Microbial Communities in the Nasal Passages of Healthy and URTD Tortoises
Ashley M. Ordorica, F. Harvey Pough, and Charles E. Deutch
Department of Integrated Natural Sciences, Arizona State University West, Phoenix AZ
PO Box 37100 Phoenix, AZ 85069-7100 Phone: (602)543-6939 FAX: (602)543-6073
Natural communities of microorganisms are believed to
• The total number of bacteria/ml in the nasal passages of desert
provide an important barrier against potential pathogens. We Strain Colony Original % Shape Gram BBL Crystal™
Color Number Stain Identification
tortoises changes seasonally, increasing from the spring to the
characterized the microbial communities in the nasal passages of summer and decreasing in the fall.
desert tortoises (Gopherus agassizii) quantitatively to determine if Ga 1 Rio 294.5.1 89 coccus + unknown
there are individual or seasonal variations, or differences between Fleece 559.6.1 26.9 • The microbial communities of both healthy and URTD tortoises
healthy tortoises and those with symptoms of upper respiratory 607.6.1 51.5 are dominated by pigmented Gram-positive cocci.
tract disease (URTD). Twelve captive tortoises at the Adobe 294.6.1 100
• Communities in URTD tortoises are more variable and complex
Mountain Wildlife Rehabilitation Center in Phoenix, AZ were than those in healthy tortoises.
divided into four groups: three healthy tortoises were sampled
• Thermal tolerances of bacteria showed optimal growth at about
monthly, three URTD tortoises were sampled monthly, three
healthy tortoises were sampled bimonthly, and three healthy Ga 2 Lemon 624.5.4 50 coccus + unknown
tortoises were sampled once. At each sampling time, the tortoises • Seven out of the twelve most frequent isolates have been
Head 559.6.4 1
were examined for general health, and both nares were probed 559.7.3 20.9 identified as Staphylococcus xylosus, Micrococcus species,
with moistened sterile swabs. The bacteria on the swabs were 294.7.2 2.8 Kytococcus sedentarius, Corynebacterium
suspended in sterile saline, serially diluted, and plated on tryptic 624.7.2 41.4 pseudodiphtheriticum, Staphylococcus vitulinus, and
soy agar medium. Total bacterial counts/ml varied from tortoise to Staphylococcus sciuri.
tortoise, increasing from May to August and decreasing from • Some of the frequent isolates tested showed resistance to
Ga 3 Sun Rise 270.6.3 18.9 coccus + Staphylococcus
September to November. Total counts were usually higher and RESULTS 270.7.5 <1 xylosus
antibiotics including Bacitracin, Rifampin, Nalidixic Acid,
more variable in URTD tortoises. The microbial communities Norfloxacin, Vancomycin, Penicillin, and Triple Sulfa.
were dominated by pigmented Gram-positive cocci, but Gram-
positive rods, coryneforms, and Gram-negative rods also were
found. The proportions of different bacteria varied from month to G roup 1 -Healthy Tortoises Ga 4 Falling 360.6.2 24.1 coccus + Micrococcus
month and were more variable in diseased animals. These studies Percent
TotalCount Star 360.7.1 10.4 species FUTURE STUDIES
suggest that a broader ecological and microbiological analysis of
these communities would be valuable. 100% 00E+08
Ga 5 Vanilla 294.6.2 <1 coccus + Kytococcus • Determine salt tolerances of bacterial isolates
1. Cream 352.7.6 8.6 sedentarius
• Determine the nutrients found in nasal passages
INTRODUCTION 60% • Confirm identity of bacterial isolates using BBL Crystal
One factor that limits the susceptibility of any organism to an 40% System and FT-IR spectroscopy of pigments
infectious agent is the presence of a community of nonpathogenic 00E+05
microorganisms at the site of infection. Indigenous bacteria in the 20%
respiratory system, the gastrointestinal system, and the urogenital 0% 1.
00E+04 Ga 6 Lemon 352.5.1 26 coccus + unknown
system are well adapted to these sites. They protect the host from
N ovem ber
Head 352.6.3 9.8
S e p te m b e r
O c to b e r
J u ly
other potentially pathogenic organisms by occupying physical sites 559.10.2 51.5
on the host tissue, by effectively consuming the available nutrients, 607.10.2 16.7 ACKNOWLEDGEMENTS
and by producing proteins and other metabolic products that are
inhibitory to other microorganisms. Little is known about the Ga 7 Squash 026.11.2 30 Coryn- +/- Corynebacterium
bacteria in the upper respiratory tract of desert tortoises (Gopherus 360.7.3 16.2 eform pseudodiphtheriticum We would like to thank Daniel Martinez, Jenny Gomes, and
G roup 2 -URTD Tortoises Albert DiOrio for their work on some of the experiments and
agassizii). These reptiles are of interest because the Mojave Desert 624.9.2 <1
population in California, Nevada, and Utah is classified as Sandy Cate of the Arizona Game & Fish Department for
Threatened by the U.S. Fish and Wildlife Service, and the Sonoran Percent TotalCount providing the captive desert tortoises. Support for this research
Desert population in Arizona is protected by state regulations was provided in part by a grant from the National Institute of
1. Health through the Minority Access to Research Careers, ASU
(Howland and Rorabaugh, 2002). One factor that has impacted
Ga 8 Cream 607.9.1 100 Coccus + Staphylococcus School of Life Sciences Enrichment Program.
these organisms is the occurrence of Upper Respiratory Tract 80% Cake 294.9.8 <1 vitulinus
Disease (URTD, Berry, 1997), which is now known to be caused by 352.9.1 43
Mycoplasma agassizii (Brown et al., 1994). Dickinson et al. (1996, 60% 811.9.1 70.1
2001) reported the recovery of bacteria from the generic groups 00E+07
1. 026.9.1 47.4
Corynebacterium, Flavobacterium, Pseudomonas, Staphylococcus, 40% REFERENCES
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J u ly
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PROTOCOL BIOLOGICAL G roup 3 - Heal Tortoi
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Fleece 559.8.2 17.5 sciuri
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•Swab both nares with sterile •Bacterial shape, size, and motility 00E+06
1. 607.10.2 16.7
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N o ve m b e r
S e p te m b e r
0.85% NaCl oxidase activities Star maltophila
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Ga 15 Angel 360.9.5 1 bacillus + unknown
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