Observations During Concurrent Anaplasmosis and Babesiosis in .pdf by liningnvp


									   may account for the removal of homologous
   transfused cells that may contain subtle foreign                              1.Love, J. l. and Wiygul, G.: Free Amino Acids In Plasma, Red
   antigens (altered cells) undetectable by existing                         Cells and Anaplasmata of Parasitized Calves. Comp. Biochem.
                                                                             Phy-ioL, 30, (1969): 923-929. -
   serologic tchniques.                                                                                       2. Darre, D. L, Wallace. W.R.and
                                                                             Dimopoullos, G. T.: Evidence of Lactate Dehydrogenase
      The apparent reduced susceptibility of HbF cells                       Anaplasma marginale. J. Blacteriol., 93, (1967): 806-810. - 3.
   to A. marginale infection opens a field of                                F. R. and Dimopoullos, G. T.: Demonstration of an Autoantibody
   interesting speculation regarding strain resistances                      Associated with Anaplasmosis. Am. J.Vet. Res., 33, (1972): 73-76.
                                                                             - 4. Schroeder, W.
   hinging on cellular components such a hemoglobin                                                F. and Ristic, M.: Blood
                                                                                                                                  Serum [-actor
                                                                             Associated with Erythrophagocytoss in Calves with Anuplusmosis.
   types, enzymatic factors, ion makeup, etc.                                Am. J. Vet. Res., 29, (1968): 1991-1995. - 5. listic, St. Lykins,
      The continuing reports of new data in the area                         D. and Morris, 11.R.: Anaplasmosis Opsonins and llernaglutinirn J.
                                                                             Etiology of Anemia. Exptl. Parasit., 31, (1972): 2-12.            in
   of immunopathology is encouraging to the research                                                                                       - 6.
                                                                             Anderson, I. L., et al.: Anaplasnla marginale: Hemoglobin Patterns
  community and with each comes a more complete                              in Experimentally Infected Young Calves. Exptl. Parasit., 32,
  understanding of the pathogenesis of the disease                           (1972): 265-271. - 7. Williams,
                                                                             Transfusions During Patent llovine E. 1. and Jones, E. W.: Blood
  through reevaluation of previously obtained data.                          29, (1968): 703-710.
                                                                                                                Anaplasmosis. Am. J. Vet. RCL,

             Observations During Concurrent

               Anaplasmosis and Babesiosis

            in Experimentally Infected Calves

 D. E. Corner, D. V.M., M.S., and L. G. Adams, D. V.M., Ph.D.
 DepartmentofPathology and the Institute of
 Tropical Veterinary Medicine
 College of Veterinary Medicine
 Texas A&M University
 College Station, Texas 77843

                            Introduction                                    Other investigatord, studying concurrent infection
    Thte common distributinn of anaplasmosis and                            in experimentally infected splenectomized calves,
 babesiosis and the high incidence of concurrent                            reported that only one of the hemoparasites
infection of cattle with Anaplasma and Babesia                              established the normal expected parasitemia (16).
species has been noted by numerous investi-                                 The invasion of erythrocytes by the second
gators (5,6,14,15,17,24,26). In many tropical                               hemoparasite, though initially substantial, decreas­
and subtropical regions, the high incidence of                              ed and occasionally ceased prematurely. It was
concurrent infection has resulted in the standard                           suggested that a non-antibody inhibiting factor
procedure of premunizing cattle against both of                             may have been produced which allowed the
the diseases (14,15,26,29). Descriptions of the                             hemoparasite with the highest established para­
clinical course and pathological manifestations of                         sitemia to prevail while further infection of
concurrent infection have been few. Simultaneous                           erythrocytes by the second organism was inhibited.
premunization against both ,naplasmosis and                                Splenectomized calves infected with B. bigemina
babesiosis was reported to produce an intense                              during the convalescent stage of anaplasmosis were
anemia (6). The severity of the anemia was                                 reported to have abnormally low Babesia para­
attributed to the additive effect of the individual                        sitemias and a prepatent period which was from 16
anemias caused by each of the hemoparasites.                               to 21 days longer than that normally observed for
Supported by linancial assistance from the Agency for International
Development and The Rockefeller Foundation. The work was done
                                                                           babesiosis (27).
at the Laboratorio de Investigaciones
                                                                              The common distribution of anaplasmosis and
Colombia, with the                    Medicas Veterlnarias I
                    support and coopcrition of The Instituto               babesiosis and the high incidence of concurrent
Colombiano Agropecuario.
                                                                           disease has indicated that a clearer understanding

of the relationship of the two disease agents during                technique using cellulose acetate membranes (4).
concurrent infection is desirable. The purpose of                   Serum glutamic oxalacetic transaminase (SGOT)
this investigation was to describe the clinical                     activity was measured by a kit procedure*
course, serological response and pathological                       modified from the original method (19). Serum
manifestations of concurrent infection in intact                    soibitol dehydrogenase (SD) activity was measured
calves experimentally infected with A. marginate                    by a modified kit procedure** adapted from an
and B. bigcmina and to study the relationship of                    original method (7). Serum biirubin concentra­
the two organisms during the concurrent infection.                  tions were determined by a modification of the
                 hi.tcniak and hicihods                             Van den Bergh procedure (8).
   Twenty-two,          male,     non-splenectomnized,                 Serum samples were screened for the presence of
Holsteii .-Friesian calves seven months of age and                  complement fixing (CF) antibodies for A.
free of previous or current Anaptusna and Babesia                   marginale by the standardized CF test for
 feon    preousedcuor         erenAntal   andmal. T                 anaplasmosis (2). The CF test employed to detect
infection were used as experimental animals. The
calves were randomly divided into four groups.                      antibodies against B. bigemina was a previously
Groups I and 11 each contained three calves and                     described modification (25) of an original method
Groupsed and        ach
                  cont              hrcalves and
                             ainedroup                              (12). All serum samples found to be positive by the
served as control groups for anaplasmosis and                       screening procedures were titrated by a micropro­
babesiosis, respectively. Groups III and IV each                    cedure technique (9). Antigen used in the CF test
contained eight calves. The calves of Group III                     for anaplasmosis was obtained from the United
were infected on day 0 with A. marginate.                           States Department of Agriculture. Antigen used in
Following recovery from acute anaplasmosis, the                     the CF test for babesiosis was obtained by a
calves were infected with efect
                       f B. bigemina cocurent
               stud th asecndar and served to                       previously reported technique using a distilled
study the effect of a secondary concurrent                          water extract of the parasite suspension (13).
infection with B. bigemina in calves recovering                       Rib bone marrow specimens were collected by
from anaplasmosis. The calves of Group IV were                      biopsy from two calves from each of the four
infected on day 0 with B. bigemina. Following                       groups of calves on days 0, 50, 90 and 120 of the
recovery from infetedwit
                 acute babesiosis, the calves were
                    argnaiA.andb~lvedto sudythe                       vestigation. Myeloid:erythroid (M:E) ratios were
infected with A. marginaeand,             rved to study the         determined.
effect of a secondary concurrent infection with A.
                        in clve rombabsiois.Two
                                 reoveing                             Trmind.concurrently infected calves from each of
marginate iGroups                                                            III and IV and a non-infected control calf
    Stabilates prepared from purified Colombian                     were euthanatized by electrocution and submitted
isolates of A. marginate and B. bigemina were used                  to gross and histopathological examinations.
 in the investigation. Each calf was infected by the
 subcutaneous inoculation of stabilate which was                                                 Results
 thawed in a water bath at 380 C for one minute.                       The clinical manifestations of infection were
 The Anaplasma inoculum contained approximately                     most severe in the concurrently infected calves, less
 9.5 x 108 marginal bodies and the Babesia                          severe in the Anaplasma controls and least severe in
 inoculum 10.5 x 108 organisms.                                     the Babesia controls. The clinical signs of infection
                                                                    consisted of increases in rectal temperature,
  The body weight of each calf was determined on                    inactivity, dull haircoats and poor body condition.
day 0 and at the termination of the investigation                   Mean total weight gains and average daily gains
and the mean total weight gains and average daily                   were     moderately       lower        in   the   concurrently
gains for each group of calves was -letermined.                     infected calves than in the Anaplasma controls and
Blood and serum samples were collected and rectal                   markedly lower than in the Babesia controls (Table
temperatures determined twice weekly from each                      1).
calf for two weeks prior to infection and for 17                       Decreases in the mean PCV were more marked
weeks following infection,                                          in the concurrently infected calves than in the
   Packed cell volumes (PCV), reticulocyte counts                   control calves (Figure 1). Mean low PCVs of 13%
and total and differential leukocyte counts were                    and 20% were observed in the calves concurently
determined by tandard hlboratory procedures                         infected with B. bi.g,,ia and A. marginate,
(23). The perc(ntage of parasitized crythrocytcs                    r(.pectively, as compared to mean low values of
'vns determined by the microscopic examination of                   27% and 22% in the Babcsia and Anaplasma
Gicmsa stained thin blood smears. Total serum                       control groups.
protein (TSP) concentrations were ascertained
using a hand held refractometer (3). Serum protein                  *Sigma Chemical Co., St. Louis. MissourL
dectrophoretic patterns were determined by a                        "Sina Chemical Co.. Si. Lous, Misouri.


                              Table 1

 initial and final wdghts with total weight Pins and averap daily 
                                                              ' .
 pins of the four groups of calves bigemin.
 Anaplame margiale and/or Babesi experimentally Infected with
                                                Group meant and                                                                  o"'*.


 standard deviations from the mean are presented.j                                                                                               ('                                   "

    cow"       ~
               8.i.i .0
               C 1- s P ,
                     w .     .161.1
                            V. 10 0 !( b .)
                            b•       1.o      Y
                                               . 10l~ 0 1 . )
                                               IV     l).t~
                                                    .g Ci.
                                                                      C 4
                                                                          .   4.2
                                                                                   .1     Itol ". O.,
                                                                                            e fl
                                                                                           ipMSI                                                        % ,q I
                                                                                                                                                         ~llf t                  ,    I

                            Il~~..* 1               2. •                ,, i~e
                                                                            •                 i       4iSe                                                -i

      II            I         hutM                   6.2)t            hill$
     322            8                                                                         O.21h0.l
                                all2                     h            14n1                "       .lJsO.­

     ,i             J i 
           24ei,   ~iP-P.                                             ft-     .6
                                                                                   4222           .264,.22                     Ie 
                    t.tl            -U*"

                                                         IHO    So.   441                                                                                         ".44.o.

    Wi                                              62                  22                        eOW

                                                                                                                    R°greZ Te          ercntde    ofpans/rted           rybrcyzs          fite   ou
                                                                                                                    goups of calves cx pea$lmenfgailyl(I inece       wihAnpamamrgnl

                                                                                                                    and/orBabes/a bigemina. Group means are presented and vertical
                                   ?' * 
                                                                           lnes represent range. Encircledletters Indicate lime ofsubcutaneous
                                ________________                                                                    nocutlation withAnaplasmna marginaale or wit/h
                                                                                                                                                                Babeuia bigenaina

                                   J ' 
                                                                            calves. A moderate monocytosis occurred in each
                                                                          of the four groups of calves.
                                      i       "The
                                              " ,,                                                                            changes which occurred in the SGOT, serum
                                                                                                                   SD and serum bilirubin concentrations were not
                                   ,                                                                               significantly different between the concurrently
             J                                                                                                     infected and the control calves.
                                              ?       ?                                                               The M:E ratio steadily decreased in each of the
                                   Z                  [,1                          ,                               four groups of calves and was substantially lower,
                                   J--            ,,,,,
                                                    ,                                   ,at                            the termination of the investigation, in the
                                  T-                                                                               concurrently infected calves than in the control
                                           ,9            1calves.                                                           The number of reticulocytes was observed
                                           .. ....
                                          . ..... .... ...
                                                        to increase moderately higher in the concurrently
                                                           infected than in the control calves.
Flgwe) . Fluctuation, in thac pocked ce/i volumes of tiae fow' 570 ups                                                Total serum protein concentrations increased
Of calves experimentally infected with Alnaplasma marginnle and/or                                                 sutailyhgerntecournlynftd
 ttabes/a bigem i a ,Group teans are presented mld vertical lines                                                   su ta ial      h g er n th co ur n ly nf t d

represent ranag.    .n7circtcd tetten iadicate time of sabcutanaeoaas                                              and in the Antaplasmna control calves th~an ;n the
inoculation! wit/a 1Aaaptazaau ,,wrgitaak or wit/a /Jabwiia lic'ahta.                                              Babe, i° controls. 'lhl             avernge               serum albumin
                                                                                                                   concentration and thre A:G ratio decreased in each
   Significant increases in the mean number of                                                                     of the four group)s of calves with little difference
infected erythrocytes were observed in the                                                                         occurring between the concurrently infected and
concurrently infected calves (Figure 2). Mean                                                                      the control calves.
maximum parasiternias of 1.0% and 2.2% were                                                                           Serum globulin concentrations increased in each
observed in the calve~s concurrently infected with                                                                 of the four groups of calves and were substantially
B. bigetnina and A. mnargitraIt, respctively, while                                                                higher in the concurrently infected and in the
the mean maximum J)arasitemias observed in)      the                                                               Anaplasmna controls than in the Ilabesia controls
Jiabe, ia and Anuluhsnia controls were 0.06%, and                                                                  (Figure 3).
1.4%, respectively.                                                                                                   Significant differences in the mean maximum
   No differences occurred in the total number of                                                                  CF antibody titers between the concurrently
leukocytes n)or in the differential leukocyte counts                                                               infected and the control calves were not observed
between the concurrently infected und control                                                                      (Figure 4). Mean maximum CF titers of 1:100 and


                                                                             calves concurrently infected with Babesia were
           •      -:150                                                              and 1:170. re. pectively.
                                                                                Gross lesions observed in the concurrently
          1:*      '. 
       4                                              infected calves which were euthanatized for
                                                                             dissection included: an excessive amount -)f
                                                        yellow fluid in the peritoneal and pleural cavities;
                                                                             moderate splenomegaly with prominent malpighian
                                                                             follicles; moderate hepatomegaly and lymph node
                                                                             enlargement; and the occasional serous atrophy of
                                                                             depot fat.
                                               I   "'--,-                       Tissue sections from the concurrently infected
                                                                             calves were grouped together by organ and
                                    ...                                      examined microscopically:
                                                                                Liver. Slight to moderate centrolobular degen­
                                                                             eration of the hepatocytcs was obsr.-rved. A slight
                                                                             to moderate infiltration of lymphocytes and
                                                                             reticulondoth'liial ccllm wm observcd in the
                                                                             peripor1d areas. The hepatic sinusoids contained
                                                    . .an 
                      excessive number of mononuchnr inflammatory
                                                                             cells and immature nucleated erythrocytes.
   iR J. 77,,onc,,,a,,,,, of alpho. hla and pi,,a s,'o,,
            c                                                                Kupffer cell hypertrophy, erythrophagocytosis and
Wiabul/s of the lor grnoti, of co'h-cx orplerhntistaly litcled 1 lh
Alnaplaiaa ,narglnak ahml/or Iah'.ia hlgncuoia. ;roup   nrm  tiare           phagocytized granules of hemosiderin were occa­
p           a
       hfrieiuld 'd
                  'rlehal !hia'n'prcinil rmuge. E'clrdeld let'er
Indlcatc linie of subaitai'us hionslollost wills Am~plame                    sionally observed.
Irdkai or witho       siia h   igent       lola.                                Kidney. An cosinophilic proteinaceous material
                                                                             was observed in Bowman's capsule and in the
                                                         lumens of the renal tubules. A slight infiltration of
                                                                             lymphocytes and plasma cells was occasionally
            ":              l        ,observed                                         in the interstitial tissues. Phagocytized
                                                                             hemosiderin granules were occasionally seen in
                                                                             macrophages in the glomerular tufts and in the
           ""    b,--                     Iinterstitium.
          IN.          ..
                        ,..                                                    Spleen.     A    moderate   to    marked    lymphoid
                                                                             hyperplasia of the malpighian corpuscles was
                                                                             commonly observed. Hyperplasia of reticu­
                                                                             loendothelial cells in the red pulp was observed in
                                                                             some    sections.     Granules    of  phagocytized
                                                                             hemosiderin were observed in the sinuses of all
                                                                               Lymph Nodes. A histological examination was
                                                                             conducted on tissue sections prepared from the
                      suprapharyngeal, prescapular, prefemoral, sternal,
                       .j]'''              .                                 bronchial, renal and mesenteric lymph nodes. The
                                                                             microscopic abnormalities observed were similar
                                          irregardhess of which of the lymph nodes was
                . 0: .0
                     M.................examined. Moderate hyperihksia of the lymphoid

Ftiprc 4. Coraplen,jtirinx .muiho-', filers of the jiurgroups of             follicles was o)served in all sections, with marked
ralhes ,xpenmnctally imJfefeld with Anaplasita mrginale and/ar               hY )perplasia hvilag observed in several sections.
Babehsm higri,' (;tup nicapitare t1'raeited and iertical lines               llyperplasia of reicinloendothclial      cells was occa­
f¢lrcsat-a rana'.    nrdekd leters indicate tsi',e of saibrulaicoiss
Innrilaii', with Analdeto marginale or with Ilabeia bhicrmina.               sionally    obs.erved.   Graniuls   of     phagocytized
                                                                             heinosiderin wcre present ir, several sections.
1:140 were ob.served in the Anaphstat controls                                  Sections of tissue.; from the lung, cerebrum,
nnd calves concurrently infected with Anaplastn,                             cera'hllum, adrenal and thyroid glands were also
respectively. The mean maximum CF antibody                                   examined. 'hllemicrorcopic alnormahities observed
titers ob.served in tl, Babesia controls and the                             were tlight and inconsistent and were not


 considered to have resulted from Infection with A.          serum globulins which occurred in the concurrent.
 margnaleor with . bigemina.                                 ly infected calves was due to an additive effect of
                       Vcussloa                              both infections with the Anaplasma infection being
    The increased severity of the efinical signs and         predominately responsible.
 the more pronounced reductions in to''.d weight                The decrease in A:G ratio which occurred in
 gains and average daily gains observed in the               each of the four groups was attributed to the
 concurrently infected calves were apparently due            increase in the gamma globulin concentration
 to the combined effects of the Anaplasma And                associated with antibody production and to the
 Babesia infections. The mild clinical signs and F.mall      decrease in serum albumin concentration.
 reductions in weight gains which occurred in the               The concurrently infected calves developed CF
 Babesia control group suggested the severity of the        antibody ters against A. marginale and against B.
 clinical disease observed in the concurrently               bigemina which were considered to be independent
 infected calves was predominately due to infection         of the concurrently infecting organism.
 with A. marginale.                                            The calves
    The severity of the anemic condition which              infectedgross lesions the same as thoseconcurrently
                                                                             were observed in the described as
 occurred in the concurrently infected calves was           occurring during nonacute anaplasmosis (11,21,22)
 considered to have resulted from the combined              or during nonacute babesiosis (11,20). The
 effects of the Anaplasma and Babesia infection             presence of transudates in the major body cavities
 with the Anaplasma organism being predominately            was considered to have been associated with a
 responsible.                                   y           decrease in the osmotic pressure of the blood due
   The significant    increase   in   the number    of      to decrease in loss albumin concentration and
                                                            Thethe moderate serumof albumin via the kidneys.
   infected erythrocytes in the concurrently infected       the presence of proteinaceous material in Bow.
   calves was an unexpected observation. Several            the psen ofd          ternal      tubul in      the
  authors reported that exacerbations of anaplas-           man's casule and in the renal tubules indicated the
   mosis may result from intercurrent infections            probable loss of albumin via the renal pathway.
  placing additional stress on the reticuloendothelial      Splenomegaly, lymph node enlargement and the
  system of the host (11,18). The ability of the            moderate to marked lymphoid hyperplasia ob­
  reticuloendothelial system to remove infecting            served in the malpighian corpuscles of the spleen
  organisms may have been moderately reduced in             and lymphoid follicles of the lymph nodes were con­
  the concurrently infected calves allowing the            sidered to be indicative of a responding reticuloen.
  secondary infecting organism to reach substantially      dothelial system.
  higher levels in the peripheral blood.                      The moderate degenerative changes observed in
     The moderate monocytosis observed in each of          the hepatocytes in the centrolobular areas of the
  the four groups of calves was considered to have         liver were attributed to hypoxia associated with
  been associated with the removal of damaged              the anemic condition of the calves. The degenera.
 erythrocytes, erythrocytic debris and infecting 
         tion of hepatocytes during anaplasmosis was
                                              previously reported to be the result of a hypoxic
     The presence of reticulocytes in the peripheral       condition resulting from anemia (1,10,22).
 blood and of nucleated erythrocytes in the hepatic           The presence of hemosiderin granules within

 sinusoids along with non-significant changes in the       macrophage in the spleen, plomerular tufts, lymph

 total and differential leukocyte counts indicated         nodes and in the Kupffcr cells ofete liver was

 the decrease observed in the M:E ratio was due to         atrotes       an the      ssee       tc     tion of

 an increase in the number of cells of the erythroid       eqthrocytes and tie subsequent accumulation of

 series. The more prolonged and severe anemic              erythrocytic debris.
 condition of the concurrently infected calves                                  Cwduiunu
 probably acted as a more intense stimulus resulting           The clinical and pathological manifestations of
 in the M:E ratio being substantially lower in the          disease were considered to have been more severe
concurrently infected than in the control calves.           in the concurrently infected culves than those
     The elhvation in '11l' observed in each of the        observed in the singularly infectid controls.
four groups of calves was attributed predominately         Infection with A. marginaleapparently contributed
to an increa.,se in the concentration of the serum         more to the severity of the cow'urrent infection
gumlnu glolbulin fraction. IncreaLses in 'TSI' and         than did infection with Bi. bigcia. Neither an
serum globulin concentrations were higher in the           inhibitory nor a synergistic effect between the
Anuplasmia controls than ini the Babesla colitruls         Infecting organisms wis evident during the
suggestiiig te     substantial Increase in 'ISI1 and       Investigation and the relationship of the organisms


during the concurrent infection was one of                             Santa Gcrtnidis Cattlc Against Piroplasma spp. and AAplama
independency.           The increased severity of the                   rm'isale. Pliipping J. Anim. Ind., 23-1/4, (1962): 1-20. - 7.
                                                                       Gerlach, U.: Zur Klinischen Bedcutung der Aktiviislmessung von
concurrent       infection       was     attributed       to    the    Sobiltdehydrogcnase In Menschlichcn lutserum. Kin. Wschr., 37,
concurrent infections being additive.                                  (1959): 93-98. Adapted by Sigma Chemical Co. as Tech. Bulletin
                                                                       No. 50-U.V., St. Louis, Mo., 1965. - 8. Gibson, R. I. and
                             Summary                                   Goodrich, G. E.: Dcterminat;on of Plasma Itilirubin; A Modified
                                                                       Van den Bergth Procedure. Proc. Soc. l.xp. Biol. Mcd., 31, (1934):
   The      clinical,     serological      and       pathological      413415. - 9. IliJalgo, R. J. and Dimopoullo% G. T.: Complement
manifestations of disease in intact calves concur-                     vi,ation Microprocr.durcs for Anaplasmosis. Am. J.Vet. Res., 28,

rently infected with Anaplasma marginale and                           (1967): 245-251. - 10. Jones, 1. W. and Brock, W. E.: Bovine
                                                                       Anaplasmosis. Its Diagnosis, Treatment and Control. J.A.V.M.A.,
Babesia bigemina were investigated. Clinical signs                     149, (1966): 1624-1633. - 11. Jubb, K.V. F. and Kennedy. P.C.:
were more severe    in the concurrently infected                       Pathology of Domestic Animals. Vol. 1, 2nd cd. Academic Press,
calves than in singularly infected controls.                           New York, N.Y., 1970. - 12. Mahoney, D. F.: Bovine Babcsiosis:
                                                                       Diagnosis of Infection by a Complement Fixation Test. Austral.
Decreases in packed cell volume, albumin:globulin                      Vet. J., 38, (1962): 4-52. - 13. Mahoncy. D. F.: Bovine
ratio, myeloid:erythroid ratio and increases in the                    Babesiosis: Preparation and Assessment of Complement Fixing
number of reticulocytcs, total serum proteins and                      Antigens. Exp. Parasite', 20. (1967): 232-241. - 14. Malson. It. A.:
                                                                       Cont11rul of Anilnsmosis, Iltabesiosis nd "hcilcrisis in Rhdcsia.
 erum gainina globulins were more proniounced ii                       hill. Off. Il. .l:i,., 64, (1965): 465A76.
                                                                                         1                             IS. Mkus, C. I'.,
tWe concturrt'lntly infected cailv.            The    cotncurrn t      Garcia, F. M. mid      )nri,   It. A.: Control ol I'l~inla~muis    in
Infections had no al,parcnt effect on the                              Viictuicla. Accliuatl/ailon, Nutrition. aoid IPrcuunition of Im11port­
                                                                       ed Cattle. Dull. Off. ht. EIplz., 64, (1965): 493-517. - 16. Orcllana,
production of complement fixing antibodies. Gross                      V. M. and Brock, W. I-.: invcsligaciones Sobre Anaplasmosis y
lesions observed in the concurrently infected calves                   Piroptasmosi. Rev. Fac. Vet. Med. y ZooS., 2, No. 4, Universidad de
included: pleural and peritoneal transudates;                          San Carlos, Guatemala, (1969): 69-75. - 17. Otte, L. and Kiltz, II.
                                                                       I!.: Tick Borne Diseases in Imported Cattle. 1. Vaccination Against
splenomegaly; hepatomegaly; and moderate lymph                         Anaplasmosis with A. centralc. i. Infection with B. argentina and
node enlargement. Histological lesions included:                       E. wernyonL Mem. Coll. Agric. Natn. Taiwan University, 9, No. 2,
moderate hepatocellular degeneration; lymphoid                         (1968): 2440. - 18. Patnaik, M. M.: A Note on Bovine
                                                                       Anaplasmosis. Indian Vet. J., 40, (1963): 655-657. - 19. Reitman,
hyperplasia in the spleen and lymph nodes; at.                         s. and Frankel, S.: A Colonmetric Method for the Determination or
hemosiderosis of the spleen, lymph nodes, liver and                    Serum Glutamic Oxalacetic and Glutamic Pyiuvic Transaminases.
kidneys. The relationship of A. marginale and B.                       Am. J. Cin. Path., 28, (1957): 56-63. - 20. Rick, R.F.: iBabesiosis.
                                                                       In Infectious Blood Diseases of Aan and Aninals. Vol. II. Ist d.
bigemina during the concurrent infections appear-                      Edilcd by D. Weiman and M. Rislic. Academic Press,. New York.
ed to      be one of independency. The increased                       N.Y., (1969): 219-268. - 21. Ristic, M.: Anaplauno-is. In
            sveryofclinical and pathological signs of
               the                                                     Infectious Blood Diseases of Alan and Animals. Vol. II, 1st ed.
severity    odited                                                            by D.Weiman and M. Ristic. Academic Prcss, New York.
disease    in the concurrently infected calves was                      N. Y., (1969): 443-542. - 22. Riqtic, M.and Sippel. W.L.: IEffcct
attributed to the          concurrent infections being                  of Cortisone on the Mechanism of Anaplasma Immunity in
additive.                                                               Experimentally Infected Calve%. II. Study of Patlholugical Changes.
                                                                        Am. J. VcL Res., 19, (1958): 44-50. -- 23. Schalm. 0. W.:
                                                                        Veterinary Hematology. 2nd cd. Lea and Febiger. Philadelphia, Pa.
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