial differential diag-
primers yielded positive results in 13/13 animals. The sensi-        nosis was leiomyoma; however, immunohistochemistry using
tivity of the G. muris primers was further evaluated by testing      antibody against the tyrosine kinase receptor KIT demonstrated
serial dilutions of plasmids containing cloned amplicons. The        strongly positive cytoplasmic labeling. This was consistent with
G. muris primers had a detection limit of 55 template copies.        a diagnosis of a gastrointestinal stromal tumor (GIST). GISTs are
The specificity of both PCR assays was confirmed by testing 17       mesenchymal tumors derived from the interstitial cells of Cajal in
fecal samples from uninfected mice as well as through sequence       the gastrointestinal tract, omentum, and mesentery. In humans,
analysis of amplicons generated from infected mice. These data       these tumors typically have activating mutations in KIT isoforms.
demonstrate that testing rodent feces with the developed PCR         Spontaneously occurring GISTs have been reported in dogs,
assays provides sensitive and specific antemortem detection of       horses, and as a case report in a rat. Because of their histologi-
G. muris infections in mice.                                         cal resemblance to leiomyomas, they may be underdiagnosed.
                                                                     KIT immunohistochemistry is generally necessary to confirm
P17 Incidence of Mycoplasma Contaminants in Cell Cultures:           the diagnosis. Differentiating these tumors from leiomyomas
Detection and Growth on Trypticase Soy Agar Containing 5%            is significant in human medicine because of the possibility of
Sheep Blood                                                          metastasis and the use of tyrosine kinase inhibitors for therapy.
                                                                     To our knowledge, this is the first report of a spontaneous GIST
GF Caviness*, JE Thigpen, J Locklear, TE Whiteside, MG Grant,        in a small laboratory animal species other than the rat.
DB Forsythe
National Institutes of Health, Research Triangle Park, NC            P19 Arthropod Detection during Screening of Sentinel Mice
Mycoplasma contaminants in cell cultures, tumors, tissues, and       J Locklear*, GF Caviness, JE Thigpen, TE Whiteside, MG Grant,
animal products can significantly impact in vitro and in vivo        DB Forsythe
studies. The Quality Assurance Laboratory routinely tests all cell
                                                                     National Institutes of Health, NIEHS, Research Triangle Park,
cultures, tumors, and animal products for viral, mycoplasmal,
                                                                     NC
and bacterial contaminants prior to animal inoculation. This
report describes the results from 2,241 specimens assayed for        Our health surveillance program is designed to detect pathogenic
microbial contaminants during the past 12 y and describes the        bacteria, mycoplasmas, viruses, ectoparasites and endoparasites
unexpected growth of some mycoplasma species on trypticase           in our laboratory animals. Sentinel CD-1 mice and F344 rats
soy agar containing 5% sheep blood (blood agar). The murine          housed in polypropylene cages with autoclaved hardwood bed-
viruses were detected using the conventional mouse antibody          ding and autoclaved NIH-31 diet were submitted for a complete
production test, serological assays, or polymerase chain reaction    microbial work-up. On physical examination, no abnormal clini-
(PCR) assay. Mycoplasmal and bacterial contaminants were             cal signs were noted. All sentinels were euthanized using carbon
detected using PCR assays or by plating the specimens on con-        dioxide. Blood, oropharyngeal swabs, and fecal samples were
ventional mycoplasma agar and blood agar. Microbial isolates         collected for microbial assays. Direct cecal preps and perianal
were identified using conventional microbial methods. Known          impression tapes were examined for endoparsites. Pelts were
mycoplasma control species were purchased from the American          examined for ectoparasites. Upon observation of the perianal
Type Culture Collection for comparative testing. The average         impression slides, 2 types of mites were detected. Murine mites
incidence of positive cell cultures was approximately 11% (255 of    were ruled out based on gross anatomical features. The first mite
2,241) during the past 12 y. The yearly incidence of mycoplasmal     species possessed a distinctive enlarged pair of pincer-like palps
contaminants in cell cultures varied from 5% (17/376) in 2000        on the dorsal claws. The second mite, which was 420 µm long
to 20% (39/196) in 1998. Surprisingly, no murine viruses were        and 320 µm wide, was creamy white, globular in shape, and had
detected in any of the cell cultures tested. An unexpected con-      a striated cuticle. The mites were diagnosed as the Cheyletus mite
firmed finding was that M. arginini, M. laidlawii, and M. pulmonis   and the American house dust mite. Mites were observed in the
formed small, visible, shiny colonies within 5 to 7 d on blood       feed bin fines, the suspected route of exposure. These species
agar. Our results confirm that mycoplasma contaminants are a         of mites are commonly found in feed warehouses where their
major problem for researchers using cell cultures; mycoplasma        food sources, such as grains, oils, and paper products, are stored.
species continue to infect a high percentage of cell cultures;       These arthropods have not been reported to cause any known
mycoplasma contaminants in cell cultures and animal products         harm to rodent colonies or research studies. It is important to
can significantly impact both in vitro and in vivo studies; and      distinguish them from pathogenic mites. Personal protective
mycoplasma species such as M. arginini, M. laidlawii, and M.         equipment should be used to protect against potential mite al-
pulmonis will grow on blood agar, which may be a useful tool in      lergens. The mite’s lifecycle requires a humid, warm environment
the detection and isolation of some mycoplasma species.              for survival. Therefore, feed storage environmental conditions
                                                                     should ensure relative humidity is below 50% and temperature
P18 Gastric Tumor in a Guinea Pig (Cavia porcellus)                  is maintained within 55 to 65 °F. Autoclaving the diet and inspec-
                                                                     tions and cleaning of feed bins and feed storage areas should be
I Bergin*, M Kiupel                                                  conducted routinely to prevent any pest infestation of laboratory
Michigan State University, Ypsilanti, MI                             animal feed products.
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

P20 Mycoplasma arginini Associated with Swollen Joints in              exposure to the rederived mice or soiled bedding. Lesions ob-
Severe Combined Immunodeficient Mice Implanted with                    served in nu/+ sentinel mice included bronchoalveolar adenoma
Tumor Cells                                                            (15/646) and epicardial mineralization (15/646). Lesions most
                                                                       commonly encountered in nu/nu mice were splenic lymphoid
J Story, J Thigpen*, K Meshaw, M Grant, L Cheatham, B Hol-
lister                                                                 hyperplasia (18/646), splenic plasmacytosis (8/646), cystic
                                                                       change of lymph nodes (12/646), bronchoalveolar carcinoma
Piedmont Research Center, Morrisville, NC                              (12/646) and acidophilic macrophage pneumonia (6/646). In
Severe combined immunodeficient (SCID) mice are more sus-              conclusion, both NU/NU and CD-1 Nude mice are acceptable
ceptible to microbial pathogens than immunocompetent mice.             sentinels for monitoring the health profile of research animals.
C.B.-17lcr Hsd-Prkcdscid mice were implanted subcutaneously            The strain-related lesions summarized in this report emphasizes
with human tumor cells and subsequently developed swollen              the importance of a thorough knowledge of the background le-
joints and hyperkeratotic dermatitis. The objective of this report     sions associated with the sentinel stock/strain used in a rodent
is to describe the microbial and histological findings observed        health program.
in these mice. Specimens of skin, intact tumors, affected hocks,
forelimbs and toes, and tumor cells were plated on mycoplasma          P22 Chronic Experimental High-fat Diet Exposure Leads to
agar and trypticase soy agar containing 5% sheep blood. M.             Coagulopathy in African Green Monkeys (Chlorocebus aethiops
arginini was isolated in pure culture on blood agar, and myco-         saebeus)
plasma agar and was confirmed by PCR assays from the intact
                                                                       K Ethun*, K Kavanagh, JR Kaplan
tumors and from purulent fluid from the swollen limbs and
toes. The skin from 5 mice was culture and PCR positive for C.         Wake Forest University, Winston-Salem, NC
bovis. The tumor cell line was PCR positive for M. hyorhinis and       High-fat diets (HDF) are often used experimentally in animal
M. arginini. Surprisingly, small, shiny, irregular colonies of M.      models to study human disease. We hypothesized that prolonged
arginini grew within 5 to 7 d on the blood agar plates inoculated      feeding of HFD can result in abnormal hemostasis after observ-
with the purulent fluid from the swollen joints and toes. These        ing 3 cases of fatal hemorrhage in nonhuman primates following
colonies were confirmed to be M. arginini by PCR procedures            routine clinical procedures. A 19-y-old male African green mon-
and sequence analysis. Swollen hocks revealed a moderate to            key (AGM) was euthanized after observing severe hemorrhage
severe pyogranulomatous inflammation involving the entire              (hematocrit [Hct] 13%) following routine venipuncture. A 13-y-
joint. There was fibrosis, destruction of bone, and pockets of         old male AGM and a 7-y-old female cynomolgus macaque died
neutrophils. Basophilic stringy material was observed in areas         due to acute anemia (Hct 17% and 18%, respectively) after initial
of inflammation of forelimbs and toes. It was concluded that           presentations of bleeding plantar xanthomas and coagulopathy
Mycoplasma arginini was associated with the swollen joints, and        characterized by melena and hematuria, respectively. All animals
was isolated in pure culture from most swollen joints. Mycoplasma      were healthy prior to presentations, but were maintained on
contaminants present a major problem to researchers using cell         HFDs containing ≥30% calories supplied by fat and supplemental
cultures and should be tested for microbial contaminants prior to      cholesterol for >1 y. The aim of this study was to screen in-house
injection into animals. This is the first report to describe swollen   AGMs (Chlorocebus aethiops saebeus) for abnormal hemostasis
joints in SCID mice due to M. arginini.                                under variable conditions of diet, disease, and age. Diabetic and
                                                                       non-diabetic AGMs (n = 21 [10 males, 11 females], 3 to 21 y old)
P21 Background Lesions in Homozygous and Heterozygous                  were divided into 2 experimental groups, including those con-
Mice of Two Stocks, Crl:NU-Foxn1nu and Crl:CD1-Foxn1nu,                suming a high-fat diet (HFD; 30% calories supplied by fat) and
Used as Sentinels                                                      those consuming a commercial low-fat chow diet (13% calories
                                                                       provided by fat) for >6 mo. There were 5 diabetics in the control
JE Thigpen*1, BA Hobbs1, CB Clifford2, MG Grant1, MB Myers1,
                                                                       group and 3 diabetics in the HFD group. Buccal mucosal bleeding
DB Forsythe1
                                                                       times (BMBT) were performed once per animal and evaluated by
1Comparative Medicine Branch, NIEHS, Research Triangle Park,
                                                                       a single blind observer. Results revealed that the consumption of
NC; 2 Charles River, Wilmington, DE                                    a HFD was associated with a 32% increase in BMBT compared
Ideally, animals used as sentinels in health surveillance programs     to those consuming a chow diet (P = 0.05). The mean BMBT for
should be free of microbial pathogens as well as background            those animals consuming a chow diet was 62.78 ± 11.016 s and
lesions which might confound results from health surveillance          99.83 ± 11.06 s for those consuming a HFD. There was a signifi-
programs or research studies. Therefore, knowledge of the              cant positive association of age with bleeding times (r = 0.56, P =
prevalence of spontaneous lesions in the age, gender, and strain       0.008). Hyperglycemic status did not affect BMBT (P = 0.8). The
selected for sentinels is critical. This report describes the spon-    duration of HFD consumption was positively associated with
taneous lesions observed over a 3-y-period in over 1200 NU/            a prolonged BMBT (r = 0.48, P = 0.03). Our results demonstrate
NU and CD-1 Nude mice. Of each stock, half of the mice were            that HFD prolongs bleeding times particularly in older animals
homozygous athymic nu/nu; the remainder were immunocom-                or following prolonged (years) of consumption.
petent heterozygous nu/+. These mice were used as sentinels
in 43 isolators housing rederived mice at Charles River. Specific      P23 Ependymal Cyst Involving the Occipital Cortex in an
pathogen-free sentinel mice were reared in isolators, then were        Asymptomatic, Adult Cynomolgus Macaque (Macaca fas-
transferred to other isolators and housed in direct contact with       cicularis)
colony mice for 2 wk, followed by 4 to 6 wk of exposure to soiled
                                                                       KP Yamada*1, SJ Popilskis2, A Garate3, M Oglesbee4, K LaPerle5
bedding. After 6 to 8 wk, sentinel mice were necropsied and ap-
                                                                       1Tri-Institutional Training Program in Laboratory Animal Medi-
propriate specimens collected for microbiology, serology, PCR,
and histopathology. No microbial pathogens were detected. All          cine and Science, Memorial Sloan-Kettering Cancer Center, The
lesions observed in the nu/nu and nu/+ sentinel mice of both           Rockefeller University, and Weill Cornell Medical College, New
stocks appeared to be strain-related, rather than resulting from       York, NY; 2Research Animal Resource Center, Memorial Sloan-
Abstracts of scientific papers 2008 AALAS National Meeting

Kettering Cancer Center, New York, NY; 3 The Animal Medical            Reducing time and manpower are also concerns, for research
Center, New York, NY; 4Department of Veterinary Biosciences,           technicians and investigators alike. A novel approach to rat re-
The Ohio State University, Columbus, OH; 5Laboratory of Com-           straint has been employed by our staff for such tasks. The rats
parative Pathology, Memorial Sloan-Kettering Cancer Center,            are guided into a conical-shaped terry cloth and held in restraint
New York, NY                                                           by rolling the edges and securing them with 2 large binder clips.
                                                                       The device has a small opening at the cranial end, which allows
An experimentally naïve, adult, male cynomolgus macaque
                                                                       for unrestricted breathing for the animal. It is also opaque and
(Macaca fascicularis) underwent a routine prestudy MRI of the
brain. The MRI revealed a diffuse, bilateral cystic lesion, in-        soft and gently molds to the animal to provide secure restraint.
volving the occipital cortex, whose content was isointense with        Most rats quickly become trained and enter the restraint readily.
cerebrospinal fluid (CSF). Clinically, the animal had normal be-       They can then be manipulated while lying quietly on one side,
havior and mentation. No observable visual deficits were noted         allowing the handler to have both hands free for equipment or
on tracking tasks, and the animal had no deficiencies in fine          animal manipulation. Holes near the rear limbs allow for access
motor control. On necropsy, the cystic structures communicated         to the saphenous veins, and the entire tail is exposed for access
with the caudal aspect of the lateral ventricles and contained         to vessels. The cloth is both inexpensive and sanitizable. Regular
clear fluid compatible with CSF by fluid analysis. Histopathol-        application of this technique by our staff has helped to decrease
ogy showed ciliated ependymal cells lining the cyst with focal         the amount of time and manpower needed to perform injection
areas of hemosiderophages within the cyst wall. This lesion is         or blood collection. In comparing 3 types of restraint methods, 9
compatible with an ependymal cyst, previously reported only            female rats were bled from the saphenous vein. Three rats were
in humans and generally considered an asymptomatic devel-              placed in each of 3 different forms of restraint: the plastic bag
opmental anomaly.                                                      decapitation cone, the bootie/shoe cover, and the terry cloth
                                                                       sewn into a cone shape. The venipunctures using the terry cloth
                                                                       restraint were completed more quickly than the other methods;
P24 Impact of Selective Alpha-2 Agonists on Blood Glucose              finishing 1 min and 7 s ahead of the bootie method and more than
Levels in the Laboratory Rat                                           5 min ahead of the decapitation cone method. The technician re-
JT Larsen, E Reardon, K Grayson*                                       ported that the cloth restraint was more secure, in that there was
                                                                       little opportunity for escape or injury to the handler or the rats.
Clinical Investigation Facility, David Grant USAF Medical Center,
                                                                       The rats demonstrated very little resistance or vocalization while
Travis AFB, CA
                                                                       in the cloth restraint, and less manipulation was necessary.
We randomly assigned 40 female Sprague-Dawley rats to 4
groups and anesthetized each with either: xylazine (5 mg/kg) +         P26 Anaplastic Large Cell Lymphoma in a Sooty Mangabey
ketamine (90 mg/kg); yohimbine (4 mg/kg) followed in 15 min            (Cercocebus atys atys)
by xylazine + ketamine; medetomidine (0.5 mg/kg) + ketamine
(90 mg/kg); or atipamezole (2.5 mg/kg) followed in 15 min by           L Collura*1, D Anderson2, JK Cohen1, P Sharma2
medetomidine + ketamine. We measured blood glucose levels              1Division of Animal Resources, 2Division of Pathology, Yerkes
(BGL) at baseline and at 15-min intervals for the 1st h, then at 2,    National Primate Research Center, Atlanta, GA
4, 8, and 24 h. In a separate experiment, 9 groups of 4 rats were
                                                                       A 13-y-old female sooty mangabey (Cercocebus atys atys) was
anesthetized with medetomidine + ketamine and had serum
                                                                       evaluated due to a large submandibular mass and decreased
analyzed for insulin and glucagon concentrations and liver
                                                                       appetite. The mangabey was serologically positive for simian
biopsies taken to determine glycogen content. In the xylazine-
                                                                       T-cell leukemia virus type 1 (STLV-1) and simian immunode-
ketamine group, BGL rose to 260 mg/dl by 1 h and fell to baseline
                                                                       ficiency virus (SIV). Generalized peripheral lymphadenopathy
by 8 h. Yohimbine partially blocked the post-xylazine blood
glucose elevation. BGL showed a much slower progression in             was also observed on physical examination. A fine needle
                                                                       aspirate of the mass was poorly cellular and did not grow any
the medetomidine-ketamine group, peaking at 2 h and remain-
                                                                       significant pathogens by aerobic bacterial culture. Biopsies from
ing elevated through 8 h. Atipamezole pretreatment prevented
medetomidine BGL increases. Serum insulin levels decreased             the submandibular mass and an inguinal lymph node revealed
                                                                       a poorly differentiated tumor, with initial consideration given
and glucagon levels increased in parallel with BGL, reaching 1
                                                                       to a diagnosis of possible sarcoma. The animal had a severe
ng/ml and 250 pg/ml, respectively. Similarly, the percentage of
                                                                       leukocytosis (79,900 with 55% lymphocytes). Due to poor prog-
liver cells stained with periodic acid-Schiff reagent that contained
glycogen fell sharply during the first 4 h after anesthesia, but       nosis, the mangabey was euthanized. Subsequently, based on
                                                                       the histologic evaluation and immunohistochemical analysis
had returned to baseline levels by 24 h. Our findings confirmed
                                                                       (vimentin positive, CD3 positive, cytokeratin negative) of the
a previous study that reported xylazine administration causes
                                                                       lymph nodes, a diagnosis of anaplastic large cell lymphoma,
hyperglycemia in rats. Medetomidine administration resulted
                                                                       a type of non-Hodgkin lymphoma, was made. This tumor has
in nearly the same BGL elevation; however, the hyperglycemia
                                                                       been reported in humans, baboons, and a chimpanzee but has
lasted much longer. BGL quickly returned to normal once the
                                                                       not been documented in a sooty mangabey. In addition, reports
rats recovered from anesthesia and resumed eating.
                                                                       of neoplasia in sooty mangabeys are infrequent.

P25 Alternative Method of Rat Restraint
                                                                       P27 Hind Limb Weakness in a Common Marmoset (Callithrix
LL Roberts*, M Dyson                                                   jacchus)
Unit for Laboratory Animal Medicine, University of Michigan,           M Batchelder*, M Sauer
Ann Arbor, MI
                                                                       Veterinary Sciences, Bristol-Myers Squibb, Wallingford, CT
Creating a comfortable confinement for animals that allows ac-
                                                                       A 3.5-y-old pair-housed male common marmoset (Callithrix
complishing tasks such as blood collection and injections while
                                                                       jacchus) presented with lameness and swelling of the left hind
minimizing stress is integral to the success of a research project.
                                                                       leg. The animal’s history included multiple minimally invasive
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

studies under isoflurane anesthesia. Physical exam revealed             P29 Characterizing a Body Condition Scoring Technique and
poor grasping ability in the left hind foot, weak reflexes, and         Scale for Rats
swelling of the left foot and ankle. Radiographs obtained the           DL Hickman1, M Swan*1, R Luksic2,1, B Bustamante1, E Paster1
following day revealed multiple abnormalities in the left tibia,
                                                                        1VA Medical Ctr, VA Medical Ctr, Portland, OR; 2College of Vet-
femur, tarsus, and phalanges consisting of radiolucencies within
the cortex and bullous periosteal densities. No lesions were            erinary Medicine, Oregon State University, Corvallis, OR
detected in other limbs, ribs, or pelvis. Subsequent radiographs        Evaluation of weight loss or gain in rodents is often problematic
revealed progression of the radiographic lesions in the left hind       for multiple reasons. Rodents are often group-housed, making
leg and the development of similar lesions in the right hind leg.       feed intake difficult to assess. Weights can vary depending on
The animal remained clinically stable during this time except           time of day, scale used, and even observer. Also, for changes
for weakness in the left hind leg. The animal did not appear            in weight to be clinically meaningful, it is necessary to have a
distressed or in pain, and was eating well. Ten months follow-          baseline weight or age-matched control rodent for comparison.
ing the initial presentation, the animal became depressed and           Body condition score (BCS) scales have been described for mul-
spent more time inside the nest box. Both hind legs were weak,          tiple species including domestic (dogs, cats, livestock), mice, and
and the animal was losing weight. The animal was euthanized,            African claw toed frogs. In this study, we used a rat model of
and both tibias were noted to be grossly abnormal at necropsy.          inherited polycystic kidney disease to characterize the applicabil-
Histopathology revealed bone marrow hyperplasia consisting              ity of the published mouse BCS scale to the rat. We have found
predominantly of granulocytic precursors and megakaryocytes,            that minor modifications to the mouse BCS scale are required
widespread cortical bone absorption, and periosteal bone for-           in overweight (BCS4) and obese (BCS5) animals as fat does not
mation. Extramedullary hematopoesis was evident in multiple             reliably deposit over the lumbar vertebrae in the rat. Specifically,
organs, and colitis was also noted. The histologic interpreta-          palpation of the dorsal fat pads over the pelvic bones is a more
tion was suggestive of several chronic myeloproliferative bone          reliable assessment of the rat BCS than palpation of the fat over
disorders in humans, but was not characteristic for any single          the vertebral column, as described for mice. The BCS technique
syndrome. This is believed to be only the second case of spon-          and scale presented here will be of use for personnel assessing
taneous chronic myeloproliferative bone disease reported in             the health of rats used in biomedical research.
nonhuman primates.
                                                                        P30 Comparison of Phlebotomy Techniques for Serial Bleeds
P28 Successful Treatment of C57Bl/6 Ulcerative Dermatitis               in Common Marmosets (Callithrix jacchus jacchus)
with Caladryl Lotion
                                                                        MB Moyer*, J Barclay, J Devorak, AM Schiavetta
ME Crowley*1, ML Delano2, SM Kirchain1
                                                                        Comparative Medicine, USAMRICD, Gunpowder, MD
1Laboratory Animal Science Center, Boston University Medical
Center, Boston, MA; 2Animal Resources at Children’s Hospital,           Veterinary technicians were assigned to collect blood samples
Children’s Hospital Boston, Boston, MA                                  for 3 time points, ranging from 10 to 45 min apart, in 16 common
                                                                        marmosets (200 to 500 g). The goal was to obtain 400 µm of whole
Ulcerative dermatitis (UD) in C57BL/6 mice and related strains          blood at each time point from the saphenous vein. Several methods
is a frustrating condition to manage in the laboratory animal           of saphenous vein blood collection were compared: use of a 1 cc
facility. Mice often become extremely pruritic ,with scratch-           tuberculin syringe, use of a cutoff butterfly catheter, and use of a
ing leading to several complications. Most of the pathology of          blood collection needle into a microvette tube or capillary tube.
UD results from secondary infections caused by the relentless           Collection devices were prepared using either liquid heparin or
scratching. Many topical and parenteral products and therapeutic        EDTA. One or more of these techniques were used on one or both
protocols have been used for years with varying success. We             legs of each marmoset. Femoral vein or tail vein phlebotomy were
have been using a therapeutic protocol for UD incorporating             conducted when all other techniques did not yield an appropriate
camphor/pramoxine/zinc (Caladryl) lotion for 2.5 y and have             sample size. Femoral phlebotomy increased the chance of large
seen a dramatic improvement in 65% of the UD cases. The first           hematomas; when used, that leg was only sampled once. The
component of the protocol is attentiveness by animal care staff,        marmosets were anesthetized using an isoflurane system (initial
who are the first to identify health cases and are critical to the      induction in an inhalant anesthesia chamber, followed by nosecone
success of this therapy. After early detection, the UD site is gently   maintenance) on a heated surface. Procedures were conducted
cleaned with chlorhexidine solution using a clean gauze and             on 4 animals simultaneously. Two technicians were assigned to
then dried with clean gauze. Caladryl is then applied in a thin         each animal: one monitored anesthesia and positioned the leg; the
layer once daily for 5 to 7 d using either a clean cotton swab or a     other performed the phlebotomy. Marmosets were prepared by
clean gloved finger. Caladryl dries the UD lesions effectively and      clipping the venipuncture site, then wiping the area with alcohol
relieves the itching. Improvement is seen in most cases within 2        to maximize vasodilation. The assistant occluded the vessel, and
to 3 d with a range up to 10 d. Healing ensues sequentially after       the sample was collected. Pressure and ice pack were then ap-
cessation of scratching, with resolution of erythema and drying         plied to the site. During recovery, technicians held marmosets
of the lesion. Treatment must continue after lesions have healed        until awake, then orally administered up to 1 ml of corn syrup.
grossly and must continue or be reinstituted if any scratching is       Upon return to their home cages, animals were provided a heat
noted. Some cases do recur several months after the initial di-         padrock in their hammock and a fleece pad on the floor. Different
agnosis. Therapy has been less successful in mice with multiple         techniques yielded different results for each technician; however,
UD lesions or advanced full skin thickness lesions. Pruritis is         the tuberculin syringe method or cutoff butterfly catheter method
a significant inciting event in many allergic and inflammatory          worked best, based on length of time to achieve blood flow
skin diseases. A major advantage of Caladryl is mitigating the          and total sample size collected. By using different phlebotomy
scratching to allow lesions to dry and heal. Many valuable mice         techniques, veterinary technicians met the study requirement
have been saved using this therapy.                                     challenges of conducting concurrent serial bleeds on marmosets
                                                                        with no adverse effects on the animals.
Abstracts of scientific papers 2008 AALAS National Meeting

P31 Topically Administered Recombinant Leptin for the Treat-          similar muscle contractions were observed in only some frogs
ment of Ulcerative Dermatitis in Zucker Lean Rats                     (2/6) with acetic acid at 15 min postinduction in the absence of
                                                                      a wiping response. AAT did not differ from baseline at 60 and
NV Gopee*, J Leakey, J Carraway
                                                                      120 min. Most animals had a depressed RR at 0, 30 and 60 min
National Center for Toxicological Research, Jefferson, AR             (100%, 83%, and 66%, respectively), and a depressed WR at 0
Zucker lean rats developed ulcerative dermatitis predominantly        and 30 min (83% and 66%, respectively). RR and WR nearly
behind the forelimbs, with isolated lesions on the head and           fully recovered 60 and 120 min, respectively. Propofol did not
behind the ears, several weeks following procurement from the         show any significant effects on HR (P > 0.05). On the other hand,
vendor. Histopathology of the skin lesions revealed moderate          propofol did induce respiratory depression (P < 0.01). The mean
to marked epidermal ulceration with neutrophilic infiltrates in       respiratory rate significantly decreased from baseline (27.8 ± 6.2
the presence of bacterial colonies, dermal necrosis of variable       breaths/min) to an average of 0.7 ± 1.6 and 0 breaths/min at 0
degrees of severity and numerous inflammatory polymorphonu-           and 15 min, respectively, and progressively returned to 19.5 ± 7.7
clear and mononuclear infiltrates, with several animals having        breaths/min at 2 h postinduction. AUC0-t was 24.07 µg min/ml
microabscesses or granulomas. Skin culture showed the pres-           and AUC to infinity (AUCinf) was 24.71 µg min/ml. The elimina-
ence of Staphylococcus aureus. Treatment with broad-spectrum          tion half-life (T1/2) was 1.18 h. In conclusion, when administered
amoxicillin/clavulanic acid per os did not resolve the skin lesions   as a single-bath immersion (88 mg/L) for 15 min, propofol does
despite bacterial sensitivity and previously reported success in      not appear to be a safe and effective anesthetic for Xenopus laevis
obese mice. Leptin is a 16 kDa circulating hormonal product of        frogs due to the narrow dose-effective window, as well as the
the ob gene and is reported to be higher in Zucker obese rats than    short duration and light anesthesia level obtained.
in lean rats. Review of the literature has reported that following
exogenous administration of leptin, enhanced re-epithelialization     P33 NHP Biodata Chart
and accelerated wound-healing were observed in obese mice.
                                                                      R Ford*
In addition, leptin alleviated the severity of the inflammation
in S. aureus-induced arthritis in rodent models. As such, a treat-    Veterinary Services, Primate Products Inc. Panther Tracks Learn-
ment regimen consisting of topical administration of 1 and 5 μg       ing Center, Immokalee, FL
recombinant rat leptin applied to the affected areas once daily       Data is very important in the world of biomedical research. It
every other day was initiated and maintained for several months.      will always determine your next step. Nonhuman primates are
Significant healing of the lesions, as evidenced by a reduction in    an important model in the biomedical industry. They are mainly
wound size and severity, was observed in the rats treated with        used for human diseases when lower-rank animals will not suf-
the higher (5 μg) dose of leptin compared with saline controls        fice, so we greatly rely on their data to determine our next step.
and lower dose (1 μg) animals. In addition, histology of the          As a technician who works with primates daily, I’ve noticed that
healing lesions was characterized by epithelial hyperplasia and       there are few pieces of information that contain important data
dermal fibroplasia adjacent to the ulcerated areas. However, due      about primates all in one place. If you have a question, you often
to the extensive nature of some of the ulcerations, healing was       find yourself riffling through pages and pages of references to
slow and prolonged and did not completely resolve in some             find an answer. I’ve designed and developed a chart that will
of the treated rats. No adverse effect on weight was observed         help answer your most common questions in one place. This
throughout the treatment period, and S. aureus was cultured           chart includes information on specific primates such as rhesus,
from the lesions following termination of leptin treatment. The       cynomolgus, African green, baboons, and chimpanzees, and
results further substantiate the potential for leptin to provide      date including hematology, chemistry, arterial blood gas, blood
beneficial therapeutic effects on the management of moderate          groups, physiology, growth curve, and dentition.
ulcerative dermatitis in Zucker rats.
                                                                      P34 Mouse Immunocomb Kit: A Quick and Easy Way to Si-
P32 Anesthetic Properties of Propofol in African Clawed Frogs         multaneously Detect Antibodies against Mouse Parvovirus,
(Xenopus laevis)                                                      Mouse Minute Virus, Murine Norovirus, and Mouse Hepatitis
S Guénette, F Beaudry, P Vachon*                                      Virus in Laboratory Mice
Veterinary Biomedicine, University of Montreal, St-Hyacinthe,         ML Wunderlich1, S Mazar2, L Campbell1, WR Shek1, RK Dha-
QC, Canada                                                            wan*1
                                                                      1Diagnostic Reagents, Charles River, Wilmington, MA; 2Biogal
The objective of this study was to determine the level of an-
esthesia attained in Xenopus laevis frogs using a propofol bath       Galed Labs, Kibbutz, Israel
administration. Thirty-three Xenopus laevis nonbreeding female        Immunocomb kit has been shown to be an effective tool to test
frogs were used. Following the determination of an appropriate        mouse sera for several important infectious agents of labora-
anesthetic dose (pilot study to show effectiveness of bath immer-     tory mice, including Sendai virus, coronavirus, and Mycoplasma
sion 88 mg/l for 15 min), the acetic acid test (AAT), withdrawal      pulmonis. Our hypothesis was that this same technology could
reflex (WR), righting reflex (WR), heart rate (HR), and respiratory   be used to detect serum antibodies against currently prevalent
frequency (RF) were used to evaluate central nervous system           infectious agents mouse parvovirus (MPV), minute virus of
depression. Pharmacokinetics of propofol were calculated fol-         mince (MMV), mouse norovirus (MNV), and mouse hepatitis
lowing blood concentration determination by tandem liquid             virus (MHV) in laboratory mice, and that this tool would perform
chromatography/mass spectrometry analyses. AAT results                with equal efficacy as the microplate ELISA and multiplexed
show that loss of the nociceptive response occurred between the       fluorometric immunoassay (MFIA) technologies. Serial dilu-
moment frogs were removed from the bath and during the fol-           tions of previously characterized standard mouse antisera and
lowing 30 min (P < 0.0001). Only at 15 min postimmersion was          nonimmune mouse serum were tested to compare the new kit
the wiping response absent. Muscle fasciculations following the       and ELISA detection limits and to assess the kit selectivity and
application of the acetic acid were observed (4/6 animals), and       diagnostic accuracy. The standard dilution at which sera are
                                                                                           Abstracts of scientific papers 2008 AALAS National Meeting

tested in the new kit is 36-fold. The ratio of the kit antiserum           Ten blood samples from 3 different colonies of mice were tested at
titers to this dilution ranged from 22 to >178. This proves that           random. Three samples were from mice known to be multiplexed
the kit sensitivity is ample for demonstrating viral seroconver-           fluorometric immunoassay (MFIA) negative for all viruses; 2
sion by mice from adventitiously infected colonies. Diagnostic             samples were from mice positive for mouse parvoviruses but
performance of the kit was assessed with serum samples from                negative for MNV; and 5 samples were from mice known to be
barrier-reared retired breeder and young adult SPF mice and                positive for MNV. We instilled 40 μl of blood from a heparin-
sera from experimentally infected mice. No false-positive or               microcapillary tube into the filter lid. Plasma was allowed to
false-negative results were obtained in 33 tests of known ELISA-           filter to the test membrane for 60 s. The lid was opened, and 150
positive sera and 128 tests of known ELISA-negative sera. The kit          μl of chase buffer was added to activate the test mechanism. The
assays detected seroconversion at the same days postinoculation            buffer saturated the test membrane, causing the plasma to flow
(DPI) as did ELISA and MFIA. These results show that the mouse             unidirectionally to a test port. The buffer resuspended a colloidal
immunocomb kit performance characteristics are equivalent to               gold marker that followed the plasma and conjugated with any
those of established and validated microplate ELISA and MFIA               antibody captured on the test or control lines. Captured colloidal
for serosurveillance of SPF colonies.                                      gold marker appeared as a red stripe of varying intensity (Rann
                                                                           scale). Tests gave positive results at the test or control lines within
P35 Biosecurity Lockout System to Influence Human Traf-                    10 min. All 5 devices detected the presence of MNV in the MNV
ficking: Ingress Control as a Component of the Prevention of               positive samples. The remaining five devices were negative for
Rodent Infectious Agent Transmission                                       MNV. Because no color developed at the test line with plasma
                                                                           positive for mouse parvoviruses, cross-reactivity did not occur.
RP Reynolds*1, SJ Pomeroy1, M Elks2, JN Norton1                            Control lines for all 10 tests were distinctly positive. Test results
1DLAR, 2Engineering and Operations, Duke University Medical                were 100% accurate for selectivity and specificity. Comparison of
Center, Durham, NC                                                         Rann scale intensity and MFIA optical density showed correlation
                                                                           in sensitivity. Results with the Fenway Direct Test demonstrated
Although there are many causes of disease transmission between
                                                                           a rapid test for MNV was feasible and could be used to detect
rodent areas in the research environment, one of the most diffi-
                                                                           infected mice and eliminate them from colonies.
cult to control is the traffic flow patterns of humans. Movement
of personnel and associated fomites from animal areas (whether
a module/suite or facility) of a lesser health status to a superior        P37 Spontaneous Case of Spina Bifida on a Mouse
health status can spread infectious agents. An application was             S Duarte-Vogel*, GW Lawson
developed where the Division of Laboratory Animal Resources
                                                                           DLAM, UCLA, Los Angeles, CA
codes the existing card access readers with a numerical level cor-
responding to the health status of the rodents in the specific area.       A spontaneous case of spina bifida occulta was found in a mouse
Individuals that enter a facility or area with rodents of a level          at our facilities. The mouse was a 22-d-old male, CD1-lpr/lpr
corresponding to lesser health status are locked out of areas of           with a “bulldog” tail that was short, thick, and had a 180-degree
a higher health status until the system automatically resets at a          curve to the right that began immediately caudal to the sacral
predetermined time. Unimpeded traffic flow can continue to occur           vertebrae. The mouse had complete bilateral rear limb paralysis
within the same level or to a level corresponding to a lesser health       with absence of superficial and deep pain as well as lack of anal
status. Currently, any restrictions incurred on one day will not           tone and severe generalized muscular atrophy. The differential
carry over to the next day due to the system reset during the early        diagnoses at this point were trauma, genetic malformation, and
morning hours, although this period of restricted access can be            spina bifida. Necropsy revealed the pelvic canal of this mouse
extended. This application also has the ability to generate reports        was extremely wide and deep because of a dorsal curve in the
of individuals who attempt to violate the prescribed traffic flow.         sacral vertebrae that began directly following the sacroiliac joint.
Training and proper notification of institutional personnel is criti-      A CT scan showed the defect in the vertebral column began at the
cal in mitigating concerns of the research staff prior to initiating the   level of the proximal thorax and continued to the coccyges. The
use of this bio-security lockout system. The support of institutional      thoracic vertebrae had partial development of the lamina and
administration is important in implementing this system and in             absence of the spinous process with exposure of the spinal cord.
setting penalties for those who violate the appropriate traffic flow.      Lumbar, sacral, and caudal vertebrae were abnormal on shape
This presentation will describe the system and its specifications,         and distribution, small, compressed, and difficult to differentiate.
other traffic flow considerations, and implementation of the system        The clinical signs and the gross and CT scan findings revealed
into the overall rodent health program.                                    spina bifida. Spina bifida is defined as failure of the neural tube
                                                                           to close during embryonic development. Different types of spina
P36 Detection of Mouse Norovirus in Less Than 30 Minutes                   bifida have been described and include spina bifida occulta,
Using a Novel Lateral Flow Test                                            spina bifida cystica (myelomeningocele), and meningocele. The
                                                                           neurological damage as well as the disability degree depends
N O’Luanaigh, CC Otte, WT Sauls, MP Goggin, BA Keniston,                   on the area of the lesion and how far down the spine it extends.
RE Hopkins*                                                                Spina bifida is commonly associated with animals with short
1BBI Research, British Biocell, Inc., Madison, WI; 2LACF, Boston           tails such as the Manx cat and the bulldog.
University, Boston, MA; 3Research-Development, Fenway Biosci-
ence, Boston, MA                                                           P38 Method for Repeated Blood Sampling from the Medial
Ten novel lateral flow test devices for detecting mouse norovirus          Saphenous Vein of Conscious Rats in a Long-term Study
(MNV) were developed to rapidly detect MNV in mouse blood                  K Koszdin1, S Frayo*2, D Cummings2
without sample preparation or instrumentation. Each device                 1Animal Research Facility, VA Puget Sound Health Care System,
had a filter lid for fractionating plasma from blood and a hous-
                                                                           Seattle, WA; 2Endocrinology and Metabolism, Veterans Affairs
ing containing a lateral flow test membrane. The membrane was
                                                                           Puget Sound Health Care System, Seattle, WA
striped with MNV antigen and anti-mouse antibody as a control.
Abstracts of scientific papers 2008 AALAS National Meeting

We developed a method for withdrawing up to 1 ml of blood from             1Division  of Animal Resources, Emory University School of
the medial saphenous vein because we needed to consistently col-           Medicine, Atlanta, GA; 2Department of Pathology, Yerkes Na-
lect at least 400 µl of blood from rats without assistance. The rat        tional Primate Research Center-Emory University, Atlanta, GA;
is restrained in a 1.5-in. diameter, 12-in. long cloth stockinet tube      3Department of Pathology and Laboratory Medicine, Emory

with one end partially closed with tape, such that the rat can push        University, Atlanta, GA
its nose out to breathe but cannot put its head through the hole.          A high incidence of gingival overgrowth occurred in a group
During placement, the front legs are pushed back to decrease the           of New Zealand White rabbits receiving daily cyclosporine
possibility of a toenail catching in the fabric. The stockinet is rolled   (15 mg/kg, IM) while on a retinoblastoma study. Initially,
up to expose the lower half of the body, and the rat is placed gently      4 rabbits presented with hypersalivation and inappetence.
on its back and restrained with the palm of one hand. One hind             Physical exam and accompanying necropsy revealed varying
limb is exposed, and the thumb and index finger of the restraining         degrees of hyperemia and moist dermatitis of the facial skin,
hand grasps this limb above the knee, straightening the knee and           and thickened gingiva in the maxillary and mandibular molar
occluding the vessel. Petroleum jelly is spread on the medial side         area. Over the next 9 mo, 10 of 18 additional rabbits presented
of the leg to visualize the vein and to prevent blood from soaking         with similar clinical signs (hypersalivation: 4/18, inappetence:
into the fur. A bright light helps with visualization. The medial          3/18, both: 3/18), prompting attempts at treatment and a clini-
saphenous vein runs along the inner thigh and branches just distal         cal investigation. In an effort to alleviate symptoms, the dose
to the knee in the shape of a wishbone. The vein is pierced 2 mm           of cyclosporine was reduced to 10 mg/kg. The dose reduction
proximal to this branch point with a 20-gauge needle. Blood can            resulted in complete resolution of clinical signs in all but 2 rab-
be collected in a capillary tube. After blood collection, the vein is      bits. These rabbits were administered azithromycin (62.5 mg PO
compressed to minimize hematoma formation. The stockinet is                once daily for 7 d), a common treatment for cyclosporine-induced
gently rolled off the rat. We were able to rapidly collect up to 1         gingival overgrowth in other species. Following dose reduction
ml of blood from a single vein in rats ranging from 350 to 660 g.          or azithromycin treatment, clinical signs resolved and rabbits
The same animals were sampled every 3 to 4 wk for over 1 y. The            remained free of symptoms throughout the remainder of the
minimum time between collections was 17 d. Using this method,              study. Fourteen rabbits were necropsied at the end of the study,
one person can reliably and repeatedly collect at least 1 ml of blood      and gingival width was measured. Although clinically normal,
from a conscious rat.                                                      the gingiva in these rabbits was grossly thickened. Rabbits on
                                                                           cyclosporine for at least 2 wk had gingiva around the molars
P39 Ensuring Normothermia during General Anesthesia                        that was significantly thicker (4.8 mm; P < 0.05) than untreated
S Luboyeski*                                                               controls (2.5 mm). Histologic analysis revealed mild to moder-
                                                                           ate gingival epithelial hyperplasia and mild inflammation of
CCM, Massachusetts General Hospital, Charlestown, MA                       the molar gingiva. Mild inflammation but not hyperplasia was
Animals undergoing prolonged imaging studies or invasive                   observed in the incisor gingiva. Gingival overgrowth is a known
surgery, or having a low percentage of body fat or a high ratio            side effect of cyclosporine administration in other species but,
of surface area to body weight may exhibit up to 8% loss of core           to the authors’ knowledge, this is the first report in rabbits. Be-
body heat during just 90 min of general anesthesia or prolonged            cause rabbits are commonly used in studies which may involve
sedation in the absence of appropriate supportive care. The resul-         systemic cyclosporine administration, clinicians are advised to
tant hypothermia routinely reduces heart and respiratory rates,            include this possibility on their differential list in cases involving
blood pressure, and cardiac output, and prolongs recovery times.           hypersalivation, facial dermatitis or inappetence.
Widely used methods to avoid hypothermia, such as circulating
warm water pads under the animal or heat lamps overhead, can               P41 Effect of Automated Cross-over Serial Blood Sampling on
be inadequate or interfere with MRI and other scanners. The effi-          Urinary Corticosterone in Rats
cacy of covering the animal’s torso and extremities with pediatric
hand and foot booties and using padded woolly blankets with                SE DeBoer*1, E Kadyszewski2, MS Kovacs1
warm water bottles during sedation and transport, was assessed             1Comparative    Medicine, 2Non-Clinical Statistics, Pfizer, Inc.,
in 4 3-y-old rhesus macaques in the course of routine veterinary           Groton, CT
support during general anesthesia for MRI scans. Animals were              Automated blood sampling provides a method for sequential
provided a circulating warm water pad only, hand and feet
                                                                           sampling of small amounts of blood in freely moving animals.
booties only, or a water pad in conjunction with booties. Some             We wanted to evaluate the utility of performing crossover pK
animals were also provided blankets and warm water bottles
                                                                           studies on the same animal using an automated blood sampler.
during sedated transport to the imaging room. All animals were             The objective of this study was to compare the effect of 1 and 2
subjected to each combination over 5 scans. Rectal temperature             serial blood collection sessions on the physiologic stress hormone
was obtained at the onset of anesthesia and at extubation. When            corticosterone in rats. A total of 15, male Sprague-Dawley rats
these novel patient comforts were applied, animals experienced
                                                                           were used in this study. Five precannulated rats were assigned
less than an average 0.5 °F body temperature difference between
                                                                           to each treatment group. Five rats received a standard pK blood
onset and end of anesthesia, compared with an average 2 to 3               collection protocol over a 24-h period, involving collection of
°F drop in body temperature with only a circulating water pad              approximately 10% of their blood volume. Five animals received
provided. In addition, recoveries were smoother and recovery               2 blood collection protocols separated by a 24-h wash-out, in-
times were reduced by 20 to 30 min. The addition of these items
                                                                           volving a total collection of approximately of 20% of their blood
has a greater impact on maintaining normothermia than current              volume. Five control animals were tethered in the automated
conventional methods alone.                                                blood sampling units but were not subjected to automated sam-
                                                                           pling. Urine was collected twice daily from each rat to measure
P40 Cyclosporine-induced Gingival Overgrowth in New Zea-                   urinary corticosterone and creatine. For single and double serial
land White Rabbits (Oryctolagus cuniculus)                                 blood sampling protocols, there was a statistically significant
S Jean*1, P Sharma2, DK Taylor1,3, DM Mook1,3                              elevation in the urinary corticosterone/creatine ratio during
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

each of the blood collection sessions in the automated sampler         tered a single subcutaneous dose of 1 of 4 different PEG-leptin
when compared to controls. Despite significant decreases in            variants, native human leptin, or vehicle control at study initia-
hematocrit in both the single and double serial blood sampling         tion. Body weight and food consumption were recorded daily for
sessions, urine corticosterone/creatine ratios returned to normal      7 d after treatment. Over that period, we saw increased weight
within 24 h of each sampling session. We determined that rats          loss and decreased food consumption in mice treated with the
recover quickly from the stress of blood collection and despite        PEG-leptin variants. Mice treated with native human leptin or
lower than normal hematocrits, evaluation of the stress hormone        vehicle control did not lose weight and had slightly increased
corticosterone indicates no residual stress effects.                   food consumption. In conclusion, we have shown that a single
                                                                       subcutaneous administration of a PEG-leptin variant in ob/ob
P42 Ambulatory Approach to Long-term Bile Collection in                mice caused increased weight loss and decreased food consump-
the Canine                                                             tion when compared to mice treated with native human leptin,
                                                                       which indicates a therapeutic advantage of these compounds
S Vessie, T Ziegelhofer*                                               when compared to commercially available compounds.
Cardiovascular/Infusion Toxicology, Huntingdon Life Sciences,
East Millstone, NJ                                                     P43 Critical Analysis of a Biosecurity Program
After presurgery bloods and jacket acclimation, each canine is         AR Santiago*
prepared for an aseptic surgical procedure during which a T-
                                                                       DVMS, UT—MD Anderson Cancer Center, Houston, TX
catheter is secured within the lumen of the common bile duct.
The terminal ends of the catheter are tunnelled through the            With the exponential growth of animal research in the Texas
abdominal wall and connected subcutaneously to 3 vascular              Medical Center comes increased concerns about biosecurity.
access ports (VAPs). For 6 wk postsurgery, animals are carefully       Maintaining the integrity of research and assuring the health
monitored for signs of pain, distress, and jaundice. Catheters are     and welfare of laboratory animal colonies require an up-to-date,
frequently flushed to ensure patency, and liver function is as-        feasible, and cost-effective biosecurity program in every animal
sessed through weekly blood collection. The occluding balloon          facility. The following major areas at our facility are described:
within the catheter is inflated, temporarily obstructing the flow      traffic control, pest control, cleaning and disinfection, host sus-
of bile into the duodenum. Animals are placed in jackets, with         ceptibility, and communication. The biosecurity program of M.D.
a bile sampling line and a cooled bile collection bulb within the      Anderson’s vivarium is current and cost effective. Occasional
jacket pocket. Simultaneously, an ambulatory pump in the op-           biosecurity breaches are attributed mainly to lack of training or
posite jacket pocket infuses a bile salts solution into the duodenal   language barrier. Human error and economic constraints are the
end of the catheter. Upon study termination, jackets are removed,      major limiting factors to an ideal program. The recommenda-
catheters are flushed with saline and the occluding balloon is         tions are inclusion of biosecurity in orientation of personnel and
deflated to allow bile flow from the gall bladder to the intestines.   investigators; translation of signs into 2 other major languages
The animals are flushed at least once every 14 d to ensure proper      spoken in the facility; enrollment in English as a second language
catheter function. Bloods are collected once per month to evalu-       courses offered at the institional level; incentive program for
ate liver values. We have successfully performed surgery on 10         compliance; termination for top 3 biosecurity measure breaches;
canines and executed several studies to completion. We believe         and the formation of an interinstitutional committee for regional
our refined, ambulatory method of long-term bile sampling is           perspective.
the most optimal. Since the model does not require tethering,
the animals are not only paired housed during non-sampling             P44 Contamination Control Mat: Solving a Hairy Situation
periods, but are also readily exercised during collection intervals,
a significant advantage to this procedure.                             A Dryman*
                                                                       University Research Animal Resources Life Sciences, University
P42.5 Increased Weight Loss and Decreased Food Consump-                of Georgia, Athens, GA
tion in Ob/ob Mice Administered PEG-leptin Variants when               Working with New Zealand White rabbits results in large
Compared to Mice Administered Human Leptin                             amounts of rabbit fur floating in the air. This fur coats the work-
M Cook*, V Kraynov, H Cho, B Kimmel, D Axelrod, J Pinkstaff,           space and personnel, aggravating allergies and any personnel
L Sullivan                                                             trying to clean the workspace after procedures using rabbits
                                                                       are complete. We run a polyclonal antibody production service
Ambrx, La Jolla, California                                            that uses New Zealand White rabbits, and sometimes an entire
This study was designed to evaluate the efficacy of 4 different        workday is spent handling rabbits in our small procedure room.
PEG-modified human leptin compounds when compared to                   After multiple procedures, the rabbit fur adheres to all horizontal
native human leptin in ob/ob mutant mice. Leptin, a 10-kDa             surfaces, along with scrubs and lab coats. Previously, our sani-
protein, is produced by adipose tissue and is thought to play a        tation method was to carefully vacuum the workspace, trying
critical role in body weight regulation. Due to leptin effects on      to suction up the fur without blowing it throughout the room,
appetite satiety, energy expenditure, and metabolism, leptin is        which met with limited success. About a year ago, we started
a viable therapeutic option for weight loss and weight manage-         using a mat on our exam tables, similar to the contamination
ment in obese patients. Initial studies evaluating the therapeutic     control flooring mats we use outside of our barrier and quar-
potential of leptin have been performed in mutant ob/ob mice.          antine rooms. Initially, the idea was to see if it would improve
These mice lack functional leptin and, as a result, have reduced       the rabbits’ welfare by preventing distress due to sliding on the
activity, decreased metabolism, and are genetically obese. We          slick metal exam table. We were excited to find a different and
hypothesized that administration of PEG-leptin would cause             entirely helpful use for the mat. The fur that once roamed our
increased weight loss and decreased food consumption when              procedure room would instead collect on the mat. We found that
compared to animals administered native human leptin. Female           those members of our staff with rabbit allergies had reduced
ob/ob mutant mice (approximately 8 wk of age) were adminis-            symptoms when we started using the mat. The mat is easy to
Abstracts of scientific papers 2008 AALAS National Meeting

care for and shortens cleanup time immensely. We would rec-                weight was measured. Average food wasted was significantly
ommend using this mat for rabbit work, and hope to try it with             (P = 1.94e-05) reduced from 26% to 0.5% with the modification.
other species soon.                                                        Food left in the ball did not vary significantly between the groups
                                                                           (P = 0.37). Each tray cost $10 in materials and took 1 h to build.
P45 Effect of Cage Change Frequency on Rodent Breeding                     Daily cleaning with running water and scheduled cage sanita-
Performance                                                                tion in the rackwasher were both easily accomplished with balls
                                                                           in situ. This simple and inexpensive modification significantly
AE Sanderson*, HM Castor, J Lohmiller, S Boutin                            reduced food waste, with the added benefit of reduced cleanup
Veterinary Services, Taconic Farms Inc., Hudson, NY                        time following use.
The Guide states that “in some instances, frequent bedding
changes are contraindicated, such as during some portions of               P47 Online Inventory Equipment Database System
the prepartum or postpartum period.” Consequently, the pro-                B Liu*, KA Wendler, CA Osina, L Sun, JJ Sharp
fessional judgment of many investigators, facility managers,
                                                                           Center of Comparative Medicine, Baylor College of Medicine,
and animal care technicians is that disrupting certain sensitive
                                                                           Houston, TX
rodents will result in breeding failures and loss of newborn pups.
We assessed breeding performance of 1 presumed sensitive rat               Equipment used in vivaria includes hundreds of racks, numer-
line and 1 mouse line to determine what effect cage change had             ous hoods, and many pieces of other equipment types. Due to
on the survivability of new litters. We selected C57BL/6NTac               frequent movement of many types of equipment among animal
mice and NTac:NIH-Whn rats based on their presumed high                    facilities in geographically separate locations, it is important to
sensitivity to parental neglect or cannibalization in response to          track the location of equipment and maintain updated informa-
cage manipulation. The study consisted of approximately 250                tion about the condition of the equipment. We have introduced a
breeding rat females and 5,700 breeding mouse females per wk               web-based online inventory database system that keeps records
and was conducted in multiple barriers at 2 geographic locations.          of equipment for animal facilities. The system is compatible with
Breeding performance was evaluated over a 22- to 25-wk period.             different computer platforms and different browsers. Each piece
Study animals were placed in 1 of 2 study groups, “change” or              of equipment is identified by an applied barcode; this barcode
“no change.” The change group had their cages changed on a                 technology ensures the integrity of data records. Data entry is
weekly basis regardless of the presence of newborn pups, and               facilitated for single items, multiples of the same item, or a large
the no change group was not changed until the following week               quantity of items. An implemented web interface allows batch
if new pups were observed. The study design was such that all              data entry. For convenience, a formatted spreadsheet can also be
environmental and caretaker factors were rigorously controlled.            uploaded from the website, which allows facility users to record
The primary factor for evaluating breeding performance was                 the information quickly for racks and cages. Users may search
comparing change with no-change pup survivability at the                   the database for equipment not in use and submit a request to
time of weaning. Data was collected on a weekly basis, captur-             have it moved to their facility or request a transfer of equipment
ing the total number of female breeders and the total number               from one facility to another. Alternatively, users may request that
of pups weaned. The production efficiency was calculated by                equipment be removed from their vivarium to the warehouse.
dividing the number of pups weaned by the total number of ac-              Equipment needing repair or to be discarded is also noted. Users
tive breeder females. The data demonstrated that there was no              can search the inventory items in the database, review the im-
statistically significant difference in the breeding performance           ages of the items, and make a request for the specific items. The
or efficiency of C57BL/6NTac mice or NTac:NIH-Whn rats based               warehouse manager will then be notified and deliver the items
on cage change alone. This outcome is contrary to the general              to the correct site. The database keeps warranty information,
belief of rodent breeding and would not support modifying                  which will be helpful in the event that service is needed for any
recommendations of the Guide stated as “There is no absolute               equipment. Equipment usage and performance information can
minimum frequency of changing bedding, but it typically varies             be obtained from archived data which can inform equipment
from daily to weekly.”                                                     replacement decisions. The database provides accessibility for
                                                                           future mobile systems such pocket PCs.
P46 Simple Modification of a Commercially Available Primate
Foraging Device Significantly Reduces Food Waste                           P48Web-based Animal Internal Transfer Systems
AS   Glenn*,   J Watson                                                    B Liu*, L Sun, JM Simonet, KA Wendler, JJ Sharp
Animal Resources, Johns Hopkins University, Baltimore, MD                  Center of Comparative Medicine, Baylor College of Medicine,
                                                                           Houston, TX
Commercially available perforated foraging balls that hang on
the outside of the cage are used at our institution for nonhuman           Internal animal transfers (room to room, PI to PI, protocol to
primate enrichment. Food treats are placed in the balls and pri-           protocol, and so forth) are one of the most important activities
mates forage for them through holes in the balls. However, 15              of an animal program. Prior to our web-based animal internal
to 55% of the treats were wasted when they fell onto the floor             transfer system, all transfer requests were submitted on paper,
and out of reach. We addressed this problem by constructing                which was slow, and there was no central database to reference
a polycarbonate tray to catch dropped food and permanently                 for status of transfers. Coordinated by the transfer agent, a series
attaching it to the bottom of each ball with 2 3-in stainless steel        of actions was required to ensure that the transfers were done cor-
bolts. Eight macaques were used in a study to assess the effect            rectly and promptly. For example, when a PI submitted a transfer
of the modification on wasted food. Balls were placed on cages             request, the transfer agent had to verify the animal protocols,
for a fixed period each day (1.5 to 5 h) in a crossover design.            retrieve health information from veterinarians, check with the
Four macaques were given the modified ball for 3 d then the                IACUC for possible compliance issues, and ask the receiving
unmodified ball for 3 d, and 4 were given the balls in reverse             facility manager to verify space availability. Once the request
order. Initial, final (left in the ball), and wasted (on the floor) food   was fully approved, the transfer agent had to contact the PI, the
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

manager of the sending facility, and the manager responsible for         were counted following 48 h of incubation at 37 °C. Mean CFU
the actual move. After the transfer was completed, the receiving         counts for clean and dirty samples from unsealed barn concrete
PI had to be informed. In most cases, the parties involved in this       floor were significantly lower than corresponding counts from
process were in separate locations. Our new web-based system             the sealed barn concrete floor. Mean CFU counts obtained after
allows all of the management steps to be handled with just a few         cleaning and disinfection were acceptable in all barn locations
clicks. Completion of an upstream step automatically triggers a          tested. Pooling of disinfectant in cracks, crevices, and pores of
notification email for next downstream steps. Notification emails        unsealed floor may allow for longer contact time compared to
for each step are automatically sent to the appropriate persons.         sealed flooring, where residual disinfectant dries more quickly
Investigators benefit from this automated system because they            or runs off into drains.
can view the real-time status of their requests. Completed transfer
requests are archived, with reporting capability available if de-        P51 Simple Environmental Enrichment Technique for Am-
sired. Validation elements have also been implemented to ensure          phibian Species
the security and integrity of data. Early use of this system shows
it to be efficient and easy to use by all parties with a high level of   CA Bluhm*, PA Stiles, A Johnson
acceptance among researchers and animal facility managers.               Animal Care, Ferris State University, Big Rapids, MI
                                                                         A common misconception is that captive reptiles are stoic,
P49 Novel Cage Flag System for Identifying Special Husbandry             adaptable, and highly tolerant of the abnormal conditions in
Requests                                                                 captivity. However, research suggests that reptiles may be less
BA Davis*, H Graninger, ND Hitt                                          tolerant and less adaptable to captive environments than was
                                                                         suspected. Because there has been limited information on ap-
NINDS/AHCS, Charles River, Germantown, MD                                propriate enriched artificial habitats, many of the current reptile
Rodent facilities are often faced with special husbandry requests        husbandry and enrichment techniques have been derived from
because of phenotype- or protocol-related issues. These requests         empirical evidence. We developed a lab-tested, simple, sustain-
can range from special feed for an entire room to varying indi-          able environmental enrichment technique for an amphibian
vidual cage care instructions. This degree of customization of           species habitat. Our previous habitat consisted of a water-filled
care can result in miscommunication and lost research data if an         36 by 36 by 24 in. square polystyrene container raised at one
efficient, accurate method to alert the animal care staff to changes     end. Rocks were placed throughout to allow the frogs rest areas.
in husbandry is not used. After trying numerous methods to               While this habitat was in use, we experienced a high incidence
identify cages where special husbandry was required, a system            of redleg and as a result a high mortality rate. A decision was
was designed where small laminated flags are clipped to cage             made to design and construct a habitat that would more closely
card holders on the individual cages. These color-coded flags            resemble that of the frog’s and turtle’s natural habitat. The end
display special instructions using a combination of the general          result was the use of a 100-gal rugged plastic livestock tank that
description of the change with specific details noted in perma-          was divided using a polycarbonate resin sheet. One side of the
nent marker. When special care is no longer required, cards are          tank was filled with river rock and pea gravel; large stones were
easily detached and wiped clear with ethanol for reuse. To make          placed on the bottom of the tank on the opposite side and the
identification of cages convenient for animal care and research          tank filled with water. In doing this, we created a dry side and
staff, several sets of each type of flag are provided in each animal     swimming hole that is approximately 12 in. deep. The frogs rest
holding room. After several revisions, we have streamlined our           on the rock-filled side and eat crickets, and swim in the water-
flag system so that it consists of 5 flags, each with our most com-      filled side. A heat lamp on a timer was placed above the tank to
mon special care and feed options, which can be checked by the           allow the frogs to bathe in the artificial sunlight. A base and lid
researcher using a permanent marker. This system has greatly             were constructed for the tank using composite decking material.
improved our ability to provide the customized care required             The base of the tank is on casters to allow easy movement of
by so many of the research projects in our facility with minimal         the tank. The lid was designed to remain in place until the frogs
additional expense.                                                      are removed or the tank is cleaned. A drain in the bottom side
                                                                         of the tank with a shut-off valve allows for easy cleaning. We
P50 Microbiological Monitoring of Sealed and Unsealed Barn               have housed up to 75 frogs at a time in this habitat with little
Floors using RODAC Bacterial Culture Plates                              or no death and have not had an incidence of redleg since. We
                                                                         also extended the same concept to housing our turtles. The tank
BF Grasperge*, J London, R Stout, D Baker                                is a 50-gal plastic stock tank, though it lacks a base and lid. A
Division of Laboratory Animal Medicine, Louisiana State Uni-             simple ramp was designed using polycarbonate sheeting and
versity School of Veterinary Medicine, Baton Rouge, LA                   indoor/outdoor carpeting secured with carpet tape to allow the
                                                                         turtles easy access to the dry area.
Sealed flooring is considered ideal for animal holding facilities
due to its impermeability to contaminants. The objective of this
study was to determine the cleanliness of sealed and unsealed            P52 Making the Transition to Biometrics
concrete floors within the Louisiana State University School of          C Kilcullen-Steiner*
Veterinary Medicine client barn before and after cleaning and
                                                                         University Animal Care, University of Arizona, Tucson, AZ
disinfection using replicate organism detection and counting
(RODAC) bacterial culture plates. The floor locations sampled            Security is paramount in biomedical research facilities. Keys and
included unsealed concrete barn aisle, sealed concrete dump-             security cards are commonly used to restrict access to the animal
ster storage area, sealed equine intensive care unit, and sealed         facility to only authorized personnel. However, these items can
equine surgery floors. For each of the 4 locations, 10 sites were        be easily shared among research personnel, and some may write
sampled immediately before and 10 min after cleaning and dis-            the access code on the card, jeopardizing facility security. To
infection. Floor sites were sampled with RODAC plates, and the           increase security, the university decided to go biometric. Optic
total numbers of bacterial colony-forming units (CFU) per plate          scanners, finger scanners, and palm scanners were investigated.
Abstracts of scientific papers 2008 AALAS National Meeting

Optic scanners were ruled out because many felt the scanners          Laboratory Animal Services, Novartis Pharmaceuticals Corpora-
could damage their eyes or make them go blind. We selected            tion, East Hanover, NJ
fingerprint scanners, despite some employees’ fears of having         The common marmoset (Callithrix jacchus) can be an invaluable
their fingerprints captured; the idea that the fingerprint was        research model. Its small size makes it particularly useful when
converted to a logarithmic equation was hard to accept. The           compound is in short supply or expensive to produce, and safety
change to biometrics took several months. New biometric cards         concerns such as those encountered when working with old
had to be purchased and issued. Fingerprint equations had to be
                                                                      world nonhuman primates are relatively nonexistent. We have
downloaded on the biometric cards and the new readers had to          used this species for approximately 10 y with success in several
be in place and functioning prior to the changeover. Having the
                                                                      types of research in both discovery and preclinical safety. The
new biometric system makes it impossible for an access card to
                                                                      small size of the marmoset allows its use with approximately
be used by someone other than the approved card holder. Usage
                                                                      1/10 the amount of compound that would be necessary when
can be tracked better with this technology, thus creating security
                                                                      working with a more commonly used model, such as the cyno-
levels to meet the needs of our users. The new system has not
                                                                      molgus macaque. Unfortunately, two aspects of the marmoset’s
eliminated all access problems; a very small percentage of us-
                                                                      physiology can present significant challenges for the researcher.
ers do not have readable fingerprints, and the system does not
                                                                      It has a high metabolism rate that makes post–blood collection
prevent “piggyback” entry. We are looking to new technology
                                                                      hemostasis difficult even though it has small blood vessels.
to remedy those problems.
                                                                      Once blood is drawn, it seems to clot quickly, often causing
                                                                      blood samples to need to be redrawn. The methods of blood
P53 The Effects of Social Housing on Diabetic Laboratory              collection and intravenous injection used at our site have been
Canines: Food Consumption, Weight, Behavior, Glucose Toler-           continually modified over the years in an effort to minimize
ance, and Insulin Levels                                              these challenges. Modifications in restraint methods, equipment,
C Coke Murphy*1, KJ Salleng1, M Scott2                                and post-collection hemostasis procedures have progressed to
1Division of Animal Care, 2Molecular Physiology & Biophysics,
                                                                      continually improve to what are considered best practices from
                                                                      an efficiency and animal welfare aspect. Today, the techniques
Vanderbilt University Medical Center, Nashville, TN                   used for these procedures with marmosets are proven to be suc-
Obesity and diabetes research models depend on monitoring             cessful; the result of many years of working with this species are
many factors, such as weight maintenance, daily food consump-         presented in this poster. Researchers will find these best practices
tion, and glucose and insulin measurement. This has historically      for injection and bleeding to be helpful in their work with the
resulted in the single housing of canines. Long-term compatible       common marmoset.
social housing has been shown to be beneficial to the physical
and psychological well-being of laboratory canines and further        P55 Quantitative Monitoring for Laboratory Animal Aller-
led to a reduction in the incidence of stereotypical behaviors.       gens
The aim of this study was to determine if social housing initi-
ated mid-study would affect experimental variables of weight,         C Zumbaugh*, C Davis
food consumption, stereotypic behavior expression, oral glucose       Eli Lilly and Company, Indianapolis, IN
tolerance test (OGTT), and fasting plasma insulin. Three adult
                                                                      Managing potential exposures to laboratory animal allergens
male hound dogs (Class B) that were part of a study on the ef-
                                                                      (LAA) is vital for maintaining a healthy work environment.
fects of a chronic high-fat diet and pancreatic insufficiency of      Quantitative air monitoring studies can be used to characterize
glucose metabolism were tested for compatibility. Dogs were
                                                                      employee exposure to LAA and assign effective control measures.
socially housed in the mornings after food bowls were pulled
                                                                      A series of studies were conducted within a large research facil-
and then separated in the afternoon to be fed. Two of the dogs
                                                                      ity to investigate the airborne levels of LAA, specifically mouse
were not compatible so 1 of the dogs alternated being paired with
                                                                      urinary protein (MUP) and rat urinary protein (RUP). A new
the other two dogs daily. After 12 mo of pair housing, 2 of the 3
                                                                      analytical technique, multiplex array technology, was established
dogs had gained weight (+2.04 kg and +1.66 kg), and the third
                                                                      to analyze the samples. After the successful completion of a pilot
had maintained its weight. Food consumption 60 d before and
                                                                      study, a comprehensive monitoring strategy was implemented to
60 d after pairing showed similar results, with 2 of the 3 dogs       collect exposure data for common rodent handling tasks. More
increasing food consumption post-pairing (+5% and +12.5%)             than 150 personal air samples and over 200 area samples were
and 1 maintaining. The dog that maintained its weight and food
                                                                      collected. The resulting data was categorized into exposure control
consumption was the control dog, so the results were as expected.
                                                                      bands of high (>50 ng/m3), medium (5 to 50 ng/m3), and low (<5
One of the 3 dogs had exhibited an atypical behavior prior to pair-
                                                                      ng/m3) based upon relative risk. The exposure control banding
ing, as evidenced by a lick granuloma. Following social housing,      system provided a data-driven framework for prioritization of
no atypical behaviors were noted and the granuloma resolved           activities intended to minimize employee exposure potential.
within 31 d of pairing. OGTT and fasting plasma insulin results       The quantitative data illustrated that tasks involving significant
were within expected range and do not appear to have been af-         bedding disturbance, such as cage changing and bedding disposal
fected by the pair housing. Compatible pair housing of captive        without local exhaust ventilation (LEV), were categorized within
canines optimizes animal welfare by reducing stress levels as         the high-exposure band. The disposal of bedding with LEV re-
shown through weight maintenance, daily food consumption,             vealed data that placed the task in the medium-exposure band,
decreased presence of abnormal behaviors, and no assignable           which confirmed the importance of source ventilation. All of the
negative effects on research, thereby refining and reducing the       area sample results taken in the hallways and corridors were below
number of dogs needed for research.                                   the limit of detection. These results assisted in the verification of
                                                                      our building ventilation capability. Current control measures,
P54 Blood Collection and Intravenous Injection in the Common          such as respiratory protection and ventilation containment, were
Marmoset (Callithrix jacchus)                                         evaluated and adjusted to align with the exposure band require-
C Maurus*, M Simkins, A Lange                                         ments. Now that tasks can be banded according to the associated
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

airborne concentration of LAA, future metrics around employee          manufacture date. During the 5-mo assessment, approximately
sensitization can be cross-referenced in order to evaluate the ef-     17,000 valves were assessed, of which, 2,300 failed (6.1% low-flow,
fectiveness of the control strategy for the reduction of LAA cases     1.4% no-flow, 3.5% leaky). The manufacturer’s target total failure
in vivarium staff.                                                     rate is 0.1%. Working together with the vendor, we determined that
                                                                       flow rate was impacted by the total number of autoclave cycles.
P56 Colors of Communication in the Veterinary Care Manage-             Failures were impacted by several factors, including residual metal
ment of a Large Rodent Population                                      shavings, silicone flashing, introduced debris or bedding and, most
                                                                       commonly, a distortion of the autoclave-rated internal diaphragm.
L Schleenbaker, DI Del Castillo*, HE Ramirez                           It was determined that the combination of heat, pressure, and
Animal Care Services, University of Florida, Gainesville, FL           steam caused the diaphragm to become water-permeable and led
                                                                       to formation of a blister. Normal flow rate often returned following
Veterinary medical management of a rodent population in a
                                                                       a period of drying out, only to reabsorb water at the next autoclave
decentralized facility requires organization and an effective
                                                                       cycle. We also established a process for rebuilding valves with the
method of communication between research, husbandry, and
                                                                       3 components susceptible to wear (shield, diaphragm, and O-ring).
veterinary staff. The proper identification of clinically sick
                                                                       To aid with future tracking of valve age, the manufacturer has cre-
and postsurgical animals, in addition to the documentation of
                                                                       ated an annual color scheme for the valve shield. A preventative
husbandry procedures performed by investigators, is of utmost
                                                                       program for water valve maintenance should include autoclave
importance in providing quality veterinary care for a larger ro-
                                                                       recalibration, prescreening of valves, and continual assessment all
dent population. Hence, the veterinary care staff has developed
                                                                       floods and clinical dehydration cases.
a silent communication system using colored cards and symbols
as the basic communication language. The cards are used by
husbandry, research, and veterinary staff to maintain an efficient     P58 Advancing and Refining Animal Welfare within an Estab-
flow of information. The following cards are primarily used in         lished Group-housed Primate Facility
our facility to document veterinary related concerns. A yellow         D Philip*, WJ Archibald, M Zubiaurre
“sick animal” card is used by the husbandry and research staff
                                                                       Preclinical Technologies, Aptuit Ltd, Edinburgh, United King-
to identify newly observed sick animals in the colony. A pink
                                                                       dom
“under veterinary care” card replaces the yellow card when
the veterinary care staff assess the animal and formulate a plan       Nonhuman primates have been used in drug metabolism, pharma-
of action. This pink “under veterinary care” card acts as the          cology, and regulatory toxicology studies for many years. In 1992,
animal’s clinical record, with all observations, treatments, and       we opened a new facility, specifically designed to meet the objec-
outcome recorded by either research or veterinary staff and is         tives of these studies, while housing the animals in social groups to
removed after resolution of the problem. A peel-off “surgery”          provide for their physiological and behavioral needs. Some of the
sticker is used to identify surgically manipulated animals and         challenges we faced were ensuring the quality and integrity of the
to indicate a postoperative check has been performed. A purple         study data, avoiding cross-contamination, and positively identify-
“under PI care” card identifies those animals in which some or all     ing individuals when in a gang cage. Simultaneously, we wanted to
husbandry procedures (food, water, cage change) are performed          further improve the animals’ environment while modifying study
by the research staff. In addition, other colored identification       designs to take advantage of the spacious living accommodation. We
and notification cards are used for identifying overcrowded            group-house all primates in large gang cages incorporating perches,
cages and notification of dead animals. As a result, the colors of     swings, puzzles, and toys. Deep litter substrate allows foraging for
documentation have made animal health–related communication            food, fruit, and supplementary mixes. Single cages are accessed from
among research, husbandry, and veterinary staff more efficient,        the gang cage by way of a hatch to allow precise post-dose observa-
therefore dramatically improving the level of veterinary care          tions. We have added acrylic sheet dividers and verandas between
provided in our rodent population.                                     cages, enabling enhanced visual contact between animals. Colored
                                                                       dyes applied on the animals’ eyebrows enable visual identification
P57 Watering Valve Life Cycle Evaluation                               when socially housed, with minimal disturbance to the animals.
                                                                       Novel methods of environmental enrichment, including swim-
DM Gonzalez*, SJ Gibson, M Leblanc, J Reuter                           ming pools, have also been introduced over time. Test compound
Animal Resources Department, The Salk Institute, La Jolla, CA          cross-contamination is avoided by careful physical separation of
                                                                       dose groups. Study procedures have been refined and adapted to
Automated watering systems provide a reliable source of ad libitum
                                                                       complement social housing, including use of vascular access ports
water to animal cages. Our facility uses reverse osmosis automated
                                                                       for infusion studies, enabling animals to remain in pairs or groups
watering for 95% of mice (approximately 14,000 cages). Racks and
                                                                       throughout dosing. We have found that we can meet the needs of
cages were brought online in 2002 and featured a quick-release
                                                                       a range of studies while housing animals in social groups. Animals
water valve that allows its removal for easy sanitation. Valves were
                                                                       are generally calm and cooperative during procedures, thereby
routinely sanitized monthly through a tunnel washer (alkaline de-
                                                                       contributing to reliable, quality study data. Later additions such as
tergent and a 180 °F rinse) and autoclaved at 250 °F for 15 min. In
                                                                       verandas and swimming pools have proved successful. This poster
the fall of 2007, we observed an increase in clinical dehydration in
                                                                       discusses our conclusion that new refinements can be incorporated
a subset of the mouse population, which prompted an investigation
                                                                       into existing facilities, further enhancing good environmental en-
of the water delivery system. Two major contributing factors were
                                                                       richment and welfare.
indentified: phenotype and decreased water flow. We sought further
guidance from the vendor and implemented an assessment plan for
measuring water output. Using manufacturer’s specifications for a      P59 Evaluation of Disposable Rodent Cage Inserts
water flow rate of 25 to 50 ml/min, we prescreened all valves. Leaky   DM Hull*
flows were identified by continual release of water after toggling.
                                                                       Research Animal Resources, Johns Hopkins University, Balti-
Low-flow, no-flow, and leaky valves were removed from circula-
tion. Unfortunately, no methods were available to identify the valve   more, MD
Abstracts of scientific papers 2008 AALAS National Meeting

Every animal facility develops occasional malfunctions of its          and contacting references and past employers can be enlightening,
cage wash equipment. This equipment can be out of service              but often only represents the aspects that the candidate wants the
anywhere from hours to weeks, depending on the severity of             potential employer to see. Interviewing the candidate can also be
the problem. A product has been developed that allows for an           helpful, but it is sometimes difficult to evaluate responses and
appropriate intracage environment to be maintained when cage           predict how the candidate will fit with the existing team. For 3 y,
wash equipment is broken for long periods of time and there is         we have been using teams and performance-based interviewing to
a need for alternative cage changing methods. Trials were con-         successfully screen potential candidates. This method of interview-
ducted in 5 ventilated mice cages. The same procedures were            ing involves asking the candidates to respond to a predetermined
followed for each cage change. These procedures consisted of           panel of questions selected to ask them to describe actions they
having the mice transferred into the disposable cage inserts, then     took with past events in their lives that meet certain criteria.
having the insert applied to the home cage, with care taken to         For example, “Tell me about a time when you had to work with
ensure proper alignment and exposure of the grommet supplying          someone you did not like. How did you handle the situation?”
air and water. The same home cage was used for 10 wk, during           The panel includes questions that require the candidates to share
which time an insert was changed every other week. These cages         positive and negative memories and experiences, with the expec-
had a variety of different numbers and strains of mice. During         tation that past behavior is predictive of future behavior. Our use
this 10-wk interval, these cages were evaluated for ease of use,       of this interview technique has allowed us to identify individuals
effects on food, bedding, grommet and wire bars, as well as the        who share our commitment to teamwork and customer service,
condition of the mice. By the end of the 10-wk test, we found          decreasing staffing turnover and increasing job satisfaction.
that the cage insert was easy to use and had no adverse affect on
the bedding, food, or health of the mice. These inserts were an        P62 Comparison of Costs Associated with the Preparation of
effective and convenient way to keep mice in a clean environ-          Nutritional Support to Multiple Rodents
ment until clean autoclaved cages are available.
                                                                       DL Hickman*
P60 A Novel Biocontainment Signage System                              Veterinary Medical Unit, VA Medical Ctr, Portland, OR
DM Mook*1,2, W Thompson1, MJ Huerkamp1,2                               Providing nutritional support to debilitated rodents is instrumental
1Division of Animal Resources, 2Department of Pathology and
                                                                       in minimizing pain and distress. Provision of nutritional support can
                                                                       be in the form of oral gavage, moistening of rodent block chow, or
Laboratory Medicine, Emory University, Atlanta, GA                     through the provision of commercial soft diets that require minimal
The use of biohazardous agents in research increased sharply           to no preparation. In this study, we evaluated the labor and material
in the past few years, as indicated in our facility by a tripling of   costs associated with the provision of moistened rodent block chow,
the number of persons oriented to our biocontainment program           powdered diet that is reconstituted with water, and a preformulated
between fiscal year 2006–2007 and 2007–2008. As is typically           nutritional supplement. Technicians recorded the time required to
the case in laboratory animal facilities, this increase in use was     prepare and provide 1 of these 3 diets in the facility animal rooms.
associated with no concurrent increase in space. In a multiple         The time per serving was estimated by dividing the total time by the
investigator room, the possibility existed for several investiga-      number of cages serviced. Four animal rooms, averaging 20 cages
tors to use various agents, and for these agents to be in use at       of animals requiring nutritional support per room, were included
different times. Users of the biocontainment suite had the right       in the study, and time was recorded over 31 d. The mean time
to be appraised of the specific risks associated with their work       required to supply moistened rodent block chow was 1.25 min/
environment, and this function was accomplished by placing             cage; the mean time required to supply reconstituted powdered
signage on animal room doors listing the agents used therein.          diet was 1.96 min/cage; and the mean time required to supply
With heavy usage, it became increasingly difficult for the animal      the preformulated diet was 0.49 min/cage. With an hourly labor
resources program to keep signage up to date. We created a novel       charge of $27/h and a cost of $0.08/disposable petri dish (required
magnetic biohazard signage system in which users changed the           for the moistened rodent block chow and reconstituted powdered
sign themselves as they added or deleted agents. A laminated           diet, but optional for the preformulated diet), we concluded that
biohazard sign was affixed to a magnetic backing, and another          the preformulated diet was the least expensive option overall, with
sign, identified as “approved agents,” was also created. Remov-        a per cage cost of $0.54 per cage. The per cage costs for moistened
able magnetic labels were generated with investigator names            rodent block chow and reconstituted powdered diet were $0.66 and
and agent names. When using an agent in a room, laboratory             $1.13, respectively. This cost differential was primarily due to the
staff retrieved the agent and PI name from the approved agents         fact that the preformulated diet did not require daily refreshment.
sign and affixed it to the biohazard sign on the door. Staff then      As our facility provides nutritional supplementation for an average
removed the magnet when they were finished using the agent.            of 200 cages per d due to experimental manipulation, selecting the
The signage system was easy to use and intuitive, was well-            preformulated diet over the moistened rodent block chow or the
received by both laboratory and animal care staff, and received a      reconstituted powdered diet represents an annual cost savings of
specific commendation from the Association for the Assessment          $8,760 and $43,070, respectively. Additionally, as the use of a petri
and Accreditation of Laboratory Animal Care, International. This       dish is not required, less waste is produced by the animal facility.
poster will share the signs themselves with our community, and
describe their rationale and use.                                      P63 Mock IACUC Meeting: A Novel Approach to Animal Care
                                                                       Staff Training
P61 Use of Performance-based Interviewing                              DM Ostmeyer*1, L Martin2, M Rosenbaum3,4
DL   Hickman*                                                          1Officeof the Vice President for Research, 2Research Integrity
Veterinary Medical Unit, VA Medical Ctr, Portland, OR                  and Compliance Review Office, 3Microbiology, Immunology
                                                                       and Pathology, 4Laboratory Animal Resources, Colorado State
Selection of employees who are a good fit with your animal care
                                                                       University, Fort Collins, CO
and use program can be quite challenging. Review of applications
                                                                                    Abstracts of scientific papers 2008 AALAS National Meeting

Animal care technicians are an integral part of the dynamic re-      are hairless, their eyes and ears are closed, the hind limbs and
search team. As primary caretakers, they must be knowledgeable       tail are stumps, and bone development is extremely immature.
regarding animal welfare regulations and governing agencies          Hind limbs begin to form between d 9 to 12.The ears opened
including the Institutional Animal Care and Use Committee            between d 12 and 15, and hair appears on the body around d
(IACUC). Technicians often receive classroom instruction about       18. The eyes begin to open on d 33. There was little variance in
the committee while preparing to take certification exams, but       the rate of development between opossum pups within a litter
they rarely have the opportunity to gain firsthand knowledge         or among litters. Additionally, male and female opossum pups
by interacting with the committee. A mock IACUC meeting              do not appear to vary greatly in size between the ages of 0 and
was designed to give the animal care staff at our institution an     90 d. We believe that these images will provide a useful guide
interactive, first-hand experience. Numerous members of the          for staging developing M. domestica pups.
animal research community enthusiastically volunteered to take
part in this exercise. The faculty for the mock IACUC training       P65 Measurement of Isoflurane Waste Anesthetic Gas Concen-
included the committee chair, 3 additional committee members, 2      tration When Using the Open-drop Technique
comparative medicine resident veterinarians, the senior commit-
tee coordinator, and the animal program coordinator. The mock        DK Taylor*
meeting was designed as a 2-part activity. The first part included   Division of Animal Resources, Emory University, Atlanta, GA
a presentation about the committee’s make-up and function,
                                                                     The open-drop technique is a commonly used method of an-
distribution of mock animal use protocols, and solicitation of
                                                                     esthetic delivery to small rodents. Operator exposure to waste
staff volunteers to serve as “student” committee members. After
                                                                     anesthetic gas (WAG) is a potential occupational hazard if this
the presentation, a mock committee of 3 technician volunteers
                                                                     method is used without a control such as a chemical fume hood.
and 7 “faculty” committee members were seated at a confer-
                                                                     This study was designed to determine if using the open-drop
ence table, with the remaining technicians in the audience. The
                                                                     technique on a surface without exposure controls generates
committee coordinator and chair each gave brief explanations
                                                                     significant WAG concentrations. Liquid isoflurane (0.1, 0.2, or
about their respective roles, and then deliberation began. The
                                                                     0.3 ml) was placed into screw-top 500-ml (3.5 in. tall) or 1000-ml
committee generated a great deal of discussion about the mock
                                                                     (6.5 in. tall) glass containers. WAG concentration was measured
protocols, and the audience interjected with several comments
                                                                     1, 2, or 3 min after liquid placement at the vessel opening and
and questions, resulting in a highly interactive and educational
                                                                     at 8 and 16 in. from the opening (operator height) using a por-
experience. The mock meeting also created an atmosphere that
                                                                     table infrared gas analyzer. A 2-factor ANOVA showed that, at
fostered positive relationships between the committee members
                                                                     the vessel opening, time was only significant factor (P < 0.0001)
and the animal care technicians. Positive feedback was received
                                                                     impacting WAG concentration when the 500-ml jar was used,
from both the students and the faculty after this training event.
                                                                     while neither time nor liquid volume were significant factors
A course evaluation was completed by the technicians in atten-
                                                                     when the 1000-ml jar was used. The WAG concentration using
dance, and they made excellent suggestions for improvement
                                                                     the 500-ml and 1000-ml containers were compared using the
which will be implemented at future mock IACUC meetings.
                                                                     Student t-test. At all liquid volumes and time points, the WAG
                                                                     concentration using the 500 ml container was marginally (P =
P64 Photographic Staging Guide for the Normal Development            0.057) to significantly (P < 0.05) greater than that for the 1000 ml
of Short-tailed Opossums (Monodelphis domestica)                     container. At operator level, WAG concentrations were at or near
D Paulus*1, SJ Karlen2, L Krubitzer1,2, CJ Clayton1                  0. Importantly, WAG concentrations at the vessel opening were
1Department of Psychology, 2Center for Neuroscience, University
                                                                     transiently as high as 662 (+168) ppm using the 500-ml jar and 122
                                                                     (+87) ppm using the 1000-ml jar, much greater than the NIOSH
of California, Davis, CA                                             60-min recommended exposure level of 2 ppm; concentrations
The grey short-tailed opossum (Monodelphis domestica) is a com-      were well below this ceiling at operator level. These results show
mon research model because it is one of the few marsupials that      that operator exposure to WAG using an open-drop technique is
can be kept and bred relatively easily in captivity. These opos-     minimal and below levels considered hazardous; however, the
sums have been used for a variety of studies, including spinal       risk of significant exposure increases as the distance between
cord and brain development, spinal cord injury and repair, skin      the operator and the jar diminishes. It also remains important
and eye cancer, genetics, olfactory communication and spatial        to properly handle the liquid agent and avoid contact with skin
navigation. M. domestica is a pouchless marsupial whose young        or mucous membranes.
are born after a 14- to 15-d gestation period. The pups migrate
to the nipples and attach for approximately 2 wk, and the pups       P66 Light Dark Studies Meeting the Demands
are weaned from the mother at 8 wk of age. The purpose of this
study was to develop a staging guide for opossums to use for         EJ Delaney*1, KL Krueger1, M Thompson2
developmental studies and animal husbandry. Photographs              1Animal Research Children’s Hospital, Children’s Hospital Bos-

were taken of the litters from 6 female opossums. Litters were       ton, Boston, MA; 2Plastic Concepts, INC., Billerica, MA
of varying size and date of birth. Imaging was done every 3rd
                                                                     Neuroscientists at our institution are evaluating the developing
d from d 0 (day of birth) to 30, every day from d 30 to 35 (the
                                                                     brain and how it is affected by environmental changes, such as
time of eye opening), every 5th d from d 35 to 60 (the age of
                                                                     alteration in light cycle. While it would be simplest to alter the
weaning), and on d 90 (prepuberty). From d 0 to 21, isofluorine
                                                                     light cycle of an animal room, the small number of animals needed
was used to anesthetize the mother so that the pups could be
                                                                     for these studies was a problem where space is at a premium in
imaged. After d 21, pups were large enough to remove from the
                                                                     our facility. In addition, the coordination of altered light cycles
mother, so one individual pup was selected at random for imag-
                                                                     between several protocols was a concern. It was determined the
ing. All photographs were adjusted for brightness and contrast
                                                                     best solution would be to design a chamber which was light-tight,
and are presented in chronological order. We found that pups
                                                                     in which light cycles could be determined by the investigator, and
changed dramatically between d 0 and d 90. At birth, the animals
                                                                     that would fit up to 8 small mouse cages. A plastics manufacturing
Abstracts of scientific papers 2008 AALAS National Meeting

firm was identified, and together with animal resources manage-          Instituto de Ciencias Básicas y Medicina Experimental Hospital
ment staff and the investigators, a design was developed. Two            Italiano de Buenos Aires, Ciudad Autónoma de Buenos Aires,
chambers per unit were created, each holding approximately 8             Argentina
small mouse cages or 6 large mouse cages. To ensure complete             Exposure and training to laboratory animal science is scarce
darkness of the interior enclosure, the doors had an overlapping         for most medical students and future researchers. One possible
gasket seal design. Small air blowers in the back of the unit pro-       reason for this is that in most biomedical careers prior training is
vided ambient air to the chambers. Temperature and humidity
                                                                         not obligatory when beginning laboratory work. Our facility re-
probes were installed, with digital readout displays to ensure           quires that people who use animals in their projects first undergo
appropriate environmental parameters were maintained for the
                                                                         training with basic contents. We designed and implemented a
animals. Air supply and return, along with the electrical con-
                                                                         program with the purpose of enhancing the adequate care and
nections, were designed with a double-wall unit to prevent any
                                                                         use of laboratory animals and thus the achievement of quality
reflective light from entering the interior chamber. It was critical
                                                                         and reliability in the expected results. It consists of an optional
that these animals experience no exposure to light, and so a very
                                                                         36-h course for 2nd-y students called Science of the Laboratory
sensitive light detection device (4 lumis of light) was placed inside
                                                                         Animal. Our aim is to sensitize the student to the harmonious
the chambers to alert users of any internal lighting by way of both
                                                                         care and use of laboratory animals, with a special emphasis on
a visual and audible alarm. A timer was installed in each chamber
                                                                         the welfare of the animal in regards to biomedical activity. The
for properly determining the duration cycle of the study. Finally,
                                                                         main idea is to educate implementing the principle of the 3 Rs of
the units were designed to keep outside noise to a minimum,
                                                                         Russell and Burch. Teachers evaluated the knowledge acquired
which is critical for most neuroscience studies. These units have
                                                                         with partial and final exams and a monograph. Through observa-
now been in place, and in constant use, for more than 6 mo. The
                                                                         tion, teachers also evaluated the practical abilities and attitudes
design ensures the animals are not exposed to even small amounts
                                                                         towards animals acquired by students. We conclude that the
of light. Investigators are extremely pleased with these units; they
                                                                         proposed objectives were satisfactorily acquired since the results
are easy to open and close even in total darkness, and can accom-
                                                                         showed that students understand the importance of a previous
modate multiple cages, allowing investigators to perform entire
                                                                         training, as much for themselves as for future collaborators.
experiments under the same environmental conditions.

                                                                         P69 Murine Retro-orbital Bleeding Under Anesthesia: An
P67 Single or Social Housing for Research Animals
                                                                         Alternative Method
E Scott*, K Ryder, DB Anderson
                                                                         R Scott, E Villalobos, A Gaudiel, R Garcia-Gonzalez, E Chua*
Animals Scientific Procedures Inspectorate, Home Office,                 Laboratory Animal Resources, Genentech, Inc., South San Fran-
Dundee, United Kingdom                                                   cisco, CA
Social housing is promulgated in virtually all legislation or guide-     This poster will outline the procedure as an alternative to using
lines for species used in scientific work. Traditional approaches        glass capillary tube or anesthesia chamber retro-orbital bleeding
using single housing should be discouraged where social housing          methods. Sharper glass capillary tubes have been replaced with
is advantageous and improves animal welfare, yet is not detrimen-        sterilized plastic pipette tips. Anesthesia chamber use has been
tal to the scientific research outcomes. In the United Kingdom,          replaced by use of sterile, flip-top plastic vials for induction by
inspectors play a role in encouraging group housing of animals
                                                                         open-drop method. The animals are also securely restrained by
and ensuring that a clear scientific, veterinary, or husbandry case is   the procedure operator at all times, minimizing head movement
made by researchers for single housing of social animals. There are      of the animal during anesthesia induction, which can cause
examples where single housing may be justified, but social housing       trauma. All materials in this procedure can be sterilized, which
options need to be explored, as they are often possible and can be       helps prevent cross-contamination between animals. In our use,
beneficial. In neuroscience, when chronic implants are used, there       this method has also aided in providing faster, more successful
is concern that interaction between animals may lead to infection        training of general animal users, because of the ease of restraint,
or implant disturbance. In the immediate postoperative period
                                                                         anesthesia induction, and avoidance of sharp objects in the ocular
single housing may be a good option, but thereafter experience           area of the animals. This method works well in larger murine
in the United Kingdom shows that social housing is possible with         facilities, while minimizing animal stress, trauma, and time spent
reduced incidence of stereotypic behavior. Telemetry animals can         by the procedure operator. There has also been a significant de-
be housed singly for short monitoring periods and then restored
                                                                         crease in hemolyzed or clotted samples due to the more conical
to their groups without disruption of social hierarchies. When           shape of the pipette tip in comparison to a capillary tube.
severe illness is expected with bacterial challenge models it may
be best to single house during the acute phase only, to allow
close and frequent monitoring. Such periods are likely to be short       P70 eCertificate: A Fully Automated Web-based Health Certi-
(hours or days) and the animal is likely to be sufficiently ill that     fication System for Rodents
interaction with peers would be unlikely. The measurement of             FR Wolf*1,2, Q Zhang2
individual feed intake or monitoring postdosing may necessitate          1Weill Cornell Medical College, New York, NY; 2 Memorial Sloan-
single housing. Husbandry measures of short-term separation at
                                                                         Kettering Cancer Center, New York, NY
feeding and dosing times, feeding systems with individual animal
recognition systems, or alternative measures of food intake, such        Sharing of unique genetically modified rodents among scientists
as body weight, have been used to reduce social isolation time           is important in biomedical research. To properly assess the risks
with no impact on study data acceptability.                              associated with the receipt of the animals at the receiving facility,
                                                                         it is essential to receive a comprehensive health report from the
P68 Laboratory Animal Science Education for Medical Stu-                 facility of origin. To facilitate this process, we developed eCertifi-
dents                                                                    cate, a web-based application that creates and distributes health
                                                                         certificates within seconds based on user input and preloaded
E Herrero*, G Dabrowski, PF Argibay
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

data. Each eCertificate consists of a PDF file with 4 pages. A cover    the innate behaviors of burrowing, tunneling, chamber grouping,
page provides general information as well as a summary of the           and a tower for them to see around their “town.” The configura-
colony’s current and historical health status, including counter-       tion of tunnels, burrows, and towers can be altered to fit any size
measures taken in cases of pathogen detection. The second page,         room and almost any number of prairie dogs. The hope was that
a pro forma invoice and health certificate, is used by customs          the prairie dogs would use the burrows for different purposes
officials when shipping internationally. The third page details the     (eating, sleeping, defecating). The tower will provide them the op-
health monitoring program and husbandry procedures used in              portunity to climb above their town and perhaps give “warning”
the room and facility of origin. The final page is a FELASA-style       or “all clear” vocalizations. We constructed the “den” from 100-gal
health report providing current as well as historic results for each    livestock troughs. Each trough is fitted with 4 6-in. wheels to allow
pathogen tested. eCertificate incorporates a number of features.        for easier sanitation and reconfigurations. The troughs are con-
Cover letters and health monitoring profiles can be custom-             nected to each other using CPVC unthreaded 6-in. diameter pipe
tailored to individual rooms, to groups of rooms, or to a specific      that serves as the tunnels. The pipe is attached to the troughs using
vivarium. Investigators have access to previous eCertificates and       CPVC unthreaded 6-in. diameter pipe fitting. The tower of each
can resend the certificate with current results with a single click.    CHEU is a cat gang-housed cage and is attached to the CPVC pipe
A search function allows the biosecurity veterinarian to find all       with CPVC unthreaded 6-in. pipe fitting without hex in center and
certificates issued for animals shipped from a specific room,           CPVC unthreaded 6-in. pipe fitting, 90 degree elbow. All exits from
facility, investigator, and so forth within a specified time frame,     the dens can be capped to prevent movement of prairie dogs. The
allowing a follow-up e-mail to be sent to selected recipients if a      troughs are covered with 48 by 96 by 0.25 in. polypropylene perfo-
change in health status is detected. The next release will collect      rated sheets which are 20% open area to allow air flow and light into
test results from a laboratory information system automatically         the troughs. It was determined that PVC would not withstand cage
updating the health report in eCertificate.                             wash temperature, and the prototype was redesigned with CPVC to
                                                                        withstand temperatures of 180 ° F. New shipments of prairie dogs
P71 How Safe Is Your Facility?                                          are scheduled to arrive within the next few months. These will be
                                                                        the first groups ever housed in the new CHEU.
F Warren*
Lab Animal Medicine, University of Delaware, Newark, DE                 P73 Animal Enrichment and Socialization in an Indoor Re-
Animal facilities have the advantage of providing everything that       search Facility
an animal needs, which includes food, water, sanitation, air, and       G Jeans*1, KD Corcoran1, JL Wheatley2, AS Fix2
lighting. The disadvantage of providing lighting is that animal         1Pharmaceuticals, 2Central      Product Safety, Procter & Gamble,
facilities are constructed with no windows that allow natural
                                                                        Cincinnati, OH
light to enter. If there was a power failure, an animal facility can
be an extremely dark place. Navigating around a dark facility           Environmental enrichment is an important element of any animal
can be not only intimidating, but dangerous. Large equipment            research program. The goals of the Procter & Gamble program
would become hazardous if personnel were trying to find their           include providing comfort and decreasing stress for the animal,
way out. In our facility, we have instituted many safety features       promoting species-specific behaviors, creating a more complex
to be used in the event of a power failure. Placing flashlights in      living environment, and enhancing animal welfare standards. All
corridors is one way. We have also found a simple, inexpensive          animals are provided enrichment unless exempted due to health
method of providing an extra measure of safety. All rooms and           reasons or scientific justification approved by the IACUC. Environ-
support areas have luminescent materials on the door frames.            mental enrichment items have been chosen for each species based
Next to the doorframe is a pen-style flashlight that can help           on actual observations of animal interactions with the items. Study
guide someone out of an animal room. We also put in place other         personnel choose species-specific enrichment items, which are
safety features, including large battery-operated flashlights in the    rotated to provide stimulation and variety. For example, rodents
office area and hallway maps showing exit doors. The hallway            are provided a variety of enrichment items (nestlets, isopads, huts,
flashlights have automatic night lights built in that light when        igloos, gumabones, nylabones, tunnels, marbles, stainless steel
power is off. Another safety feature of the hallway flashlights         bells) on a rotating basis. Dog enrichment includes a variety of toys,
is that they make a beeping sound after 3 min when power has            treats, and various socialization activities (grooming, brushing,
been cut off, which aids in locating them.                              nail trimming, person-to-dog and dog-to-dog socialization, and
                                                                        clicker training). In addition, we provide a large indoor play area
P72 Cynomys Housing and Enrichment Units                                using child-based playground structures and setup a mock living
                                                                        room using common household furnishings. Dogs quickly learn to
G Langham*1, K Kelly1, C Walls2, N Powell1                              anticipate their socialization experience and these experiences have
1Animal  Resources Branch, Centers for Disease Control and              greatly facilitated the adoption of dogs from our research programs.
Prevention, Atlanta, GA; 2 Four Seasons Environmental, Inc,             In addition, these rooms have become a center of activity for the
Monroe, OH                                                              animal care and veterinary staff, providing an additional positive
                                                                        benefit to facility personnel. Based on our experiences, a broadly
The use of prairie dogs (Cynomys spp.) in infectious disease research
                                                                        applied environmental enrichment program across all species im-
has increased due to their susceptibility to monkeypox, bubonic
                                                                        proves the welfare of the research animals, the quality of the research
plague, and hepatitis. Cynomys have a complex social system and
                                                                        environment, and job satisfaction for employees.
housing has posed a difficult challenge. Other facilities have tried
housing them in storage bins or on the floor. We were dissatisfied
with these housing methods, and decided to construct a Cyonmys          P74 Enhanced Indoor Housing for Research Dogs
housing and enrichment unit (CHEU) that is easily sanitizable,          GL Jeans*1, DA Fryer1, KW Casey1, CR Bohne1, GL Calihan1,
mobile, and promotes as many species typical behaviors as possible      AS Fix2
in a captive environment. A group housing scheme was developed          1Pharmaceuticals, 2Pet
that provides quarantined prairie dogs the opportunity to exhibit                                Care, Procter & Gamble, Lewisburg, OH
Abstracts of scientific papers 2008 AALAS National Meeting

Enhanced indoor housing for our research dogs was a key interest           shows that allergen exposure can be drastically reduced by use
among our personnel and stakeholders. In response, we bench-               of ventilated caging, by handling the cages always with filter top
marked with other research institutions and collected input from           on, and by changing cages only under laminar flow conditions.
our staff to determine how best to optimize our kennels and meet           In the cage wash area, a 32-fold reduction was achieved using an
our animal welfare goals. The goals were to improve our existing           inexpensive dust extractor. Further studies will be necessary to
kennel wards; enhance our animal welfare standards; provide op-            elucidate whether this allergen reduction is sufficient to prevent
portunities for the dogs to be group-housed, yet maintain ability          mouse-related allergic sensitizations. A striking result was that
to be periodically single-housed; create a more visually appealing         relevant Mus m 1 levels were detectable even at an indirectly
and open kennel environment; and promote increased dog-to-dog              exposed work area like a PC work desk.
and human-to-dog interaction. After researching kennel options
with various vendors, we decided upon a modification of our                P76 Quantitative Culture and ATP Detection Used to Validate
existing design. The new panels were shorter and included side-            Rodent Water Bottle Change Intervals
access gates, visual barriers, and raised benches. Additionally, we
added sound baffling panels to the wards, which dramatically               H Tobin*, NH Shomer
reduced the reverberant noise levels. After installation of the            Merck Research Laboratories—Boston, Boston, MA
modified kennels, the dogs quickly adapted to the new design
                                                                           Changing water bottles is labor intensive, poses an ergonomic
and are now predominantly pair-housed. The combination of
                                                                           hazard for husbandry staff, and increases biosafety risks to the
the new kennel design and installed sound baffling has resulted
                                                                           mice as it requires handling the bottles more frequently. We
in quieter dogs who appear more content and no longer exhibit
                                                                           investigated the feasibility of extending the water bottle change
undesirable behaviors. The dogs all use their raised platforms and
                                                                           interval from 7 to 14 d for singly housed, specific pathogen-free
can easily see what is happening throughout the ward because of
                                                                           mice. We hypothesized that water quality in 14-d-old water
the increased visibility. The flexibility of the side access gates allow
                                                                           bottles from singly housed (SH) cages would be at least as good
us to house dogs based on compatibility and research study needs.
                                                                           as the quality in 7-d-old cages from group-housed (2 to 5 per
In addition, shorter kennel walls invite human-to-dog interaction.
                                                                           cage) mice. Based on the current accepted 7-d change interval for
In our experience, these redesigned kennels met our research
                                                                           group-housed (GH) mice, we set the GH contamination level at 7
needs, provided optimal housing for our dogs, and increased job
                                                                           d as our acceptance criterion. Water in our facility was autoclaved
satisfaction for employees.
                                                                           municipal tap water with undetectable levels of free chlorine after
                                                                           autoclaving. Mice tested were free of pathogens and common
P75 Effective Strategies to Diminish Critical Allergen Exposure            opportunistic pathogens, including Pseudomonas. Water bottles
while Handling Laboratory Mice                                             of GH and SH mice were tested at predetermined intervals by
H Riedesel*2, J Krause1, E Hallier3, S Turowski3, A Heutelbeck3            aseptically removing a 1-ml water sample, vortexing, and testing
1Animal Facility, Max-Planck-Institute of Experimental Medicine,
                                                                           using a luminometer to estimate organic contamination levels.
                                                                           Quantitative cultures were performed using nutrient agar plates
Goettingen, Germany; 2Animal Facility, Helmholtz-Centre for                incubated for 3 d at 37 °C. At 24 h postplacement, water bottles
Infection Research, Braunschweig, Germany; 3Occupational                   from SH and GH mice averaged 95 ± 210 colony-forming units
Medicine, Georg-August-University, Goettingen, Germany                     (CFU)/ml (n = 19) and 489 ± 1778 CFU/ml (n = 14), respectively.
Allergy caused by laboratory mice is a common occupational                 At 7 d, SH and GH mice averaged 35518 ± 152843 CFU/ml (n =
health hazard for personnel. We evaluated the exposure of mouse            19) and 96412 ± 241625 CFU/ml (n = 14), respectively. At 14 d,
allergens in the air under different working conditions when               SH mice averaged 126320 ± 257137 CFU/ml (n = 19). Average
handling mice. With on-the-person and stationary air sampling              luminometer counts for SH and GH mice at 24 h postplacement
methods (flow: 2.5 to 3 l/min, 8 µm cellulose nitrate filter), inhal-      were 1 ± 0 (n = 19) and 62 ± 230 (n = 14), respectively. At 7 d, SH
able and respirable dust was collected over several day shifts and         and GH mice had counts of 992 ± 2020 (n = 19) and 12612 ± 22870
during night in animal rooms, cage wash, and areas with a PC               (n = 14), respectively, and at d 14 SH mice had an average count
work desk. Work areas with different conditions were included,             of 11639 ± 35127 (n = 19). Plate counts and organic contamina-
such as open cages compared with individually ventilated cages             tion levels at 14 d for SH mice were not significantly different
(with positive or negative pressure) and technical equipment               (P = 0.73 and 0.92, respectively) from our acceptance criterion
(changing cages with or without a laminar-flow hood). After                (contamination levels in GH mice at d). Based on this data, we
extraction of the dust samples the level of mouse allergen Mus             extended our water bottle change interval for SH mice to 14 d.
m 1 was determined using a commercially available ELISA. In
the laboratory rooms with individually ventilated cages, over              P77 Work-related Mouse Allergens in the Home Environment
the measuring time mean levels of Mus m 1 were detectable be-              of Animal Technicians
tween 0.55 ng/m3 (positive pressure) and 0.31 ng/m3 (negative
pressure), in rooms with open cages up to 12.97 ng/m3. Without             J Krause*1, H Riedesel2, E Hallier3, S Turowski3, A Heutelbeck3
using a laminar flow hood for cage change, Mus m 1 levels up to            1Animal Facility, Max-Planck-Institute of Experimental Medicine,

17.55 ng/m3 were measured compared to 0.67 ng/m3 when using                Goettingen, Germany; 2Animal Facility, Helmholtz-Centre for
a laminar flow hood. Higher allergen levels were detected in the           Infection Research, Braunschweig, Germany; 3Occupational
cage wash area near the disposal station for soiled bedding. The           Medicine, Georg-August-University, Goettingen, Germany
air here showed 84.93 ng/m3 Mus m 1. By installing a commercial
                                                                           Laboratory animal allergy is a common occupational health
dust extractor, levels of Mus m 1 could be lowered to 2.59 ng/
                                                                           hazard for research scientists and animal technicians. Risk fac-
m3. Levels up to 7.16 ng/m3 were detectable on indirect exposed
                                                                           tors include genetic and environmental components. The aim of
work areas, such as a PC work desk. Under most working con-
                                                                           this study was to evaluate the airborne mobility characteristics
ditions, relevant mouse allergen levels were detectable. Up to
                                                                           of mice allergens and their transmission into the homes of ani-
20 times higher Mus m 1 level could be found in animal rooms
                                                                           mal technicians. Dust samples were vacuumed from car seats,
with open cages compared to rooms with ventilated caging. This
                                                                           carpets, kitchen chairs, and mattresses of animal technicians’
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

private dwellings. Eight technicians working in 4 workplaces            P79 Creating a Safer Environment for Laboratory Animal Han-
of different allergen exposure levels (specific pathogen-free           dlers Anesthetizing Rodents in Induction Chambers
[SPF] and conventional rooms equipped with open compared                JB Hoff*
with individually ventilated cages) participated in this study.
Depending on the housing unit different types of personal pro-          Center for Intgrative Genomics, University of Michigan, Ann
tective equipment was used, including sterile gowns with or             Arbor, MI
without face mask, hair cover, and gloves. Technicians working          Recent research indicates laboratory animal personnel using
in the SPF barrier unit passed a shower lock and changed their          induction chambers to anesthetize rodents may be placing
clothes twice. Technicians of the conventional unit did not shower      themselves at risk of exposure to unknown levels of volatile
and changed their gowns only once. Showering after work was             anesthetics. We hypothesize risk can be greatly reduced by per-
optional; however, every employee used 2 separate lockers for           forming a simple modification to an existing rodent anesthesia
private and for work clothing. All animal rooms were ventilated         station. An isoflurane vaporizing system was used to deliver a
with HEPA-filtered, conditioned air at a temperature of 22 ± 1 °C       measured amount of isoflurane and oxygen into an induction
and 55 ± 5% relative humidity, with 14 air changes per hour. No         chamber. Rats were anesthetized in the chamber by administer-
study participant had pet rodents at home. After extraction of          ing 5% isoflurane and oxygen at a rate of 1 l/min continuously
the dust samples, the level of the mouse allergen Mus m 1 was           for 60 to 90 s. Isoflurane and oxygen were turned off before
determined using a commercially available ELISA. In the settled         opening the chamber to retrieve the animal. With additional
dust of the private dwellings of the animal handlers, the Mus           tubing and a Y connect, we attached the blower end of a small
m 1 levels ranged between 0.29 and 2.46 µg/g (mean 1.3 µg/g)            hand-held vacuum in the gas line directed towards the chamber.
in cars, 0.006 to 0.18 µg/g (mean 0.081 µg/g) in carpets, lower         A stopcock was used to stop the anesthetic gas from backing up
than detection limit up to 0.81 µg/g in mattresses, and 0.028 to        into the line or to allow the air from the blower to flow into the
0.19 (mean 0.07 µg/g) on kitchen chairs. Our results indicate           induction chamber, blowing most of the volatile gas out of the
that the private home may be an important location for exposure         chamber before opening. We compared isoflurane concentration
to work-related allergens from mice. This should be taken into          levels in the environment surrounding the induction chamber
consideration when developing an overall occupational health            with and without the benefit of the blower. A halogenated gas
intervention strategy. We observe the tendency of mouse al-             monitoring badge was placed near the opening of the induction
lergens to stick to skin, hair, and clothes, thus giving allergens      chamber for a period of 150 min. In that time, 6 rats were anes-
the opportunity to be carried from work into private areas. This        thetized, as described above. Isoflurane concentration reached
can only be prevented by strict separation of the working and           6.7 ppm. The following day, a new halogenated gas monitoring
living environments.                                                    badge was placed in the same vicinity as the first for 135 min.
                                                                        Six rats were anesthetized. With the added benefit of blowing the
P78 Fur Mite Treatment 101: Labor, Time Management, and                 volatile anesthetic out of the induction chamber before opening,
Project Coordination                                                    concentrations were reduced to 1.7 ppm. The system described
                                                                        here reduces exposure to potentially harmful volatile anesthet-
J Pinkney1,2, D Smith2, T Burkholder*2
                                                                        ics by 75%. Our system is unique because few rodent anesthetic
1   SoBran, Bethesda, MD; 2 NIH/DVR, Bethesda, MD                       stations allow the operator to flush isoflurane from the induction
Fur mite eradication is known to be a challenge in large trans-         chamber before opening. It is easy to obtain, affordable, and
genic rodent colonies. Considerations when making a plan of             adaptable to permanent and portable anesthetic systems.
action include the size of the colony or number of cages to be
treated, ease of administration, choice of chemical or drug for         P80 Washroom Efficiency: A Pilot Study
treatment, and the coordination of animal care staff. Our past
                                                                        JM Traylor*, GR Robinson
facility treatment protocols required a time commitment of 10
to 20 h per wk from the technical staff for up to a year per room.      Comparative Medicine, Pfizer Inc., Groton, CT
When developing our treatment plan for fur mite eradication,            Washrooms are essential to the operation of an animal research
efforts were taken to ensure the plan would provide the most            facility and are busy places that have to meet the needs of the
effective care while striving to minimize the amount of labor and       scientific and husbandry staff. Historically, the washroom staff
time involved in providing the treatment. Animals were screened         was sometimes comprised of contract personnel with limited
to confirm suspected fur mite presence based on 1 mouse that            experience in laboratory animal science. To mitigate this lack
had been exported from the colony. Once confirmed positive, the         of experience, more experienced husbandry technicians would
room was treated with ivermectin in water at specific intervals.        help when time was available. They did so by assisting with
Results from retesting indicated eradication of mites was possible      equipment cleaning and preparing caging for their assigned
with 2 mo of treatments on alternate weeks and follow-up testing        rooms. This process created numerous trips in and out of the
for 6 mo after treatment. This presentation will explain several        washroom, which led to decreased efficiency and potentially
factors that were considered in the development of the treatment        compromised biosecurity. A pilot study was undertaken at one
plan initiated in this facility. Additionally, it stresses the impor-   of our facilities to improve the washroom traffic patterns, equip-
tance of strategic planning, teamwork, and cooperation across           ment maintenance, inventory management, and the reduction
departmental lines. It also further discusses the background            of biosecurity risk. We staffed the washroom with 4 experienced
and support information, the advantages and disadvantages of            husbandry technicians and implemented a menu system to ad-
the program, diagnostic techniques and frequency of testing,            dress the organization, inventory, and biosecurity issues. This
and the overall outcome of the program used to combat such              system is a service provided by the washroom personnel to fill
a novel topic. Lastly, this presentation can also be useful when        the scientists’ and husbandry technicians’ caging requirements.
combating other treatable common laboratory animal conditions           The use of the menu system has decreased the daily clean-side
when looking into treatment timelines and strategic planning            entries of husbandry staff by a minimum of 64 times. The system
according to testing results.                                           has taken 14 husbandry technicians’ hands off of the equipment
Abstracts of scientific papers 2008 AALAS National Meeting

stocks, enabling better resource management. This tool has also        attention behaviors (being at the front of their cages and watch-
allowed for proper equipment rotation by the washroom staff            ing the monitor) when that video was played; some NHPs also
and has improved accountability and ownership. Organizational          displayed lip smacking when monkeys in a similar laboratory
maintenance and biosecurity has also improved and continue             environment appeared on the screen. The other 3 videos elicited
to be preserved.                                                       much smaller percentages of monkeys watching them (range:
                                                                       10% to 40%). One conclusion drawn is that NHPs may respond
P81Neonate Scooper                                                     to video scenes with which they are more familiar.

J Burgess*
                                                                       P83 Castration Eliminates Conspecific Aggression in Group-
Animal Facility—E.A.M, Trudeau Institute, Saranac Lake, NY             housed Male Surveillance Mice
Neonatal mortality can be a problem in mouse production fa-            JL Lofgren*, SE Erdman, C Wong, R King, T Chavarria, A Discua,
cilities. In breeding colonies, one of the goals is to keep stress     JG Fox, K Maurer
levels as low as possible. After cage changing, females with
                                                                       Division of Comparative Medicine, Massachusetts Institute of
young pups show high stress levels. These females will pick up
                                                                       Technology, Cambridge, MA
their pups and move around the cage franticly. Neonates will
be left randomly about the cage, some left uncared for and oth-        Microbiological surveillance is an essential component of murine
ers cannibalized. The stress of the female is seen immediately         health maintenance. To adequately ensure the pathogen status
after cage changing and can continue for up to 1 h. With some          of mice used in our health-surveillance program, we use in-
modification to a simple food scoop, we are able to easily and         house–bred CD-1 mice. To make optimal use of mice from this
quickly move an entire nest of pups during cage changing. A            colony, thereby reducing overall animal use, we use the female
200-ml polypropylene scoop was modified to place a 90-degree           progeny for recipient animals in our core transgenic facility
bend in the open portion of the scoop. The bend was made by            and the male progeny in our surveillance program. Veterinary
gently heating the scoop with a propane torch, creating the 90         technicians noted that there was a great deal of fighting among
degree bend by pressing the scoop against a solid surface and          cohoused male surveillance mice that was not remediated
then quickly cooling the scoop in cold water. The modified scoop       through environmental enrichment. We hypothesized that early
withstands autoclaving to 121 °C. The neonate scooper is used          castration may eliminate fighting in male mice. As such, male
where neonates (0 to 12 d) are present. Moving the entire nest         mice were castrated prior to 2 mo of age and then released into
at once eliminates handling of individual pups and appears to          the surveillance program. Castration was performed using an
produce less stress on the female. By moving the nest, the scent       open technique under general chemical anesthesia, and mice
stays on the pups, there is no manual pressure on the pups, and        were given buprenorphine for postoperative pain. Veterinary
less time is spent in breeder cage changing. Use of the neonate        technicians recorded fighting in castrated and non-castrated male
scooper decreases the time needed for cage cleaning and pro-           surveillance mice during their weekly rounds to determine any
duces higher neonate survival.                                         benefit. Castrated mice recovered from the surgery within 24 h
                                                                       with no noted negative effect. Over a 3-mo period, the overall
P82 Video Enrichment Preferences of Laboratory Macaques                prevalence of fighting in intact male cages was 54% (12 of 22
(Macaca mulatta)                                                       cages) compared with 0% (0 of 16 cages; P = 0.0019) in castrated
                                                                       cages. Additionally, 11 intact male mice were separated and 7
JN Camacho*, J Tsui                                                    were euthanized due to fighting. Castration served as both a
CCM, Massachusetts General Hospital, Charlestown, MA                   refinement by eradicating pain and stress resulting from fight-
                                                                       ing and a reduction in animal numbers by allowing the males
Video is a common means of environmental enrichment for
                                                                       from the breeding colony to be used as surveillance mice. If
laboratory nonhuman primates (NHPs). Other studies indicated
                                                                       only females from the colony were to be used, the size of the
that NHPs appear to prefer to watch something on a screen as
                                                                       breeding colony would need to be doubled and the majority of
opposed to nothing at all. However, there is still much unknown
                                                                       male progeny would be euthanized. In conclusion, castration is a
about which video topics may be more effective in promoting
                                                                       minimally invasive, safe, humane, and rapid method to eliminate
species-specific behaviors and minimizing apparent boredom
                                                                       conspefic aggression in male CD-1 surveillance mice.
in captivity. We performed basic observational assessments to
determine if adult M. mulatta and M. fasicularis exhibited prefer-
ences for different video subjects, involving a total of 14 monkeys.   P84 Canine Home Environment Acclimation Program: A
Each animal’s normal behavior was established during routine           Conditioning Program to Better Prepare Research Dogs for
husbandry procedures and interaction with animal care staff over       Adoption
several days in the absence of video enrichment; other enrich-         JL Wheatley*, GL Jeans, KD Corcoran, AS Fix
ment modalities (for example, contact with conspecifics, audio
                                                                       Procter & Gamble Company, Cincinnati, OH
enrichment, nutritional variety, manipulanda) were unchanged.
Four videos were played: NHPs in a naturalistic setting, a nature      Many canines retiring from research service are deemed eligible
video involving no NHPs, a children’s education video, and an          for adoption. However, without the proper intervention, a dog
AALAS video tutorial on proper occupational safety practices           raised solely in a research institution may have difficulty adjusting
when working with NHPs. Each video was played 10 to 12 times           to life outside the facility. In an effort to better prepare these dogs
between 0800 and 1600 on a given day, while NHP behavior was           for a successful transition to a home environment, we describe an
observed for 20-min increments 4 times throughout the day. A           acclimation program implemented at the Procter and Gamble Co.
different video was played on each of 4 consecutive d. Time and        This program includes a specific acclimation and training plan for
reaction were compared for each animal directly observing each         each dog with exposure to environments and stimuli likely to be
video to determine if different behavior resulted. Nonhuman            encountered postadoption. Our goal is to address any existing
primates appeared to prefer the human tutorial video more              behavioral or socialization issues, introduce housebreaking prin-
than the other videos because 70% of macaques displayed direct         cipals, and familiarize dogs to common household items prior to
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

adoption. The artificial home environment resides in an unoccu-         staff management and performance. Bioluminescence systems im-
pied lab within our facility and includes a television, refrigerator,   mediately calculate adenosine triphosphate (ATP) levels, indicating
radio, telephone, floor fan, couch, dinette, mirror, and area rugs.     the amount of organic material present on surfaces. At our facility,
Acclimation to the home environment, along with other interven-         we used a bioluminescence device to identify which management
tions such as leash walking and obedience training, are provided        factors contribute to enhanced work performance. We assessed
based on individual animal needs. We have found this program            sanitization of animal room floors, fencing, and pens under 4 condi-
to be beneficial and enriching not only for the canine subjects, but    tions: blinded (baseline), non-blinded (supervision), non-blinded
also for the animal care and veterinary staff members who have          with 48 h delayed feedback and delayed accountability (required
participated. The program has been used for many dogs that have         to resanitize and retest), and non-blinded with immediate feedback
been placed in adoptive homes. The success of these adoptions           and immediate accountability. Our results indicate that without staff
has been increased by exposing the dogs to the home environ-            knowledge of testing (blinded), there was no significant difference
ment and household items prior to their leaving the facility. We        between pre- and postsanitization values. This suggests that without
advocate an acclimation program for canine adoption candidates          supervision, the staff were not effectively performing sanitization
as a way to improve the transition to a home environment and to         procedures. When the staff were aware of testing (supervised),
increase job satisfaction for employees working in a laboratory         pre- and postcleaning values were significantly different (t = 1.90,
animal research setting.                                                P < 0.05), but only 60% of the measured areas reached our pass cri-
                                                                        teria of greater than 75% reduction in surface ATP levels. Delayed
P85 Supplemental Care for ERCC-1–deficient Mice                         feedback and delayed accountability had little effect on sanitation
                                                                        efficacy. However, immediate feedback and immediate accountabil-
J Mensch*                                                               ity cohorts demonstrated significantly improved performance (t =
SoBran, Inc., Rockville, MD; OLAS, NIAAA/FLAC, Rockville,               2.95, P < 0.01). Also, in the immediate feedback and accountability
MD                                                                      condition, 100% of the tested areas reached a pass score of greater
                                                                        than 75% reduction in ATP. In conclusion, bioluminescence can be
ERCC-1–deficient mice tend to be runted at birth and die before
                                                                        both an effective and inexpensive vivarium management tool. By
weaning. Knockout of the ERCC-1 gene in mice results in a shorter
                                                                        using bioluminescence to collect quantitative assessments, and by
life span and the mice display signs of premature aging. At our
                                                                        providing real-time feedback and accountability to staff, supervisors
facility, these strains accurately portray these problems, along
                                                                        can improve employee performance. Furthermore, because surface
with degeneration of the hind limbs. The litters born from ERCC-1
                                                                        ATP values are obtained before and after sanitization procedures,
-/- deficient females were broken down into 3 categories based on
                                                                        bioluminescence reduces the amount of direct supervision required
genotype. The wild type mice resembled normal mouse weights at
                                                                        during sanitization, increasing supervisors’ time management and
weaning (15 g at 21 d). The heterozygous mice tend to be slightly
                                                                        efficiency.
smaller at weaning age (7 to 8 g), and the knockout mice were very
small (3 to 5 g) at weaning age. Our weaning policy indicates that
7 d post weaning, the animals are given extra supplements such a        P87 Double-mouse Perch Designed as an Enrichment and
fruits (apples and oranges) and gelatin containing whey protein.        Life-saving Device
Incidentally, the heterozygous and knockout mice struggled to           KA Phillips*, G King, A Ibanez, L Altholtz, CV Johnson
mature, and survival was not expected. Researchers required these
                                                                        Laboratory Animal Resources, Merck & Co., Rahway, NJ
animals to be weaned early (14 d) and performed an AAV1 injection
on the brain. Postsurgical knockouts recovered much slower than         Rodents housed on racks with automatic watering are widely
the heterozygous animals subjected to the same procedure. In order      used in the research community. Although there are many ben-
to increase their survival probability, the animals were provided       efits to this type of housing, occasionally cage flooding due to a
with additional surveillance and extra nutritional supplements          leaky valve occurs. A novel double-mouse perch was designed
beyond the 7 d postweaning period. These supplements consisted          as an enrichment device and provides elevated shelter in the
of fruits, gelatin fortified with whey protein, a nutritional drink,    event of a cage flooding. It is completely constructed of stain-
bacon-flavored soft pellets, and a special-needs dough diet. With the   less steel and is suspended from the wire grid of a mouse cage
supplemental care in place, we found that the animals gained weight     lid without compromising floor space. The upper level indents
more rapidly and their overall body condition did not degenerate        over the bottom level giving it a stair-step, climbing functional-
as quickly. One particular mouse was given a supplementary brain        ity that provides 2 levels of activity for enrichment opportunity.
injection of AAV1 and ERCC-1 gene which posed to reverse the            The perch allows for full viewing of the mice through the cage
effects of the knockout genotype. The mouse’s presurgical weight        during daily observations and can easily be sanitized through
was 4.30 g; it continued losing weight postsurgery, until reaching      the rack washer. It also provides aid for small and aged mice in
3.25 g. After our postsurgical treatment, its weight increased to       accessing food and water within the cage, as well as increased
3.30 g and it appeared much healthier than the previous knockout        vertical space. The functionality of this novel device was tested in
animals injected only with AAV1 injection. Combined with the            an observational study. A total of 10 mice, 5 per box, were housed
constant supplemental care and ERCC-1 injection our test animal’s       in a small ventilated mouse box (7.5 by 13.75 by 5.5 in.) and were
condition remained stable and healthy.                                  given 1 wk to acclimate to the perch. During the acclimation pe-
                                                                        riod, mice were observed interacting with the perch by actively
P86 Use of Bioluminescent Technology as a Novel Husbandry               climbing on both levels. After the acclimation period, the mice
Staff Management Tool                                                   underwent 2 separate simulations of an actual cage flooding.
                                                                        The first simulation was a pilot study in which warm water was
KM McDonald*, PA Chalmers, JT Newsome                                   slowly introduced into the cage using a flexible tube. This was
Division of Laboratory Animal Resources, University of Pitts-           done to monitor their response to the rising water and interaction
burgh, Pittsburgh, PA                                                   with the double mouse perch. The second simulation study was
                                                                        done to mimic an actual cage flooding from a leaking sipper tube.
Bioluminescent technology is often used in vivariums to verify the
                                                                        In both simulations, all 10 mice safely climbed to elevated shelter.
efficacy of santiation procedures, but it can also be used to enhance
Abstracts of scientific papers 2008 AALAS National Meeting

They were moderately wet and huddled together on the upper              over a 12-mo period. As a control, biofilm from rack manifolds
level of the perch. From these studies, we have determined that         in a facility with no additional water treatment (municipal wa-
this device is a well-used form of enrichment and an effective          ter passed through a 5-μm filter) was also sampled. At periodic
life-saving platform during accidental cage flooding.                   intervals, biofilm samples were obtained from the inside of rack
                                                                        manifold piping using sterile swabs. Biofilm samples were as-
P88 Use of a Remote, Real-time Infusion Monitoring System               sayed for total bacterial population by a rapid assay for microbial
                                                                        ATP. Results showed minimal biofilm growth in 4 racks sampled
K Pickel*, J Resendez                                                   from the facility with RO-DI-UV–treated water for up to 12 mo
Infusion, MPI Research, Mattawan, MI                                    of use. In contrast, the 3 racks from the control facility showed
                                                                        substantial biofilm growth between 3 and 6 mo of use. Median
The use of percutaneous indwelling catheters is a widely ac-
                                                                        RLU/swab values for the RO-DI-UV facility were 11.8, 24.0, and
cepted method of administering pharmaceutical compounds
                                                                        19.3 for mo 6, 9, and 12, respectively, compared to 90.0, 203.0,
into systemic circulation. Vascular access is readily established
                                                                        515.0, and 759.0 for mo 3, 4, 5, and 6 in the control facility. These
using this type of catheter, but long-term patency is often dif-
                                                                        results suggest that the combined processes of reverse osmosis,
ficult to maintain. Consequently, surgically implanted vascular
                                                                        deionization, and ultraviolet filtration are effective in limiting
catheters are often used when longer duration administrations
                                                                        biofilm growth in AWS for up to 12 mo. Facilities that provide
are required. As this infusion model has become more established
                                                                        drinking water conditioned in this manner may be able to reduce
in the preclinical arena, so has the acknowledgment that continu-
                                                                        the frequency of sanitization and sterilization to once per year,
ous intravascular administration can present unique challenges
                                                                        which would mitigate the cost of purchase and installation of
compared with other more standard routes. Consideration must
                                                                        the water treatment system.
now be given to the technical requirements and equipment
necessary to successfully conduct this type of study (such as
tethers, animal jackets, infusion pumps, surgical capabilities,         P90 Comparison of Two Foraging Devices for the Psycho-
and aseptic technique). Longer administration durations are             logical Well-being of Caged Cynomolgus Primates (Macaca
accomplished by using syringe or ambulatory infusion pumps              fascicularis)
either intermittently for repeated doses over the course of a day       J Howard*, K Pyle, M Liddle, C Noyes, E Kirchner, H Byrnes, C
or weeks, or continuously over a period of consecutive days. The        Grant, C King, K Trotta
logistics required to maintain and monitor the infusion over the
course of a typical study often result in increased labor hours         Veterinary Science Technology, SUNY Delhi, Delhi, NY
or staffing. This operational impact may result in a less efficient     In maintaining compliance for the psychological well-being of
overall infusion program. To address this issue, MPI Research           caged cynomolgus monkeys (Macaca fascicularis), environmental
has developed a monitoring system capable of continuously               enrichment through the use of foraging devices has been accepted
evaluating the performance status of each infusion pump. An             in our industry. This study compares a widely used, commercially
in-house software program was created that communicates di-             available foraging device that is hung outside the cage with a
rectly with each pump; receives specific, real-time operational         new prototype foraging/enrichment device that is offered inside
information regarding the infusion progress; and remotely alerts        the cage. The prototype is made of polypropylene plastic with a
technical staff of situations or interruptions, such as catheter line   2.5-in. hollow spout connected to a 2.75-in. hollow ball. Peeled
occlusions or a low battery, that may compromise the infusion.          baby carrots were used as a treat in this device. This study used 9
Use of this system allows for a more timely response to pump            monkeys ages 1 to 27 y old, individually and pair housed. For a
alerts, which is critical to the ultimate success of the planned        4-wk period, 10-min direct observations of the animal’s interaction
administration. Since the monitoring system was implemented,            with the foraging devices were performed 3 times per d, 3 d per
the number of technical staff and time required to maintain an          wk. Interactions were recorded as discrete events on an ethogram
infusion study has decreased, and the ability to successfully ad-       to evaluate the animals behavior in the categories of agonistic,
dress infusion interruptions has significantly improved.                human-directed, affiliative, non-social, exploratory (including
                                                                        environment and foraging device) and enrichment. At 1100, the
P89 Effect of Reverse Osmosis, Deionization, and Ultraviolet            foraging devices were loaded and presented to the monkeys. The
Filtration on Biofilm Development in Automated Watering                 test objects were left in or on the cages and were not refilled for the
Manifolds                                                               1300 and 1500 10-min ethogram evaluations. Treat removal from
                                                                        the available device was significantly easier than treat removal
KM Hugunin*, CJ Maute, RC Dysko                                         from the prototype because the treat was visible and could be eas-
Unit for Laboratory Animal Medicine, University of Michigan,            ily grasped through the holes. The prototype required substantial
Ann Arbor, MI                                                           manipulation to get the carrot lined up with the spout in order
                                                                        to obtain the treat. The opacity of the material also made it dif-
Providing drinking water to laboratory rodents through auto-
                                                                        ficult for the primate to see the treat inside, therefore providing
mated watering systems (AWS) has become commonplace over
                                                                        extended enrichment. The new prototype foraging/enrichment
the last decade. Previously, our laboratory found that biofilm
                                                                        device performed statistically better (1-tailed t tests were used with
increased significantly over a 6-mo period within automated
                                                                        α = 0.05) than the widely used, commercially available foraging
watering manifolds provided municipal water (2.8 to 3.3 ppm
                                                                        device in the areas described.
monochloramine) passed through a 5-μm filter. The effects of
various water purification treatments on biofilm growth within
AWS have not been independently examined. The newest vi-                P91 Commercial Modular Housing Facilities for Ossabaw
varium at our institution provides municipal water that has been        Miniature Swine with Metabolic Syndrome
treated by reverse osmosis (RO), deionization (DI), and ultra-          KM Pohle*, LD Jefferson, JW Wenzel, M Sturek
violet filtration (UV) using AWS. The combined effect of these 3
                                                                        Cellular and Integrative Physiology, Indiana University Purdue
processes on biofilm growth in AWS over time was investigated
by sampling biofilm from rack manifolds in use in this facility         University Indianapolis, Indianapolis, IN
                                                                                         Abstracts of scientific papers 2008 AALAS National Meeting

This presentation describes the husbandry facilities used suc-            replacing toe clipping, toe tattooing can be used as a reliable
cessfully for Ossabaw miniature swine over the past 4 y. Main             back-up to ear tagging or notching. It is also significantly less
guidelines for care include easy-to-clean surfaces, daily pen             expensive than installing a transponder.
washing and excrement removal, space, temperature, and
light:dark cycle. Ossabaw miniature swine are housed in 3                 P93 Disposable Nonhuman Primate Puzzle for High-contain-
separate commercially produced modular swine housing units                ment Labs
located near Purdue University. Each building is equipped with
2 60,000 btu natural gas heaters and 2 12-in. and 2 24-in. fans for       K Kelly*, N Powell
convective cooling. The units house the entire Ossabaw miniature          Animal Resources Branch, Centers for Disease Control and
swine colony, including breeding boars; open, gestating, and              Prevention, Atlanta, GA
lactating sows; suckling piglets; weaned piglets; young adult pigs
                                                                          Some primates are used for the advancement of public health
that have not yet been assigned to research projects; and pigs that
                                                                          with regards to infectious diseases. A few of these are located
are on research protocols being fed excess calorie, atherogenic
                                                                          in high-containment laboratories that deal with emerging in-
diets to induce obesity, metabolic syndrome, and heart disease.
                                                                          fectious diseases at BSL3/4. Enrichment items that enter these
Stainless steel gating, fiberglass outer walls, and slatted flooring
                                                                          laboratories do not come back out once the study has ended;
enable rapid, efficient daily sanitation. All buildings maintained
                                                                          they are discarded. The disposable enrichment puzzle costs
animal holding room average monthly temperatures between
                                                                          pennies to construct, does not require cleaning, and allows
61 °F (low limit) and 81 °F (high limit) guidelines over a 1-y
                                                                          primates located in BSL3/4 the opportunity to express species-
data collection period, despite the variable Indiana temperature
                                                                          specific behaviors that they would otherwise not be able to. The
fluctuations, which exceeded the limits on all days. The modular
                                                                          bulk of the device is made from cardboard plotter paper tubes.
housing units exceeded the low-temperature limit on a mean of
                                                                          One tube is approximately 3.5 in. in diameter and cut to a 6-in.
only 9 (8%) of the days that the exterior temperature was below
                                                                          length. The other tube is 2.5 in. in diameter and cut to a 10-in.
the low limit (111 d). The high-temperature limit was exceeded
                                                                          length. The smaller diameter tube comprises the inside of the
a mean of only 17 (6%) of the d that the exterior temperature
                                                                          puzzle and nestles inside the larger tube. Holes are cut into both
was above the high limit (288 d). Effective sanitation, space, and
                                                                          tubes to allow access to treats that will be later hidden within.
temperature maintenance of the animal housing area makes these
                                                                          Two lengths of plastic chain are cut to around 10 to 11 in. in
commercially produced modular swine housing units acceptable
                                                                          length. These pieces of chain will be attached at the end of the
housing for biomedical research swine.
                                                                          tubes through holes cut on both sides of the tube. Another link
                                                                          of larger plastic chain is used to hold the length together. This
P92 Benefits of Toe Tattooing as a Method of Identification               single link is used by cutting a small gap on its side and holds
KJ Diven*                                                                 the entire device to the front of the primate cage. The inner tube
                                                                          is filled with shredded paper and treats. The primates must align
Animal Care and Use Committee, John Hopkins University,                   the holes to have access and then have a fun time shredding the
Baltimore, MD                                                             cardboard. The puzzle device was tested on a room of 8 singly
Small rodent identification can be problematic when using vari-           housed adult male rhesus macaques at BSL2. These primates
ous popular techniques. When rodent pups are young they are               had used a similar PVC device and were aware of the necessary
too immature to ear tag, ear notch, or implant a transponder.             mechanics required. Most of the contents were removed within
Historically, the solution to this problem was to toe clip. This          15 to 30 min, and the tubes themselves were a continued source
involves the removal of the first bone of certain toes. The Guide         of interest for several days as the cardboard could be shredded.
for the Care and Use of Laboratory Animals limits its use and states,     Some primates had destroyed the device by the next morning;
“toe-clipping, as a method of identification of small rodents,            others were still working on theirs 2 d later. These devices could
should be used only when no other individual identification               be used for any level of containment, but were specifically de-
method is feasible and should be performed only on altricial              signed to be used in a high-containment lab. These labs will soon
neonates.” Other rodent identification methods also have their            be coming online at CDC, and this device will be given its first
drawbacks. Ear tags, if not properly sterilized and placed, can           true trial run in the coming months.
cause irritation to the ear pinna and be pulled out. Ear notching
uses a numbering code that is limited to the hundreds. When               P94 Wild-caught Virginia Opossums (Didelphis virginiana)
animals fight, the ear can be ripped, resulting in a loss of the tag or   Thriving in a Research Environment
disruption of the pattern. Even excessive grooming can cause the
pattern to be unreadable or the tag to be removed. Toe tattooing          L Andrews*, W Beckman, KJ Thompson, E Paster, DL Hickman
is accomplished with simple tools and requires limited practice           Veterenary Medical Unit, VA Medical Center, Portland, OR
time. The tools needed include a sterile lancet, green tattoo paste,
                                                                          This presentation will describe our experiences in developing and
something suitable to hold a dab of the paste, and an alcohol
                                                                          implementing a regiment to support sustained health, provide
wipe. Pups (1 to 14 d old) are cupped in the hand; older animals
                                                                          environmental enrichment, and acclimate wild opossums to a
are put in a restrainer. The lancet tip is dipped in the tattoo paste
                                                                          research setting. The wild-caught opossum is widely regarded
and the desired toe or combination of toes are poked with the
                                                                          as a difficult and possibly dangerous animal. Handling, restrain-
lancet. The skin must be punctured to introduce the paste into
                                                                          ing, and transporting these misunderstood marsupials presents
the skin, leaving a mark. The spot is then gently wiped with the
                                                                          a unique set of challenges to the animal care staff and investiga-
alcohol pad to remove excess paste, and the pup is returned to
                                                                          tors. These essential techniques, if not done properly, can result in
its dam. A green dot is visible on the toe that has been tattooed.
                                                                          great physical harm to the handler and the opossum. Using food
This dot will persist for the life of the animal. Toe tattooing is an
                                                                          as enrichment helps determine each individual opossum’s palate,
excellent replacement for toe clipping because it is less invasive
                                                                          helps maintain health and promote weight gain, and builds a re-
and uses the same coding system. Therefore, transitioning from
                                                                          lationship with the technician. The demeanor and personality of
one to the other does not disrupt recordkeeping. In addition to
                                                                          each animal can be determined through these daily interactions.
Abstracts of scientific papers 2008 AALAS National Meeting

Through the use of proper handling techniques and positive en-         (P = 0.001), were observed between the means of the samples
richment, some opossums are able to be tamed to the point where        that were taken after the installation of the HEPA filtration (15
procedures and treatment administration are considerably less          CFU/m3) to that of samples taken before the installation of the
stressful on the animals and technicians. Learning to understand       HEPA filtration (36 CFU/m3). This poster presents the findings
attitude and behavior trends through working with animals on           of a study to investigate the efficacy of a portable HEPA filtration
an individual basis increases a technician’s ability to provide        unit installed in the operating theatre.
appropriate care. By improving the animal’s environment, the
productivity and quality of the research is enhanced.                  P97 Cage Color Preference for Maintenance and Breeding of
                                                                       Rats
P95 Use of a Hand-held Computer Device to Increase Efficiency          DR Almeida, VL Amaral, VB Rosa, M Cordini, M Frajblat*
in Observations of Single-housed Nonhuman Primates
                                                                       Department of Health Sciences, Universidade do Vale do Itajaí,
LM Mello*, KL Potter                                                   UNIVALI, Itajaí, Brazil
Animal Resources, Lovelace Respiratory Research Institute,             Animal well-being is a fundamental aspect of laboratory animal
Albuquerque, NM                                                        science. The conditions under which animals are maintained have
As part of our enrichment program, all single-housed nonhuman          an impact in the scientific data collected. Light is essential for the
primates (NHPs) have their object enrichment rotated weekly.           rat circadian rhythm; it also regulates reproduction and other
The enrichment technicians, or a trained designee, monitor the         systems. The objective of this study was to test rats’ preferences
NHP’s reaction to the object as well as making behavioral ob-          between white or black cages and to investigate if the black cage
servations. Previously, these observations were made on printed        has an effect on the reproduction and growth rate of these animals.
forms that included animal identification and location, with the       The preference study was performed with 8 male and 8 female
observer filling out by hand the object offered, the NHP’s reac-       Wistar rats divided in groups of 4 and maintained in 2 propyl-
tion to the object, and the animal’s behavior. Even though the         ene cages (black and white, 49 by 34 by 16 cm) connected with a
object enrichment and the animal’s reaction to the enrichment          transparent tube. A total of 8 observations were performed during
had preestablished codes, the process was very time consuming,         a 48-h period when the location of the animals and the nest was
with observations of approximately 150 animals requiring 6 to 8        recorded. The cages were cleaned, inverted, and new set of data
h. There were also issues of contamination when the enrichment/        was collected. Feed and water consumption and fecal production
observation paper forms were brought out of the animal rooms.          were recorded. Four groups of 6 males and 6 females (total of 48
Legibility was also a problem for this handwritten recorded data.      animals) were maintained in a white or black cage for 13 wk from
To help increase efficiency of all observations department wide,       weaning and body weight was recorded weekly. Fifty breeding
the decision was made to begin using a computer-based program          pairs were divided into white and black cages and reproductive
for recording and reviewing data using hand-held computer              data was recorded for 6 mo. This study as approved by the institu-
devices and wall-mounted computers in the animal areas. The            tion ethical committee. Nests were built in the black cages in all
enrichment observation forms on the computer listed the NHP’s          tests. During the 8 observations, an average of 89.5 ± 1,1% of the
identification and location, as well as gave the user the ability to   time the animals were in the black cages. The water consumption
use a default object item for the room, with drop-down menus for       and fecal production was higher in the white cage (201.5 ± 15.5
the reaction and behavior codes, and a space to type in comments.      compared with 145.5 ± 6.5 ml and 331.5 ± 16.5 compared with
The use of the hand-held device decreased the time required to         159 ± 5.0 fecal pellets for white and black cages, respectively).
complete these observations by almost half, now taking only 3          However, a higher food consumption was observed in the black
to 5 h. Also, the device can be sprayed with disinfectant when         cage (81.0 ± 21.1 compared with 154.3 ± 11.8 gr for white and
leaving the animal room, reducing the risk of contamination.           black cages, respectively). No differences were observed in the
This device has increased animal observation efficiency, reduced       reproductive parameters observed (8.6 ± 0.3 compared with 8.1
the potential for contamination, and can be viewed by anyone           ± 0.3 pups/litter and 5.7 ± 0.2 compared with 5.8 ± 0.3 births/6
who has access to our web-based data system, which reduces             mo for white and black cages, respectively). The results of this
our paper usage. In addition, enrichment technicians now have          study show a strong preference of rats for black cages. Also, the
more time to develop and evaluate novel enrichment ideas.              use of black cages did not influence the breeding performance
                                                                       or the growth rate. These observations suggest that these cages
P96 Managing Air Quality in a Large Animal Operating The-              could be used to improve the well-being of the animals without
atre                                                                   affecting its production.

L Schofield*, D Matthews
                                                                       P98 Systematic Approach to Solving Breeding Colony Prob-
Hercus Taieri Resource Unit, University of Otago, Dunedin,             lems
New Zealand
                                                                       MC Rosa*, D Perez, M Sabol-Jones, ND Hitt
Sheep and pigs are routinely used in our large animal operating
                                                                       AHCS, NIH/NINDS, Bethesda, MD
theatre for cardiovascular and orthopaedic survival research
studies. Despite careful attention to standard aseptic techniques      The increasing use of transgenic and knockout mouse models
used in infection control, wound contamination can occasion-           has resulted in more researchers maintaining their own breeding
ally occur. Our experimental surgical facilities were originally       colonies. Because many researchers are not familiar with mouse
designed without the provision of HEPA filtration. With the            reproductive biology and breeding colony management, vivari-
advent of new technology, the use of readily portable equipment        um staff members are being consulted on these topics on a more
may offer significant advantages in the overall management of          frequent basis. This type of consultation can be time-consuming
infection control. Airborne bacterial samples were taken during        for vivarium staff, but provides an opportunity to demonstrate
orthopaedic surgical procedures both prior to and after the instal-    the value of their unique background and expertise to directly
lation of a portable HEPA filtration unit. Significant differences     contribute to the success of research projects. The most common
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

obstacles encountered when advising researchers on breeding           complex and diversified environmental enrichment (EE) that
colony issues are incomplete colony histories and compliance          promotes well-being and alleviates stress-related abnormal
with recommendations. To improve the efficiency of our breed-         behaviors and health problems. Despite their positive impact
ing colony consultations, we have developed a system which            on research, the use of enrichment strategies in neuroscience is
maximizes our ability to gather pertinent strain and breeding         often limited by concerns about safety, cost, and damage to head
colony background and to monitor compliance with recom-               implants. In conjunction with planned facility renovations, we
mendations. To obtain the necessary background information,           designed independent EE cubicles that promote arboreal, object-
we have developed a questionnaire consisting of breeding- and         oriented, and curiosity behaviors while ensuring the safety of
strain-related questions for the researcher to complete. Once we      implanted macaques. We hypothesized that such units would
have this information, we meet with the researcher to clarify de-     improve well-being and have positive impact on stereotypic and
tails and to identify the researcher’s perceptions of the problems.   feeding behaviors, activity levels, obesity, and interaction with
After analyzing the information, a breeding plan is provided          staff. Each EE cubicle measures 36 by 8 ft, was made of stainless
to the researcher and our services are offered to initiate the        steel frames and panoramic 0.5-in. laminated glass windows,
program. We offer to manage the colony for a trial period after       and was supplied with HVAC support and floor drains. Units
which a written report with our findings and recommendations          were paired with a removable divider and positioned between
for continuing maintenance of the colony is provided. Using           2 holding rooms to increase access and visual contact between
this system we have been able to identify specific problems and       animals. Each unit was equipped with multiple, sanitizable, non-
improve breeding colony production for several transgenic lines.      toxic enrichment devices and various destructible enrichment
This system has resulted in the researchers receiving practical       materials. Devices were sanitized after each use and rotated
training on breeding colony management and has increased the          every week to maintain novelty and increase stimulation. Each
satisfaction of the vivarium staff as well.                           animal was given 1 to 2 rotational time slots of 2 to 3 h per wk.
                                                                      Animals were easily trained to transfer to and from their cages
P99 Enhancements to the Canine Socialization and Training             to the units in transport cages. After 1 y of operation, no injuries
Program                                                               have been observed in implanted macaques. More importantly,
                                                                      we observed increases in time spent in species-typical activities
M Huntsberry*                                                         and overall activity level, as well as decreased stereotypical
SoBran, Bethesda, MD; NIH/DVR, Bethesda, MD                           behaviors, appetite disorders, and abnormal social behaviors.
                                                                      EE cubicles also improved the quality of social interaction with
Dogs on long-term research studies benefit from an organized
                                                                      staff. Increased activity and decreased hyperphagia, combined
program of socialization and obedience. Learning basic com-
                                                                      with implementation of a balanced diet resulted in weight loss
mands increases the manageability of the dogs for care staff and
                                                                      in the majority of animals. The observed improvement in animal
veterinarians. A formal program such as this has been executed
                                                                      well-being and health demonstrate that these EE cubicles offer
by a dedicated behavior staff at my facility. From the beginning,
                                                                      significant benefits to neuroscience macaques with no obvious
handlers have met once a week as a group with a behaviorist
                                                                      risks to head implants.
for “canine class.” Classes cover positive early human social ex-
periences for puppies, providing environmental enrichment for
all dogs in the form of agility-related games and teaching basic      P101 Use of Luminometers as an Alternative for Evaluating
commands such as sit, stay, down, and loose-leash walking. In         Animal Housing Sanitation
the last several months, our canine socialization and training        M Abernathy*
program has expanded to include, among other things, talks on
                                                                      Borgess Medical Institute—Medical Device Research Laboratory,
canine behavior. Topics taught consist of canine body language
                                                                      Portage, MI
indicative of fear, aggression, and affiliation and the appropriate
human reaction to assuage a tense situation or reward a happy         The use of contact plates for evaluating animal housing sanitation
dog. Additionally, handlers participate in individual monthly         can be a lengthy process, especially if a positive result leads to re-
meetings with the behaviorist allowing for refinement of canine-      testing. Effective sanitation of animal housing is a critical function
handling skills without the distraction of several other dog/         in a research laboratory and plays a role in how a facility allocates
handler pairs. A systematic procedure for pairing dogs to be          space for animal housing. The use of luminometers can decrease
housed together has also been implemented along with a formal         the time taken to verify that effective sanitation has occurred.
treatment program for rehabilitating fearful animals arriving         Luminometers measure the light emitted when luciferase reacts
from other facilities. As a whole, the canine program at NIH          with adenosine triphosphate (ATP). Luciferase is an enzyme that
provides both human and dogs with supplemental enrichment,            uses ATP to catalyze the oxidation of luciferin, which in turn emits
positive experiences with one another, and individual atten-          light. ATP is created by all living organisms and can be used to
tion. Opportunities such as these contribute to alleviating and       assess biological activity. A swab that contains enzymes to break
possibly preventing the development of abnormal behaviors             down the cell wall of organisms and release ATP can be used as the
resulting in mentally, in addition to physically, healthy subjects    mechanism to sample a surface for living organisms. The ATP col-
for research.                                                         lected can then be coupled with the appropriate enzymes, resulting
                                                                      in a measureable light reaction that can be quantitated by a lumi-
P100 Novel, Safe Environmental Enrichment Unit for Rhesus             nometer in relative light units. The resultant relative light units
Macaques (Macaca mulatta)                                             are directly proportional to the ATP in that swabbed area. Given
                                                                      the level of ATP, one could assess the biological activity present
M Watson*, A Ramirez, DM Gonzalez, J Reuter, M Leblanc                using this technology. If a region of a dirty pen was sampled and
Animal Resources Department, The Salk Institute, La Jolla, CA         resultant relative light units compared to the relative light units of
                                                                      the same region sampled after sanitation, then a reduction in bio-
Neuroscience research with macaques requires normal behavior
                                                                      logical activity could be calculated. A passing or failing reduction
with minimal extraneous variability to produce valid, accurate,
                                                                      in biological activity could be assigned based on what value was
and reproducible models. These animals models benefit from
Abstracts of scientific papers 2008 AALAS National Meeting

effective in preventing the clinical manifestation of disease due to   involved water restriction protocols in which NHPs receive a
bacterial contamination. The practice of using a luminometer could     set volume of water each day; if the NHP removed its sipper
then be used to evaluate the effectiveness of sanitation as quick      tube, there was no way to know if or how much of its allotted
as the sanitation could be performed. The use of luminometers as       water was consumed. Lastly, this made for potential safety
an alternative for evaluating animal housing sanitation is efficient   hazards due to the floor being slick when wet, and also created
and economical. Using this method as compared to contact plates,       a general mess in the animal room. Our solution was to cut
our facility obtained sanitation results 94% faster.                   2-mm slits into the bottle end of the sipper tube, approximately
                                                                       1 mm apart. These created sections of metal that were then bent
P102 Novel Approach for Providing Rodent Soft Food Using               backwards and fanned out horizontally to create a lip. When the
a Semi-automated Watering Valve                                        tube was inserted into the stopper from back to front, it could
                                                                       no longer be pulled all the way out. Since modifying all of our
M Campbell*, K Brayton, S Nigra, KE Saunders                           sipper tubes, not one has been removed by an NHP, even though
Comparative Medicine, Oregon Health & Science University,              animals have been observed trying. This simple modification
Portland, OR                                                           has helped maintain a clean, safe working environment for all
                                                                       staff. But more importantly, it has ensured that all NHPs have
In an attempt to use resources efficiently, we looked for novel
                                                                       uninterrupted access to water or are receiving accurate amounts
ideas to reduce labor cost. Studies at our facility determined
                                                                       when on restriction protocols.
that providing soft food to rodents incurred significant labor
costs. Although most of our rodents receive water by way of
an automatic watering system, there are occasions where the            P104 Refined Method of Cage Changing for Nonhuman Pri-
veterinary or research staff requests that softened food be sup-       mates
plied to animals. Consideration was given to using city water          L Teixeira, ME Graffam*, S Sleboda
provided through the tap in each rodent room to soften rodent
pellets. This solution was deemed unacceptable since the water         CCM, Massachusetts General Hospital, Charlestown, MA
supplied through the automatic watering system was treated,            Nonhuman primates (NHPs) continue to serve as valuable
reverse osmosis water, and the city water came directly from open      animal models for biomedical research. Because animal facility
surface water supplies. It was determined that providing water in      space is so expensive to build and maintain, and with biomedical
bottles that were filled with treated, reverse osmosis water from      research funding becoming tighter, many animal care facilities
a bottle-filling station, wrapped, autoclaved, and transported to      are looking for ways to get more done in less space. The conven-
rooms where it could be mixed with rodent pellets was an ac-           tional NHP cage change procedure uses a jump box to transfer
ceptable solution. We found, however, that a significant amount        animals from dirty cages to clean ones. The obstacle we faced
of time was spent washing the bottles, filling them, autoclaving       was that the size of the rooms did not allow the jump box and a
them, and transporting them. In order to reduce the amount of          clean cage rack to fit in the room with the dirty cage. The solution
time technicians spent preparing and transporting the water            we developed proved to be a safe, easy, and efficient alternative
bottles and to ensure that the animals were getting water that         method to jump boxes. We intended to eliminate the jump box
was consistent throughout the study, we devised a system to tap        by pushing the dirty and clean cages together so their doors
into the automatic watering lines at the room level. We attached       lined up, open the cage doors directly across from each other
air valves (the type commonly used with air compressors) to a          and have the NHP jump directly into the new cage. But with
quick disconnect flexible water hose. This flexible water hose         the cages so close there was no way to open both doors in a way
was attached to the automatic watering systems in the room with        that protected the worker and was not threatening to the NHP.
a quick disconnect fitting. These hoses are the same type used         Therefore, we devised a pulley system that enabled one to open
to provide water to the rodent racks. The installation of these        both top and bottom cage doors easily from a safe distance. This
air valves and hoses was simple and inexpensive and required           method of cage changing proved to be quicker and resulted in
little modification to the existing automatic watering system.         a safer environment for the staff and an easier transition for the
Importantly, the use of semi-automatic water valves provided a         NHP. We recommend this method in a room of any size because
significant cost savings by reducing the labor and time involved       of its ease, safety, and animal acceptance. It allows animal care
in the administrations of soft food to rodents.                        staff to change a 4-cage bank by having only to move the bank
                                                                       a few feet while putting less stress on the animals.
P103 Solution to Sipper Tube Removal by Nonhuman Pri-
mates                                                                  P105 Increased Breeding Efficiency in Mice Using Miniature
                                                                       Treats
L Teixeira, ME   Graffam*,    S Sleboda
                                                                       Y Sun, J Wang, ME Graffam*
Center for Comparative Medicine, Massachusetts General Hos-
pital, Charlestown, MA                                                 CCM, Massachusetts General Hospital, Charlestown, MA
Nonhuman primates (NHPs) continue to be valuable animal                Mice continue to be invaluable animal models in the advance-
models for biomedical research. But their high level of intelli-       ment of biomedical research, with unique strains maintained
gence sometimes presents unique challenges in providing routine        internally for individual investigator ’s study requirements.
care. A common problem we faced was laboratory macaques                These unique strains are often found to be difficult breeders,
removing the sipper tubes from drinking water bottles, even            produce small litter sizes, and experience high mortality rates
though the bottles were affixed to the outside of their cages.         amongst neonates. Cross-fostering is one solution, but it also
An animal would pull on the sipper tube until it slid out of the       leads to increased expense due to the extra cages and animals
rubber stopper, resulting in emptying its contents onto the floor.     maintained. Difficulties encountered at our facility with one
This created several problems, the most serious being that a NHP       particular breeding colony included low pregnancy rate, small
could be without water for extended periods of time (up to 17          litter size, and high pup mortality rate, primarily due to aban-
h if staff had already left for the day). Another major concern        donment. The colony experienced only 20% of all mated females
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

giving birth. Litters were generally small, consisting on average        Hypothermic mice are generally awake; therefore, external
of 4 pups, and abandonment by dams was 100%. In an effort to             warmers only were considered to avoid possible ingestion of
improve production, miniature treats were trialed by the animal          devices placed inside the cage. Devices tested were 250W heat
care staff, with each breeding female being fed 1 ball (1 g) 1 to        lamps; microwaveable gel packs, electric kennel heating plates;
2 times per wk. The results were significant in both increases           and reusable, chemically activated heating pads. Internal cage
in pregnancy rate and litter sizes at weaning and in decreases           temperature was measured with a high-low thermometer on top
in neonate mortality associated with dam abandonment. Of 30              of the bedding in microisolation cages with filter-top lids, at an
females, 23 gave birth (57% increase), with litter sizes averaging       ambient temperature of 70 to 72 °F. Covered cages were placed
6 pups (50% increase), and only 4% of dams abandoning their              directly on top of microwaveable gel packs, chemical pads, heat-
offspring (96% decrease). The combination of high biomedical             ing plates, or with the lid 4 in. under a heat lamp. Heat lamps
research costs and industry-wide funding reductions makes it in-         were the fastest, heating to 100 °F in 5 min and overheating to
creasingly vital for investigators to use the most efficient practices   120 °F in less than 15 min. Lamps required constant monitor-
available. With each mini-treat costing $0.02 ($20/1000-count            ing to ensure they did not overheat the cage, and also required
bag), this treatment proved to be a low-cost solution to docu-           a free electrical outlet and significant storage space. An 8- by
mented breeding difficulties.                                            12-in. microwavable pad could heat the cage interior to 90 °F in
                                                                         10 to 20 min and maintain temperature for approximately 1 h.
P106 Asexual Reproduction Rates of Box Jellyfish (Carybdea               However, microwaveable pads required the most preparation
marsupialis) Increase in Circular Tanks                                  time (4 min heating at 1000 W, plus travel time), and required
                                                                         heat-resistant gloves to handle. The electric kennel heating plate
MS Eckerd*1, VJ Martin2                                                  could not reach 90 °F within 20 min, though after 30 min it was
1Animal Facility, 2Department of Biology, Appalachian State              able to maintain 91 to 95 °F indefinitely. It required an electrical
University, Boone, NC                                                    outlet for operation. Chemically activated pads (2 6- by 9-in.
                                                                         pads or 1 8- by 12-in. pad) warmed the cage to 90 °F within 20
Box jellyfish (Carybdea marsupialis) are currently being studied
                                                                         min, and maintained 90 to 97 °F for over 1 h. They require less
for a variety of interests, ranging from the evolutionary history
                                                                         than 30 s activation time, do not require a microwave or electri-
and structure of photoreceptors to anti-venom and protein-
                                                                         cal outlet for deployment, cannot overheat, and require minimal
based toxins. The economic impact of recent increases in jellyfish
                                                                         storage space. The pads could be reconstituted by autoclaving in
blooms worldwide underscores the need to understand the life
                                                                         a water-filled rat cage on a liquid cycle. Reusable chemical pads
cycle and habitat preferences of the box jellyfish. In order to
                                                                         offered the best combination of user and animal safety, heating
obtain meaningful data on growth rates, reproduction rates,
                                                                         speed, and convenience.
and optimal living conditions, a population has been established
and cultured long-term in the laboratory. The box jellyfish are
housed in covered, large finger bowls, Petri dishes, and tempered        P108 Harmonization of a Global Vision for Laboratory Animal
glass dishes. Culture conditions include growing animals in real         Medicine: The International Association of Colleges of Labora-
seawater at 19 °C and feeding them brine shrimp once a week,             tory Animal Medicine
followed by a water change of the dishes. No attempts are made           PV Turner*1, WJ White2, PJ Baneux5, K Bayne4, J MacArthur
to regulate water temperature or light/dark cycles. Under these          Clark3, I Alvarez Gomez de Segura6, T Kurosawa7, KJ Lee8, J
conditions, animals asexually bud off polyps and jellyfish. Box          Park9
jellyfish polyps were seeded into round aquaria made of clear            1Department of Pathobiology, University of Guelph, Guelph, ON,
plastic with a filter system. The round design of the system
                                                                         Canada; 2 Charles River, Wilmington, MA; 3Animals Scientific
eliminates corners which can trap jellyfish while providing more
                                                                         Procedures Inspectorate, The Home Office, London, United
surface area for polyp attachment than do the culture dishes.
                                                                         Kingdom; 4 AAALAC, International, Frederick, MD; 5Center
The hypothesis tested is that asexual reproduction rates of box
jellyfish (polyp and jellyfish formation) will increase in the circu-    for Comparative Medicine, Northwestern University, Chicago,
                                                                         IL; 6Veterinary Teaching Hospital, University Complutense,
lar tanks with continuous water movement and water filtration
                                                                         Madrid, Spain; 7The Institute of Experimental Animal Sciences,
when compared to budding rates of animals in tempered glass
                                                                         Osaka University Medical School, Osaka, Japan; 8Center for
dishes. Results indicate that budding rates of polyps and jellyfish
increased in the round aquaria when compared to budding rates            Biomedical Research, University of North Dakota, Grand Forks,
                                                                         ND; 9College of Veterinary Medicine, Seoul National University,
in the tempered glass dishes. As the circular tank environment
                                                                         Seoul, South Korea
more closely resembles the natural habitat of the jellyfish than
does that of the rectangular tempered glass dishes, an increased         Trained, competent laboratory animal veterinarians play a critical
budding rate was predicted.                                              role in refinement of research animal care and use. Laboratory
                                                                         animal medicine has existed as a veterinary specialty in North
P107 Comparison of Methods to Rewarm Hypothermic Mice                    America for more than 50 y, and several other formal colleges
                                                                         of laboratory animal medicine have been developed in different
NH Shomer*, H Berenblit                                                  regions around the world in the past decade. Although laboratory
Laboratory Animal Resources, Merck Research Labs Boston,                 animal medicine is an international field, it has been difficult to
Boston, MA                                                               address relevant global issues with a consistent voice because
                                                                         of the lack of a common global platform for communication.
Cage floods are a common cause of accidental morbidity and
                                                                         Recently, the International Association of Colleges of Laboratory
mortality in research rodents. Even minor floods that do not
                                                                         Animal Medicine (IACLAM) was formed by ACLAM, ECLAM,
result in drowning can cause death due to hypothermia. Prompt
                                                                         JCLAM, and KCLAM to enable the specialty to have broad in-
warming is an important first aid procedure for wet mice. We
                                                                         ternational representation. The mission of IACLAM covers four
compared methods to warm cages to a target temperature
                                                                         primary areas, including: certification of veterinary specialists
(90 to 100 °F), considering animal and operator safety, speed
                                                                         (harmonizing international standards and sharing best practices
of warming, convenience of use, and storage space required.
Abstracts of scientific papers 2008 AALAS National Meeting

among existing colleges); education of veterinarians (encourag-       while no biological endpoints are available. We recently devised
ing training at various levels, including training in developing      a simpler and reliable approach to determine if animal care
countries); dissemination of information relevant to the field        staff and investigators are practicing proper cage and animal
(delivering veterinary expertise in international discussions);       handling techniques. Gnotobiotic mice free of aerobic bacteria
and serving as research partners (promoting the veterinary            were maintained in sterilized cages, provided irradiated feed
role in refinement). An additional benefit of this international      and acidified (sterile) drinking water, and opened or changed
partnership is that demographic analyses of laboratory animal         only in an appropriate laminar flow hood after decontaminating
veterinary specialists may be conducted at a global level. Results    gloves and forceps with chlorine dioxide before handling sterile
of a recent IACLAM survey conducted within each college have          equipment or mice. Fresh fecal pellets were collected from these
pinpointed critical international pipeline trends of veterinary       mice and cultured in tryptic soy broth for 24 h, then swabbed
specialists in this field; identified employment sector statistics,   onto 5% sheep blood agar plates and incubated 48 h aerobically.
which may be useful for lobbying for training support at in-          Any bacterial growth from these cultures indicated a break in
ternational levels; and enabled identification of subspecialists      technique because the intracage environment was originally free
within the field who may be able to represent the community           of aerobes. Such mice remained gnotobiotic for many months
internationally, as needed. As globalization trends continue, IA-     when proper technique was used. Conversely, aerobic culture
CLAM is expected to play a key role in supporting initiatives to      results were used to institute additional training in addition
train veterinarians in this field, as well as in the development of   to monitor cages and supplies for sterility. These gnotobiotic
global standards for veterinary care of animals used in research,     mice may simultaneously serve as routine pathogen sentinels,
testing, and education.                                               thereby reducing costs even further while better monitoring
                                                                      colony management.
P109 Improving the Efficiency and Accuracy of Animal Health
Reporting by Means of Personal Digital Assistant Scanners             P111 Adoption as an Alternative Destiny for Retiring Research
and Bar Codes                                                         Dogs
R Scott, E Villalobos, A Gaudiel, R Garcia-Gonzalez*, E Chua          AS Fix3, JL Borger3, RS Cooper*1, G Jeans1, B Rupert3, MA Tet-
                                                                      rick3, JL Wheatley2
Laboratory Animal Resources, Genentech, Inc., South San Fran-
cisco, CA                                                                          Product Safety, 3Pet Care, Procter and Gamble,
                                                                      1BTF, 2Central

In order to provide better monitoring of clinical health issues       Lewisburg, OH
in a large animal facility, our department has rolled out a PDA-      Adoption is a viable destiny alternative for animals involved
based tracking system. The in-house bioinformatics department         in some, but not all, types of research service. We describe an
developed a real-time application within our already existing         adoption program for dogs implemented at our company. This
Laboratory Animal Systems and Reports (LASAR) database,               program included a set of principles for adoption, with inclusion
which allows interfacing clinical case information with order-        and exclusion criteria; an adoptability classification system based
ing, protocol, and other sensitive information. In the past, case     on thorough medical and behavioral assessments; a specific ac-
information was tracked by a mixed system of paper records            climation and training plan for each dog; and a screening and
and spreadsheets in a shared folder, which required many points       approval process for prospective owners. Adoption candidates
of manual data entry. With the rapid growth of our company’s          were dogs from a wide age range and service history. Dogs
mouse population, this system became inadequate in handling           received complete preadoption assessments and were grouped
the increasing number of clinical health cases. The new system        according to the adoptability classification system. This helped
has significantly improved the clinical monitoring efficiency         identify areas to address during preadoption acclimation and
and accuracy, while also decreasing response time to urgent care      training. Specific training or other interventions were provided
cases. Most human data entry errors have been eliminated due to       based on individual needs, including leash training and exposure
the use of automated data entry through PDA barcode scanning          to environments likely to be encountered postadoption. A num-
and automatic exportation of data into the real-time database.        ber of dogs were moved through this program and successfully
This results in all data being stored in one centralized database,    placed in homes. Implementation of this program required the
which allows for better searching and reporting options.              collaborative efforts of staff from the veterinary, animal welfare,
                                                                      and facility operation areas. As a way to improve the quality of
P110 Using Gnotobiotic Mice as Technique Sentinels                    life of retired research dogs and decrease the number of animals
                                                                      euthanized in nonterminal research, we advocate broader con-
RS Sedlacek*, DM Jarrell, SM Niemi                                    sideration of active intervention programs and adoption as an
CCM, Massachusetts General Hospital, Charlestown, MA                  alternate destiny for these animals.
Widespread use of microisolation cages has dramatically reduced
infections in rodent colonies in a cost-effective manner by shift-    P112 Biohazardous Waste Disposal Cost Savings
ing the effective barrier from the animal room or vivarium to         RF Lariviere*, GR Robinson
the cages themselves. As evidence, it has been our experience
                                                                      Comparative Medicine, Pfizer inc, Groton CT
that unwanted infections from mice injected with contaminated
cell lines or mice with previously undetected pathogens are           Properly disposing of animal waste is vital to an animal research
contained to only those cages, while adjacent static or ventilated    vivarium. Normal animal waste and Class II biohazardous
microisolation cages on separate experiments remain protected         animal wastes are commonly produced in the animal research
if proper technique is used. Continuous protection of rodents         environment. These wastes consist of soiled bedding and ani-
from infection requires strict adherence to maintaining sterility     mal feces and urine, and are typically in polycarbonate animal
of cages and supplies. Currently the only methods to detect poor      boxes. Normal conventional animal waste is removed by dump-
cage handling technique involve fluorescein powder or other           ing the box contents into a vacuum bedding dump station. The
chemical tracers and expensive equipment to assay samples,            waste is vacuumed directly into a specialty dumpster for in
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

state incineration. Biohazardous waste must be autoclaved in           for service charges to investigators as activity coordinates with
a vacuum-type sterilizer/autoclave. Once the boxes have been           normal husbandry routines.
sterilized, the waste is hand scraped under a protective hood and
deposited into a biohazard cardboard box that is lined with 2 red      P114 Use of Flashlights in Old World Nonhuman Primate
biohazard bags which have a weight limit of no more than 40            Health Monitoring
lbs. The biohazard boxes are then sent to a Midwestern state for
disposition. The costs associated with disposal of these wastes        S Savane*
were significant enough to thoroughly investigate our standard         SoBran, Bethesda, MD; NIH/DVR, Bethesda, MD
operating procedures and determine if a change could be made.
                                                                       Health monitoring is a critical element in managing a nonhuman
Our purpose was to determine if there would be cost savings in
                                                                       primate facility. Effective monitoring ensures timely identifica-
diverting the sterilized biohazardous waste into the normal waste
                                                                       tion and reporting of animals needing veterinary care. Personnel
stream. After working directly with the Environmental Health
                                                                       performing health checks monitor animals for illness, injuries,
and Safety Department, it was determined that the biohazardous
                                                                       or abnormalities twice daily. This task can be challenging in an
waste could in fact be diverted into the regular waste stream if
                                                                       environment where nonhuman primates are housed using quad-
certain procedures were followed. The autoclave must be tested
                                                                       cage units. The Guide for the Care and Use of Laboratory Animals
after every 40 h of service, the autoclave must reach 270 °F (130
                                                                       recommends light at the cage level should be approximately 325
°C) at 27 lbs/in.2 gauge pressure for no less than 45 min, and it
                                                                       lux at 1 m above the floor, which may not provide sufficient light
must be validated. With all these requirements being met, the
                                                                       for adequate observation of animals in the lower-level cages of
autoclaved animal bedding is now dumped directly into the
                                                                       quad-cage units. Additionally, while some animals come to the
vacuum bedding dump station along with the normal animal
                                                                       front of the cage during observations, others tend to hide in the
waste. The result is that there is a 92.5% reduction in cost as well
                                                                       back of the cage where light transmission is significantly reduced.
as 7 to 10 working h saved per work week.
                                                                       Bringing these animals to the front of the cage using a squeeze bar
                                                                       can be very stressful for the animal and can potentially interfere
P113 Humane, Efficient, Safe, Cost-effective Rodent Euthanasia         with the observation of the animal and the physical environment.
Chamber                                                                Facility management evaluated the use of flashlights to observe
S Gibson*, S Berry, B Lawless, M Tengco, M Leblanc, J Reuter           animals in lower-level cages to determine its usefulness in al-
                                                                       lowing animal care staff to observe animals without the use of
Animal Resources Department, The Salk Institute, La Jolla, CA          squeeze bars. Tests were performed to determine the length of
Population management in modern vivaria requires efficient and         time required for care staff using flashlights compared to those
humane methods for euthanizing large quantities of rodents.            not using flashlights to find 5 ml of an experimental substance
Unique solutions such as modified ventilated rack systems,             placed in the cage. Results showed that the length of time to
refrigerator units, or expensive commercial devices have ef-           find and report the experimental substance by staff not using
fectively used CO2 to meet these requirements. While greatly           flashlights exceeded, by as much as 24 h, the time required for
streamlining operations, each is with limitations including large      those using flashlights. Our conclusion is that the use of flash-
expense, obscured views, or obligation to servicing contracts. Our     lights in performing daily health checks in a nonhuman primate
facility has historically used single-cage CO2 euthanasia, which is    facility enhances the quality of observation and the detection of
time consuming, labor intensive, and cost-inefficient. We sought       potential health problems.
an affordable in-house method to address growing demands
while optimizing cost and labor and without compromising               P115 Optimizing Timed Pregnancies in Genetically Engineered
animal well-being or staff safety. Working with institutional          Mice
facilities, we designed a novel, easy-to-maintain, transparent
acrylic euthanasia chamber (17 by 27 by 33 in.) that has a total       SL Mader, N Libal, R Yang, SJ Murphy*
capacity of 16 standard mouse shoe box cages. It contains 2            Anesthesiology and Peri-Operative Medicine, Oregon Health
rows separated by a barred shelf to allow free flow of gases,          and Science University, Portland, OR
and closes with an easy access doubled-latched hinge door. It’s
                                                                       Many departments are developing mouse breeding cores to supply
equipped with a 0.5-in. CO2 entry valve, a 1.5-in. air intake valve,
                                                                       researchers with genetically engineered mice (GEMs). Such cores
a 0.25-in. exit vent port, and a 1.5-in. exhaust valve connected to
                                                                       can provide other services like timed pregnant GEMs. Due to the
house exhaust. The chamber is directly connected to house CO2
                                                                       variety of GEMs, strategies are needed to optimize GEM-specific
lines with a regulating valve, eliminating the need for portable
                                                                       timed pregnant protocols. Our study objective was to evaluate preg-
tanks. We installed a wall-mounted CO2 room monitor/alarm
                                                                       nancy indices (plug rate, pregnant plugged rate, overall pregnancy
for added safety. The chamber minimizes animal distress by
                                                                       rate, litter number and size, body weight) between a homozygous
reducing animal handling, using home caging, and producing
                                                                       soluble epoxide hydrolase knockout (KO, n = 84 females) and a L7-
rapid loss of consciousness through a steady flow of CO2. The
                                                                       tau-green fluorescent protein transgenic (TG, n = 69 females) mouse
materials cost approximately $800. Development challenges
                                                                       colony in order to optimize our timed-pregnant protocols. Estrous
included achieving steady CO2 dispersal, moderation of force-
                                                                       cycles were synchronized by exposing group-housed females to
ful house exhaust, identifying port entry locations, and safety
                                                                       male urine-soaked bedding for 4 d, thus taking advantage of the
assessment. Variables effecting cost included material, chamber
                                                                       Whitten effect. Timed matings were set up by placing 2 synchro-
dimensions, and availability of house CO2 lines. Maintenance is
                                                                       nized females with a male for 48 h. Females were checked daily
minimal and includes standardized cleaning and decontamina-
                                                                       for the presence of a vaginal plug while males were present. The
tion procedures with a chlorine dioxide-based sterilant followed
                                                                       day a plug is found was considered to be embryonic day 1 (E1).
by ethanol. Modification of work flow and husbandry practices,
                                                                       We weighed females upon removal of males and then weekly for 2
in conjunction with the new chamber, increased our capacity (16
                                                                       wk to confirm pregnancy. Plug rates were 68% and 26%, pregnant
cages), reduced required effort (10 technicians reduced to 1) and
                                                                       plugged rates were 49% and 78%, and overall pregnancy rates were
time (10 h/d decreased to 2 h/d), and has eliminated the need
                                                                       39% and 49%, respectively, for the KO and TG colonies. Average
Abstracts of scientific papers 2008 AALAS National Meeting

litter size for 1st (4.9 ± 0.3, n = 30) and 2nd litter (5.0 ± 1.4, n = 2) was   1Research Animal Resources, 2Research Animal Resources, Johns

similar for the KO colony but was increased for the TG colony (1st              Hopkins University, Baltimore, MD
litter, 6.5 ± 0.3, n = 27; 2nd litter 9.6 ± 0.8, n = 7). In the KO colony,      This abstract outlines the transformation of investigator-main-
pregnancy (n = 33 mice) increased % baseline body weight at E7                  tained satellite facilities at our facility. The chain of events that
(105 ± 1%) and E14 (126 ± 1%) compared to nonpregnant mice (n =                 led to the transformation, the key persons that contributed to the
51; E7, 102 ± 0%; E14, 103 ± 1%). Pregnant mice (n = 34) in the TG              transformation, and an outline of the actions taken to complete
colony also showed an increase in % baseline body weight at E7                  the transformation will be described in detail. The university
(108 ± 1%) and E14 (142 ± 2%) compared to nonpregnant mice (n                   was confronted with the need to address inconsistencies in the
= 35; E7 and E14, 102 ± 1%). Based on plug and pregnant plugged                 animal care practices that occurred among some of the satellite
rates, we will refine our protocols by limiting male exposure to 24             animal housing facilities in our program. Key personnel within
to 48 h for timing pregnancies and using weight measurements at                 Research Animal Resources (RAR) and the Animal Care and Use
least 1 wk after male removal for early pregnancy confirmation, as              Committee (ACUC) were able to devise policy and procedures
plug detection was not a reliable pregnancy indicator in our KO and             to improve the satellite facilities’ operation and function. These
TG colonies. Further refinements are planned based on litter size               policies and procedures were put in place to meet the standards
and number such as using nulliparous and monoparous females                     outlined in the Guide for the Care and Use of Laboratory Animals,
in our KO and TG colonies, respectively.                                        as well as provide a means for maintaining these standards.
                                                                                To explore these events, we visited these satellite facilities and
P116 Can Continuous Harem Breeding Be Successful in Small                       investigated the role that they played in this transformation. We
Shoebox Cages and Still Meet Regulatory Expectations?                           examined them before and after the transformation and analyzed
N Libal, SL Mader, R Yang, SJ Murphy*                                           what it took to complete this change. The policy and standard
                                                                                operation procedures that were created specifically for this
Anesthesiology and Peri-Operative Medicine, Oregon Health                       program were also explored in detail. Lastly, we explained the
and Science University, Portland, OR                                            current procedures and plans that have been devised to main-
A continuous harem breeding strategy would exceed the recom-                    tain these standards as a long-term goal. With the cooperation
mended space for group-housed mice when litters are present in a                of investigators and staff, we were able to make a vast number
small shoebox cage with a breeding harem. However, regulations                  of improvements to the satellite facilities’ operations. We were
do allow space allocations to be modified to address issues like                able to reduce the number of satellite facilities by moving some
postnatal care and group housing of social species likely to share              activities to central facilities, a clean/dirty exchange has been
space. We received IACUC approval for a housing exception to                    created between labs and central facilities, investigators have
allow continuous harem breeding in small shoebox cages (total                   corrected maintenance and cleanliness problems, and animal
floor space 67.6 in.2 [436 cm2]) for departmental mouse breeding                housing has been designated to single specific locations in the
colonies so as to improve postnatal care and litter survivability.              labs. Through such actions, as well as the dedication of our in-
Data was collected for 10 mo using a customized breeding colony                 vestigators, we have enhanced the animal care and use program
database from the following 6 knockout colonies: Akt1, soluble                  at the investigator-maintained satellite facilities.
epoxide hydrolase, P450 aromatase, estrogen receptor α and/
or β. We evaluated colony performance indices (litter number/                   P118 Use of Soft Hydrothermal Processing to Improve and
harem, litter size, preweaning losses, incidence of death and                   Recycle Bedding for Laboratory Rodents
euthanasia for female breeders, health and behavioral problems)
as recommended by the Guide for each colony and overall to                      T Miyamoto*1,2, N Kasai1, Z Li2, T Kibushi1, N Yamasaki2
ensure the adequacy of continuously housing breeding harems                     1 Institute for Animal Experimentation, Graduate School of

and their litters in our facilities cages. Litter number/harem                  Medicine, Tohoku University, Sendai, Japan; 2 Graduate School
was 5.6 ± 0.2 (n = 198 harems). Litter size was 6.8 ± 0.1 (n =                  of Environmental Studies, Tohoku University, Sendai, Japan
1103 litters). Preweaning pup loss was 28.0 ± 4.1%. Incidence
                                                                                Bedding is one of the common means of improving well-being
of euthanasia and death for female breeders (n = 385) was 8.3 ±                 and the environment of laboratory animals. It also influences the
2.5%. Common health problems seen in female breeders were                       accuracy of the experimental results. In addition, a large amount
dystocia (2.2 ± 1.0%), uterine prolapse (0.5 ± 0.4%), animals found             of used bedding containing excrement is produced as the medical
dead of undetermined cause (1.7 ± 1.0%), and skin problems
                                                                                waste from animal facilities of life science institutes. On the other
(1.3 ± 0.9%). No fighting was observed in any harem cage. Litter
                                                                                hand, the high temperature and pressured dry steam has potential
number/harem, litter size, and preweaning losses in individual
                                                                                for promoting organic reactions, such as extraction and drying
knockout colonies are comparable to what is reported for these                  with soft hydrothermal processing. We developed a groundbreak-
colonies in the literature or anecdotally by colleagues using pair              ing system to improve fresh bedding and to recycle used bedding
breeding or removal of pregnant mice from harem cages. Colony
                                                                                by applying a soft hydrothermal processing. The system removes
performance indices for all colonies collectively are similar to or
                                                                                both harmful organic components and aromatic hydrocarbons
at the high end of that observed in other facilities which do not
                                                                                that can affect animals’ metabolism. The purpose of the present
use continuous harem breeding for genetically engineered mice.                  study is to evaluate the effects of soft hydrothermal processing on
Our individual and collective colony performance indices and                    the beddings of laboratory rodents. Soft hydrothermal processing
the lack of fighting in harem cages suggest that in our continu-                removed 68% to 99% of the predominant aromatic hydrocarbons
ous harem breeding system, neonatal and adult mice housed in
                                                                                in softwood chip bedding (spruce) treated at 0.35 MPa and 140 °C
small shoebox cages have adequate space to address reproduc-                    for 120 min (“improved bedding”). In addition, 59.4% to 99.0% of
tive, social, and individual needs.                                             predominant harmful organic compounds derived from excrement
                                                                                were also removed in used softwood chip bedding (spruce) treated
P117 Transformation of Investigator-maintained Satellite                        at 0.45 MPa and 150 °C for 60 min (“recycled bedding”). The soft
Facilities                                                                      hydrothermal treatment increased the number of acidic functional
SP Williams*1,2, L Barnes1,2                                                    groups on the bedding surface, thus increasing its adsorption
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

of ammonia gas. Harmful substances such as microorganisms,            consider when identifying personality traits, which include gen-
heavy metals, and pesticides decreased below the detection limit.     der, upbringing, education, and health. The temperament theory
Then we examined effects of reproduction with these beddings in       is not an exact science; however, when the staff at our facility
mice. Body weight of male and female mice and reproductive data       considered all the variables including each individual’s inherited
(litter size and total number of offspring) showed no significant     temperament, they had a good perspective of themselves and
differences among different bedding groups. The results showed        the people they work with. By creating an optimal atmosphere
that the improved and recycled bedding are safer for laboratory       in the workplace, the manager will be able to generate more
rodents and has the potential to ameliorate conditions in primary     teamwork and attain more productivity.
and secondary enclosures (such as cages and animal rooms) used
for maintaining laboratory animals. This process has the potential    P121 Light Intensity Comparisons between the Macroenvi-
to be one of the most advanced techniques in improving softwood       ronment and Microenvironment of Various Caging Systems
bedding, economizing through the recycling of used bedding and        Housing Albino Laboratory Animal Species
reducing by 30% the bedding waste from animal facilities.
                                                                      VT Lea*, SJ Young, AE Kraycirik
P119 Rodent Bedding Comparison Study                                  LASW, sanofi-aventis, Bridgewater, NJ
T Putro*, PA Straeter                                                 According to the Guide for the Care and Use of Laboratory Animals,
                                                                      light can affect the physiology, morphology, and behavior of
Veterinary Sciences, Bristol-Myers Squibb, Princeton, NJ
                                                                      animals used in research. Potential photostressors include in-
A large percentage of the operating expenses accrued in maintain-     appropriate photoperiod, photointensity, and spectral quality
ing a rodent facility involves animal bedding. The predominant        of light. During an AAALAC site visit, the site visitors recom-
bedding used at our institution is an alpha cellulose material        mended that we reevaluate light intensity levels in some of our
that provides favorable benefits such as defined material com-        animal holding rooms. We conducted extensive testing of the
position and dust-free, sanitary environments for the rodents         light intensity levels within cages housing albino rabbits, guinea
and personnel. However, the alpha cellulose material is more          pigs, mice and rats. All intensity levels were measured using the
expensive than other bedding, which increases expenditures.           Extech Instruments Model #401025 foot candle/lux meter. An ini-
Other bedding alternatives, such as corn cob or a bedding blend,      tial lighting intensity reading was taken in the center of an empty
may be more affordable, but potentially compromise the health of      animal room at approximately 1 m off the floor. Multiple types
the rodent colony and possibly the integrity of the research data.    of caging were tested, as were multiple cage positions through-
As a result, a study was designed and executed to compare the         out the room. Different types of bedding were also evaluated to
microbial analysis of pre- and postautoclave bedding of 3 types:      determine if bedding impacted light intensity. Measurements
alpha cellulose, corn cob, and an alpha cellulose-corn cob 50/50      were taken at various positions on the rack in the microenvi-
blend. Eighteen samples (9 preautoclave and 9 autoclaved), along      ronment and the macroenvironment. Results were compared to
with 6 control samples were evaluated. Microbial cultures were        the recommended range of 130 to 325 lux at cage level, which
analyzed from a 10-g homogenized slurry of each bedding type,         is sufficient for animal care and does not cause clinical signs of
and organism counts and identifications were performed. Results       phototoxic retinopathy in albino rats. During our testing, we
of the preautoclave samples indicated the corn cob bedding had        found no significant changes in readings with the different types
the highest quantity of bacterial and fungal organisms, while the     of bedding used; however, cage positioning had a positive affect
alpha cellulose bedding presented with the least. All autoclaved      on the readings. Moving the cage away from direct light reduced
samples yielded no microorganisms.                                    the intensity readings. One alternative to help regulate light in-
                                                                      tensities in animal rooms is to rotate cage positions away from
P120 Comparative Analysis of Diversity in Temperaments                direct light. Data collected outside the acceptable ranges were
Commonly Associated with Animal Technician Models                     reviewed by management. Results of this study led to corrective
                                                                      actions to provide a higher-quality environment for the animals
VR Riojas*, JA Kundert, TK Marcotte, RE Arens, TS Frerck, DW          used in drug discovery research.
Harn, CM O’Rourke
Animal Resources Center, Montana State University, Bozeman,           P122 Effect of Directed Changes to Business Processes on Time
MT                                                                    to Approval for Animal Use Protocols
Maintaining a team atmosphere in an animal facility with a large      W Moseley*1, DM Ostmeyer2, L Martin1, J Owiny2
or diverse staff can seem daunting. The typical challenges associ-    1Research Integrity & Compliance Review, 2Vice President for
ated with this type of situation can be simplified by identifying
                                                                      Research, Colorado State University, Fort Collins, CO
each technician’s personality traits and innate characteristics. We
have seen dramatic changes in our staff meetings by educating         Institutional Animal Care and Use Committees (IACUCs) are
ourselves on the temperaments of others in order to understand        charged with oversight of the animal care program. Part of that
them better. We each took a personality test based on Hippo-          role involves review of proposed animal activities. Virtually all
crate’s 4 basic temperaments. The Sanguine is charming, but can       institutions have developed forms and processes to aid review
be unproductive. The Choleric is very productive, but tends to be     of animal protocols. Based on perceived problem areas, our
intense and unsympathetic. Melancholies are compassionate, but        IACUC and administrative office have instituted changes to
moody. The Phlegmatic temperament is cool, calm, and collected,       protocol application forms and review processes in a phased
but can be too leisurely. Each individual possesses a blend of all    manner over the past 12 mo. The changes include the addition of
4 of these traits. For this study, fabricated people conducting a     university veterinarian prereview of select protocols, extensive
staff meeting as models to distinguish the differences in these       administrative prereview of all protocols prior to committee
temperaments and identify the strengths and weaknesses of             review, incremental changes to the A-100, and additional PI
each were used. It is important to point out that each subject        training. The goal of this study was to determine whether the
is an individual and completely unique. There are variables to        changes instituted over the past year have reduced the time
Abstracts of scientific papers 2008 AALAS National Meeting

between submission of animal use protocols and final approval.        1CCM,   Massachusetts General Hospital, Charlestown, MA;          2

Using 2006 as a baseline, we compared the number of protocols         Digital Paradigms, Inc, Boston, MA
submitted in 2006, 2007, and the first 4 mo of 2008 to determine      Per diem charges for husbandry services is the most common
if our incremental changes have had any effect on protocol            means of recovering lab animal care costs in academic research
turnaround time. In 2006, a total of 267 protocols were reviewed      institutions. Many institutions are using bar codes and scanning
and approved; 219 protocols (82%) were reviewed and approved          technology to more efficiently label and track cages for per diem
within 30 d, while 48 (18%) took longer than 30 d to review and       billing as well as for animal health, breeding, and regulatory
approve. In 2007, a total of 270 protocols were reviewed and ap-      compliance purposes. As lab animal care becomes more sophisti-
proved; 233 (86%) of those protocols were approved within 30 d,       cated and expensive while federal funding of biomedical research
and 37 protocols (14%) took longer than 30 d to approve. During       declines in real dollars, maximizing cost recovery through per
the first 4 mo of 2008, 112 protocols were reviewed, and 97 (87%)     diem revenues becomes even more important. Historically,
were approved within 30 d, while 15 protocols (13%) took more         our department at MGH performed cage census on Mondays,
than 30 d to approve. Additionally, the mean number of days to        Wednesdays, and Fridays. For billing purposes, the system was
approval for each of the 3 periods has decreased from 18.49 to        programmed to “fill in” for the non-census days if a cage was
14.62, to 14.22 d. Although it appears from the data on the first     scanned on the 1st and the 3rd d. Taking census more frequently
4 mo of 2008 that we may have reached a point of diminishing          was presumed to be an additional and costly labor expense, with
returns, the overall data indicate that the incremental changes       no net revenue gain. Because of the dynamic census in some of
in the application form and review processes implemented thus         our animal facilities, in which animals are coming and going on
far resulted in improved time to approval.                            a daily basis, this presumption was tested. Rodent cages in one
                                                                      facility were scanned every weekday for 1 mo and the resultant
P123 Comparison of Two Hydration Gel Products for Use in              daily census and revenue were calculated. The same data were
Shipping of Guinea Pigs (Cavia porcellus)                             then used to simulate what the census and per diem revenue
Y Luo*, GB Mulder, RR Morin                                           would have been using the traditional 3-d approach. Performing
                                                                      census 5 d per wk increased monthly per diem revenue 3%. On
Charles River, Raleigh, NC                                            an annual basis at our institution, this extrapolated to a $225,000
Guinea pigs are generally considered to adapt poorly to changes in    increase in revenue for the department. Bar code scanning for
food presentation and to novel food items. During transportation,     census averages 34 cages per min, so the resultant additional
it is common to provide animals with water replacement gels to        labor cost for scanning cages an additional 2 d per wk consumed
aid hydration. The goal of this study was to investigate preference   only 10% of the additional revenue gained. This yielded a return
of guinea pigs for 2 different water replacement gels. The primary    on labor of 900%, creating a low cost:high yield improvement to
difference between the products was the presence or absence of        increase animal care revenues simply.
high-fructose corn syrup (HFCS). This study also evaluated the
effect of the sweetener ingredient on palatability to this specie.    P125 Characterization of Western Equine Encephalitis Virus in
Fifty-two male Hartley guinea pigs (Crl:HA) weighing 330 to 400       the Syrian Golden Hamster (Mesocricetus auratus)
g were used. While 10 of the animals served as control for clinical
chemistry and CBC, the other 36 guinea pigs were divided into 2       AL Olsen*1,2, J Morrey2
groups of 18 animals. Each group was supplied with either sweet-      1Laboratory  Animal Research Center, 2Animal, Dairy, and Vet-
ened or unsweetened gel product and transported within filtered       erinary Sciences, Utah State University, Logan, UT
crates overnight to a remote facility and subsequently held within
                                                                      It was hypothesized that intraperitoneal inoculation of Syrian
the shipping crates for an additional 3 d. Assessment of animals
                                                                      Golden hamsters with Western Equine Encephalitis Virus (WEEV)
after arrival included clinical chemistry, CBC, and body weights.
                                                                      did not routinely cause viral encephalitis, but rather resulted in
In the group supplied unsweetened gel, animals consumed 99.5%
                                                                      a nonneuronal systemic disease, similar to a disease phenotype
of supplied gel (124 g/d/animal). In contrast, the consumption
                                                                      reported in humans infected with similar alphaviruses. It was
of HFCS-sweetened gel was 63.6% (82 g/d/animal). The average
                                                                      further hypothesized that virus infection produced secondary
body weight losses were 2.7% and 23% for the unsweetened and
                                                                      effects, such as overwhelming systemic inflammation, which con-
sweetened gels groups, respectively. The values for hematocrit,       tributed to the high mortality and rapid death observed. Finally,
and serum sodium, chloride, total protein, and albumin were sig-
                                                                      it was hypothesized that WEEV retained the ability to induce
nificantly higher (P < 0.05) in the animals supplied sweetened gel
                                                                      viral encephalitis if animals survived the systemic disease, or if
as compared to the animals provided unsweetened gel. Follow-up
                                                                      the virus was introduced directly into the central nervous system
preference testing was performed by providing 6 singly housed
                                                                      (CNS). Hamsters inoculated intraperitoneally with WEEV exhibit
male guinea pigs both water and HFCS-flavored water and moni-         a unique disease phenotype as compared to that reported for mice
toring the consumption of both fluids for 5 d. Animals consumed
                                                                      inoculated with WEEV. Hamsters displayed increased sensitivity
approximately 20% more unflavored water than water sweetened          to WEEV, showing high rates of mortality and death by 96 h post-
with HFCS. The results of this study indicated that Crl:HA guinea     virus inoculation (hpi), even when inoculated with relatively low
pigs provided an unsweetened hydration gel consumed greater           doses of virus. Furthermore, hamsters did not display overt signs
amounts of gel, showed reduced body weight loss, and decreased        of neurological disease. Experimental results found that splenic
clinical pathologic indicators of dehydration than animals pro-       necrosis was a consistent pathological finding in WEEV-inoculated
vided a HFCS-sweetened gel product. In addition, the apparent         hamsters, with an absence of neuropathological changes prior to
preference for the unsweetened gel over the sweetened gel may         death at 96 hpi. A potential role for the inflammatory response in
have been due to decreased preference, or aversion, for HFCS.         the disease process was shown by increased serum concentrations
                                                                      of tumor necrosis factor-alpha in WEEV-inoculated animals, and
P124 Maximizing Per Diem Revenues through More Frequent               significantly improved survival when virus-inoculated animals
Cage Census                                                           received anti-inflammatory therapy. Evidence for the encephalitic
Y Sun*1, M Moore2, M Depathy1, J Joncas1                              capability of WEEV in hamsters was provided by the observation
                                                                                    Abstracts of scientific papers 2008 AALAS National Meeting

that a portion of those animals that survived beyond 96 hpi due      at 0 min to 162.1 ± 14.1 mg/dl at 48 h) and isoflurane (from 72.29
to anti-inflammatory therapy displayed overt signs of neurologic     ± 7.83 to 133.3 ± 17.5 mg/dl at 60 min). Both anesthetics increase
disease and histopathological evidence of CNS necrosis. Intro-       serum glucocorticoids and catecholamines and contribute to fat
duction of WEEV directly into the CNS resulted in rapid death        mobilization. However, decreases of serum cholesterol concentra-
and similar neuropathological lesions. Inoculation of WEEV into      tions in the group H (from 66.9 ± 5.78 to 40.4 ± 6.6 mg/dl at 24 h)
Syrian Golden hamsters provides a unique disease phenotype           may result from production of free radicals during biotransfor-
which mimics certain characteristics of related alphavirus infec-    mation of halothane. A decrease in serum albumine (from 3.88 ±
tions in humans.                                                     0.13 to 3.2 ± 0.15 g/dl at 72 h) and total protein levels (from 5.96
                                                                     ± 0.12 to 4.9 ± 0.2 g/dl at 72 h) occurred in the group H. It has
P126 Rabbit Model of Alzheimer’s Disease: Valid from Neu-            been shown that halothane appeared to exert inhibitory effects
ropathological to Cognitive Levels                                   on synthesis of albumine and total proteins. Serum potassium
                                                                     levels significantly decreased in both groups (4.2 ± 0.3 mmol/L
A Agelan*1, L Del Valle3, L Bright1, M April1, DS Woodruff-          at 72 h in group H and 3.9 ± 0.2 mmol/l at 60 min in group I).
Pak2                                                                 There was a significant decrease in serum calcium levels in the
          Laboratory Resources, 2Psychology, 3Neuroviology,
1University                                                          group I (12.3 ± 0.6 mg/dl at 72 h) and a significant increase in
Temple University, Philadelphia, PA                                  serum phosphorus (from 4.69 ± 0.23 to 7.6 ± 0.8 mg/dl at 120
                                                                     min) and chloride concentrations (from 105.32 ± 1.33 to 110.3 ±
Supplementing a rabbit’s diet with 2% cholesterol alone or with
                                                                     1 mmol/l at 1 min), probably due to a transient reduced renal
a trace amount of copper created neuropathological changes that
                                                                     function immediately after isoflurane. Caution is required in
resembled those seen in Alzheimer disease (AD). AD model rab-
                                                                     interpreting data on serum biochemical parameters from rabbits
bits were impaired in eyeblink classical conditioning, a form of
                                                                     anesthetized with halothane or isoflurane.
learning severely impaired in AD. Our aim was to replicate AD
rabbit model neuropathology, test eyeblink conditioning in this
model, and determine if galantamine would ameliorate impaired        P128 Heart and Respiratory Rate, Rectal Temperature, and
conditioning. Rabbit chow with 2% cholesterol and drinking           Glucocorticoid Concentrations after Ketamine-medetomidine
water with 0.12 mg/l copper sulfate were administered for 10         Administration in New Zealand White Rabbits (Oryctolagus
wk. Control rabbits received normal food and water. Rabbit           cuniculus)
brains were probed for neuropathology. AD model rabbits had          M Martínez-Mateos1, A Gonzalez Gil*1, P Lorenzo García1, C
significant neuronal loss in frontal cortex, hippocampus and cer-    Pérez García2, I Díez Prieto2, J Illera1
ebellum. Changes in neurons in the hippocampus were consistent       1Fisiología Animal, Universidad Complutense de Madrid, Facul-
with neurofibrillary degeneration and cytoplasmic immunore-
                                                                     tad de Veterinaria, Madrid, Spain; 2Patología Animal (Medicina
activity for beta-amyloid and tau. Next, AD model rabbits were
                                                                     Veterinaria), Universidad de León, Facultad de Veterinaria,
injected daily with vehicle or 3.0 mg/kg galantamine and tested
                                                                     León, Spain
on 750 ms trace and delay eyeblink conditioning. Galantamine
improved eyeblink conditioning significantly over vehicle. The       The objective of this investigation was to determine the effects of
AD rabbit model has validity from neuropathological to cognitive     the anesthetic combination ketamine-medetomidine on heart and
levels and offers a promising addition to the available animal       respiratory rate, rectal temperature, and serum glucocorticoid
models of AD. Galantamine ameliorated impaired eyeblink              concentrations in New Zealand White rabbits. All the rabbits
conditioning, extending the validity of the AD rabbit model to       were clinically healthy prior the study. Ten animals maintained
treatment modalities.                                                under conventional conditions (12:12 light:dark cycle; 20 to 22
                                                                     °C; 50% to 55% relative humidity; 10 to 15 air changes/h) were
P127 Clinical Biochemical Response after Inhalant Anesthetics        anesthetized with ketamine (15 mg/kg, IM) and medetomidine
in New Zealand White Rabbits (Oryctolagus cuniculus)                 (0.25 mg/kg, IM). Blood samples were obtained from the central
                                                                     ear artery using 23-gauge needles at 6 time points: before injec-
A Gonzalez Gil*1, G Silván1, C Pérez García2, I Díez Prieto2, A      tion (0 min) and 10, 30, 60, and 120 min and 24 h postinjection
Martínez3, J Illera1                                                 of the anesthetics. The Departmenth of anesthesia was moni-
1Fisiología Animal, Universidad Complutense de Madrid, Facul-        tored by using the pedal withdrawal, ear pinch, and righting
tad de Veterinaria, Madrid, Spain; 2Patología Animal (Medicina       reflexes. Serum cortisol and corticosterone concentrations were
Veterinaria), Universidad de León, Facultad de Veterinaria, León,    measured by competitive enzyme inmmunoassay. An adequate
Spain; 3 GlaxoSmithKline R&D Spain, Madrid, Spain                    level of surgical anesthesia was observed for about 1 h, with no
                                                                     reaction to pedal and ear pinching. Heart rate was significantly
The effects of anesthetics on biochemical parameters can confuse
                                                                     decreased at 10 to 120 min when compared to baseline (time 0)
the interpretation of laboratory results. In order to characterize
                                                                     rate (from 255 ± 9.14 at 0 min to 197 ± 5 beats/min at 120 min)
the variations of selected biochemical parameters after inhalant
                                                                     and respiratory rate decreased at 10 to 120 min (from 109.25 ±
anesthetics, 20 New Zealand White rabbits were assigned to 2
                                                                     8.73 at 0 min to 60 ± 5.88 breaths/min at 30 min), probably due to
groups (n = 10): group H (halothane) and group I (isoflurane). The
                                                                     the medetomidine component of the mixture. It has been shown
induction of the anesthesia was achieved by a face mask (3.5%
                                                                     that medetomidine is a respiratory and cardiovascular depres-
halothane and 4.5 % isoflurane in oxygen) and were intubated
                                                                     sant. Rectal temperature was significantly decreased at 120 min
for the maintenance of the anesthesia for 30 min (1.5% halothane
                                                                     when compared with the initial levels (from 38.95 ± 0.15 to 37.27
and 2.5% isoflurane in oxygen). Serum cholesterol, triglycerides,
                                                                     ± 0.24 °C). Serum cortisol concentrations increased at 10, 30 and
albumine, total bilirubin, sodium, potasium, chloride, calcium,
                                                                     120 min when compared to baseline concentrations (from 3.09 ±
and phosphorus concentrations were measured by the Hitachi
                                                                     0.73 to 16.91 ± 3.58 ng/ml at 10 min). It is probably that ketamine
747 autoanalyzer. Blood samples (2 ml) were obtained from the
                                                                     produced the inhibition of GABA receptors and consequently
central ear artery before induction, 1, 10, and 30 min and 1, 2,
                                                                     the release of ACTH and cortisol. Although a small increase was
24, 48, and 72 h after intubation. Significant increases in serum
                                                                     observed at 10 to 30 min, serum corticosterone concentrations
triglycerides levels occurred after halothane (from 88.94 ± 13.78
Abstracts of scientific papers 2008 AALAS National Meeting

did not significantly change after injection of anesthesia when             artery catheter for sampling. Following 2 recovery d, mice were
compared with the baseline levels. The ketamine-medetomidine                placed on an automated blood sampler with a unique movement-
anesthetic combination provided an adequate level of surgical               responsive caging system. Concurrently, the stylet was removed
anesthesia and increase cortisol but not corticosterone secretion.          from the implanted guide and replaced with a brain microdialysis
Respiratory and cardiac depression effects have to be considered            probe perfused with artificial CSF at a rate of 1.5 µl/min. Follow-
for any experimental procedure.                                             ing a 24-h acclimation period, 2 predose dialysate samples were
                                                                            collected. The mice were dosed with 1 mg/kg of carbamazepine
P129 Canine Model of Relatively Sustained Atrial Fibrillation               into the jugular vein catheter. Nine blood samples (5 µl) were
Induced by Rapid Atrial Pacing and Phenylephrine                            collected from each mouse over 4 h, as well 10 dialysate samples
                                                                            (25 min/sample). Carbamazepine was detected in both the blood
A Kijtawornrat*1,2, BM Roche2, RL Hamlin1,2                                 and dialysate samples. Mice remained healthy throughout the
1TheOhio State University, Columbus, OH;              2   QTest Labs, Co-   study, demonstrating the efficacy of the surgical method. This
lumbus, OH                                                                  combination of sampling techniques within a single mouse serves
                                                                            as a useful model for obtaining multiple streams of information
Atrial fibrillation is a common arrhythmia with important mor-
                                                                            while reducing interanimal variability.
bidity and mortality. Limitations in studying both mechanisms
and therapy arise for paucity of models that possess a high
yield, are not costly, and are sustained long enough to make the            P131 Effects of Doxorubicin on Renal Function in the Gerbil
necessary observations. This model is based upon the thought                (Meriones unguiculatus)
that atrial fibrillation requires heterogeneity of repolarization,          M Peña1, I Diez1, M García1, MJ Cano1, M Ríos1, AM Montes2, C
that distribution of vagal fibers is heterogeneous in the atria,            Gutiérrez2, CC Perez*1
and that when stimulated by elevation of systemic arterial                  1Departamento  de Medicina, Cirugía y Anatomía Veterinaria,
pressure, atrial fibrillation will persist. We postulated that rapid
atrial pacing together with phenylephrine infusion will induce              Universidad de León, León, Spain; 2Departamento de Medicina
relatively sustained atrial fibrillation for at least 40 min in dogs.       y Cirugía Animal, Universidad de Murcia, Murcia, Spain
Dogs were anesthetized with morphine-chloralose because this                Doxorubicin administration in rats is a recognized model of the
permits nearly intact autonomic control. Systemic arterial pres-            nephrotic syndrome observed in humans, while administration
sure was elevated approximately 75 mmHg and mean heart                      in the gerbil leads to a more moderate proteinuria in the 1st wk,
rate was decreased approximately 9 beats/min with a constant                which tends to settle down from d 21. The aim of this work was to
infusion of phenylephrine, 2 μg/kg/min. The right atrium was                establish the effect of doxorubicin administration on the different
paced for 20 min at 40 Hz. Atrial fibrillation was sustained in             renal function parameters in the gerbil (Meriones unguiculatus) in
dogs receiving phenylephrine but terminated within seconds in               a long-term study. Twenty conventionally housed gerbils, aged
normotensives. In dogs receiving phenylephrine infusion, 50%                4 mo (10 males and 10 females), were used. They were housed
of the dogs maintained atrial fibrillation for > 40 min, and 50%            in the conventional area of the Central Laboratory Animal Facili-
returned quickly (<19 s) to sinus rhythm. In conclusion, atrial             ties of the University of León. A control group (n = 10) and an
fibrillation may be maintained for at least 40 min following ces-           experimental group (n = 10) were organized, including 5 males
sation of rapid atrial pacing in dogs with hypertension produced            and 5 females in both groups. In the animals belonging to the
by phenylephrine infusion.                                                  experimental group, doxorubicin (a single dose of 4 mg/kg) was
                                                                            injected intravenously into the femoral vein. Blood and 24-h urine
P130 Combinatorial Pharmacology in Mice: Integration of                     (collected from a metabolic cage) were obtained. Plasma calcium,
Brain Microdialysis and Automated Blood Collection in a                     phosphorus, magnesium, urea, creatinine, and total proteins and
Single Subject                                                              urinary calcium, phosphorus, magnesium, creatinine, and protein
                                                                            excretion were measured. Animals were sampled at 0, 14, 28, 42
C Rohde*1, R Guinn1, R DeGraw1, SN Katner1, BP Solomon2                     and 56 d. Plasma creatinine remained stable, and plasma urea
1In Vivo Research, 2Bioanalytical Research, Bioanalytical Systems,          increased only in the final sampling. Plasma phosphorus increased
Inc, West Lafayette, IN                                                     from the second sampling, but its fractional excretion remained
                                                                            stable. While hypocalcemia was observed only in the final sam-
In recent years, mice have become an extremely attractive model
                                                                            pling, calcium fractional excretion was high since d 28. Magnesium
in drug research. Their small size makes them suitable candidates
                                                                            remained stable throughout the study, but its fractional excretion
for early discovery studies where very little drug is available, and
                                                                            increased from d 42. Normoproteinemia was observed, but 24-h
the ability to genetically modify mice adds to their appeal in the
                                                                            urine protein excretion increased in the final sampling. These
research community. An additional trend in drug discovery has
                                                                            data reveal that the Mongolian gerbil is less sensitive than the rat
been a major focus on central nervous system (CNS) active drugs.
                                                                            to the administration of doxorubicin and that the mechanisms of
For this drug type, it is useful to look at drug concentration and
                                                                            renal adaptation are able to maintain the renal function during a
effect in both the blood (systemic levels) and the brain (target
                                                                            reasonable period in this species.
tissue levels). The ability to obtain both types of data from a
single mouse will reduce the number of subjects needed for such
research and improve the quality of the data by reducing inter-             P132 NIH/rnu: Are They All Created Equal? A Comparison of
animal variability. This study sought to develop a surgical model           Taconic and Charles River Rat Strains
that allowed the simultaneous collection of blood and brain                 CJ Andresen*
dialysate samples in individual mice following an intravenous
                                                                            Comparative Medicine, Pfizer, Groton, CT
dose of carbamazepine. Eight male CD-1 mice were anesthetized
with an intraperitoneal dose of ketamine/medetomidine and                   Preclinical oncology research relies on nude rodent xenograft
implanted with a guide cannula in the caudate putamen. After                models, where human tumor cells are implanted in nude mice
4 d of recovery, mice were anesthetized using isoflurane, and               or rats. Most commonly used are nude mice; however, using
implanted with a jugular vein catheter for dosing and a carotid             rats presents some advantages. Historically, our lab has used the
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

NIH/rnu rat strain from 2 suppliers, Taconic (Tac) and Charles        for Helicobacter, when compared to fecal pellets stored at 4.0 °C.
River (CR), to create xenograft models. Some phenotypic and           A comparison of multiple processing techniques demonstrated
performance differences were observed between the 2 sources           that nucleic acid isolated from the supernatant of low-speed
which prompted us to look closer at the history and phenotype         centrifuged slurries is an effective method for detecting both
of the animals. The athymic hairless mutation in rats was first       Helicobacter and MNV from the same sample. This study supports
observed in 1953 at the Rowett Research Institute in England.         that feces should be shipped and stored at 4.0 °C if used for pre-
The National Institutes of Health (NIH) nude rat was developed        paring slurries for Helicobacter and MNV PCR testing (although
in 1979–1980. Taconic source rats were obtained from the NIH          ambient temperature is also acceptable for MNV). However, all
in 1981 and were derived by hysterectomy in 1987 and again in         evaluated storage temperatures up to 2 mo are adequate when
1998, designation NTac:NIH-Whn. The Charles River source rats         feces are placed directly into lysis buffer prior to extraction.
were obtained from the NIH in 2001 and cesarean rederived,
designation Crl:NIH-Foxn1rnu. The nude rat is athymic and             P134 Molecular Characterization of a Newly Indentified Rat
T-cell–deficient, with a normal complement of B and NK cells.         Parvovirus
To test our hypothesis, we performed flow cytometry analy-
sis of lymphocyte subsets in 3 strains of 6- to 8-wk-old male         C Wan*1, Y Liao1, Y Chang1, M Wang2
rats: Charles River CD-1, Charles River homozygote nude rat           1VeterinaryMedicine, National Taiwan University, Taipei, Tai-
Crl:NIH-Foxn1rnu, and Taconic homozygote nude rat NTac: NIH-          wan; 2Laboratory Animal Center, Tzu Chi University, Hualien,
Whn, and tumor take-rate/growth studies in the 2 nude strains.        Taiwan
Data demonstrated that normal CD rats have significantly higher
                                                                      One new isolate of rat parvovirus was identified and char-
values for NKT, T, CD8+, CD4+, CD25+ and RT1B+ than either
                                                                      acterized by our laboratory. The isolate was obtained from a
strain of homozygote nude rats, while both strains of homozy-
                                                                      laboratory rat colony (RPV-1c). In this study, the DNA sequence
gote nudes have higher B and NK cell values than normal CD
                                                                      and the predicted viral capsid protein sequence of this isolate
rats. CR source nudes have significantly higher NKT, T, CD8a+,
                                                                      were determined and compared to previously characterized
CD4+ and RT1B+ levels than Tac rnu, while Tac source nudes
                                                                      rodent parvoviruses. When the DNA sequence of this new RPV
have significantly higher NK levels than CR. We also found that
                                                                      isolate was compared to other rodent parvoviruses, RPV-1a was
take-rates and growth rates for tested tumor cell lines are higher
                                                                      the most similar to this new RPV with 98% identity. Nucleotide
in Taconic source rats. In conclusion, NIH/rnu source rats differ
                                                                      homologies of the new RPV isolate with Kilham rat virus (RV),
between vendors, and researchers should consider the applica-
                                                                      H-1 virus, and rat minute virus (RMV) were lower (71% to
tion or particular model before choosing a supplier.
                                                                      72%). The regions of the genome previously determined to be
                                                                      important in transcription, mRNA processing and translation of
P133 Evaluation of Feces Shipping Conditions and Sample               prototypic rodent parvoviruses were highly conserved between
Preprocessing for PCR Detection of Helicobacter and Murine            this new isolate and RPV-1a. In summary, the new RPV isolate
Norovirus                                                             has been identified which are closely related to RPV-1a, but
CL Perkins *1, KA Sellitto 1, JL Latchaw 1, AL Kelleher 1, ME         distinctly different from other previously characterized rodent
Vigliotti2, KS Henderson1                                             parvoviruses.
1Research Animal Diagnostic Services, 2Genetically Engineered

Models and Services, Charles River, Wilmington, MA                    P135 Alternate Rat Pharmacokinetics Methodology Valida-
                                                                      tion
The sensitivity of the polymerase chain reaction (PCR) is often
associated with the amplification and detection portion of the        C Nunes*1, X Shen1, B Wang1, S Xu2, S Zheng2, J Wang2, I Ca-
process; however, detection of an agent may be affected by            podanno1, W Ferguson1, BJ Doerning1, LH Cohen2, X Tong2, C
shipping conditions or sample processing prior to nucleic acid        Johnson1
isolation. Fecal pellets are common samples used for screening        1Lab   Animal Resources, 2DMPK, Merck & Co., Rahway, NM
rodents for enteric viruses and pathogens. The steps between
                                                                      The evaluation of novel pharmaceutical compounds using rodent
sample collection and nucleic acid are not standardized, nor are
                                                                      pharmacokinetics screens is common within pharmaceutical
the effects on agent detection clear. We investigated the impact
                                                                      drug discovery. Single- or double-line surgically vascular can-
of storage and shipping temperature (–20 °C, 4.0 °C, ambient,
                                                                      nulated rats are routinely used for rodent pharmacokinetics (PK)
and 37 °C) over time (2 h, 1 wk, 2 wk, 1 mo, 2 mo) on agent
                                                                      compound screens. Surgical preparation time, non-naïve state
detection for dry feces placed directly in lysis buffer. Because
                                                                      of animal, and cost of surgically cannulated animals led us to
fecal slurries are often used as a method for pooling samples,
                                                                      evaluate the feasibility of an alternate procedure for intravenous
subsequent investigations were used to evaluate the impact of
                                                                      (IV) dosing and blood sample collections from PK rats that
temperature and time on slurries and to compare different pre-
                                                                      would reduce our reliance upon surgically prepared animals.
processing methods (direct lysis, slurry, and slurry supernatants).
                                                                      This study was conducted to make 2 side-by-side comparisons.
Nucleic acid was evaluted by quantitative real-time fluorogenic
                                                                      First, we compared manual jugular vein dosing and tail blood
PCR for murine norovirus (MNV) or Helicobacter. When dry
                                                                      sample collection with the current method of IV dosing using
feces was added directly to lysis buffer, Helicobacter target copy
                                                                      a cannulated femoral vein and blood collection from a femoral
numbers decreased by approximately 80% after 1 wk, regardless
                                                                      artery catheter. Second, we compared PO dosing and tail blood
of storage temperature; however, the copy number remained
                                                                      collection with the current method of blood collection using a
fairly consistent for the remaining time points. Conversely, the
                                                                      cannulated femoral vein. Compounds were selected for side-
results for slurries indicated a 2 log10 decrease in Helicobacter
                                                                      by-side dosing comparisons based on known difference in PK
target copy number when stored at –20 °C as compared to 4.0 °C.
                                                                      and physical properties to investigate if the alternate dosing and
Results from a follow-up experiment demonstrated that target
                                                                      collection methods offered comparable PK profile data. The rats
copy numbers were substantially reduced for both agents when
                                                                      were manually restrained, dosed IV in the jugular vein and per os
feces was stored at –20 °C, –70 °C and at ambient temperature
                                                                      (PO) by oral gavage, and whole blood was collected by tail snip.
Abstracts of scientific papers 2008 AALAS National Meeting

Results of side-by-side studies showed that there was no signifi-       with heat-killed ETEC on days 6 and 12 of gestation. Postparturi-
cant difference (P > 0.01) in the PK profiles between alternate and     tion colostrum samples were collected from each dam’s mammary
conventional dosing and collection methods. The estimated cost          glands using a milking device especially design for this purpose.
savings in the elimination or reduction of cannulated rats in our       Blood samples were drawn from each dam’s orbital sinus. The rat
facility is significant, with potential reduction of up to 75% of       antibody responses to ETEC were analyzed with Western blot and
current expenses for vendor-supplied surgically cannulated rats.        enzyme immuno-assay (EIA) to determine the specificity and titers
In summary, the implementation of alternate PK methodology              of antibodies in the colostrum and sera. Whole cell extracts of swine
is a significant cost and time savings initiative, but feasibility at   ETEC isolates were prepared for electrophoresis in 12.5% SDS PAGE
this time is under consideration due to the skill set proficiencies     gels. Western blots were probed with sera and colostrum from each
required to reliably jugular dose the animals.                          dam as well as the sera from the offspring. The antibody response
                                                                        in rats challenged with ETEC compared with the antibody response
P136 Technique Refinement for Tail Tip Biopsy Procedures for            of rats challenged with saline showed an increased specificity to the
Genotyping Mice                                                         swine ETEC antigens. Antibody titers measured by EIA also indicate
                                                                        a significant increase in the antibody titers in rats challenged with
CC Sem*, S L’Italien, G Ma, H Daly, L Jackson                           ETEC antigens. The Sprague Dawley rat is an efficient laboratory
Biogen Idec, Inc., San Diego, CA                                        animal model for the study of the colostral antibody response to
                                                                        ETEC antigens of swine origin.
The most commonly used technique for genotyping genetically
engineered mice (GEM) is the tail tip biopsy. At many institutions,
this procedure is performed by using a sterile blade to amputate        P138 Effect of Selenium and Selenoprotein on Brain Function
the tip of the tail, and then using direct pressure and electro-        in Transgenic Rats
cautery or procoagulants to ensure hemostasis. In an attempt to         D Hwang*1, J Sin2, S Yim2, C Kim2, B Kim2, S Shim2, S Lee2, C
refine these procedures, we used a glass bead sterilizer (275 °F)       Bae2, J Woo2, J Cho3, K Chae2
to heat the end of a sterile, #21 scalpel blade for approximately       1College of Natural Resources & Life Science, Pusan National
30 s before amputating the tip of the tail. Our hypothesis was
                                                                        University, Miryang-si, Gyeongsangnam-do, South Korea; 2De-
that simultaneously cutting and cauterizing the tissues by this
                                                                        partment of Laboratory Animal Resources, National Institute
method would lead to rapid hemostasis, a reduction in tissue
                                                                        of Toxicological Research, Korea FDA, Seoul, South Korea;
damage and postprocedural inflammation at the amputation                3Multidisciplinary Technology Center, Hoseo University, Asan,
site, and a reduction in the technician time necessary to perform
                                                                        South Korea
the procedure. One hundred GEM, 4 to 6 wk of age, were used
for this study. All mice were anesthetized with isoflurane. Tail        Selenium (Sel) is widely distributed throughout the body, but is
tip biopsies were conducted on 50 mice using a sterile, room            particularly well maintained in the brain. It has been reported
temperature scalpel blade followed by direct pressure and elec-         that changes in selenium concentration in the brain are associated
trocautery to ensure hemostasis. Tail tip biopsies on the other 50      with abnormal brain functioning in various brain diseases such as
mice were conducted using a sterile, heated scalpel blade only.         Alzheimer disease and brain neoplasia. In this study, to investigate
All tail tip biopsy procedures were performed by the same tech-         whether selenium treatment would affect brain function, we ex-
nician. The duration of time to complete the procedures for each        amined the phosphorylation level of functional proteins including
group was recorded. Daily clinical observations were conducted          Tau, PKC, and MAPK in the Sel-treated SelM Tg rats. The CMV/
for 5 d following the procedure, to include scoring the degree of       GFP-hSelM Tg (SelM Tg) rat used in this study were developed
inflammation at the amputation sites. In the heated blade group,        by microinjection of CMV/GFP-hSelM fusion gene into fertilized
hemostasis was rapidly achieved in all mice, the degree of tissue       rat eggs and showed a high antioxidant status in various tissues.
damage at the time of amputation was noticeably reduced, and            SelM Tg rats were treated with sodium selenite for 3 wk and the
the incidence of postprocedural inflammation at the amputation          level of related proteins were detected with western blot analyses.
site was significantly reduced as compared to the group where           Based on the results of the present study, we concluded that the
electrocautery was used. Even though the heated blade method            phosphorylation of Tau protein was significantly changed in the
removed the step of using electrocautery, there was no significant      ser 404, ser 396, and ser 202 site of cortex and hippocampus region.
difference in the time it took to perform the two techniques. In        The analysis of PKC phosphrylation showed that PKC-alpha,
conclusion, we found that the heated blade technique described          delta, and zeta were key proteins to phosphorylate Tau protein.
here represents an animal welfare refinement.                           Sel treatment decreased the activity of alpha and gamma-secretase
                                                                        producing Abeta-42 amyloid peptides as well as increase alpha-
P137 Rat Animal Model for the Study of Colostral Antibodies             secretase degrading Abeta-42 amyloid peptides. The microglial cell
                                                                        harvested from SelM Tg rat showed the high level of cell viability
C Walden*, A Byers, S Hurley, J Allen                                   compare with that of Non-Tg rats. Therefore, our results suggested
Animal Science, North Carolina A&T State University, Greens-            that the SelM expression and Sel concentration play a important
boro, NC                                                                role the maintain of brain function and signal pathway, and the
                                                                        SelM Tg rat can serve as an animal model studying the biological
A laboratory animal model was developed to study the antibody
                                                                        function of selenoprotein and selenium on the brain disease.
response of Enterotoxigenic E. coli (ETEC) of swine origin in the
colostrum and sera of the Sprague Dawley rat. ETEC is an eco-
nomically important bacterial infection in neonatal swine. Neonatal     P139 In Vivo and In Vitro Validation of Stored Homologous
swine are protected at birth from ETEC infections by passive an-        Red Cell Transfusion in Pigs
tibodies transferred in the mother’s colostrum. The study of these      S Biagini, A Rosário, DS Camila*, C Tanikawa, L Azevedo, P
antibodies and their impact on infections in neonatal swine is made     Aguirre
difficult by the gestation period, logistics, and overall cost. The
                                                                        Institute of Learning and Research, Sirio Libanês Hospital, São
Sprague Dawley rat was developed as an animal model for these
studies. Specific pathogen-free rats were challenged intramuscularly    Paulo, Brazil
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting

This work aimed to evaluate the feasibility of using stored red        basement membrane. The cause of the decrease in blood glucose
blood cells in pigs in order to further validate a model of blood      is still unclear but may be related to the level of Plasmodium
cell transfusion. One pig (50 kg) was submitted to a controlled        membrane protein GPIs (glycosylphosphatidylinositols) and
hemorrhage of 1033 ml of blood. The blood collected produced           the balance between Th1/Th2 responses. These results also
2 unities of red blood cells that were preserved for 13 d under        corroborate the “hygiene hypothesis” which postulates that
controlled temperature. Validation of these unities in vitro           the increased incidence of autoimmune diseases (such as dia-
was carried out at the day of the collection and at the 13th d of      betes) in developed countries is result of decreased exposure
storage through the evaluation of volume, level of hemoglobin          to pathogens.
(Hb) and hematocrit (HT), hemoglobin concentration, hemolysis
determination, sodium and potassium concentrations, glycemia           P141 Evaluation of Two Pachymeters for Corneal Measurement
and pH. After 13 d, in vivo hemolysis was evaluated through            in Rabbits (Oryctolagus cuniculus) and Guinea Pigs (Cavia
chromate-marked red cells (0.7 ml of radioactive sodium chro-          porcellus)
mate [Na251CrO4] for 100 g of red cells). Five animals (38 to 58
kg), including the animal previously submitted to hemorrhage           DN Earnest*, G Hamilton, MA McLaughlin
were anesthetized and received 2.5 ml/kg of marked red cells           DIS-Pharmacology Screening, Alcon Research, Ltd, Fort Worth,
(0.412 MBq/kg). Blood was collected at 5 min, 10 min, 1, 3, 6, 9,      TX
12, and 24 h, and these samples were counted on an automatic
                                                                       The link between corneal thickness (CT) and intraocular pressure
gamma counter. After 24 h, the animals were sacrificed and the
                                                                       (IOP) has driven the need to measure CT in animal research. Pa-
spleens analyzed by scintillography. Transfusion units had low
                                                                       chymeters measure corneal thickness, and most of those available
volumes (187.1 ml and 144.7 ml at 13 d) and low concentrations of
                                                                       are designed for human clinical use. We evaluated 2 pachymeters
total hemoglobin (41.73 g/U and 31.85 g/U at 13 d) as compared
                                                                       for use in the research environment and also established in-house
to human standards. In vitro validation demonstrated indexes
                                                                       reference CT values for 2 strains of rabbits and a strain of guinea
of hemolysis below 0.1% and similar electrolyte concentrations
                                                                       pigs. Reliability and comparability were evaluated for the Accu-
when compared to human data. In vivo results demonstrated
                                                                       tome AccuPach V Pachymeter (measuring range 300 to 999 μm)
that the percentage of chrome remaining on red blood cells was
                                                                       and the Sonagage Corneo-Gage Plus Pachymeter (measuring
over 70% up to 24 h of observation. Spleen analysis did not show
                                                                       range 200 to 999 μm) in adult New Zealand albino (NZA) rabbits,
significant accumulation of the radioisotope. Our results show
                                                                       Dutch Belted (DB) rabbits, and Hartley outbred (HOB) guinea
that red blood cell storage in pigs does not induce significant
                                                                       pigs. Machines were evaluated for ease of use (based on portabil-
hemolysis up to 14 d. These data demonstrate the feasibility of
                                                                       ity, set-up time, and learning time), reliability and repeatability
studies that evaluate the effects of stored red cell transfusion
                                                                       (prolonged use for 2 h), and comparability (same eyes measured
in pigs.
                                                                       by both machines). Conscious animals were restrained manually;
                                                                       2 drops of topical anesthetic were instilled, and 3 measurements
P140 Plasmodium chabaudi chabaudi CR in Non-obese Diabetic             were taken on each eye and averaged. Both machines were easy
(NOD) Mice                                                             to use, being portable, quick to set up, and having a sharp learn-
DR Machado*1, AA Guaraldo1, BC Vidal2                                  ing curve. The reliability of both machines was excellent, with no
1Parasitology, 2Cell
                                                                       malfunctions for prolonged sessions of 2 h constant use. Corneal
                       Biology, Unicamp, Campinas, Brazil              thickness measurements were 388 μm ± 4.0 (AccuPach V) and
Malaria is a parasitic disease that kills over a million people each   386 μm ± 3.8 (Corneo-Gage) in the same 44 NZA eyes; 368 μm ±
year. In Brazil, 99% of malaria cases occur in the Amazon Basin.       3.4 (AccuPach V) and 362 μm ± 3.5 (Corneo-Gage) in the same
Plasmodium chabaudi is an important organism for studying sev-         26 DB eyes. The AccuPach V could not measure CT in guinea
eral aspects of malaria because of its usefulness as a substitute      pigs since average CT was found to be less than 300 μm. With
model for Plasmodium falciparum. Non-lethal P. c. chabaudi CR          the Corneo-Gage, CT in 78 guinea pigs was 217 μm ± 0.9. The
strain is the autocontrol in such studies. Hypoglycemia is a com-      Accupach V and Corneo-Gage pachymeters are easy, accurate,
mon symptom of murine malaria and of P. falciparum malaria in          and robust enough to use in the research environment. Normal
humans. Type 1 diabetes is an autoimmune disease that destroys         CT values have been established for NZA and DB rabbits and
the insulin-producing beta cells, and non-obese diabetic (NOD)         HOB guinea pigs.
mice provide a good model of human type 1 diabetes. In this
work, we examined the course of type 1 diabetes in NOD mice            P142 Effect of Sub-tenon Injections on Intraocular Pressure in
infected with P. c. chabaudi CR. SPF female NOD/Uni mice were          Dutch Belted Rabbits (Oryctolagus cuniculus)
inoculated intraperitoneally with red blood cells infected with
P. c. chabaudi CR. Blood glucose was measured by using Roche           DN Earnest*, S Yacoub, MA McLaughlin
Accu-Chek Active kits in samples of tail blood collected every         DIS-Pharmacology Screening, Alcon Research, Ltd, Fort Worth,
other day for 11 d postinfection. The level of parasitemia was         TX
expressed as the percent of infected erythrocytes based on cell
                                                                       Progressive therapy for glaucoma will include routes of drug
counts of tail blood smears stained with May-Grünwald-Giemsa.
                                                                       administration that allow sustained contact with the eye or
The mice were classified according to the severity of their dia-
                                                                       sustained delivery of active drug from a device (such as a
betes: moderate (300 to 500 mg/dl) or severe (>500 mg/dl). The
                                                                       suspension or tablet). No published accounts were found for
control group consisted of BALB/c/Uni mice. The mortality
                                                                       sub-tenon drug delivery for intraocular pressure lowering in
was recorded for healthy and infected NOD/Uni mice. Blood
                                                                       rabbits. Several parameters were investigated to define a model.
glucose decreased at the height of parasitemia, around the 5th
                                                                       Intraocular pressures (IOP) in female Dutch Belted rabbits (n =
d of infection. Infected diabetic mice survived longer after the
                                                                       5 to 7) were measured with an applanation pneumatonometer
onset of diabetes than the non-infected controls. Histologically,
                                                                       after light corneal anesthesia with topical 0.1% proparacaine.
the main renal lesions included glomerular edema and necrosis,
                                                                       Animals were then anesthetized with isoflurane to effect. Using a
cell infiltration, tubular atrophy and thickening of the glomerular
                                                                       30-gauge needle on a sterile 1 ml syringe, various volumes (25, 50,
Abstracts of scientific papers 2008 AALAS National Meeting

100, and 250 µl) and placements (2 mm from and perpendicular         rabbit diets have consisted primarily of pellets with an alfalfa
to the limbus, 2 mm from and parallel to the limbus, and 4 mm        base. Exotic animal veterinarians now recommend low-protein,
from and parallel to the limbus) of 3 vehicle depots (mannitol,      high-fiber diets consisting primarily of timothy hay and timothy-
saline, and BSS Balanced Salt Solution) under the tenon capsule      based pellets, rather than alfalfa. An evaluation of the available
were investigated. Following recovery from anesthesia, IOP           commercial diets for laboratory rabbits found that all pellets,
measurements were taken at 2, 4, 6, 8, and 24 h postinjection.       even the high-fiber varieties, are alfalfa-based. Because alfalfa
There was an unexpected pronounced volume effect on IOP;             is higher in calcium than timothy, alfalfa-based pellets contain
the greater the volume, the greater the IOP reduction (at 4 h        1.2% calcium, which is higher than even the published lactational
postdose): 250 µl = –13.0%; 100 µl = –9.9%; 50 µl = –5.7%, 25        requirement of rabbits (0.75%). Timothy pellets, on the other
µl = –5.9%. Contralateral, untreated eyes also responded with        hand, contain about half the calcium of alfalfa-based diets. Our
reduction in IOP with the maximum effect 2 h earlier than in         hypothesis was that rabbits fed an alfalfa-based pelleted diet
treated eyes: 250 µl = –11.8%; 100 µl = –3.0%; 50 µl = –11.1%; 25    would have increased serum calcium resulting in increased
µl = –6.0%. IOP in treated eyes returned to baseline within 24 h;    renal pathology compared to those rabbits fed a timothy-based
in contralateral eyes, it returned by 4 h. The type of vehicle did   pelleted diet. One group of 6-mo-old rabbits was fed a high-
not influence outcome. Rabbits, whether through pseudofacility,      fiber, alfalfa-based diet (control), while another group was fed,
consensual neurological or barometric response, or some other        upon arriving at our research facility, a commercially available
cause, respond to sub-tenon injection with lowering of IOP both      timothy-based diet that is not currently used for laboratory
ipsilateral and contralateral in the absence of any pharmaco-        species. Urine and blood was subsequently collected monthly
logical agent. Care must be taken when using this model that         for calcium evaluation. We found that both urine calcium and
parameters are standardized. A separate control (vehicle-treated)    serum calcium levels remained within the normal published
group of animals is essential.                                       ranges for both timothy and alfalfa groups. Comparisons of
                                                                     average serum calcium levels over time revealed no significant
P143 Endpoints for Mouse Subcutaneous Tumor Models: Re-              difference between the control and experimental groups. There
finement of Current Criteria                                         was also no significant difference in the overall urine calcium
                                                                     levels between the timothy and alfalfa groups. However, when
E Paster1, K Villines2, DL Hickman*1                                 serum calcium levels were initially examined in the experimental
1Veterinary Medical Unit, VA Medical Ctr, Portland, OR; 2College     group after switching from a alfalfa-based to a timothy-based
of Veterinary Medicine, Oregon State University, Corvallis, OR       diet, a difference was found that later normalized as time pro-
                                                                     gressed. Our data suggests that a timothy-based diet does not
Currently accepted methods of humane endpoint determination
                                                                     lower serum or urine calcium over time; however, when a new,
for subcutaneous tumor growth models in mice include tumor
                                                                     lower calcium diet is fed, there can be a change in the initial
size (generally maximum size of 1 cm), presence or absence of
                                                                     serum calcium level.
ulceration, and changes in body weight. Because tumor mass
may mask pathologic weight loss, this study explored the use
of body condition scoring (BCS) as an additional criteria to bet-    P145 Infectivity of Giardia spp. in BALB/c Nude Mice In-
ter assess morbidity in these animals. To complete this study, 2     oculated with Cysts of Effluent Wastewater after UV Light
different tumor models were assessed: a skin papilloma model         Disinfection
with multiple tumors produced after exposure to an appropriate       L Santos1, R Franco2, A Guaraldo2,3, R Cantusio Neto4, D Al-
carcinogen, and a subcutaneous melanoma study where a single,        ves*3
bulk-producing tumor was produced after subcutaneous injec-          1Laboratory of Advanced Oxidation Processes—Faculty of Civil
tion of tumor cells. Animals were evaluated 3 times per wk for
                                                                     Engineering, 2Department Of Parasitology—Institute Of Biology,
body weight, BCS, tumor number, size of the largest tumor (if        3CEMIB—Multidisciplinary Center for Biological Investiga-
multiple), and general clinical presentation (such as grooming,
posture, and responsiveness to stimuli). Data analysis revealed      tion on Laboratory Animal Science, UNICAMP—University
                                                                     of Campinas, Campinas, Brazil; 4Laboratory of Microbiology,
that BCS was significantly more reliable than body weight as an
                                                                     SANASA—Society for Water Supply and Sanitation, Campinas,
indicator of anorexia when correlated with a moribund clinical
                                                                     Brazil
condition. However, tumor growth in these models was found
to be well tolerated. Tumors over 1 cm in diameter did not result    Giardia spp. is a protozoa of public health interest that causes
in a decrease in BCS. Instead, we found that BCS consistently        gastroenteritis in humans and other animals, and is very common
did not decline until the tumor became ulcerated or interfered       in developing countries. In the last decades, outbreaks have been
with mobility because of location on or near a limb. Based on        recorded around the world, and the domestic sewer is suspected
the conclusions of this study, our IACUC has updated its tumor       to be the source of river contamination. In the city of Campinas in
evaluation guidelines to allow tumors of up to 2 cm in diameter,     southeast Brazil, giardiosis is endemic; the pathogen is detected
as long as mobility is not decreased, there is no ulceration, and    at high concentrations in effluent wastewater, which is a poten-
the mouse maintains a BCS of 3. The results of this study will be    tial reservoir for transmission. The Wastewater Treatment Plant
of interest to investigators and animal care staff working with      (WWTP) Samambaia in Campinas used a system of activated
similar studies.                                                     sludge for sewage treatment and UV light for disinfection of ef-
                                                                     fluent wastewater. In order to evaluate this disinfection process
P144 Does Feeding an Alfalfa-based Diet Predispose Labora-           in inactivating cysts of Giardia spp., 2 samples types were selected
tory Rabbits to Hypercalcemia and Renal Disease?                     for study: effluent wastewater without UV disinfection (EFL) and
                                                                     effluent wastewater with UV disinfection (EFL+UV). BALB/c
E Paster, DL Hickman*                                                nude immunodeficient mice were intragastrically inoculated
Veterinary Medical Unit, VA Medical Center, Portland, OR             with a mean dosage of 14 cysts of Giardia spp. collected from
                                                                     either EFL or EFL+UV. All animals inoculated with Giardia spp.
Available literature on rabbit nutrition offers conflicting advice
                                                                     cysts developed the infection, but the animals inoculated with
on feeding our laboratory lagomorphs. Historically, acceptable
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

UV-exposed cysts released cysts in the feces with a lower average     the animals were tested in the system under the same conditions.
concentration than that animals inoculated with cysts without         We were able to show significant differences (P < 0.05) between
UV disinfection. Trophozoites were also observed in the animals       the baseline and the 4-h samples post-carrageenan injection
of both groups. These findings suggest that cysts exposed to UV       in the following parameters: swing time, stance/swing ratio,
were damaged, but were able to cause infection.                       stance time, % stance/stride, stride length, paw area, stride
                                                                      time, % swing/stride, % propulsion/stride, % braking/stride,
P146 SCID Mice as Adjuncts to Rodent Sentinel Health Moni-            propulsion time, % braking/stance and % propulsion/stance.
toring Programs                                                       We validated that the system is a reliable, accurate and repeat-
                                                                      able system for detecting changes in gait related to acute pain.
D Gettys*, CL Franklin, LK Riley, R Livingston                        We have also demonstrated the repeatability in both interday
University of Missouri, Columbia, MO                                  and intraday testing, as well as the lack of effect of anesthesia
                                                                      on gait dynamics.
Pathogenic microorganisms can have many adverse affects on
rodents used in biomedical research. As a result, the health
status of contemporary rodent colonies is monitored regularly.        P148 Use of m-CT to Assess Anatomical and Structural Features
Researchers rely on sentinel programs as a measure of colony          of Proximal Femurs in Commonly Used Laboratory Animals
health, but there are published reports and abundant anecdotes        ER Berryman*, C Bagi, MR Moalli
of sentinel program failure. Our hypotheses were that certain
                                                                      World Wide Comparative Medicine, Pfizer Global Research and
agents do not readily transmit to sentinel animals and that
                                                                      Development, Groton, CT
incorporation of immmunodeficient (SCID) mice into sentinel
programs would improve detection of these agents. For this            The physical characteristics of bone include the morphology, di-
study, soiled bedding from mice infected with a number of             mensions, and distribution of apatite crystals that forms cortical
pathogens was transferred to cages containing 3 sentinels (n =        and cancellous bone structures. The skeleton is a mechanically
5 cages) or 2 sentinels and 1 SCID mouse (n = 5 cages). Every 2       optimized biological system whose composition and organization
wk, sentinel mice were evaluated for exposure to multiple patho-      reflects the functional demands made upon it differing between
gens by serology, fecal PCR, oral bacterial culture, and perianal     various species. Hydroxyl apatite accounts for approximately
tape tests. The infectious pathogens tested for included: murine      65% of bone weight, provides most of the strength and stiffness
norovirus, mouse parvovirus, minute virus of mice, Helicobacter       of bone, and allows for use of radiological technologies to assess
spp., Mycoplasma pulmonis, Pasteurella pneumotropica, and Theiler     bone mass and structure. Development of imaging technologies
murine encephalomyelitis virus. All viral pathogens readily           such as microcomputed tomography (m-CT) has markedly im-
transferred to most sentinel mouse cages. However, bacterial          proved our ability to assess structural parameters of cortical and
agents, including Helicobacter spp. and Pasteurella pneumotropica,    cancellous bone. In the current study, we sought to determine
were rarely transmitted to sentinels, and Mycoplasma pulmonis         and describe differences in cortical and cancellous bone structure,
did not transfer to any sentinel cages. The addition of SCID          bone mineral density and morphology (geometry) at proximal
mice did not result in increased detection of the latter patho-       femur (hip) between most commonly used laboratory animals
gens. This study supports published and anecdotal reports that        including mice, rats, rabbits, dogs, and nonhuman primates. This
Helicobacter spp. and Pasteurella pneumotropica are not reliably      work enhances our understanding on bone physiology across lab
detected by sentinel programs and demonstrates that inclusion         animal species and should enable scientists to select appropriate
of SCID mice does not improve their detection. Because of this,       species for research involving skeleton including metabolic bone
alternative methods of monitoring for these agents, such as test-     diseases, bone fragility or skeletal toxicology. Preliminary data
ing of colony animals, should be considered in design of health       demonstrated cross-species differences in cancellous bone struc-
monitoring programs.                                                  ture, including trabecular number, thickness, plate separation,
                                                                      and connectivity. Bone mineral density was greatest in rodents,
P147 Digital Gait Analysis Validation in CD Rats Using the            followed by dog, nonhuman primate, and rabbit. Differences in
DigiGait System                                                       geometry and structure of the femoral head, neck, and midshaft
                                                                      were demonstrated in 2- and 3-dimensional analysis that may
ER Berryman*, C Bagi, R Harris, C Andresen                            indicate adaptation to the mechanical loads that each species
World Wide Comparative Medicine, Pfizer Global Research and           experiences during normal cage activity.
Development, Groton, CT
The gait analysis system we tested combines a motorized tread-        P149 Development of an Ambulatory Continuous Intravenous
mill, high-speed digital imaging, and broadband video storage,        Infusion Method in the Nonhuman Primate
and provides visual assessment of gait dynamics in rodents and        EA Lambert*, JR Hynick, JJ Setser, TD Morris, WR Miller, CP
quantitative analyses of gait that may be related to alterations      Chengelis
in genes, disease, and drugs that affect coordinated motion. The
                                                                      Toxicology, WIL Research Laboratories, LLC, Ashland, OH
advantage of the system is that we may conduct noninvasive,
longitudinal studies and obtain quantitative assessments of           As recent industry demands have broadened, a need for the de-
response to therapy and correlate the data with traditional post-     velopment of an effective ambulatory continuous infusion model
mortem endpoints. The main goal of this study was to validate         in nonhuman primates has increased. Most current continuous
the system for use in the evaluation of movement/gait dynamics        infusion models involve jacket and tether systems connected
in the published carrageenan rat model of pain and inflamma-          to syringe pumps that are externally attached to the cage. Our
tion. Following IACUC review and approval, test subjects were         objective was to develop a viable ambulatory infusion model in
run on the treadmill system for 5 to 10 s to obtain at least 6 step   nonhuman primates that resulted in an accurate administration
cycles, and video images were obtained for analysis. The animals      of a test article continuously while allowing freedom of move-
were then anesthetized, and the right stifle was injected with        ment and socialization within the cage, as well as manipulation
either saline or carrageenan. Two hours and 4 h after injection,      outside of the cage. A study was conducted in 4 phases using a
Abstracts of scientific papers 2008 AALAS National Meeting

total of 24 cynomolgus macaques, 2 animals of each sex during         in contrast to conditioned cats, the Class B cats exhibited wide
the first 3 phases and 6 animals of each sex during the last phase.   variations in their coagulation parameters and indigenous inhi-
All animals were catheterized in a femoral vein with a 3 to 6 Fr      bitions of coagulations. Furthermore, Class B vendor cats may
Cath-in-Cath system and fitted with infusion jackets. Infusion        not be optimal for vascular or cardiovascular research due to the
pumps and syringes were used to continually infuse saline over        variations in the haemostatic parameters.
a 28-d period. The syringes were weighed and changed 3 times
weekly during the first 2 phases, 5 d a wk during the third phase     P151 Helicobacter Detection in Murine Fecal Samples in Nine
and 7 d a wk during the fourth phase. Specific endpoints, such        Diagnostic Laboratories Using PCR
as clinical pathology parameters, equipment-related issues, dose
accuracy, and macroscopic examination of the infusion site, were      E Mahabir*1, C Mörth1, M Brielmeier1, K Markoullis1, W Beisker2,
evaluated to determine the effectiveness of this model. Twenty-       J Needham3, H Balzer4, LE Bater5, A Bleich6, A Deeny7, J Dix5,
two of the 24 animals placed on study successfully completed the      K Jacobsen1, A Lorenz8, M Mähler6, W Nicklas9, JD Phipps5, KE
28-d infusion period. There were 2 incidences of localized inflam-    Seidel10, M Seidelin11, MF Toft11, A Tomlinson7, P Wilhelm1, J
matory reactions resulting in removal of these 2 animals from         Schmidt1
study, but no significant alterations in clinical pathology. Dose     1Department of Comparative Medicine, 2Institute of Toxicology,

volume accuracy for phases 1, 2, 3 and 4 were 93.5%, 96.6%, 96.0%     Helmholtz Center Munich—German Research Center for Envi-
and 97.8%, respectively, with an overall accuracy of 96.5%. Over      ronmental Health, Neuherberg, Germany; 3 The Microbiology
the course of the study, the number of incidences of equipment-       Laboratories, North Harrow, United Kingdom; 4Department of
related issues decreased, from 9 events for 4 animals in the first    Molecular Biology, Vet Med Labor GmbH, Division of IDEXX
phase, to 8 events for 12 animals in the fourth phase. All of these   Laboratories, Ludwigsburg, Germany; 5LAS Diagnostics, Glaxo-
issues were correctable, allowing animals to be maintained on         SmithKline, Herts, United Kingdom; 6 Institute for Laboratory
study without being compromised. As a result, a refined con-          Animal Science and Central Animal Facility, Hanover, Germany;
tinuous infusion model in nonhuman primates was successfully          7Molecular Biology Laboratory, Harlan UK, Hillcrest, United

developed, in which saline was accurately administered up to 28       Kingdom; 8 GIMmbH, Wildenbruch, Germany; 9Microbiologi-
d with a low rate of equipment issues, but still allowing freedom     cal Diagnostics, German Cancer Research Center, Heidelberg,
of movement and socialization among animals.                          Germany; 10Institute of Microbiology and Hygiene, Technical
                                                                      University Munich, Munich, Germany; 11Taconic Europe, Silke-
P150 Coagulation Profiling of Class B Vendor Cats with Refer-         borg, Denmark
ences to Serine Protease Inhibitors                                   Since Helicobacter infections in mice may influence the results of in
E McGeehan*1, K Wielgus1, F Azarafrooz1, LM Cera1, R Bues-            vivo experiments the Federation of European Laboratory Animal
ing1, J Fareed2                                                       Science Associations (FELASA) recommended investigations
1Comparative Medicine Department, 2Department of Pathology,
                                                                      for Helicobacter spp. during regular annual health monitoring.
                                                                      Helicobacter diagnostic is highly dependent on methods that are
Loyola University Medical Ctr, Maywood, IL                            specific, sensitive and rapid. Currently, the polymerase chain
Class B Vendor cats (Felis catus) are commonly used in biomedi-       reaction (PCR) is the preferred diagnostic tool, but results ob-
cal research; however, the laboratory parameters, including the       tained in different laboratories from aliquots of the same fecal
hemacologic and clinical chemistry, widely vary between each          samples may vary considerably. To determine the PCR methods
cat due to initial health status. A systemic study on the plasmatic   used for detection of murine Helicobacter spp. in fecal pellets and
coagulation profile and serine protease inhibition is not reported.   to compare their sensitivity, their specificity, and their repro-
It was hypothesized that due to the health status of these cats       ducibility, a ring study involving 9 commercial and academic
the hemostatic system is also compromised. To address this            European laboratories was designed. The study was conducted
hypothesis, 4 groups of Class B vendor cats (n = 42) were ini-        over a 3-y period and consisted of 4 rounds during which fecal
tially profiled for prothrombin time (PT), partial thromboplastin     samples were analyzed. The initial round revealed up to 3 log
time (aPTT), Heptest, and thrombin time (TT). The antiprotease        differences in detection of Helicobacter. The sensitivity, specific-
activities were also measured in terms of the anti-Xa, anti-IIa,      ity and reproducibility of testing improved considerably during
heparin cofactor II (HCII), and antithrombin III (ATIII) using        the 4 rounds of analyses. After completion of the 4 rounds of
specific chromagenic substrate methods. Along with the Class          analysis, determination of the number of Helicobacter genomes
B vendor groups, 1 additional group, comprising 10 clinically         in fecal pellets from naturally infected mice by the organizing
healthy cats which were habituated at a veterinary research           laboratory revealed that each method used by the participating
facility for 6 mo, were used as a control group. Citrated blood       laboratories was sufficiently sensitive to reliably detect Helico-
samples were drawn from all groups and centrifuged to obtain          bacter in naturally infected mice.
plasma samples. The samples were analyzed in groups using
standard clotting assays (PT, aPTT, Heptest, TT and antiprotease      P152 Increased Hemorrhagic and Antithrombotic Activities of
activity). The ATIII and HCII activities were measured using          Contaminated Heparin as Studied in an Animal Model
commercially available chromagenic substrate assays. All results
were compiled as groups, with applicable statistical methods          E Litinas*1, E McGeehan2, W Jeske1, D Hoppensteadt1, J Fa-
used to analyze the data. The Class B vendor cats did not show        reed1
any variation in the PT and TT assays, and only weak anti-Xa          1Department of Pathology, 2Comparative Medicine Department,

activity (>10%); however, there was a wide variation in the aPTT      Loyola University Medical Center, Maywood, IL
(12.2 to 49.9 s) and Heptest (20.8 to 51.7 s) assays between the
                                                                      The USFDA recently recalled heparin preparations contaminated
2 groups. The HCII activities measured by Dermatin Cofactor
                                                                      with hypersulfated chondroitin sulfate (HCS) due to reported
also showed wide variations which were largely dependent on
                                                                      adverse events and deaths. Although the adverse events were
the individual groups (range = 50% to 150%). In the ATIII as-
                                                                      reported to be associated with the activation of the contact sys-
say, similar trends were also noted. These studies suggest that
                                                                      tem by HCS, no data on the anticoagulant and antithrombotic
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

bleeding effects of these contaminated heparins is available.          P154 Antitumor Efficacy and Safety of Pteris semipinnata Linn
To address the in vitro anticoagulant profile and the bleeding         Extracts in Chemical-induced Liver Cancer in Mice
and antithrombotic actions of heparin, a contaminated heparin          GG Chen*, J Leung, JH Yip, NM Choi, PS Lai
(CH) and a noncontaminated heparin (CFH) were compared
in standard laboratory methods. Both heparin preparations              The Chinese University of Hong Kong, Hong Kong, China
were compared in equigravimetric concentration and dosages;            Pteris semipinnata Linn (PsL), is a traditional Chinese herbal
coagulation profiles of both agents were carried out in normal         medicine which has been used to subside lumps and to improve
pooled rat plasma. The hemorrhagic effects of both groups were         a variety of inflammatory conditions. We have shown that the
tested in a rat tail transaction model in individual groups (n = 6).   extracts of PsL, such as 5F and 6F, are able to inhibit the growth
In the same groups, the antithrombotic effects were measured           of a number of tumors in cell culture experiment. However, its
using a jugular vein clamping model. Ex vivo analysis of blood         efficacy and safety in in vivo tumor models are unknown. In
samples were carried out to determine the pharmacodynamics             this study, we have established a mouse model of liver cancer
differences between the 2 agents. In the in vitro profiles using the   induced by diethylnitrosamine (DEN), tested the efficacy of 5F
partial thromboplastin time and antiprotease assays at potency         on the growth of tumor and monitored the side effects. DEN was
adjusted levels, the 2 preparations were comparable; however,          administered by intraperitoneal injection to 12-d-old juvenile
the contaminated product showed slightly stronger activity. In         C3H/HeJ mice to induce HCC. At wk 17, 5F was given after
the hemorrhagic model, both agents produced a dose-dependant           DEN administration; mice were sacrificed 12 wk later. We found
(2 to 5 mg/kg) prolongation of bleeding time compared to sa-           that 5F (12 to 48 mg/kg) significantly reduced the number of
line. At the higher dose (5 mg/kg) the contaminated heparin            the tumors in the liver in a dose-dependent manner. It also ap-
produced much stronger bleeding times (67.8 ± 1.8 min CH               peared to markedly decrease the total volume of liver tumors.
compared with 57.1 ± 1.9 min CFH). Similar trends were noted           These findings are in agreement with results obtained in the cell
in the antithrombotic model where a higher number of jugular           culture study of 5F. Thus, the data have indicated that 5F is able
clampings were needed in rats treated with CH (7.6 ± 0.8 clamps        to kill liver tumor cells in vitro and in vivo. We did not observe
CH compared with 6.0 ± 0.2 clamps CFH). Ex vivo analysis of            convulsion, sedation, lack of grooming, loss of righting reflex,
plasma also revealed a stronger anticoagulant/antiprotease             increased or decreased respiration, gross ascites, signs of dehy-
affect of the contaminated product. This data suggests that the        dration, lethargy, emaciation, or weight loss during the period
presence of HCS as a contaminant enhances the in vivo bleeding         of treatments. Serum markers also showed no significant change
and antithrombotic effects of the recalled product. Therefore,         in the markers related to toxicity in liver and kidney. However,
besides the nonanticoagulant effects, the CH may also modulate         hyperactivity occurred in some treated mice. In conclusion, 5F
the bleeding and antithrombotic efficacy of heparin, thus having       can effectively inhibit the development or growth of liver cancer
significant drug/drug interactions.                                    in mice and does not generate significant side effects. Therefore,
                                                                       5F is a promising anti-liver cancer agent.
P153 Further Analysis of LCMV M1 Isolate
F Ike*1, K Sakai3, T Mizutani3, K Takimoto4, F Bourgade2, J            P 1 5 5 Ta c o n i c C 5 7 B L / 6 J B o m Ta c , C 5 7 B L / 6 N Ta c , a n d
Jaubert2, X Montagutelli2, A Kajita1, H Nakata1, A Yoshiki1, S         C57BL/10SgSnAiTac Mice Do Not Carry the Nnt Mutation
Morikawa3                                                              JL Seymour1, MA Young1, GW Bothe*1, AV Perez2
1BioResource Center Experimental Animal Division, RIKEN,               1R&D, 2Veterinary      Services, Taconic, Hudson, NY
Tsukuba, Japan; 2The Central Animal Facilities, The Institut
Pasteur, Paris, France; 3Department of Virology 1, 4Division of        Nicotinamide nucleotide transhydrogenase (NNT) is a trans-
Experimental Animal Research, National Institute of Infectious         membrane protein that catalyzes the interconversion of NADH
Diseases, Tokyo, Japan                                                 and NADPH in the mitochondria. Under conditions of oxidative
                                                                       stress NADH will be converted to NADPH, which in turn re-
We isolated lymphocytic choriomeningitis virus (LCMV M1                generates reduced glutathione. Studies suggest that the presence
isolate) from wild-derived mice originally caught in Austria.          of NNT is essential for mitochondrial defense against oxida-
M1-infected cells were recognized by mouse anti-LCMV Arm-              tive stress and for normal cellular metabolism. Furthermore,
strong serum. But homology analyses on nucleoprotein (NP)              mitochondrial dysfunction has been implicated in a number of
nucleotide and amino acid sequences indicated that there was a         degenerative diseases, including Parkinson disease, Alzheimer
genetic distance between M1 and already-known LCMV strains,            disease, and diabetes. Recently, several reports have documented
which makes it difficult to create PCR primers for these types         that C57BL/6J (B6J) mice carry a 17.8 kbp deletion in the Nnt
of viruses. On the contrary, the recently identified LCMV-like         gene including exons 7 to 11. We decided to screen the “Black”
Dandenong virus showed a high homology with M1 on NP                   substrains, C57BL/6JBom (B6JBom), C57BL/6NTac (B6NTac),
amino acid sequence (97%). To know the genetic location of M1          and C57BL/10SgSnAiTac (B10Tac) for this mutation. Two sets
among LCMV, we determined full nucleotide sequences of S-              of primers were used. The first set amplifies 312 bp in intron
segment and L-segment of M1 and compared these with those              6, a region that is deleted in mutant mice. B6JBom, B6/NTac,
of already-reported LCMV. RNA of M1-infected Vero E6 cells             B10Tac, DBA/2NTac (DBA/2), and B6D2F1/Tac (B6D2F1) show
was extracted and transcribed to cDNA using random hexamer             the presence of the 312 bp band. The control C57BL/6J strain
primer. Nucleotide sequences were determined and homology              received from The Jackson Laboratory did not amplify the 312
was analyzed by Mega4 program. Amino acid homology between             bp band, indicating the absence of the intron 6 DNA sequence.
M1 and Dandenong virus was very high on NP, GPC protein,               A second set of primers was used to amplify across the dele-
Z protein, and L protein. LCMV M1 isolate seemes to belong to          tion breakpoint, giving a 547 bp product if the DNA carried the
the new group of LCMV as indicated on LCMV-like Dandenong              deletion. The only amplified band is the one found in C57BL/6J
virus. Dandenong virus was isolated from fatal transplant re-          mice. Interestingly enough, B6JBom does not present an ampli-
cipients. Adequate, thorough LCMV surveillance is necessary            fied band. The amplification of intron 6 and the absence of the
for mice and other animals, including hamsters.                        breakpoint amplified band indicate that B6JBom does not present
Abstracts of scientific papers 2008 AALAS National Meeting

the deletion of exons 7 to 11 of the Nnt gene. The B6JBom strain        and intestines from mice contaminated with S. muris were PCR
was obtained through Taconic Europe in 1988. The original stock         positive, whereas feces and intestines from mice contaminated
was received from The Jackson Laboratory in 1971. Therefore, we         with other common protozoa but not S. muris were PCR negative.
can assume that the Nnt mutation arose at The Jackson Labora-           The assay was also found to have high sensitivity, detecting as
tory after 1971. We can confidently say that C57BL/6JBomTac,            few as 6 template copies per microliter of diluent. Importantly,
C57BL/6NTac and C57BL/10SgSnAiTac do not carry the 17.8                 all mice positive for S. muris on postmortem exams were also
Kbp deletion in Nnt.                                                    positive by fecal PCR. Moreover, S. muris was detected by PCR
                                                                        in mice negative by postmortem examination but from colonies
P156 Injection Variability Observed with Syringe Pumps                  known to be infected. Because S. muris loads have been reported
                                                                        to decrease with age, we adapted the PCR to a quantitative format
GM Herrera*1, BD Hare1, SH Kojtari1, KR Zurn1, SI Dworkin2              and assessed protozoal loads in feces of naturally infected mice of
1Med Associates, Inc., Saint Albans, VT; 2Psychology, Jacksonville      several ages. Fecal loads of S. muris ranged from 1.4×105 template
State University, Jacksonville, AL                                      copies per fecal pellet in a 4-wk-old mouse to 1.4×102 template cop-
                                                                        ies per fecal pellet in an 18-wk-old mouse, confirming that fecal
The use of motor-driven syringe pumps in biomedical research is
                                                                        loads decreased with age. In conclusion, the PCR assay developed
wide ranging, from studies of the direct intracranial infusion of
                                                                        promises to be a highly specific antemortem diagnostic assay with
drugs to rapid fixation of tissues for neurobiological assessments.
                                                                        higher sensitivity than existing postmortem tests, especially in
One of the most common uses of motor-driven syringe pumps
                                                                        animals harboring low levels of S. muris.
is for evaluating the effects of intravenous drug administration.
Specifically, drug self-administration studies allow nonhuman
subjects to control the intravenous delivery of drugs typically         P158 Footpad Sweating Is Related to Cardiovascular Auto-
abused by humans. The results of these studies are used to              nomic Responses in the Dog
schedule drugs and evaluate behavioral and neurobiological              H Yamamoto*1, Y Arasaki1, M Can2, E Hamade3, H Yokoi3, C Tsut-
effects of these drugs as well as assess potential treatments for       sumi1, S Ohtsuka1, E Hori2, K Umeno2, A Fujiki2, H Nishijo1,2
drug abuse. These studies involve training a subject to engage in       1Life
                                                                            Science Research Center, Toyama, Japan; 2Graduate School
a lever press or other response that results in the activation of a
                                                                        of Medicine and Pharmaceutical Science for Research, The
motor-driven syringe pump. The dosage of the drug infused is
                                                                        University of Toyama, Toyama, Japan; 3 Life Care Giken Inc,
a function of the concentration of the drug placed in the syringe
(mg/ml) and the time the pump is activated (approximately               Toyama, Japan
1 to 5 s). While the average amount of drug infused during a            To investigate the relationship between stress and sweating from
session tends to be very accurate, we have noticed that there is        the dog footpads, cardiovascular autonomic functions (heart
considerable variability in the dosage delivered for individual         rates, heart rate variability) and sweating volume in unstressed
injections. We investigated the involvement of syringe con-             and stressed conditions were analyzed in dogs. Sweating vol-
struction, syringe size, and pump activation duration on the            ume was measured as transepidermal water loss (TEWL), which
variability of consecutive infusions. To conduct this study, we         was semi-quantified using skin patches including color-reacting
used polypropylene and polycarbonate syringes in 20 ml and              papers impregnated with cobalt chloride. The skin patches were
10 ml sizes and a single-speed syringe pump (3.3 rpm). Each             stuck to the footpad; the color of the skin patches changed in
syringe was tested using 10 injections with 1-, 3-, 6-, and 10-s        response to water in sweat from the footpads. This simplified
injection times. No significant differences were found in injec-        system enabled us to assay TEWL from conscious dogs. In the
tion variability based on syringe type (P > 0.05 polypropylene          first experiment, electrocardiograms (ECGs) and TEWLs were
compared with polycarbonate). Injection variability does, how-          recorded from laboratory beagles. The same experiment was re-
ever, vary significantly as a function of injection time. With 20       peated again after the dogs were allowed to freely move around
ml syringes, injection volumes vary by 17%, 9%, 6%, and 4% for          for about 20 to 30 min. Finally, ECGs and TEWL were also mea-
1-, 3-, 6-, and 10-s injections, respectively. Using 10ml syringes      sured from the anesthetized beagle dogs. Both heart rates and
the pattern is similar, with injection volumes varying by 28%,          TEWL were lower after the laboratory beagles were allowed to
11%, 3%, and 4% for 1-, 3-, 6-, and 10-s injections, respectively. In   relax for 20 to 30 min. These parameters further decreased when
order to minimize the impact of injection-to-injection variability      they were anesthetized. Furthermore, spectral analysis of heart
when using motor-driven syringe pumps, longer injection times           rate variability indicated that TEWL was positively correlated
(>3 s) are advisable whenever possible.                                 to heart rates and sympathetic nervous activity, while TEWL
                                                                        was negatively correlated to parasympathetic nervous activity
P157 Development of a Fecal PCR Assay for Spironucleus                  in the laboratory beagles. In the second experiment, the TEWL
muris                                                                   was indexed by visual inspection according to the color table,
                                                                        while arousal or stressed levels were indexed according to the
G Jackson*, R Livingston, LK Riley, CL Franklin                         behavioral criteria using domestic dogs under various stressed
Research Animal Diagnostic Laboratory, University of Missouri,          and non-stressed conditions. These 2 indices were significantly
Columbia, MO                                                            correlated. The results suggest that sweating from the footpads
                                                                        in dogs might corresponds to human sweating from the palms,
Spironucleus muris is a protozoan of the intestinal tract in mice,
                                                                        and that the present simplified method using color-reactive skin
rats, and hamsters. While its effect on the health of animals and
                                                                        patches is useful to assess degrees of stress in dogs.
its capacity to confound research are debated, it is often included
in rodent health monitoring programs. Diagnosis usually relies on
postmortem examination of the intestinal tract for trophozoites.        P159 Migration of Ascaris suum Larvae in Mice
In this study, we sought to develop a PCR-based antemortem              HZ Naghashyan*, OV Shcherbakov
diagnostic test with improved sensitivity and specificity. A PCR
assay targeting the 16S-like rRNA gene of S. muris was designed         Veterinary Medicine, Armenian State Agrarian University, Ye-
and optimized. The assay was found to be highly specific as feces       revan, Armenia
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

Research of Ascaris suum larvae migration in mice has a great           air or ground transportation has negligible impact on the tumor
importance, because mice serve as an experimental model for             growth kinetics in the A431 xenograft mouse model. Therefore,
study of early migration in pigs, as well as for syndrome of            the importation of tumor-bearing mice is a viable alternative to
visceral larva migrans in humans. Many authors deal with the            creating xenograft models in house.
problem of ascarides larvae migration in uncommon hosts, but
some aspects of migration remain controversial, and not well            P161 Withdrawn
studied. According to literature data, A. suum larvae penetrate
the mouse cecum and colon and reach the liver by 4 h postinocu-
                                                                        P162 Comparison of Diagnostic Methods for Surveillance of
lation. As for other authors, larvae appear in liver at days 2 to 7
                                                                        Helicobacter Species Infection in Mouse Colonies
postinoculation. Taking into account this information, we aimed
to study migration of A. suum larvae in laboratory mice. For this       JL Lofgren*, A Potter, M Fortin, NS Taylor, MT Whary, JG Fox
purpose, 10 adult males (weight, 20 g) were orally infested with        Division of Comparative Medicine, Massachusetts Institute of
1000 to 5000 eggs. Mice were euthanized by ether every other            Technology, Cambridge, MA
day postinfection and dissected with larvae detection by one of
the following methods: compression with dyeing by 1% solution           Helicobacter species naturally infect research mice and may con-
of methyl-violet, compression with clearance by 1% solution of          found experimental studies; thus, surveillance for helicobacter
lactic acid, or digestion in artificial stomach juice. Results show     infection status should be a component of murine health moni-
that the most informative and efficacious method for larvae             toring. Although choice of diagnostic method should be based
detection appears to be compression with clearance by 1% so-            on available sample type, technical requirements and cost, assay
lution of lactic acid. It allows detect maximal number of larvae        limitations of sensitivity and specificity are critical to obtaining
in the internal organs of mice. The first free larvae are detected      reliable results. We compared genus-specific PCR based on a 1200
in the gastrointestinal tract, mainly in stomach and cecum, on          base-pair 16s RNA product, culture, and Helico-Stix Test strips
d 2 after infestation. On d 4, larvae are detected in mesenteric        for detection of Helicobacter spp. infection using feces collected at
lymph nodes, liver, and lungs, and quantity of migrating larvae         necropsy from 34 sentinel CD-1 mice. Sentinels were housed for
in gastrointestinal tract is maximal. Larvae were detected in the       6 mo in 31 rooms, representing 5 facilities, and were exposed to
internal organs up to 10th d postinoculation. Therefore, our re-        weekly dirty bedding transfer from colony mice of varying ge-
search shows that A. suum larvae migration in uncommon hosts,           netic and SPF health status. Fresh fecal pellets from each sentinel
namely mice, is rather longer than described in literature, and         (n = 8 to 10) were solubilized in 1 ml of laboratory-grade distilled
does not depend on inoculating eggs dosages. This circumstance          water for the Helico-Stix Test assay. Several additional feces were
may be considered in clinical and laboratory practice when early        frozen at –20 °C for subsequent DNA extraction and at –70 °C in
migration phase of ascariasis is studied.                               30% glycerol broth for subsequent microaerobic culture at 37 °C
                                                                        on blood agar and antibiotic-selective media with and without
                                                                        filtration through a 0.45 μ filter. Nine of 34 fecal samples were
P160 Impact of Transportation on Murine Xenograft Tumor
                                                                        PCR-positive for helicobacter, and helicobacter isolates were re-
Models
                                                                        covered by culture from 7 of these 9 PCR-positive samples. Only
H Liang*, R Driscoll, K Harbert, LA Conour                              4 of the 9 PCR-positive samples were positive when screened
Discovery Services, Charles River, Wilmington, MA                       for helicobacter infection using the Helico-Stix Test assay. The
                                                                        Helico-Stix Test had a preponderance of false positive outcomes;
Xenotransplantation of human tumors into immunodeficient                20 PCR-negative samples were strip-positive. In comparison to
rodents is a frequently used animal model for assessing the             PCR, these results indicate that the sensitivity and specificity of
efficacy of antineoplastic therapies. Among them, the subcu-            culture were 78% and 100%, and for Helico-Stix Test 44% and
taneous xenograft model is the most common due to technical             20%, respectively. Thus, genus-specific PCR was the most sensi-
ease of tumor cell implantation and tumor growth monitoring,            tive diagnostic method for testing fecal samples from sentinel
permitting high-throughput assessment of compound efficacy.             mice and can be confirmed by culture when indicated.
Depending on the particular cell line, tumor growth rate can
vary in the live animal model. Time required for tumor growth
                                                                        P163 Laser-assisted IVF and Modified ICSI Markedly Improve
prior to therapeutic testing results in associated losses in valuable
                                                                        the Efficiency of IVF in C57BL/6J Mice
animal housing space, personnel time, and laboratory resources.
To address these resource constraints, many pharmaceutical and          J Mkandawire*1,2, T Chavarria1, P Qi1, A Discua1, A Viana1, SE
biotechnology companies outsource the creation of xenograft             Erdman1, JG Fox1, KJ Maurer1
tumor models with import of the animals in house for compound           1Division of Comparative Medicine, 2Center for Cancer Research,
screening. To date, however, the impact of shipping stress on           Massachusetts Institute of Technology, Cambridge, MA
tumor-bearing mice and the tumor kinetics in vivo has not been
reported in the literature. To determine whether air or ground          Assisted reproductive technologies (ART) are an essential com-
transportation influences tumor growth, we have evaluated               ponent of a modern mouse husbandry program. Of all ARTs, in
the NU/NU (Crl:NU-Foxn1nu) mouse model subcutaneously                   vitro fertilization (IVF) has perhaps the most potential because
implanted with A431 human epidermal carcinoma cells. At 4 or            only a single male is necessary to provide a large number of pups.
8 d post–tumor cell injection, the mice were randomly divided           Unfortunately, in some inbred strains of mice IVF is not highly ef-
into 3 groups and either shipped by air or by ground or remained        ficient. Recently, we tried two modifications of IVF to see if we could
in-house. Travel time for air and ground transportation was 20 h        increase the effectiveness. Specifically, we performed laser-assisted
and 30 h, respectively. In addition to daily clinical observations,     IVF and a simplified version of intracytoplsamic sperm injection.
animal body weight and tumor mass were measured 2 times                 All studies described herein used wild-type C57BL6/J oocytes
per wk for 5 wk. Our data showed no statistically significant           and sperm. Laser-assisted IVF was performed using a zyclone to
differences in animal body weight and tumor volume between              make 2 to 3 holes in the zona pellucida of eggs. Ablated oocytes
groups. Our study suggests that shipping stress sustained during        were co-cultured with sperm in human tubal fluid (HTF) for 5 h,
                                                                        washed, and cultured overnight in KSOM at 37 oC in 5% CO2. Modi-
Abstracts of scientific papers 2008 AALAS National Meeting

fied intracytoplasmic sperm injection (ICSI) was conducted using          1Comparative Medicine, 2Pharmacology, Pfizer, Inc, Saint Louis,

simple manual microinjection of sonicated sperm heads deposited           MO
within the oolema with gentle, positive, micropipette air pressure.       Intranasal (IN) dosing has long been the prominent technique
A sperm microinjection micropipette was calibrated in-house for           used when targeting the lungs. Different dosing methods can
easy aspiration of sperm cells and entry past the zona and oolema         affect systemic delivery of compounds, inhibiting the lung in-
barriers while averting oocyte lysis. After injection of sperm in fluid   flammatory process and costing valuable time. An innovative
holding medium (FHM), eggs were transferred to KSOM and incu-             method of delivery is modified intratracheal (ModIT) dosing
bated overnight at 37 oC in 5% CO2. Standard IVF was conducted            which historically has been used with much less frequency.
as described for laser assisted IVF without ablating the zona prior
                                                                          Specifically, mice are anesthetized with isoflurane and verti-
to incubation with sperm. Fertilization rates for all procedures
                                                                          cally suspended on a bar by the upper incisors. The tongue is
were scored by analyzing the amount of eggs that proceeded to
                                                                          extended by lined forceps, a finger is placed over the nostrils,
the 2-cell stage following overnight incubation. Both laser-assisted
                                                                          and 50 µl of compound is pipetted onto the back of the tongue.
IVF and the simplified ICSI method produced significantly greater
                                                                          Air is blown into the face of the animal prior to the release of the
fertilization rates (55% and 46%, respectively) compared to standard
                                                                          tongue, causing the animal to inhale; this reaction is unique, as
IVF (2%; P < 0.0001). In conclusion, both laser-assisted IVF and the
                                                                          there is no discovered documentation explaining this occurrence.
modified ICSI technique are simple, rapid methods to increase the
                                                                          We examined 2 in vivo dosing techniques—intranasal (IN) and
fertilization rate of IVF in inbred mice.
                                                                          ModIT—to evaluate and compare methods for time and resource
                                                                          management, dosing variability measured by lung dispersion
P164 Withdrawn                                                            and deposition, and efficacy using compounds. Compound
                                                                          efficacy was determined by dosing BALB/C mice (ModIT and
P165 Commercially Available Corn-cob Bedding Naturally                    IN for fluticasone, PF-xxx, and ± LPS controls) 1 h prior to IN
Contaminated with Zearalenone Significantly Advances the                  LPS challenge with a takedown 4 h post-LPS. Inflammatory cell
Time of Vaginal Opening in Immature CD-1 Mice                             counts (total cells and PMNs) in BAL fluids were analyzed by
                                                                          flow cytometry. Separately, compound dispersion was examined
JE Thigpen*, E Padilla-Banks, G Kissling, GF Caviness, TE Whi-
                                                                          using injection of coummasie blue dye (50 µl) IN and ModIT with
teside, J Locklear, MG Grant, DB Forsythe
                                                                          endpoints of 30 min and 4 h. After euthanasia, the lungs were
National Institutes of Health, Research Triangle Park, NC                 harvested and observed for dispersion of dye in the interstitial
We reported that corn-cob bedding contains the mycoestrogen               lung tissue. Data points (in min) were also taken to determine
zearalenone (ZEN). This report determines the concentration               the time elapsed/dosing group. Animals dosed with IN and
of ZEN in twelve different types of corn-cob bedding, and its             ModIT compounds showed equivalent efficacy in the number of
effects on time of vaginal opening (VO) in CD-1 mice. Liquid              lung infiltrates for each compound. ModIT coumassie blue dye
chromatography tandem mass spectrometry was used to mea-                  groups displayed a higher dispersion in the lungs with less dye
sure the concentration of ZEN and alpha-Zearalenol in samples             in the stomach or intestines. Variability within the ModIT group
of plasma, urine, feed, and bedding spiked at 0, 5, or 10 ppb/g.          was minimal in contrast to the IN groups, and was supported by
Upon arrival, mice were given a phytoestrogen-free diet and               necropsy images. Plotted dose time per animal varied, revealing
reverse osmosis/deionized water ad libitum. Pups were weaned              that ModIT has a 40% to 50% quicker dose time per animal in
at PND 15, and 15 mice per group were randomly assigned to                comparison to ModIT. Ultimately, ModIT dosing was proven to
control or test bedding. Body weights at weaning were similar             be equally efficacious, have a quicker dosing time, and provided
among the groups. Mice were weighed weekly and on the day of              less variability in lung dispersion.
VO and observed daily for VO. Plasma and urine samples were
collected on PND 22 and PND 34. Low, medium, or high levels               P167 Characterization of the Charles River GK Rat (GK/To-
of ZEN were detected in corn-cob bedding from all vendors. The            hiCskCrljCrl)
time of VO was significantly (P < 0.05) advanced in mice exposed          K Harbert*1, ST Chou2, R Driscoll1, L Barbeau1, D Mendonca1, H
to high levels of ZEN (4202 to 5284 ng/g) as compared to mice             Liang1, C Clifford3, LA Conour3
exposed to low levels of ZEN (<500 ng/g). The time of VO was
                                                                          1 DiscoveryServices, 2Discovery Services/GEMS, 3Research
advanced, but not significantly in mice exposed to medium levels
of ZEN (1900 to 2200 ng/g). The time of VO was significantly              Models and Services, Charles River, Wilmington, MA
(P < 0.05) advanced in mice exposed to bedding spiked with                The Goto-Kakisaki Rat (GK Rat) is a well-established model of
10,000 ng/g of ZEN as compared to controls. Concentrations of             non–insulin-dependent, nonobese Type II diabetes mellitus (T2DM).
ZEN and alpha-zearalenol in the plasma and urine of the mice              Distinct substrain colonies have been maintained around the world
were correlated with the concentration of ZEN in bedding. We              since the model’s introduction in the 1970s and have demonstrated
concluded that ZEN contaminated corn-cob bedding can expose               similar levels of glucose intolerance and impaired insulin response.
mice to ZEN by contact, ingestion, or aerosol exposure, and               Some of these source colonies are known to differ from each other in
can significantly impact time of VO, an estrogen-sensitive bio-           pancreatic morphology and insulin level. Currently, we use a sub-
marker. This is the first report to determine that corn-cob bedding       strain of GK Rat (GK/TohiCskCrljCrl) for internal and sponsored
naturally contaminated with ZEN can significantly advance the             studies. We have performed an in-Departmenth characterization of
time of VO in CD-1 mice and has the potential to impact other             this substrain, comparing it to data reported in the literature. Both
hormonal endpoints.                                                       male and female GK rats demonstrate a similarly reduced growth
                                                                          rate and average body weights compared to Wistar (Crl:WI) con-
P166 Comparison of Intranasal Dosing and Modified Intra-                  trols. As expected, the GK/TohiCskCrljCrl rats have significantly
tracheal Dosing in BALB/c Mice: Compound Efficacy, Time                   elevated fasting blood glucose levels (P < 0.0001) and an altered
Management, and Compound Dispersion                                       response to oral glucose tolerance test at as early as 8 wk of age.
                                                                          In addition, the baseline blood glucose is significantly higher in
J Daller*1, JD Blom2
                                                                          male GK/TohiCskCrljCrl rats than female cohorts at 16 wk of age
                                                                                       Abstracts of scientific papers 2008 AALAS National Meeting

(P < 0.02). Indirect blood pressure measurements and pathological       Oregon National Primate Research Center, Beaverton, OR
assessment at 11 and 22 wk of age do not indicate signs of cardiac      Positive reinforcement training (PRT) is a type of operant condi-
disease. As previously reported, the male GK/TohiCskCrljCrl rats        tioning in which the trainer reinforces desired behaviors (such
also maintained a significantly lower pulse rates (323 ± 68 bpm, P      as coming to the front of the cage) by rewarding the animal
< 0.05 at 22 wk). Unexpectedly, the male GK/TohiCskCrljCrl rats         when it performs the target behavior. PRT increases the animal’s
demonstrate proteinuria (>200 mg/dl) compared to both Wistar            control over its environment and reduces the stress associated
controls and female cohorts at as early as 9 wk of age (P < 0.001).     with procedures (such as blood draw or injection) and as such is
Histopathological examination of the pancreas showed chronic            beneficial to the well-being of the animal. While there are many
diffuse pancreatic islet degeneration and regeneration, hemosi-         benefits associated with training, it often involves a large initial
derosis, and islet cell anisocytosis. However, examination of the       input of time resources, which may preclude some facilities from
kidney did not reveal any significant changes in relation to diabetic   using PRT. Finding alternate training methods that can make the
nephropathy at up to 26 wk of age. Our findings maintain that the       process less time consuming would be of great value. Because
GK/TohiCskCrljCrl rats have similar diabetic characteristics as         some primates are known to imitate the actions of others, one
those found in established substrains published in the literature.      potential alternate technique is to have subjects watch conspecif-
The source of sex-specific proteinuria in young male rats suggest       ics being trained to perform various tasks. In the present study,
early glomerular injury and filtration deficits, and is under further   we examined whether observing others would reduce training
investigation for serving as an indicator for monitoring the develop-   time in adult, male rhesus macaques (Macaca mulatta). Monkeys
ment of nephropathy associated with this diabetic model.                (n = 12) were housed in individual cages located in a single room.
                                                                        We used PRT techniques to train the monkeys to present a thigh
P168 Effects of Feed and Water Deprivation on Japanese Quail            at the front of the cage and remain stationary for injection. We
(Coturnix japonia)                                                      first trained 6 “demonstrator” monkeys, all housed on the same
KL Arora*, B Biswas                                                     side of the room, for approximately 4 mo. We then trained the
                                                                        other 6 monkeys (“observers”) using the same techniques. The
Department of Veterinary Science, Fort Valley State University,         observers were located directly across from the demonstrators.
Fort Valley, GA                                                         For both groups, training sessions were 5 to 10 min, 2 to 3 times
This study resulted from an accidental deprivation of feed and          per wk. Training was considered successful when the task was
water in Japanese quails over 1 weekend. Apparently, the birds          performed on command during 3 consecutive training sessions.
didn’t exhibit any adverse effect, and accepted feed and water          While we were able to train all 12 monkeys to present their
readily when offered, and appeared satisfied in a couple of min-        thigh at the front of the cage, it took the observers about 25%
utes. This experiment was planned to address adverse effect of          fewer training sessions than the demonstrators to complete the
deprivation on physiological parameters. Six adult males and six        task (P < 0.01). Additionally, significantly more observers than
females (layers) of uniform size and weight were deprived of both       demonstrators allowed needle insertion (P < 0.05). These results
feed and water for 20 h or 40 h and the effects on body weight,         suggest that observing others being trained may help to reduce
hematocrit, plasma proteins, egg production, and shell quality          overall training time in macaques.
were studied. At the start of experiment, the birds were weighed
and blood collected from the wing vein for evaluation for sugar,        P170 Locomotor Activity in Rhesus Macaques (Macaca mulatta)
PCV, and plasma proteins. Blood sugar was measured using a              on Long-term Calorie Restriction
glucometer, PCV using hematocrit tubes and plasma proteins
using a refractometer. After fasting, the birds were weighed daily      K Hagelin*1, J Young1, J Mattison2, D Ingram3
for 9 consecutive d and the blood was evaluated on d 1 and 3.           1Division of Veterinary Resources, NIHAC SoBran, Inc., Pool-

After 20-h deprivation, the males lost 7% (118.3 g compared with        esville, MD; 2Laboratory of Experimental Gerontology, NIA,
109.5 g) and females 13% (141.1 g compared with 122.4 g) of their       Poolesville, MD; 3Nutritional Neuroscience and Aging Laborato-
body weight (P < 0.05). After restoring feed and water, the males       ry, Pennington Biomedical Research Center, Baton Rouge, LA
recovered 96% and females 93% of their prefasting weights by d
                                                                        Decline in locomotor activity is a robust feature of aging that
3. The blood glucose decreased (P > 0.05), and PCV and plasma           has been documented in both humans and nonhuman primates.
proteins increased (P < 0.05) during fasting, then returned to          The effects of long-term calorie restriction (CR) on aging and its
normal levels by d 3. Females resumed laying after skipping 1
                                                                        impact on behavioral processes are still being investigated. Age
or 2 d. After 40-h deprivation, males lost 12% (123.2 g compared
                                                                        and diet effects on locomotor activity were evaluated in a group
with 107.9 g) and females 25% (144.6 g compared with 107.6 g) of
                                                                        of healthy young and aged male and female rhesus monkeys
their body weight (P < 0.05). After restoring feed and water, the       involved in a long-term longitudinal study of CR. All monkeys
males recovered 94% and females 90% of their prefasting weight          are provided a highly nutritious diet, with the CR group receiv-
by d 3, with full recovery by d 9. Eggs were smaller, and egg shells    ing 30% less than that provided to comparable age, gender, and
lacked proper pigmentation. The PCV increased during fasting            body size controls (CON). CR was initiated when the monkeys
(48.9% compared with 42.9%; P < 0.05) and returned to prefasting        were juveniles, adults, or aged. At the time of data collection,
level after d 3. Results indicated that water and feed deprivation      monkeys were grouped as either Young (CON: n = 28, age = 15
could seriously alter the physiochemical parameters, with the ef-       to 21 y; CR: n = 25, age = 15 to 27 y) or Old (CON: n = 7, age = 28
fects increasing with prolonged fasting; these, in turn, could affect   to 35 y; CR: n = 6, age = 32 to 37 y); monkeys had been on CR for
experiments in progress. Furthermore, the data could also be used       16 to 18 y. Locomotor activity was analyzed using noninvasive
as a resource for animal care committees if faced with protocols        activity monitors that were attached to a primate collar. These
that include bird feed and water deprivation.                           monitors directly record activity counts by analyzing the occur-
                                                                        rence and intensity of motion through omnidirectional sensors.
P169 Watching Conspecifics Being Trained Helps Rhesus Ma-               All activity was collected in a monkey’s home cage over a period
caques (Macaca mulatta) Learn Faster                                    of 24 h for 7 consecutive d. Because the activity levels of rhesus
K Mueller, K Moore*, A Maier, K Coleman                                 monkeys are highly dependent upon the circadian cycle, activ-
Abstracts of scientific papers 2008 AALAS National Meeting

ity data was analyzed for 24-h, day, and night periods. Young           The GE Lunar PIXImus II (PIXImus) is used for dual energy
CON monkeys were significantly more active than Old CON                 x-ray absorptiometry (DEXA) in laboratory mice. However, the
monkeys for the 24-h (R2 = 0.12, P = 0.04) and Night (R2 = 0.12,        image acquisition area is only 80 by 65 mm, limiting the animal
P = 0.04) periods, while Young CR monkeys were significantly            size and weight (<50 gm) for which it can accommodate. The GE
more active than Old CR monkeys for the Day (R2 = 0.15, P = 0.03)       Lunar Prodigy (Prodigy) is an FDA-approved imaging modality
and Night (R2 = 0.37, P = 0.003) periods. In terms of diet effect,      for human use and validated for animals weighing over 200 gm.
Young CR monkeys were significantly more active than Young              When using these 2 densitometers, a diagnostic gap therefore
CON monkeys, regardless of time of day. Conversely, Old CON             exists for evaluation of obese mice and young rats for which no
monkeys were significantly more active than Old CR monkeys.             validated guidelines exist. A study was conducted to evaluate
To gain a better understanding of the effect of CR on aging and         animals weighing less than 200 gm using the Prodigy. Twenty-
its associated health benefits, locomotor activity will be examined     three adult, mixed-sex C57BL/6NCrl (n = 9) and Crl:CD-1(ICR)
every 3 y. In addition, we hope this longitudinal assessment will       (n = 14) mice with body weight ranging from 19 g to 41 g were
provide additional information regarding whether or not the age         used. Each animal was scanned using both densitometry models,
in which CR is first introduced negatively or positively impacts        and values for bone mineral density (BMD), bone mineral con-
various behavioral functions.                                           tent (BMC), fat, lean, and total tissue recorded. For the Prodigy,
                                                                        an average value was calculated from a minimum of 3 scans on
P171 Evaluation of Three Catheter Maintenance Schedules on              each mouse. Each data point was plotted against the animal’s
Patency of Jugular Vein Catheters in CD Rats                            body weight to generate best-fit linear regression lines. Despite
                                                                        good correlation between body weight and DEXA data, 2-tailed
KL Walter*1,2, Y Luo3, Y Luo3, T Fisher4, GB Mulder4                    Wilcoxon matched pairs test showed a statistically significant dif-
1HarleysvilleVeterinary Hospital, Harlerysville, PA; 2Virginia-         ference (P ≤ 0.03) for all 5 parameters measured. Except for total
Maryland Regional College of Veterinary Medicine, Virginia              tissue, the slopes of best-fit lines do not differ significantly for
Tech, Blacksburg, VA; 3Charles River, Raleigh, NC; 4Charles             the parameters, suggesting that the inaccuracy may be attributed
River, Wilmington, MA                                                   to software differences or inappropriate calibration standards. It
                                                                        is possible to calculate for a variation factor for data correction,
Surgical placement of jugular vein catheters (JVCs) in rats is a
                                                                        but validation will require comparison of body composition by
common procedure to allow central venous access in conscious
                                                                        chemical carcass analysis. Despite inaccurate data, repeat mea-
animals. Recommendations for catheter maintenance in rats, such
                                                                        surements taken on individual animals using the Prodigy proved
as frequency of catheter flushing and types of lock solution used,
                                                                        to be consistent. In addition, there is no significant difference in
are variable between institutions and generally based on limited
                                                                        the values calculated for mice scanned either as an individual or
scientific evidence. We conducted a study investigating the opti-
                                                                        as a group, with individual bodies analyzed as a region of interest
mum maintenance schedule for rats with JVCs. Sixty male Crl:CD
                                                                        (P > 0.05). We do not recommend the Prodigy for quantitative
(SD) rats weighing 225 to 250 g were used; 30 with polyurethane
                                                                        analysis on rodents weighing less than 200 g without calibration
(PU) JVC catheters and 30 with PU-silastic (Blended) JVC catheters
                                                                        adjustment. However, the consistency of data suggests its poten-
originating from 2 different Charles River commercial barrier sur-
                                                                        tial use as a high-throughput tool for detecting general bone and
gical facilities. Animals were randomly assigned to 1 of 6 groups
                                                                        body composition differences between animal groups.
(n = 10) based on surgical facility of origin, catheter material, and
maintenance frequency (catheter flushing every 3 (Q3d), 5 (Q5d)
or 7 (Q7d) d after surgery). The day of surgery was considered d 0.     P173 Effect of Buprenorphine Analgesia on Growth in Neo-
Catheters were considered patent if blood was successfully with-        natal Mice
drawn. Treatment consisted of weighing each animal, aspiration of       K Vail*1, A Echeverry2, R Adkins2, J Zaias1
the lock solution (50% heparinized dextrose) and blood, followed        1Division
by flushing with physiologic saline and replacement of lock solu-               of Veterinary Resources, 2Immunology, University of
tion. Catheter patency rates for treatment groups were 100% on the      Miami, Miami, FL
first assessment for Q3d and Q5d, and 80% for Q7d. At d 14, 70%         Experimentation and handling of neonates can often incite ma-
of Q7d catheters remained patent and at d 15, 65% of Q5d catheters      ternal cannibalism or neonatal mortality from lack of maternal
remained patent. Thirty percent of Q3d catheters remained patent        care. Therefore, studies involving neonates must be carefully
at d 15. At the conclusion of the study, d 21 (Q7d) and d 20 (Q5d),     done in order to obtain accurate results and ensure that the
35% and 55% of the catheters remained patent, respectively. Fifteen     results obtained are from the experimental intervention rather
percent of Q3d catheters remained patent on d 21. No significant        than handling or anesthetic effects. Further complicating experi-
difference in patency rates between catheter types (blended com-        mentation with neonatal rodents is the issue of addressing pain
pared with PU) were found for the Q5d and Q7d treatment groups          or distress in neonates. Anesthesia and analgesia for surgery or
(P > 0.966). For the Q3d treatment group, PU catheter performance       procedures involving neonatal rodents are challenging. Scant
was significantly improved (P = 0.007) compared to the blended          information exists on the metabolism and biotransformation of
catheters. In summary, patency rates were greater than 80% for each     analgesics in neonatal rodents, and the routes of administration
treatment group from d 3, 5, or 7 through d 10. Longer-term patency     of analgesics to suckling neonates are more limited as compared
rates were greatest for catheters flushed every 5 or 7 d.               to adults. The purpose of this study is to determine whether there
                                                                        are untoward effects of administering subcutaneous analgesia
P172 Evaluation of the GE Lunar Prodigy Densitometer for                to young pups. The study assessed the effects on weight gain
Use in Mice                                                             of subcutaneously injected buprenorphine to neonatal BALB/c
                                                                        mice both uninfected and infected with a pathogen (Yersinia en-
KL Walter*2, ST Chou1, E Ahearn1, D Mendonca1, H Liang1, LA             terocolitica) over a 2-wk period. We weighed neonates beginning
Conour1                                                                 on postnatal d 7 to post natal d 23 and administered buprenor-
1Charles River, Wilmington, MA; 2 Harleysville Veterinary Hos-          phine (0.05 mg/kg) twice daily for 14 d beginning on postnatal
pital, Harleysville, PA                                                 d 10. There were 3 treatment groups each for uninfected and
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

infected animals: 1 group was handled only (weighed twice             nonsurgical approach to fight this disease, our group developed
daily); the second group was injected twice daily with PBS; and       a canine model to mimic human gastric emptying delay. The
the third group was injected twice daily with buprenorphine at        model increased the sensation of stomach fullness, decreasing
12-h intervals. No mortality was encountered. Weight gain did         caloric consumption, which caused eventual weight loss in
not differ between the noninfected groups as analyzed by 1-way        the dog. An endoscopic procedure was developed for pylo-
ANOVA. However, buprenorphine had an inhibitory effect on             ric restriction (PR) using sutures to narrowing the pylorus. To
growth in the neonates infected with Yersinia enterocolitica rela-    conduct these experiments, 16 male dogs weighing between 13
tive to the handled-only and control mice. In addition, we found      and 18 kg were randomized into 3 groups. Group 1 underwent
that injecting mice subcutaneously affected the spleen weights        the actual PR technique (8 dogs), group 2 were shams (4 dogs),
compared to handled-only or infected-only mice. These findings        and group 3 were controls (4 dogs). Gastric emptying was mea-
suggest there might be synergistic effects between pathogens          sured in all dogs using the octanoic acid gastric motility breath
and analgesics that may alter development of neonatal mice.           test (GMBT). Dogs in all groups were monitored daily for food
This information should permit investigators to make educated         intake and weight gain or loss.The PR dogs decreased their
decisions regarding the effects of analgesics alone or in combina-    food consumption by a mean of 48% (P < 0.002). At the same
tion with infectious agents, and of potential handling or injection   time, the sham and control dogs showed an 8% increase in food
stress in neonatal mice.                                              consumed, resulting in a 56% difference between the PR-sutured
                                                                      dogs and the sham and control groups of dogs (P < 0.0002).
P174 Percutaneous Intrasalivary Gland Dosing Technique in             There was a delay in gastric emptying between the presuturing
Rats                                                                  baseline and first postsuturing gastric emptying tests by 14% (P
                                                                      < 0.02). In the sham and control dogs, there was an increase in
JG Tontini1, LM Serfilippi*1, DR Pallman1, CV Mujer2, CB Spain-       gastric emptying between the presuturing baseline and suturing
hour2                                                                 gastric emptying test by 18% (P = 0.08).We found a significant
1Department of Laboratory Animal Medicine, 2Department of             delay in the PR dogs’ gastric emptying compared with the sham
Toxicology, Calvert Laboratories, Inc., Olyphant, PA                  and control dogs’ gastric emptying. PR dogs lost 13% of their
                                                                      initial body weight (P < 0.03). Sham and control dogs gained
Intrasalivary gland injections have been an area of increased
                                                                      14% of their initial body weight. There was a 26% difference
interest recently. This route has become important in develop-
                                                                      in the change in weight (P < 0.0002) between the PR dogs and
ment of treatments for sialorrhea as well as delivery of local
                                                                      the sham and control dogs. Significant statistical difference was
radiation and potential gene therapy. Difficulties with this dos-
                                                                      demonstrated in all parameters (weight loss, gastric emptying,
ing technique are the ability and necessity to isolate the gland
                                                                      and food consumption) in PR dogs compared with sham and
effectively to ensure that the dose is placed into the gland and
                                                                      control dogs. This canine model proved to be a reproducible
not the contiguous tissues. Different modalities have been used
                                                                      model for an effective cure of human disease. The dog model
to assist in the attainment of this goal, such as ultrasound or
                                                                      was suitable enough to achieve our goals of using a permanent
surgical exposure of the gland. However, ultrasound requires
                                                                      (but reversible) nonsurgical procedure to induce weight loss by
specialized equipment, is costly, and requires extensive train-
                                                                      restricting the pylorus.
ing, while surgical exposure is associated with risk and is not
conducive to repeat dosing since it induces scar tissue formation
at the site. Our goal was to develop a noninvasive percutane-         P176 Assessment of Chemotherapy-related Fatigue in Mice
ous technique in the rat model which is easy to use and would         MA Ray, R Trammell, S Ran, LA Toth*
eliminate the need to use these other methods. Sprague-Dawley
rats were anesthetized using isoflurane and placed in dorsal          SIU School of Medicine, Springfield, IL
recumbency with necks in dorsoflexion. The area over the sub-         The frequent occurrence of fatigue in cancer patients and survi-
mandibular salivary glands was shaved and swabbed with 70%            vors and the negative impact of this problem on quality of life
isopropyl alcohol. Using atraumatic tissue forceps, each gland        and clinical outcome underscore the need to identify mechanisms
was elevated, then isolated and a 0.1-ml injection of Evan blue       that underlie fatigue, with a view toward developing treatments.
was made through the skin into the glands’ parenchyma. The rats       To further that goal, we developed a model for quantifying fa-
were then euthanized by carbon dioxide asphyxiation and the           tigue and determining its mechanisms during chemotherapy in
glands were harvested. Glands were cross-sectioned to determine       mice. Female BALB/cJ mice were implanted with a telemetry
accuracy of the technique by noting the location of the Evan blue     device that transmitted information on both core temperature
dye, intraglandular compared with periglandular. With practice,       and horizontal locomotor activity. Mice were also given access
the success rate of delivery of dye to the submandibular gland        to a running wheel in the home cage. Activity, wheel running,
using this technique appears to be over 90%, which supports           temperature, food intake, and body weight were monitored be-
the claim that percutaneous salivary gland dosing may be an           fore, during, and after administration of paclitaxel (10 mg/kg IV
effective noninvasive alternative to deliver test article to the      daily for 5 sequential d; n = 9 per group) either as the standard
submandibular salivary glands.                                        formulation prepared in cremophor EL vehicle (Taxol) or as the
                                                                      albumin-embedded nanoparticle formulation nab-paclitaxel
P175 Canine Model of Obesity Using a Minimally Invasive               prepared in saline (Abraxane). Taxol and Abraxane both reduced
Pyloric Restriction                                                   horizontal locomotor activity and wheel running in mice. With
                                                                      either drug, mice showed essentially normal amounts of activity
L Bright*1, K McIntyre1, M Salvatore1, A Agelan1, L Miller2, A        during the first 2 h of the dark phase, indicating the ability and
Vegesna2, M April1                                                    initial desire to run during the active phase. However, during
1UniversityLabratory Animal Resources, 2Gastroenterology,             the 5 d of drug administration, activity fell below normal for
Temple University, Philadelphia, PA                                   both measures during the remainder of the dark phase, with
                                                                      further reductions during the first week after chemotherapy.
Obesity is a worldwide pandemic disease and a major concern
                                                                      Mice treated with Abraxane resumed normal amounts of dark-
in the US healthcare system. In search of a noninvasive, quasi-
Abstracts of scientific papers 2008 AALAS National Meeting

phase activity during wk 2 after treatment, whereas mice treated      data to support or refute these claims. In this study, the effects of
with Taxol remained depressed until wk 4 after treatment, con-        perioperative administration of buprenorphine, an opiate anal-
sistent with the greater toxicity of Taxol and/or its cremophor       gesic, and carprofen, an NSAID, were evaluated for effect on the
vehicle. Assessment of potential causes of fatigue revealed that      development of the CCI model of neuropathic pain. Thirty male
during periods of fatigue, mice did not show anemia, elevated         Sprague Dawley rats underwent chronic constriction of the left
serum concentrations of proinflammatory sleep-modulatory              sciatic nerve or sham surgery. Test groups received perioperative
cytokines, or disturbed sleep. However, they did show mild            treatment with buprenorphine (0.05 mg/kg SC twice daily for
general debilitation (weight loss, anorexia, and hypothermia)         2 d), carprofen (5 mg/kg SC once daily for 2 d), or subcutane-
and mild neuromuscular impairment. Thus, the combined as-             ous saline twice daily for 2 d. Animals were tested at days –1,
sessment of running wheel activity and horizontal locomotor           3, 7, 14, and 21 postsurgery for mechanical allodynia, thermal
activity suggests that mice treated with taxane chemotherapy          hyperalgesia, and incapacitance. The percentage of body weight
develop fatigue.                                                      change in each group was measured for 3 d following surgery
                                                                      and weekly thereafter. Statistical analysis by 2-way repeated
P177 Acclimation to Changes in the Light:Dark Cycle in Mice           measures ANOVA demonstrated significant difference (P < 0.05)
                                                                      between the carprofen-treated and control group, as evidenced by
CA Bosgraaf, LA Toth*                                                 higher withdrawal thresholds, higher latency times, and increased
Southern Illinois University School of Medicine, Springfield, IL      weight-bearing. There was no difference at any time point between
                                                                      the buprenorphine-treated and control group in any of the behav-
Accomplishing research goals can sometimes be facilitated by
                                                                      ioral tests. No difference in the percentage of body weight change
changing the light:dark (L:D) cycles of the experimental animals.
                                                                      was demonstrated between the carprofen and control groups,
For example, performing sample or tissue collection during
                                                                      while percentage reduction in body weight was significantly
the dark phase may be logistically easier if the animals are ac-
                                                                      greater in buprenorphine-treated rats compared to other groups
climated to a reverse light cycle that allows their dark phase to
                                                                      on days 3 and 7. These results provide evidence that perioperative
occur during normal work hours. However, prior to experimental
                                                                      treatment with buprenorphine can be used without significantly
use, animals subjected to such changes must be permitted suf-
                                                                      impacting behavioral tests in the rat CCI model.
ficient time to adapt physiologically and behaviorally to the
new light:dark rhythm. To determine the minimal time likely
to be required for such adaptation, we subjected individually         P179 Interference of the Clomiphene Citrate in BALB/c/Uni
housed male BALB/cByJ mice (n = 13) to a 12 h delay in dark           Females Estrous Cycle
onset, followed by resumption of a reversed L:D pattern. For          A Salgado, M Corat*, VL Dias, A Gimenes, L Passos
14 consecutive d, mice were evaluated continuously for core
                                                                      CEMIB/UNICAMP, Campinas, Brazil
body temperature and horizontal locomotor activity by using a
telemetry device implanted in the abdomen. In addition, mice          Most animal facilities that work with mouse embryos usually use
had continuous access to a running wheel in the home cage. After      superovulation to maximize the number of embryos for use in
7 d during which light and dark onset occurred at 0900 h and          embryo banks, strain decontamination, transgenesis, and other
2100 h, respectively, the light phase was extended by 12 h, such      techniques. This protocol is important into foundation colonies,
that dark onset did not occur until 0900 h, with light and dark       where the number of breeding pairs are limited and expensive.
onset thereafter maintained at 2100 h and 0900 h, respectively.       Usually pregnant mare serum gonadotropin (PMSG) is used for
Mice showed reduced locomotor and running wheel activity for          follicular stimulation and for synchronization of estrus. Due to
approximately 24 h after the L:D change occurred. However, both       the expense and delivery time of this hormone to South America,
activities then returned to basal levels, with the normal circadian   we decided to explore alternative methods of follicular stimula-
pattern of maximal activity during the dark phase. During the         tion. Our objective was evaluate the use of clomiphene citrate
baseline period, peaks and nadirs of core temperature occurred        to influence the ovulation of BALB/c/UNI mice females. All
during the dark and light phases, respectively. After the time        the animals used in this work were specific pathogen-free and
change, temperature peaks were reduced for approximately 2            were supplied and housed in a sterile environment in ventilated
to 3 d. Thereafter, normal rhythms resumed, although the circa-       racks. Female mice were dosed by gavage with clomiphene cit-
dian temperature range appeared greater than during the basal         rate (0.250, 0.375, 0.500, or 0.750 mg). On d 3, females received
period. Taken together, these data indicate that the behavioral       10 UI of human chorionic gonadotropin (HCG) and were bred
(activity) and physiological (temperature) stabilization of mice      with males. On d 1.5 of gestation, we collected the embryos (2-
to drastic (12 h) changes in light cycles occurs within 72 to 96 h.   cell stage) and checked their integrity. This methodology was
However, because other parameters (such as hormone secretion)         compared with superovulation by PMSG and natural ovulation.
may nonetheless adapt more slowly, longer acclimation periods         Clomiphene citrate outcomes were promising. The synchroni-
may be warranted.                                                     zation of estrus with PMSG was highest, but with clomiphene
                                                                      citrate synchronization was significantly higher than control,
P178 Evaluation of Perioperative Analgesia on Behavioral Tests        which did not receive any hormone stimulus. Our results show
in the Chronic Constriction Injury Neuropathic Pain Model             that the number of viable eggs was the same between groups,
                                                                      although the number of collected embryos was higher in the
LE Clarey*, C Huang, L Jackson                                        group stimulated with PMSG. We conclude that clomiphene
Biogen Idec, Cambridge, MA                                            citrate is an acceptable alternative to PMSG for reproductive
                                                                      studies, including those related with embryo banks.
Surgical models of neuropathic pain are used to evaluate candi-
date therapeutic agents for clinical use. Investigators have been
reluctant to provide perioperative analgesia in surgical models       P180 Evaluation of Different Ovariectomy Techniques on the
of neuropathic pain, citing concerns that the anti-inflammatory       Induction of Experimental Menopause in Rats
properties of analgesics would affect model development or con-       M Guzman-Silva*1, HN Henriques2, AB Carvalho3, C Pollastri3,
found interpretation of behavioral tests. There is little published   JS Pantaleão3
                                                                                      Abstracts of scientific papers 2008 AALAS National Meeting
1Pathology, 2Biomedical   School,   3Postgraduation    Program in      P182 Multiphase Surgical Approach for the Evaluation of
Pathology, UFF, Niteroi, Brazil                                        Active and Passive Fixation Pacing Leads for Telemetered
                                                                       Electrocardiograms in the Dog
Many researchers use ovariectomy in rats for induction of meno-
pause and evaluation of varied aspects related to the lack of sex      MG Taschwer*
hormones or the effect of hormonal replacement. Details of the         Department of Laboratory Animal Medicine, Covance Labora-
technique and the success of ovariectomy is not always reported.       tories Inc, Madison, WI
Two different ovariectomy techniques were compared and evalu-
ated through vaginal cytology at the time of surgery and after         The use of pacing leads in conjunction with telemetery trans-
30 d. Bilateral oophorectomy was performed in 35 adult female          mitters has become a popular method for the acquisition of
Wistar rats (222 ± 19 g) under xylazine and ketamin anesthesia.        relatively artifact free electrocardiograms (ECG). This study
In one group (OVX, n = 20), ovaries identified at laparotomy           employed a multiphase surgical approach to compare the per-
were excised; in the other group (OVX+PF), ovaries plus about          formance of active and passive fixation intracardiac (IC) pacing
1 cm of surrounding periovarian fat were removed. Vaginal              leads. Four purebred beagles were used per IC fixation type. In
smears were fixed in 95% alcohol, stained (Papanicolaou), and          Phase I pacing leads were introduced into the right ventricle
classified by microscopy as trophic (estrus, metestrus, diestrus,      through the jugular vein and guided into position by fluorogra-
proestrus) or atrophic (anestrus). At the time of surgery, all rats    phy and ECG signal. The distal ends of the leads were tunneled
showed trophic smears. One month later, estrus vaginal citology        into the subcutaneous tissue layer over the right lateral thorax.
was found in 2 OVX rats; all OVX+PF rats showed anestrus vagi-         Surgical phase II was performed 1 wk later. In this phase, a small
nal citology. Effective menopause status was achieved in 100%          incision was made to expose the distal IC connector and con-
OVX+PF and in 90% OVX. No statistical difference was found             nect it to a sterile transmitter for evaluation of the ECG signal.
(Z statistic), but biological implications are evident since animals   Candidates for transmitter implantation were selected based
still showing sex hormonal influence can not be used in any kind       on this evaluation. The final surgical phase required 6 IC-lead–
of forward research where the menopause condition is required.         implanted animals to receive telemetry transmitters. Results
The refinement introduced in the ovariectomy technique led to          indicated 2 of the 4 passive fixation pacing leads shifted position
improved effectiveness and increased the number of research            within 1 wk post implantation, creating less desirable ECG mor-
subjects available under menopause status.                             phologies. One of the 4 passive leads dislodged completely 4 wk
                                                                       post implantation. In contrast, all 4 active fixation pacing leads
                                                                       remained in their implanted positions allowing these animals to
P181 Species Variation in the Heparin Contaminant-mediated
                                                                       be chosen for transmitter implantation and further testing. The
Generation of Kallikrein
                                                                       multiphase surgical approach employed two minor procedures
M Clark*1, E McGeehan2, O Iqbal1, C Adiguzel1, J Cunanan1, D           prior to a major surgery. This approach provided an intermit-
Hoppensteadt1, J Fareed1                                               tent step to verify good IC lead placement and signal quality
1Department of Pathology, 2Comparative Medicine Department,            after 1 wk of healing. Although this intermittent step required
Loyola University Medical Center, Maywood, IL                          additional anesthesia and time, doing so confirmed that the ani-
                                                                       mal selected for implantation with the telemetry device would
One of the major pathopysiologic responses related to the adverse      be a viable candidate for test and would not require corrective
reactions due to recalled contaminated heparins is attributed to       surgery to remove and reimplant the IC lead. In addition, study
the generation of kallikrein, leading to the formation of brady-       time schedules would not be disrupted for corrective surgery.
kinin. The generation of kallikrein is reported to be due to the       We conclude that the performance of the active fixation lead,
deliberate adulteration of heparin by hypersulfated chondroitin        with respect to maintaining its implanted position, was superior
sulfate. The contaminated heparins were found to contain up to         to the passive fixation lead. The multiphase surgical approach
30% of this contaminant on a weight basis in heparin. The re-          provides the advantage of evaluating the IC lead position and
ported hypotensive effects of the contaminated heparin and the         ECG quality prior to implantation of the telemetry unit.
contaminant were found to be species dependent. To further in-
vestigate the variations in the contaminant-mediated generation
of kallikrein in different species, citrated plasma samples from       P183 Establishment of DMN-induced Liver Cirrhosis Model
human, dog, rat, cow, rabbit, and monkey were supplemented             in NOD/SCID Mouse for Evaluation of the Efficacy and Safety
with contaminated heparin (CH) and contaminant-free heparin,           of Human Mesenchymal Stem Cells
the isolated heparin contaminant (HC), and hemisynthetic hy-           M Hyon *1, E Kwon 1, H Choi 1, M Jang 1, K Cho 1, J Che 1, B
persulfated chondroitin sulfate (HCS-1 and HCS-2). In human            Kang1,2
plasma, all agents produced varying degrees of concentration-          1Department of Experimental Animal Research, Clinical Research
dependent activation of kallikrein formation (range, 0.3 to 0.7/15     Institute, Seoul National University Hospital, Seoul, South
min). In the monkey plasma, the activity was somewhat weaker           Korea; 2 Collage of Medicine, Seoul National University, Seoul,
(0.13 to 0.36/15 min). The generation of kallikrein was found to       South Korea
be higher in dog plasma (0.27 to 0.65/15 min). However, in rat
plasma relatively weaker activity was observed (0.12 to 0.18/15        Recently, many studies have suggested the potential of human
min). Interestingly, no activation of prekallikrein was noted in the   mesenchymal stem cells (hMSC) to differentiate into a hepatocyte-
cow plasma. Various agents produced different degrees of activa-       like lineage. In this study, we tried to establish an animal model
tion with the rank order of HC > HCS preparations > CH. CFH            of dimethylnitrosamine (DMN)-induced liver cirrhosis in NOD/
showed the least activity. These variations in the activation of       SCID mouse for evaluation of the efficacy and safety of cell therapy
the contact system in different species may be responsible for the     such as human mesenchymal stem cells. NOD/SCID mice were
differences in the observed hemostatic and vascular responses.         treated with DMN (1, 5, or 10 mg/kg IP) for 3 consecutive d per
These observations should be considered while screening the            wk for 5 wk and sacrificed at week 3, 4, 5, and 6 after first DMN
effects of drugs and agents with vasoactive actions.                   injection. In the 10 mg/kg group, most mice were critically ill or
                                                                       died. In the 5 mg/kg group, there were obvious clinical signs such
Abstracts of scientific papers 2008 AALAS National Meeting

as abdominal distention and piloerection. However, there was no        mechanisms and interventions of diabetes and “diabesity” (dia-
difference between the control group and the 1 mg/kg group in          betes with obesity). Automated clinical chemistry analyzers have
clinical findings. Based on serum biochemistry for evaluation of       been used for clinical chemistry and glucose monitoring of rats
the liver function, ALT, AST, ALP, and total bilirubin in the 5 mg/    and larger species, but blood volume requirements often in excess
kg group were significantly increased and albumin was decreased        of 100 µl, and the time to process serum and plasma for typical
compared with the 1 mg/kg or control group. In histopathological       diagnostic clinical chemistry instruments preclude practical use
findings in the liver, there were central venous necrosis, hemor-      with mice. Hand-held glucometers are used for diabetic patients.
rhage, and fibrillar septa formation in the 5 mg/kg group at 4 wk.     They are gaining popularity for mouse studies, but comparisons
The extracellalar matrix (collagen), iron, and glycogen deposition     of different instruments and validation against clinical chemistry
were confirmed by Masson trichrome (MT), Prussian blue (PB) and        analyzers for mouse specimens are difficult to find. We compared
periodic acid Schiff (PAS) staining, respectively. We also found       2 glucometers that require < 10 µl of whole blood, Accu-Chek
degenerative changes of renal structure and splenomegaly in the        Compact Plus and OneTouch Ultra2, with each other and against
5 mg/kg group. In conclusion, treatment of NOD/SCID mice               our VetACE clinical chemistry system for glucose measurement.
with 5 mg/kg of DMN for 4 wk can be used to create a pertinent         We collected 33 blood samples between 1100 and 1130 h, over a
liver cirrhosis model and can be useful to evaluate the efficacy and   2-wk period by cardiac puncture of adult mice of various age,
safety of cell therapy such as human mesenchymal stem cells.           sex, and strain. Fresh blood specimens were tested immediately
                                                                       on each glucometer. The remaining blood was separated into
P184 Sleep Deprivation Impairs Acquisition, Consolidation,             EDTA tubes (to be tested 15 min later) and serum separator
and Retrieval of an Ethical Discriminative Avoidance Task              tubes. Sera were tested within 2 h of collection. Data were ana-
in Rats                                                                lyzed to compare the instruments and condition effects (whole
                                                                       fresh blood, whole blood EDTA, and serum) on each specimen
TA Alvarenga1, ML Andersen*1, CL Patti2, RH Silva3, MB Calza-          using 2 tailed paired t-test. Significant and consistent differences
vara2, GB Lopez2, R Frussa-Filho2, S Tufik1                            (SD < 0.05) were found between glucometers both immediately
1Psychobiology, 2Pharmacology, Univ Fed Sao Paulo, Sao Paulo,          after blood extraction and after 15 min at room temperature
Brazil; 3Physiology, Univ Fed Rio Grande do Norte, Rio Grande          in EDTA. Whole fresh blood results from ACCU-CHEK were
do Norte, Brazil                                                       similar to serum results of our analyzer. Results from OneTouch
                                                                       were slightly but significantly higher than the analyzer and
The plus-maze discriminative avoidance (PM-DAT) is a new ani-
                                                                       ACCU-CHEK. EDTA blood results from both glucometers were
mal model of learning and memory that provides simultaneous
                                                                       significantly lower than the analyzer. Both glucometers yielded
information about anxiety. This behavioral paradigm complies
                                                                       consistent results that are comparable to the chemistry analyzer.
with ethical protocols, since there is no foot shock as imposed
                                                                       The low cost, cage-side operation, small volume requirement,
by classical avoidance tests. As sleep deprivation promotes
                                                                       and rapid results of these glucometers make them a practical tool.
important behavioral alterations in both humans and rodents,
                                                                       However, results from different instruments, or from different
the aim of the present study was to investigate the effects of
                                                                       specimen types may be difficult to compare. Protocols should
paradoxical sleep deprivation (PSD) for 96 h on the learning and
                                                                       be developed to standardize specimen handling and testing to
memory processes in rats submitted to the plus-maze discrimina-
                                                                       optimize utility.
tive avoidance task (PM-DAT), which simultaneously evaluates
learning, memory, anxiety, and motor function. Four experiments
were performed in which rats were submitted to: I) posttraining        P186 Effects of Blood EDTA Saturation on Selected Mouse
combined with pretest PSD; II) posttraining or pretest PSD; III)       Hematology Variables
pretraining PSD or paradoxical sleep (PS) rebound (24 h); and          NA Forbes*, CF Brayton
IV) pretest PS rebound. Posttraining PSD combined with pretest
PSD induced memory deficits, an anxiolytic-like behavior, and an       Molecular and Comparative Pathobiology, Johns Hopkins School
increase in locomotor activity. Both posttraining PS-deprived and      of Medicine, Baltimore, MD
pretest PS-deprived groups showed memory deficits; however,            Ethylenediaminetetraacetic acid (EDTA) is a widely used antico-
only the pretest PS-deprived animals presented anxiolytic-like         agulant for blood submission for complete blood count (CBC).
behavior and increased locomotor activity. In experiment III,          Blood tubes contain the appropriate amount of dry (powder)
pretraining PS-deprived rats showed learning and memory defi-          or liquid anticoagulant for the recommended blood volume as
cits, anxiolytic-like behavior, and increased locomotor activity. A    indicated on each tube. Overfilling blood collection tubes results
24-h sleep recovery period after the PSD abolished the learning        in insufficient chelating of calcium and blood clots. Underfilling
and memory deficits, but not anxiety and locomotor alterations.        tubes is reported to result in cell shrinkage and potentially al-
Finally, sleep rebound did not modify acquisition (experiment          tered blood cell parameters and morphology. Many mouse blood
III) and retrieval (experiment IV). This study strengthened the        submissions to our laboratory are less than 20% of the recom-
critical role of sleep (but not sleep rebound) in all the steps of     mended volume for the anticoagulant tubes (40 to 70 µl of blood
learning and memory formation. In addition, it suggests that PSD       in a 500-µl tube). The purpose of this study is to determine the
effects on acquisition and consolidation do not seem to be related     optimal blood volume for EDTA K2 to have confidence in RBC
to other behavioral alterations induced by this procedure.             parameters and CBC results. Cardiac blood samples obtained at
                                                                       euthanasia from 40 adult mice of various ages, sexes, and strains
P185 Glucometer Validity in Mouse Research                             were used to investigate the effect of underfilling anticoagulant
                                                                       blood tubes on CBC parameters obtained with our Hemavet
NA Forbes*, CF Brayton                                                 950 impedance-based blood analyzer. Becton Dickinson 500 µl
Molecular and Comparative Pathobiology, Johns Hopkins School           Microtainer EDTA K2 tubes filled with 50, 100, 175, 250, and 500
of Medicine, Baltimore, MD                                             µl of blood (10%, 20%, 35%, 50% and 100% of the recommended
                                                                       tube volumes, respectively) were analyzed by our Hemavet 950
Blood glucose monitoring is essential to various metabolic
                                                                       instrument, and results were examined by paired t-test. At 10%
studies, especially those of genetic, dietary and therapeutic
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

and 20% of recommended tube volume, hematocrit, WBC, RBC,                separate from the video recording devices. New technology in
and Hb were found to be significantly lower than those with              telemetry which uses time-synchronized video cameras seems
100% recommended volume of 500 µl (P < 0.05). At 35% recom-              to be the solution. Here we describe the use of the synchronized
mended volume, WBC and Hb were significantly lower than                  video function of our validated telemetry acquisition and analy-
at 100% recommended volume. At 50% recommended volume,                   sis system to identify behavior and positional-associated artifacts.
there were no significant differences between values (P > 0.05).         ECG, blood pressure, and body temperature were acquired from
In conclusion, within the parameters of this study relevant to           5 telemetered cynomolgus monkeys (Macaca fascicularis). ECG
submissions to our laboratory, multiple CBC parameters may be            and blood pressure waveforms and body temperature were ex-
impacted by underfilling of EDTA K2 anticoagulant tubes. We              amined in correlation with the time-synchronized videos. Four
currently use and recommend 100-µl anticoagulant tubes, filled           animals exhibited T wave morphological changes associated with
to at least 50% and not overfilled, for mouse blood submissions          positional changes. Noisy blood pressure waveforms were noted
to our laboratory. There are several options for anticoagulants,         in all animals when they scratched with their leg containing the
blood-collecting tubes, and the analyzers, and it is the responsi-       pressure catheter (motion-related artifact). Wet-dog–shaking
bility of investigators to use appropriate protocol and techniques       behavior observed in all animals caused characteristic fluctua-
for optimal and valid results.                                           tions in both ECG and blood pressure waveforms (myogenic and
                                                                         motion-related artifacts). Body temperature was not affected by
P187 New Method for Determining Urine Frequency in Rats                  the animals’ behavior/positioning. Based on our observations,
                                                                         animal behavior and position are important factors to consider
NN Nikolaidis*, R Guinn                                                  in the analysis of telemetry data. A time-synchronized video
R&D, Bioanalytical Systems, Inc, West Lafayette, IN                      system allows for accurate identification of physiological artifact
                                                                         in telemetry data.
Urine is often collected in pharmacological studies in order to
analyze metabolic compounds. These analyses often require
that urine remain chilled during collection. Measurements of             P189 Mouth Cell Collection Device for Newborn Rodents
urine volume and urinary frequency can also provide useful               NK MacLennan*1, Y Zhang1, B Huang1, K Eastman1, M Bedernik1,
data during these experiments, especially in animal models with          LL McCabe1, E McCabe1,2
altered urination, such as diabetes or overactive bladder. While         1 Pediatrics/Human Genetics, 2 Bioengineering, UCLA, Los
measuring urine volume over time is relatively simple, tracking
                                                                         Angeles, CA
urinary frequency can be more difficult. We developed a simple
method to automatically track urine frequency in rats during             For efficient and accurate genotyping of transgenic and knockout
urine collection, while keeping the urine chilled for further me-        mice, the ability to reduce pain and suffering and to obtain DNA
tabolite analysis. Using metabolic cages and funnels, urine was          early in life are critical. We have developed a novel method to
collected into scintillation vials placed into a chilling chamber,       sample buccal cells from neonatal mice to obtain DNA. Our
which chilled the urine to 0 °C. A temperature probe was placed          mouse mouth cell collection process includes an oral speculum
into the vial, and the temperature of the interior was recorded by       and collection device which enables rapid extraction of enough
a computer once every 10 s. Each urinary event caused a transient        DNA for up to 50 PCRs from each buccal sampling. This cell
increase in vial temperature. Eight male Sprague Dawley rats             collection device fills a clear need for buccal sampling from
were placed in metabolism cages. Urine was collected for 24 h to         neonatal mice, greatly facilitating research in mouse models of
provide baseline frequency and volume data. The rats were then           human disease. We have data to support a decreased incidence
injected intraperitoneally with 1 mg/kg oxybutynin (a bladder            of cannibalism of pups by mothers with use of this device.
smooth muscle inhibitor), and urine was collected for another 24         Eliminating the pain and distress caused by traditional cell collec-
h. After a 24-h washout period, the rats were injected intraperi-        tion procedures lessens the potential for confounding variables
toneally with 20 mg/kg furosemide (a diuretic), and urine was            between control and experimental animals. In conclusion, our
again collected for 24 h. Oxybutynin did not significantly affect        mouse mouth cell collection process can be applied to very small
urinary frequency. However, furosemide caused an immediate               animals for which there exists no current device.
increase in urinary output. Both urinary frequency and overall
urine volume increased in furosemide-treated animals. The                P190 Prevalence of Bordetella hinzii in Laboratory Mouse
temperature tracking method clearly indicated the time of each           Facilities in Japan
urination event, and distinguished urinary events as close as 90 s
apart. This method allows urinary frequency to be tracked easily         N Hayashimoto*, M Yasuda, T Ishida, S Kameda, K Goto, A
during a urine collection study, and can provide additional data         Takakura, T Itoh
during metabolic studies.                                                Central Institute for Experimental Animals, Kawasaki, Japan
                                                                         Bordetella hinzii is commensal in the respiratory tracts of fowl, and
P188 Identification of Behavior- and Position-related Artifacts          has been previously shown to have some pathogenic potential
in Nonhuman Primate Telemetry Data Using a Time-synchro-                 in immunocompromised humans. Isolation of B. hinzii from
nized Video Telemetry System                                             nonhuman mammalian species remains extremely rare. Recently,
N Crosby*, Y Mori, K Maulding, J Montalbano, D Moskowitz, E              we isolated B. hinzii from the trachea and lungs of a C57 BL/6
Bou-Abboud, S Meyer                                                      mouse showing sneezing with a chattering noise. The agent also
                                                                         induced respiratory disease in the same way as in the initial case
Scientific Services, SNBL USA, Ltd., Everett, WA
                                                                         in ICR and NOD-SCID mice in our experimental study. Although
In nonhuman primate telemetry studies, abnormal data can be              the etiological role of B. hinzii in mice was suggested by our
observed due to physiological artifacts. Animal behavior and             study, the prevalence of B. hinzii in laboratory mouse facilities
position may cause physiological artifacts ,and should be consid-        has never been reported, and the actual figure is still unknown.
ered in telemetry data analysis. It can be difficult to identify these   The objective of this study is to reveal the prevalence of B. hinzii
artifacts using the traditional telemetry configuration, which is        in laboratory mouse facilities in Japan. A survey was performed
Abstracts of scientific papers 2008 AALAS National Meeting

on a total of 6,040 mice from 1,020 facilities (5,254 mice from 890      Central pain has been well documented in humans and occurs
facilities in university research institutes and 786 mice from 130       with lesions all along the neuraxis involving neural structures of
facilities in pharmaceutical companies) in Japan for microbiologic       the spinothalamocortical pathway. However, most animal models
monitoring at the Central Institute for Experimental Animals             of central pain are based on spinal cord injury. The objective of this
from April 2007 to January 2008. The detection of B. hinzii was          study was to evaluate pain perception in rats following an intrac-
performed by culture of tracheal swabs using horse blood agar,           erebral hemorrhage in basal ganglia. Twelve male Sprague-Dawley
and suspected colonies were identified by phenotypic tests and           rats weighing between 175 and 250 g were used. Two groups of 6
PCR using B. hinzii specific primers designed for this study. In         animals were used to evaluate thermal sensitivity (Hargreave test)
university research institutes, 88 out of 5,254 mice (1.6%) derived      to a progressive intensity of light set at 35 or 50% (groups 1 and 2,
from 24 out of 890 facilities (2.7%) were positive. On the other         respectively) of the maximal output. Following 3 d of behavioral
hand, no B. hinzii-infected mice were found in 130 pharmaceuti-          testing (baseline values), rats were injected intracerebrally with
cal companies surveyed. The overall prevalence of B. hinzii in           0.5 U collagenase Type VII in the right caudoputamen nucleus,
mouse facilities in this survey was 2.4%. No clinical symptoms           inducing a hematoma. They were then tested for the next 7 to
and no gross lesions were observed in any B. hinzii-infected             9 consecutive d. With the light intensity set at 35%, no changes
mice, but moderate sinusitis and mild rhinitis were observed             were observed following surgery when compared to presurgical
in all selected 14 B. hinzii-infected mice free from other known         baseline results. However, at 50% of maximal output, reaction
pathogens by histopathology. Our survey provided evidence for            times were significantly faster on days 3, 4, 5, 6 (P < 0.0001) and 7
the prevalence of B. hinzii in laboratory mouse facilities in Japan,     (P < 0.006). Rats returned to presurgical baseline reaction times on
and indicated the pathogenicity of B. hinzii in naturally infected       days 8 and 9 (P = ns). Histopathology showed that the lesion was
mice. Based on these findings, B. hinzii should be eliminated from       located mainly in the caudoputamen nuclei; however, the internal
laboratory mouse facilities.                                             capsule and the thalamus were also affected. In conclusion, a tran-
                                                                         sient central pain was induced in rats with a collagenase-induced
P191 Quantitative Comparison of Ovarian FSH receptors                    intracerebral hemorrhage of the caudoputamen nuclei most
among Six Mouse Strains                                                  probably associated with lesions of the internal capsule as well
                                                                         as the thalamus. These results suggest that animal models used
O Suzuki*                                                                for studies other than pain, and with lesions affecting structures
Laboratory of Experimental Animal Models, National Institute             along the spinothalamocortical pathway, should be evaluated for
of Biomedical Innovation, Ibaraki, Japan                                 pain perception, as this may have an impact on the interpretation
                                                                         of the results.
Superovulation induction by gonadotropin treatment is widely
used in reproductive experiments. Previously, I showed that the
differential response among various mouse strains to gonadotro-          P193 Spontaneous Tumors in VEGFP-GFP/FVB and Tie2P-GFP/
pins is still a serious problem for practical applications. In order     FVB Transgenic Mice
to elucidate the mechanism underlying the strain difference in           P Huang*, M Dawson, R Lanning, RK Jain, D Fukumura
superovulation induction, I planned to compare the FSH-receptor
                                                                         Radiation Oncology, Massachusetts General Hospital, Boston,
in various mouse strains qualitatively and quantitatively. I have
                                                                         MA
already reported that no difference was found in the FSH-receptor
amino acid sequences among 12 mouse strains. In this study, I com-       The phenomena in our previous observations indicated that the
pared ovarian FSH-receptor mRNAs and proteins among mouse                presence of green fluorescence protein (GFP) reporter trans-genes,
strains quantitatively. Total RNA (5 mice per strain) and proteins       such as VEGFP-GFP and Tie2P-GFP, may affect the incidence of
(4 mice per strain) were extracted from the ovaries of 4-wk-old fe-      spontaneous tumors as well as their histopathological and biological
males of 6 strains: C57BL/6Cr, A/J, DBA/1, DBA/2, C3H/HeJ, and           characteristics in transgenic mice with FVB/N background. In this
BDF1. The FSH-receptor mRNA was quantified using quantitative            study, a total of 94 retired transgenic mice breeders (14 males and
real-time RT-PCR, with normalization by the amount of GAPDH              80 females; 37 of VEGFP-GFP/FVB and 57 of Tie2P-GFP/FVB mice)
mRNA. The amounts of FSH-receptor (approximately 70 kDa)                 from our defined-flora mice facility were observed during their
and β-tubulin (approximately 55 kDa) proteins were measured              natural lifespan. Our preliminary results showed the incidence of
using Western blot analysis, visualizing band fluorescence with          spontaneous tumors was 71.4% (10/14) in males and 57.5% (46/80)
two rabbit antibodies against mouse FSH-receptor and β-tubulin,          in female transgenic mice. Primary tumors found in the males were
respectively, as primary antibodies, and a fluorescent-labeled sec-      3 liver hepatocellular carcinomas, 2 lung alveolar-bronchiolar (AB)
ondary antibody against rabbit IgG. The amounts of FSH-receptor          tumors, 2 lymphomas, and 3 others. In females, pathologic examina-
proteins were normalized with those of β-tubulin proteins. Neither       tion revealed 27 mammary tumors, 3 liver hepatocellular tumors, 3
transcripts (P = 0.54) nor proteins (P = 0.29) differed significantly    lung AB tumors, 3 fibrosacomas, 3 lymphomas, and 7 others. Com-
among strains judged using 1-way ANOVA (P > 0.05). Therefore, it         pared to our wild-type FVB/N retired breeds, these transgenic mice
is unlikely that the strain difference in the response to gonadotropin   demonstrated a significantly lower incidence of spontaneous lung
treatment is due to differences in the structure or abundance of         AB tumors (P < 0.01) and a higher rate of liver tumors (P < 0.01).
ovarian FSH-receptors. As the next target, LH-receptors should be        Examination by confocal microscopy found that GFP was expressed
compared among mouse strains to elucidate the mechanism of the           strongly in fibrosarcoma cells (arose in VEGFP-GFP/FVB), or in
strain difference in superovulation induction.                           adenocarcinoma stromal cells (arose in VEGFP-GFP/FVB and Tie2P-
                                                                         GFP/FVB mice). Several of these tumors have been transplanted
P192 Transient Central Pain following Collagenase-induced                into the 6- to 8-wk-old transgenic and wild-type FVB/N mice. When
Intracerebral Hematoma in Rats                                           these isografts grew in secondary FVB/N mice, the fibrosarcomas
                                                                         continuously expressed GFP in their tumor cells; the adenocarcino-
H Orlando, C Savard, P Vachon*                                           mas brought their own stromal cells (such as endothelial cells) to the
Veterinary Biomedicine, University of Montreal, St-Hyacinthe,            new hosts, and these stromal cells could express GFP and survive
QC, Canada                                                               for more than 2-wk postimplantation. These histopathological and
                                                                                     Abstracts of scientific papers 2008 AALAS National Meeting

biological characteristics of spontaneous tumors arising from the     mutations in Lep/Lepr are rare in the human population, the
transgenic mice are clearly different than those of spontaneous tu-   ZDSD/Pco diabetic (type II) rat model has been designed and
mors in wild-type FVB/N mice. In particular, a significantly lower    developed without these mutations. This rat model was initiated
incidence of spontaneous lung AB tumors in the transgenic mice        by crossing a diet-induced obese (DIO) rat that was derived
was observed. Further studies to better understand the differences,   from the Crl:CD(SD) strain (exhibiting insulin resistance and
such as lung AB tumor incidence, within these spontaneous tumors      polygenic obesity) with a homozygous lean ZDF/Crl rat (which
in transgenic and wild-type FVB/N mice are warranted.                 will express beta cell failure with the Leprfa/Leprfa genotype).
                                                                      Selective inbreeding for obesity and diabetes (19 generations) has
P194 Acute Oxidative Injury Promotes Venous Endothelial               resulted in a strain that exhibits these characteristics without the
Dysfunction without Thrombogenesis in a Rodent Model                  Leprfa mutation. At 6 wk of age the ZDSD males average 217 g,
                                                                      compared to 187 g for the obese ZDF/Crl-Leprfa male rats. Only
RM Moore*1,2, A Hawley2, D Farris2, S Wrobleski2, R Sigler3, D        males develop diabetes, with glucose levels averaging >500 mg/
Myers2,1                                                              dl. The strain also expresses high insulin levels before the onset
1 Unit for Laboratory Animal Medicine, 2 Jobst Vascular Re-           of diabetes (approximately 5 ng/ml), which decrease with the
search Laboratories, Section of Vascular Surgery, University of       progression to overt diabetes to <1 ng/ml. This rat also exhib-
Michigan, Ann Arbor, MI; 3 Walker Downey & Associates, Inc.,          its hyperlipidemia with triglyceride levels of >600 mg/dl and
Verona, WI                                                            cholesterol levels of >100 mg/dl. Micro-albumin excretion is >75
                                                                      mg/d. The majority become diabetic between 15 and 21 wk of age
Oxidative stress has been associated with many cardiovascular
                                                                      on Purina 5008. Dietary manipulation can reliably prompt and
diseases, including endothelium dysfunction. Little is known
                                                                      synchronize the onset of diabetes in animals that are not already
regarding the role of oxidative injury to venous endothelium,
                                                                      diabetic by dietary supplementation with corn syrup (30% solids
and we sought to achieve this using a rodent model. We hy-
                                                                      in an extra water bottle) or a change to Purina 5015 or Research
pothesized that venous endothelial dysfunction subsequent
                                                                      Diets D12468 to synchronize the onset of diabetes. Treatments
to oxidative injury would activate adhesion molecules, thus
                                                                      with metformin (150 mg/kg twice daily); rosiglitazone (3 mg/
promoting venous thrombosis in a rat model. Sprague Dawley
                                                                      kg twice daily) and exenitide (1 µg twice daily) prevent the
rats (n = 72) underwent photochemical injury to the inferior
                                                                      onset of diabetes in this model. In summary, the ZDSD/Pco rat
vena cava (IVC) by injection of Rose Bengal dye (50 mg/kg, IV)
                                                                      is an important new model that expresses obesity and diabetes
and green light laser (540 nm) exposure. Experimental groups
                                                                      without apparent alterations in the leptin pathway. This novel
included controls, shams (Rose Bengal, but no laser), and ani-
                                                                      model appears to more closely mimic the human condition, than
mals that underwent oxidative injury. Groups were evaluated
                                                                      current models, since it is obese and develops diabetes without
at baseline, 1 h, and 1 d post-oxidative injury (POI). Vein well
                                                                      leptin pathway defects. It is thus likely to become more useful
inflammatory cell counts per high power field, gene array (fold
                                                                      than other rat models in testing anti-diabetic and anti-obesity
change relative to aldolase), real-time reverse transcriptase
                                                                      compounds for human obesity and diabetes.
polymerase chain reaction (expression relative to β-actin), tis-
sue inhibitor of metalloproteinase 1 (TIMP-1) levels (pg/mg
total protein), and tissue factor activity (pM/mg total protein)      P196 Alternative Method for Intrathymic Injections in Mice
were evaluated. Polymorphonuclear cells significantly increased       R Blair*2, SL Hoogstraten-Miller1, K Mueller2, P Schwartzberg2
in the 1-h group compared with controls (26 ± 13.5 compared           1Office
with 6.6 ± 4.6 cells/5HPFs, P < 0.05). Gene array and real-time              of Laboratory Animal Medicine, 2Genetic Disease Re-
polymerase chain reaction (RT-PCR) identified angiotensinogen         search Branch, NIH/NHGRI, Bethesda, MD
(AGT) and L-selectin as genes upregulated due to injury, while        The thymus is an organ located in the anterior portion of the
metalloproteinase-14 (MMP-14) was downregulated. Plasma               thoracic cavity, just behind the sternum. The thymus is the site of
TIMP-1 levels significantly increased at 1 h (P < 0.05) and 1 d (P    development of T cells, which are a critical part of the immune
< 0.01) compared with controls POI. Vein wall TIMP-1 levels at        system, required for fighting infection. The widespread use of
1 d were also elevated compared with controls (P < 0.01). Tissue      transgenic and gene-targeted mice has allowed expansion into
factor activity was significantly decreased in both the 1-h (1.95     many important areas in the study of T cell development. Because
± 0.5 compared with 4.0 ± 0.8 pM/mg total protein, P < 0.05)          of its crucial role in immunity, the thymus is a frequent target of
and 1-d (1.5 ± 0.3 compared with 4.0±0.8 pM/mg total protein,         experimental manipulation in the quest for understanding various
P < 0.01) POI groups compared with shams. Oxidative injury to         immune processes in these animals. Our lab wanted to perform
the rat vena cava produces moderate endothelial dysfunction,          intrathymic injection with fluorescein isothiocyanate (FITC) to
characterized by altered gene expression and inflammatory cell        locally label thymocytes and follow the fate of developing T cells
populations. This study is the first to demonstrate that focused      as they exit the thymus and enter the peripheral circulation in
oxidative injury to venous endothelium alters cellular transcrip-     mouse models of primary immunodeficiencies. Intrathymic injec-
tional properties without acute thrombogenesis in this model.         tion has traditionally been performed using surgical procedures
                                                                      to open the chest cavity for direct visualization and injection into
P195 ZDSD/Pco Rat: A Model for Diabetes, Metabolic                    the thymus. This technique subjects the mice to significant pain
Syndrome and Obesity without Leptin or Leptin Receptor                and distress, which can alter T cell development and potentially
Mutations                                                             increase mortality. Intrathymic injections can also be performed
                                                                      by blind injection into the thoracic cavity. However, the success
RG Peterson*, KL Coy, Y Wang, CR Stauss, KA McAndrews, W              rate for this technique is usually low. In order to improve suc-
Yeh                                                                   cess rates with nonsurgical intrathymic injections, our lab has
PreClinOmics, Inc, Indianapolis, IN                                   performed intrathymic injections in isoflurane-anesthetized mice
                                                                      using ultrasound (US)-mediated guidance. The success rate us-
The purpose of this study is to develop an obese-diabetic rat
                                                                      ing this technique has been high, as the needle and injectate can
model that does not have monogenic mutations such as those
                                                                      both be visualized with the US probe as they enter the thymus.
that produce leptin (Lep) or leptin receptor (Lepr) defects. Since
Abstracts of scientific papers 2008 AALAS National Meeting

Complications have been very rare and include the accidental          University, New York, NY; Research Animal Resource Center,
laceration of an intrathoracic vessel, which resulted in the eutha-   Memorial Sloan-Kettering Cancer Center and the Weill Medical
nasia of the mouse; this complication occurred in 1 mouse out         College of Cornell University, New York, NY
of approximately 85 injected to date. Moreover, it is important       Myobia musculi (MB) and Myocoptes musculinus (MC) are 2 of
to note that even though this mouse was euthanized, we were           the most common fur mites affecting laboratory mice. Detection
able to see the immediate hemorrhage on US and euthanize the          can be difficult due to low levels of infestation or inadequate
mouse while it was still anesthetized, thereby avoiding any pain      monitoring programs. During a recent outbreak, we tested the
or distress. Our results suggest that use of US-mediated guidance     sensitivity of our sentinel monitoring program to detect mites. We
is a refinement that can help reduce morbidity, distress and animal   compared skin scrapes (SS) from 3 different sites (head, back, and
usage in experiments involving intrathymic injection.                 ventrum) and found that mites were most prevalent on the head.
                                                                      Contact sentinels, Tac:(SW), tested positive for either MB or MC
P197 Efficacy of Ad Libitum Antibiotics as Countermeasures            within 2 d of exposure to infested mice (C57BL/6NTac). We then
against Acute Radiation Syndrome in C57BL/7 Mice                      exposed naïve Crl:CFW(SW) mice to different concentrations of
A Plett1, R Crisler-Roberts*2, H Chua1, N Nguyen1, ES West1, KJ       mite contaminated/clean bedding (100/0%, 50/50%, 25/75%,
Kohlbacher1, CH Sampson2, CA Johnson2, BE Juliar3, BM Katz3,          2.5/97.5% and 0/100%; groups A through E, respectively).
LJ Brossia2, CM Orschell1                                             Each group consisted of 1 cage with 2 mice. SS were performed
                                                                      weekly on every animal until both mice per cage tested positive
Department of Medicine, 1Division of Hematology/Oncology,             on 2 consecutive wk. Groups A, B, and C tested positive on wk
2Laboratory Animal Resource Center, 3Division of Biostatistics,
                                                                      1, 2, and 3, respectively. Group D first tested positive on wk 7.
Indiana University School of Medicine, Indianapolis, IN               All groups remained positive the following week after the first
Our laboratory has developed a murine model of the hematopoi-         positive test. Group E remained negative. As mite eggs are per-
etic syndrome of the acute radiation syndrome (HS-ARS) for            manently attached to the hair shafts, they persist on the animal
efficacy testing of potential radiomitigators. Since first-line       after successful treatment, potentially confounding posttreat-
treatment after irradiation includes antibiotics and wetted feed,     ment confirmatory testing. Four BALB/cAnNCrl, Crl:CFW(SW),
feed and water consumption were evaluated in irradiated mice          and Crl:CD1(ICR) mice were colored with hair dye and changes
to gauge how to use these as vehicles for antibiotic delivery. To     in coat color were monitored to determine the rate at which
this end, consumption of water from sipper tubes was followed         mice shed fur. After 5 mo, all mice had shed most, but not all of
after exposure of C57BL/6 mice to the LD50/30 dose of 137Cs           the dyed hair. In summary, soiled bedding sentinels are able to
(67 cGy/min). Among our significant findings, we discovered           detect fur mites within a 2-mo testing cycle; the head needs to
that healthy 20 g mice consume approximately 4 ml of acidified        be included as a sampling site, and egg-positive SS taken after
water/d; that by d 1 postexposure, consumption is reduced by          treatment need to be interpreted cautiously.
30% and continues to decrease to about 1.5 ml/d over the next 3
wk; that overall consumption of water from sipper tubes decreased     P199 Toxicologic and Efficacy Testing of Ivermectin-impreg-
by approximately 50% when wetted chow is available; and that          nated Feed for Murine Fur Mite Eradication
consumption of acidified water containing ciprofloxacin after irra-
                                                                      RJ Ricart Arbona*1,2, N Lipman1,2, FR Wolf1,2
diation was similar whether grape flavoring or sugar were added
for increased palatability. With this background, we compared         Tri-Institutional Training Program in Laboratory Animal Medi-
the efficacy of 100 mg/kg/d of ciprofloxacin (Cipro), a broad-        cine and Science, Memorial Sloan-Kettering Cancer Center, The
spectrum fluoroquinolone antibiotic clinically used in neutropenic    Rockefeller University, and the Weill Medical College of Cornell
humans, to that of 125 mg/kg/d of the tetracycline doxycycline        University, New York, NY; Research Animal Resource Center,
plus 300 mg/kg/d of the non-absorbed aminoglycoside neomycin          Memorial Sloan-Kettering Cancer Center and the Weill Medical
(Doxy+Neo) in irradiated mice (n = 912 total mice). Mice were         College of Cornell University, New York, NY
provided 0.67 mg/ml Cipro in sipper tubes and wetted chow or
                                                                      Fur mite outbreaks have been a persistent problem in labora-
2 mg/ml Neo in drinking water plus wetted chow containing
                                                                      tory mouse colonies. Multiple treatment protocols have been
0.0625% Doxy. Thirty-day survival of mice exposed to a range of       published. All are either labor intensive, expensive, or unreli-
doses (775 to 900 cGy, n = 633) was increased in mice provided        able. During a recent outbreak with Myobia musculi (MB) and
Doxy+Neo (P = 0.005), but only marginally in mice provided            Myocoptes musculinus (MC) in a large colony (approximately
Cipro (P = 0.061) compared to untreated mice. Antibiotics tended      30,000 cages), we searched for a treatment modality that would
to increase survival in females to a greater extent than in males.    prove safe, efficacious, not labor intensive, and cost effective. We
Mean survival time (MST) of decedents was increased by 4 to 5 d       compounded and tested irradiated rodent feed impregnated with
in both antibiotic groups (P < 0.0001), although Cipro increased      3 different concentrations (12, 24, 48 ppm) of ivermectin (IVM).
MST more than Doxy+Neo (P = 0.031). These data demonstrate            Postproduction analysis by ESI-MS revealed an approximately
efficacy of antibiotics provided ad libitum in feed and water, and    40% loss of IVM during the manufacturing process. An 8-wk
define important parameters required for the development of           toxicity study was conducted to determine the highest dose that
murine models of HS-ARS that can assess effectiveness of coun-        resulted in the least adverse effects on a C57BL/6NTac mouse
termeasures against radiation.                                        breeding colony. No adverse clinical effects were noted, except
                                                                      1 of 64 adult mice in the 48 ppm feed group developed tremors
P198 Diagnostic Considerations in Murine Fur Mite Detec-              and ataxia. The number of litters, pups born or pups/litter did
tion                                                                  not differ significantly between groups. Pup mortality did not
RJ Ricart Arbona*1,2, N Lipman1,2, FR Wolf1,2                         differ between the control and 12 ppm groups, but increased to
                                                                      77% and 100% in the 24 ppm and 48 ppm groups, respectively.
Tri-Institutional Training Program in Laboratory Animal Medi-         No significant differences between groups were detected by
cine and Science, Memorial Sloan-Kettering Cancer Center, The         CBC, serum chemistry, body weight and gross necropsy. Plasma
Rockefeller University, and the Weill Medical College of Cornell      drug concentrations plateaued (concentration peak was dose
                                                                                    Abstracts of scientific papers 2008 AALAS National Meeting

dependent) at d 7 and decreased to undetectable levels 6 to 9        stress indicators. Metabolism excretion balance studies can in-
d following discontinuation of treatment. We then determined         volve the single housing of rats up to 168 h. The welfare of rats
the efficacy in eradicating mites from a small colony of MB and      in laboratory conditions is important to ensure physiological
MC infested mice using the 12 ppm feed. Based on estimated           and behavioural parameters fall within acceptable ranges. In
mouse food consumption and production-induced drug loss, the         order to assess the level of stress response (physiological and
ingested dose was approximately 1 mg/kg. A 1-wk treatment            behavioral parameters) of the singly housed rat an investigative
schedule was sufficient to eradicate both MB and MC as evident       study was conducted in which group-housed and singly housed
from weekly testing for up to 21 wk. We have determined a            (5 males and 5 females per group) rats were clinically observed
relatively safe dose of IVM-medicated feed to effectively and        daily. Feces was collected at 24-h time points, up to 168 h. Fecal
efficiently eradicate fur mites from a large mouse colony.           glucocorticoid hormone and metabolite analysis is being used
                                                                     as a tool by more animal researchers in the laboratory to provide
P200 Homologous Collagen Implant to Repair a Tracheal Wall           a noninvasive biomarker as an indication of the stress response.
Segment                                                              By subjecting the collected feces samples to solid phase extrac-
                                                                     tion to remove impurities from the matrix, and then using liquid
F Villegas-Álvarez, A González-Maciel, RE Soriano * , B              chromatography-tandem mass spectrometry analysis, it can be
Pérez-Guille, JF González-Zamora, L Padilla-Sánchez, R Reynoso-      seen that the level of stress response in the singly housed rat
Robles, A Ramos-Morales, E Zenteno Galindo, A Pérez-Torres           is higher than the group-housed rat. Levels of glucocorticoid
Experimental Surgery, Instituto Nacional de Pediatria, Mexico        hormone in the singly housed rat is much higher than the levels
City, Mexico                                                         seen in the group-housed rat.
Tracheal stenosis might lead to tracheal malfunction, which
is potentially life-threatening. Among other treatments, the         P202 Transmission of a Bordetella bronchiseptica between
bioprosthesis has been suggested as a tracheal substitute. This      Rabbits and Guinea Pigs
kind of prosthesis self-integrates to the respiratory epithelium     S Pritt*1, RM Bunte2, D Sinha3, E Harvill3
without collapse, inflammatory response or secondary stenosis to     1 Covance Research Products, Denver, PA; 2University Laboratory
maintain tracheal function. The goal of this study was to evaluate
                                                                     Animal Resources, University of Pennsylvania, Philadelphia, PA;
a polyester prosthesis mesh covered with homologous collagen         3Department of Veterinary and Biomedical Science, Pennsylvania
as tracheal wall segmental replacement (TWSR). We conducted
                                                                     State University, University Park, PA
a single-blind study using randomly selected adult Wistar rats.
The prosthesis was created from a homologus group that was           Bordetella bronchiseptica is a Gram-negative coccobacillus consid-
implanted with a subcutaneous polyester mesh. The control            ered to be a veterinary pathogen infecting a number of species
group had no surgical procedure. In the experimental group,          with several strains of varying virulence. While rabbit-to-guinea
we implanted the prosthesis to replace a segment of the anterior     pig transmission is postulated through direct transmission,
tracheal wall. Clinical, histomorphological, and immunochemi-        fomites, or facility ventilation, it has not been demonstrated.
cal postimplant evaluations were performed at 4 different times.     This study examined the transmission of B. bronchiseptica (rabbit
Rats in the experimental group presented with a few respiratory      strain of low virulence) between rabbits and guinea pigs. Twelve
malfunctions during the first week. The intraluminal area and        B. bronchiseptica-free guinea pigs (6 males and 6 females) were
thickness of the bioprosthesis wall in the experimental group was    housed in a conventional facility also housing rabbits colonized
higher (P = 0.001) than the control group at 4 mo; this increased    with B. bronchiseptica. These guinea pigs were isolated in one
thickness was due mainly to angiogenesis, without evidence of        room, and personal protective equipment (hair cover, gloves,
fibrosis or chronic inflammation. At the beginning of the study,     fresh lab coat, shoe covers, surgical mask) was required for entry.
undifferentiated cells were predominate on the implant surface;      Personnel entered the guinea pig room before entering other
however, in the last 2 mo, the epithelial and ciliated cells were    animal rooms. During the study period, no animals became
similar to the controls. At 4 mo postimplant, no histomorpho-        clinically ill. At the end of 30 d, cultures of the nasal cavity, tra-
logical or functional differences were found between groups. In      chea, and lung were negative for B. bronchiseptica. Another 12
conclusion, collagen-polyester homologous prosthesis as TWSR         B. bronchiseptica-free guinea pigs (6 males and 6 females) were
in rats was able to produce proliferation of normal respiratory      inoculated with 5,000 CFU per nostril of B. bronchiseptica upon
epithelium and maintained tracheal function without collapse,        arrival. During the study period, no animals became clinically
inflammatory response, or secondary stenosis.                        ill. At the end of 30 d, 11/12 (92%) animals had positive nasal
                                                                     cavity cultures, while 8/12 (67%) had positive tracheal cultures
P201 Comparison of Fecal Biochemical Stress Indicators in            for B. bronchiseptica. No organisms were retrieved from lung
Group-housed and Singly Housed Rats                                  cultures. Histopathology revealed mild to moderate inflamma-
                                                                     tion in most animals, with a lymphoid infiltrate in nasal cavities
SE Wall*                                                             and heterophilic inflammation in the trachea. This study dem-
Rodent Toxicology, Covance Laboratories Ltd, Harrogate, United       onstrates that guinea pigs may be maintained up to 30 d, with
Kingdom                                                              special precautions, in conventional facilities also housing rabbits
                                                                     colonized with a B. bronchiseptica strain of low virulence and not
It is widely recognized that commonly used laboratory rats in
                                                                     become colonized. Direct inoculation with the strain will result in
general display fewer signs of stress and experience enhanced
                                                                     colonization and inflammation of the nasal cavities and tracheas
welfare when housed in social groupings, compared to in isola-
                                                                     of guinea pigs, but no clinical disease or lung lesions.
tion. Historic stress assessments at our facility have included
quantification of glucocorticoid hormone levels in the blood,
obtained by venipuncture. Developments in analytical tech-
niques have enabled the monitoring of glucocorticoid hormone
in the feces of experimental animals, providing a less invasive
and more readily available means for monitoring biological
Abstracts of scientific papers 2008 AALAS National Meeting

P203 Optimization of Locking Catheter Ports for Rats                    lated and sentinel mice at 6 wk. The cage lid with the partitioning
                                                                        barrier we developed is very useful when sentinel animals are
S   Pawl*,   J Agate, CM Dugan
                                                                        used in the quarantine of laboratory mice, such as genetically
Toxicology Services, Covance Laboratories, Madison, WI                  modified or immunodeficient mice, because sacrifice of valuable
The use of a heparin-dextrose solution to lock catheterized             genetically modified mice should be avoided, and because im-
ports (hep-lock) in rats is common practice in a nonclinical set-       munodeficient mice cannot produce serum antibodies.
ting. Due to the large volumes of animals requiring hep-lock, it
became necessary to evaluate and streamline this process. The           P205 Assessment of RNA Polymerase B Gene Compared with
goal was to evaluate and maximize the amount of time between            16S Gene as a Target for PCR-based Detection of Pasteurella
hep-lock procedures while minimizing the risk of failing to es-         pneumotropica
tablish patency. Different lengths of time between flushing and         VS Dole*, LA Banu, RD Fister, KS Henderson
replacing the hep-lock were evaluated. Replacing the hep-lock
daily, every other day, and every 3, 4, and 6 d resulted in 100%        Research Animal Diagnostic Services, Charles River, Wilming-
patency. The remaining groups tested every 2 and 5 d, which             ton, MA
resulted in 97.6% and 90.9% patency, respectively. The ease of          Currently, DNA-based differentiation of Pasteurella pneumotro-
establishing patency was also evaluated. Harness removal and            pica from other Pasteurellaceae involves PCR amplification of
time required to complete the hep-lock were the determining             the 16S gene region, although some attention has been given to
factors for the optimal procedure. Replacing the hep-lock daily         the RNA polymerase B (rpoB) gene. To assess whether the rpoB
and every other day resulted in no instances of harness removal         gene sequence may serve as an additional diagnostic target,
and took approximately 1 min per animal. All other intervals            partial rpoB gene sequences of various P. pneumotropica culture
resulted in harness removal for less than 20% of animals; how-          isolates were determined. P. pneumotropica culture isolates were
ever, the time required varied. Replacing the hep-lock every 2,         initially identified by microbiological and biochemical techniques
3, and 4 d took approximately 1.40 min per animal; replacing            and further characterized as Heyl or Jawetz biotypes using
it every 5 d took approximately 2.3 min per animal. Although            a 16S-based fluorogenic nuclease PCR (fnPCR) assay. Partial
results showed that performing the lock every 6 d was possible,         sequencing of the rpoB gene (540bp) and the 16S gene (680bp)
this took longer per animal, and it was determined that 2 to 4 d        was done following PCR amplification. The rpoB sequences of
was optimal for maintaining patency and lowering time needed            4 out of 5 isolates identified as Heyl by 16S-based fnPCR assay
per procedure.                                                          were 100% identical to each other. Similarly, rpoB sequences
                                                                        of the 5 isolates identified as Jawetz by our 16S based fnPCR
P204 Lid with Barrier for Partitioning Cage for Laboratory              assay were 99% to 100% identical to the rpoB Jawetz sequence
Animals                                                                 in the GenBank (AY314034). However, alignment of rpoB gene
                                                                        sequences indicated that it will be challenging to develop an
T Kokubo*, T Nishikawa, S Matsushita
                                                                        rpoB-based conventional two primer PCR (gel-based) or fnPCR
National Institute of Radiological Sciences, Chiba, Japan               assay to distinguish between the Heyl and Jawetz biotypes,
Genetically modified laboratory mice are frequently transferred         since the sequence variability between these two biotypes is not
among research institutions. When these mice are quarantined,           clustered in any particular region of the rpoB sequence. Also,
sentinel animals are used to indirectly monitor the health of the       the current sequence information available for the rpoB gene
mice in order to avoid having to sacrifice the valuable geneti-         (our sequencing data and the 324 Pasteurellaceae sequences in
cally modified mice. Recent data on the health monitoring of            the GenBank) does not indicate any conserved regions to dis-
laboratory mice indicate that opportunistic infections are more         tinguish P. pneumotropica from other members of Pasteurellaceae
common than severe contagious infections. During quarantine,            family. In contrast, a total of 1427 16S Pasteurellaceae sequences
test animals and sentinel animals should be placed in the same          are available in GenBank, and the sequence information indicates
cage because opportunistic infections often exhibit low levels          that amongst Pasteurellaceae, the 16S gene is more conserved
of transmissibility. However, it is important to prevent fighting       than rpoB. However, unique island clusters of differentiating
among the animals. Thus, we developed a new cage lid made of            nucleotides in the 16S gene can be exploited for specific PCR
a stainless steel wire screen, with a screen barrier for partitioning   detection of P. pneumotropica. Complete sequencing of the rpoB
a cage into compartments. To evaluate the effectiveness of the          gene from a larger number of isolates from various members of
cage lid with the partitioning barrier, the transmission of cilia-      Pasteurellaceae family may eventually be useful to further char-
associated respiratory (CAR) bacillus was tested. A total of 40         acterize the Pasteurellaceae family.
specific pathogen-free (SPF) mice were used. Sixteen mice were
inoculated intranasally with the SMR strain of the CAR bacillus         P206 Oral Transmission of Experimental Chagas Disease by
under light ether anesthesia. Two inoculated mice were placed in        Açaì Pulp Contaminated with T. cruzi Y Strain
one compartment, and 3 uninfected sentinel mice were placed in
                                                                        VL Dias*1, RL Barbosa2, AR Salgado1, A Gimenes1, M Corat1, AM
the other compartment. Each cage was covered with the lid we            Guaraldo2, LA Passos1
developed. At weekly intervals, the 5 mice were sacrificed by ex-
                                                                               Laboratory—CEMIB, 2Parasitology—IB, UNICAMP,
                                                                        1Genetic
sanguination under anesthesia and were examined. Using a PCR
assay, the 16S rRNA gene from the CAR bacillus was detected             Campinas, Brazil
from swab samples of the larynx and the respiratory organs of           Because of its socio-economic impact, Chagas disease (T. cruzi) is
both inoculated and sentinel mice at 4 wk. Histopathologically,         the most important parasitic infection in South America. Despite
CAR bacilli were shown on the surface of the epithelium of the          the progresses in vector transmission control, new specific cases
respiratory tracts of both inoculated and sentinel mice at 4 wk.        showed the oral route as an important factor for disease dis-
Typical bronchopneumonia and peribronchitis were observed               semination. Recent outbreaks that arose in north Brazil indicate
in both inoculated and sentinel mice at 5 wk. Serologically, IFA        likely contamination of açaí pulp, a highly nutritional fruit that
antibodies against the CAR bacillus were detected in both inocu-        serves as a major food source for the poor people of this region.
                                                                                        Abstracts of scientific papers 2008 AALAS National Meeting

However, no data has been reported regarding outbreaks of T.            compared with baseline). During fluid resuscitation, MAP
cruzi in people through consumption of açaí pulp. Thus, in this         increased to 95 ± 5 mmHg (P = NS compared with baseline)
work, we evaluated the survival of T. cruzi in the açaí pulp and        and PETCO2 increased to 33 ± 3 mmHg (P = NS compared with
the potential for oral contamination through açaí pulp consump-         baseline). Landrace/ Large White swine are suitable models for
tion. Tripomastigotes (1×105) of T. cruzi Y strain were extracted       hemorrhagic shock. Various parameters of acute blood loss can
from the blood plasma of infected mice. The plasma was mixed            be recorded in this model.
at a 1:4 proportion in the açaí pulp and kept at room temperature
for 28 h. The vitality of the parasite was checked at 6, 8, 12, 24,     P208 Assessment of QTc Formulas in Conscious Dogs with
and 28 h; it was observed directly in 5 μl of mix with trypan blue      Pacing-induced Heart Failure
on optical microscopy and in the eluate obtained from a forced
straining. Parasite movement was classified as greatly active           A Kijtawornrat1,2, Y Panyasing*1,2, JL Lolly2, JJ Schmidt2, Y Nishi-
(GA), active (AT), and slow (S). The results demonstrated that          jima2, BM Roche2, RL Hamlin1,2
at 6 h, 100% of parasites were GA; at 28 h, 67% were GA and             1The Ohio State University, Columbus, OH; 2 QTest Labs, LLC,

33% were S. To confirm the hypothesis of oral transmission, 100         Columbus, OH
μl of T. cruzi mix was administered either intraperitoneally, per
                                                                        Drugs given to people with naturally occurring diseases have been
os, or by gavage to 10- to 12-wk-old immunodeficient female
                                                                        shown to be more toxic than when given to those without disease;
mice (CB17scid/Uni); plasma from infected animals was used
                                                                        therefore, it appears reasonable to search for potential toxicity in a
as a positive control and negative control animals received only
                                                                        model possessing a pathophysiological substrate known to sensitize
fruit pulp. Animals were observed for 40 d, and infection was
                                                                        humans to toxicity. Since a search for potential effects of drugs on
confirmed by the Brener method. Death was noted at d 14 for
                                                                        ventricular repolarization is so critical, models possessing heart
positive control animals dosed intraperitoneally and by gavage
                                                                        failure (HF) have become increasingly popular to investigate poten-
and at d 18 for those dosed per os. For the test group we ob-
                                                                        tial toxicity. Prolongation of heart rate (HR) corrected QT interval
served death at d 24 in animals dosed per os; no negative control
                                                                        (QTc) is one of the surrogate markers for potential torsadogenicity
animals died. These results showed the pulp does not interfere
                                                                        of a test article. This study was conducted to determine which cor-
in the parasite’s capacity to cause disease; rather, it seemed only
                                                                        rection method best accounts for the effects of changes in the RR
to delay infection for 6 to 8 d when orally administered.
                                                                        interval on the QT interval of conscious, sling-trained dog with
                                                                        HF. Bipolar, transthoracic ECGs were obtained from 18 conscious
P207 Developing a Hemorrhagic Shock Model in Landrace/                  dogs at baseline and after 12 wk of rapid ventricular pacing. RR
Large White Swine                                                       and QT intervals were measured and QTc were calculated using 3
T Xanthos, X Balkamou*, S Konstantinos, L Papadimitriou, I              common algorithms (Bazette, Fridericia, and Van de Water). After
Dontas                                                                  12 wk of pacing dogs with HF confirmed by reduced LV shortening
                                                                        fraction had significantly slower HR (P < 0.01) and significantly
Experimental Surgery and Surgical Research, University of               longer QTc (P < 0.01) when compared to baseline. The following
Athens, Athens, Greece                                                  regression equations were related QTc to RR: QTc(V) = 239.4–2.68
Shock following acute blood loss is a daunting medical emer-            RR, r2 = 0.00 for the healthy dog; QTc(F) = 258.4–0.01 RR, r2 = 0.01
gency. Various organs are affected due to the compromised               for the dogs in HF. The Van de Water method demonstrated least
oxygen delivery. The aim of the present study was to describe           dependency of QTc on RR interval in conscious, healthy dogs,
the model of hemorrhagic shock developed in our department.             but the Fridericia method was best for correcting dogs in HF. In
Twenty Landrace/ Large White swine with an average weight               conclusion, the rate-corrected QT formula of Fridericia appears
of 19 ± 2 kg were included in the present study. The animals            to be useful as a biomarker for assessing effects of test articles on
were premedicated with ketamine hydrochloride (10 mg/kg),               ventricular repolarization in dogs with HF.
midazolam (0.5 mg/kg ) and atropine (0.05 mg/kg). Anesthesia
was induced with propofol (2 mg/kg) cis-atracurium (0.15 mg/            P209 Food Restriction Changes Physiological and Pathological
kg) and fentanyl (2 mγ/kg). Intubation followed and the swine           Parameters in Zucker Diabetic Fatty Rats
were ventilated with FiO2 21% and with a tidal volume 12 ml/
kg. The respiratory frequency was adjusted in order to maintain         Y Kirihara*1, M Takechi1, A Ago1, S Kaidzu2, Y Notsu3, Y Saito4,
PETCO2 = 35 to 40 mmHg. The left carotid artery was surgically          M Ito5, T Yamada1
prepared, and an arterial catheter was advanced in the thoracic         1Experimental  Animals, Center for Integrated Research in Sci-
aorta. The right internal jugular vein was also surgically pre-         ence, 2Ophthalmology, 3Medical Technology attached to School
pared, and a Swan-Ganz catheter was inserted for measurements           of Medicine, 4Anesthesiology, Shimane University, Izumo, Japan;
of the right atrial pressures. The animals were left to stabilize for   5Technical Center, Charles River Japan, Aikougun, Japan

30 min following surgical manipulations. Afterward, 50% of the
                                                                        ZDF rats (Charles River, Japan) are a disease model of human
total blood volume was drained over a period of 20 min. Animals
                                                                        type 2 diabetes. It is well known that dietary restriction is the
were left in shock for 90 min. Lactated Ringer solution (20 ml/kg)
                                                                        first therapy for human diabetes. However, there is no data about
was then infused over 20 min. Animals were monitored continu-
                                                                        effects of food restriction in ZDF rats. Therefore, we examined the
ously for a period of 4 h. The animals were then euthanatized
                                                                        effects of 80% food restriction on physiological and pathological
with 2 g of pentobarbital. All animals were routinely examined
                                                                        parameters in ZDF rats. We also examined the relationship of
postmortem to record any underlying pathology and for organ
                                                                        ZDF rats to diabetic retinopathy and neuropathy. We used ZDF/
harvesting. Mean arterial pressure (MAP) was 100 ± 12 mmHg
                                                                        CrlCrlj-Leprfa/Leprfa (Fatty) rats for a diabetic model and ZDF/
following surgical manipulations. PETCO2 was 35 ± 4 mmHg.
                                                                        CrlCrlj-?/+ (Lean) rats for control. We divided the rats into four
Hemoglobin was 10.3±0.8 mg/dl. Following the hemorrhagic
                                                                        groups: group A (n = 7), Fatty rats given free-feeding; group B (n
phase MAP declined to 45 ± 12 mmHg (P < 0.01 compared with
                                                                        = 7), Fatty rats given 80% food restriction; group C (n = 8), Lean
baseline), PETCO2 declined to 26 ± 3 mmHg (P < 0.05 compared
                                                                        rats given free-feeding; group D (n = 8), Lean rats given 80%
with baseline) and hemoglobin was 6.6 ± 0.9 mg/dl (P < 0.001
                                                                        food restriction. When rats were 6 wk old, food restriction was
Abstracts of scientific papers 2008 AALAS National Meeting

started. Every day, we measured food consumption of groups            P210 Monitoring of Infectious Bacteria in Poultry
A and C, then gave 80% of that amount of food to groups B             D Sullivan*, A Byers, W Willis, J Allen
and D the next day. The experiment was conducted for 30 wk.
Body weights were measured once a week. We analyzed GOT,              Animal Science, North Carolina A&T State University, Greens-
GPT, T-Cho, HDL-Cho, LDL-Cho, TG, BUN, glucose (Glu), and             boro, NC
glycated albumin (GA). Groups A and B were given a glucose
tolerance test (GTT). To evaluate pain thresholds, a paw pressure     Determination of the health status of the flock and offspring has
(PP) test was used. We also measured electroretinogram (ERG).         traditionally been accomplished by analysis of serum antibod-
Statistical analysis was performed using ANOVA with repeated          ies. This study compared the antibacterial activities of chicken
measures, followed by a Scheffe test. Body weights of the groups      antibody in egg yolk compared with serum in the detection of
given food restriction were significantly lower than those of the     Salmonella typhimurium, Campylobacter, and Salmonella enteritidis
groups fed ad libitum. There were no significant differences          for monitoring the health of flock. Salmune, a live avirulent strain
between groups C and D in laboratory data. Group A showed             of Salmonella typhimurium, is typically given to birds at d 1 and
significantly higher To-Cho, LDL-Cho, BUN, Glu, and GA than           wk 6, 10, and 18 to vaccinate the flock against S. typhimurium
the other groups. Group A also showed significantly higher Glu        and S. enteritidis. Samples of egg yolk and sera were collected for
in the GTT than that of Group B. Group A indicated significant        5 mo from 15 broiler breeder hens previously vaccinated with
lower pain thresholds compared to the other groups on the PP          Salmune. Antibody specificity to these 3 bacteria was determined
test. There were significant differences between Fatty rats (groups   by western blot assay. The nitrocellulose membranes were probed
A and B) and Group D in the ERG data. However, all rats in            with purified IgY antibody isolated from the serum and egg yolks
Group A developed cataracts by wk 30. The results indicated           of each hen. At wk 27 and 36, all 15 hens had IgY titers in their
that food restriction changed physiological and pathological          serum and egg yolks specific for S. typhimurium, S. enteritidis and
parameters in ZDF rats. ZDF rats might be a useful model for          Campylobacter. These studies show that antibodies to specific bac-
diabetic retinopathy and neuropathy.                                  teria present in both serum and egg yolk samples from vaccinated
                                                                      hens can be detected by western blot assay. The analysis of egg
                                                                      yolk can be a rapid and effective alternative method for screening
                                                                      flocks for the presence of antibodies to specific bacteria.