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Bovine Fetal Encephalitis and Myocarditis Associated with Protozoal Infections

B. C. Barr, M. L. Anderson, P. C. Blanchard, B. M. Daft, H. Kinde and P. A. Conrad
Vet Pathol 1990 27: 354
DOI: 10.1177/030098589002700508
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Vet Pathol 27:354-361 (1990)

Bovine Fetal Encephalitis and Myocarditis Associated with

Protozoal Infections
B. C. BARR,M. L. ANDERSON,
P. C. BLANCHARD,
B. M. DAFT,H. KINDE,AND P. A. CONRAD
California Veterinary Diagnostic Laboratory System and
Department of Veterinary Microbiology and Immunology,
University of California, School of Veterinary Medicine, Davis, CA
Abstract. Bovine fetuses submitted to the California Veterinary Diagnostic Laboratory System were evaluated during a 2-year period (1987 to 1989) for the presence of multifocal necrotizing nonsuppurative encephalitis, nonsuppurative myocarditis, or tissue protozoa. Eighty-two of 445 (1 8%) fetuses submitted met these
histologic criteria. Fetuses were from 54 dairy and two beef herds located throughout the state. In 17 fetuses
(2 1Yo) protozoa were found in fetal tissues. Protozoa were found in brain parenchyma of ten fetuses (12%), in
endothelial cells in four fetuses (5%), in cardiac myofibers in one fetus (lYo), and were associated with endothelial
cells in two fetuses (2%). In most fetuses there were no significant gross pathologic findings other than autolysis.
While aborted fetuses were from 3 to 9 months gestation, the majority were between 5 and 7 months gestation.
They were submitted year round, but more were seen in the fall and winter months. Additional salient histologic
features included portal nonsuppurative hepatitis, focal hepatic inflammation and necrosis, and focal nonsuppurative myositis. Nonsuppurative inflammation was also found in decreasing frequency, in the adrenal medulla,
kidney, mesentery or abdominal fat, placenta, and lung. In two fetuses (Nos. 1 and 2), the location and
morphology of the protozoa were compatible with Sarcocystis spp. The identity of protozoa in the remaining
15 fetuses is unknown. The histopathologic changes in these 82 fetuses and the presence of protozoa in 2 1Yo of
the fetuses suggest these abortions are due to fetal protozoal infections.

Key words: Bovine; encephalitis; fetus; myocarditis; protozoa.

Numerous sporadic cases of bovine protozoal abor- seen in abortions of sheep due to T. gondii,8J6and
tion or congenital disease are reported in the litera- similar changes have been found with Sarcocystis and
t ~ r e . ~1~15~17-19,24-26,29
-l
Reports of multiple bovine abor- Neospora infections in the bovine
Pathologists at the California Veterinary Diagnostic
tions due to protozoal fetal infections, however, are
i n f r e q ~ e n t .Sarcocystis
~ ~ , ~ ~ species have been identified Laboratory System have recognized a significant numor presumed to be the causative agent in a large number ber of bovine abortion cases with a similar pattern of
Toxoplasmosis is a well- encephalitis, accompanied by a nonsuppurative myoof these
carditis.
documented cause of abortion in sheep and g o a t ~ . ~ , ~
, ~ ~ - The purpose of this study was 1) to identify
and review all bovine fetuses with these histopathologl 4 9 l 6 In cattle, however, reports of confirmed abortions
due to Toxoplasma gondii are infrequent, and the sig- ic changes received by the laboratory during a 2-year
nificance of this protozoan as a cause of abortion in period; and 2) to determine if there is an association
cattle has been q~estioned.~
Recent evidence has sug- between these changes and protozoal infections.
gested that a new protozoal organism, Neospora caMaterials and Methods
ninum, is also capable of producing fetal infection and
From June 1, 1987 to May 3 1, 1989, a total of 445 bovine
abortion in the bovine.10J1,29,32
fetuses
were submitted to the California Veterinary DiagHistopathologic changes associated with fetal pronostic
Laboratory
System, located at Davis, Petaluma (now
tozoal infections share unique characteristics that are
closed), Tulare and San Bernardino. All fetuses with a dioften useful in making presumptive diagnoses of proagnosis of encephalitis, myocarditis, or presumptive prototozoal infections. For example, Toxoplasrna gondii can zoal infection were collected. One pathologist re-examined
cause inflammation in any tissue, but the most frequent all tissue sections from these fetuses by light microscopy. The
or striking inflammatory changes are found in the fetal criteria established for selecting a case in this particular study
3 ~ 1 partic6
placenta, brain, and striated muscle. L , 4 ~ 1 2 ~ L In
group were 1) the presence of encephalitis with multifocal
ular, multifocal nonsuppurative encephalitis, with fo- necrosis or focal vascular nonsuppurative infiltrates and a
cal necrosis and gliosis are well-documented changes nonsuppurative myocarditis, or 2) the identification of pro354

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Bovine Fetal Encephalitis and Myocarditis

tozoa in fetal tissues. Cases with any other etiologic diagnosis
were omitted from the study. The presence of protozoa and
any significant inflammatory changes were recorded from
each case. The date of submission and the location of the
cases in this selected group were recorded, and the clinical
histories were reviewed. Estimates of the gestational age of
fetuses were determined by one of three methods: crownrump length,* breeding dates, or estimates made by veterinarians. Estimates of gestational age based on crown-rump
length were as follows: 13-2 1 cm = 3 months; 22-3 1 cm =
4 months; 32-43 cm = 5 months; 44-57 cm = 6 months;
58-67 cm = 7 months; 68-85 cm = 8 months; and greater
than 86 cm = 9 months.
Abomasal contents, liver, and lung were cultured on blood
agar, and MacConkeys agar at 37 C in 5-10/o CO, for a
minimum of 48 to 72 hours. Abomasal contents and liver
or lung were cultured on media selective for Campylobacter
containing polymyxin B sulfate, trimethoprim, and vancomycin, at 37 C in 5% oxygen, 10% CO,, and 85% nitrogen
for 5 to 7 days. Abomasal contents and lung were cultured
for Brucella abortus on selective agar for 7 to 10 days33with
and without crystal violet, or by incubation on blood agar
for 7 to 10 days. Darkfield examination of abomasal contents,
Gram stains of abomasal, liver, and lung impression smears
were performed on several cases. Direct fluorescent antibody
tests (National Veterinary Services Laboratories, Ames, IA)
for Leptospira spp. were run on kidney impression smears
in most of the cases.
Total immunoglobulin G levels and titers to infectious
bovine rhinotracheitis (IBR) virus and bovine virus diarrhea
(BVD) virus were determined by serum neutralization and
titers to Leptospira spp. (6 serovars) by microagglutination.
In many cases, fetal titers were determined to Parainfluenza
3 virus by hemagglutination, bluetongue virus by agar gel
immunodiffusion, and B. abortus by buffered acidified plate
antigen. Titers to T.gondii were determined either by latex
agglutination or by modified direct agglutination3 (Toxoscreen DA, bioMerieux, Charbonnieres-les-Bains Cedex,
France) on blood or fetal fluids from a total of 44 fetuses.
Sera from cows were checked routinely for titers to IBR,
BVD, Leptospira spp. (6 serovars), and B. abortus. In two
herds, multiple paired dam sera (1 3 cows and 4 1 cows bled,
respectively) were collected. Paired sera were tested for IBR,
BVD, Leptospira spp. (4 serovars), B. abortus, and T. gondii.
The following serologic tests were performed on fetal fluids
collected from the body cavities of 69 fetuses for serology:
total fetal immunoglobulin G levels in 62 samples and antibody titers to T. gondii in 44 samples, IBR in 63 samples,
BVD in 51 samples, Parainfluenza 3 virus in 46 samples,
bluetongue virus in 57 samples, 6 serovars of Leptospira spp.
in 56 samples, and B. abortus in 27 samples.
For viral isolation organ pools of lung, liver, spleen, kidney, thymus, brain, and, when submitted, placenta, were
homogenized, and diluted in a viral transport medium, centrifuged, and the supernatant was inoculated into cultures
with Madin-Darby bovine kidney cells. Cultures were incubated in 5% CO, at 37 C for 5 to 7 days. Direct fluorescent
antibody tests for noncytopathogenic BVD were performed
whether or not cytopathic effects were observed. Cultures
were checked for hemadsorption with bovine red cells to

355

screen for Parainfluenza 3 virus. Direct fluorescent antibody

tests for IBR and BVD were run on frozen sections of lung
or liver, adrenal, and spleen in 45 fetuses.
Chlamydia1 cultures were performed on tissue pools or
placenta from 24 fetuses. Tissues were placed in a chlamydia1
transport medium,28 homogenized, centrifuged, the supernatant inoculated onto McCoy cell cultures, and incubated
for 5 days. Cultures were stained with a monoclonal conjugate to Chlamydiapsittaci (California Integrated Diagnostics,
Berkeley, CA).
In 58 fetuses, ocular aqueous humor was tested for nitrates
by ion exchange chromatography.
Multiple tissues were sampled and placed in 10% neutral
buffered formalin, fixed for 24 hours, and processed and
embedded in paraffin using routine procedures. Five micron
sections were stained with hematoxylin and eosin. The tissues
and the number of fetuses from which they were examined
included liver (77), kidney (79), heart (82), spleen (61), lung
(go), thymus (66), brain (82), gastrointestinal tract (57), adrenal gland (54), skeletal muscle (64), and lymph node (33).
Placenta was examined when submitted (19). Sections stained
with periodic acid-Schiff and Giemsa stains were also examined on some tissue sections.

Results
Eighty-two ( 18%)bovine fetuses that were submitted
met the criteria listed in the Materials and Methods.
These 82 fetuses were received year-round, although
approximately half of the cases were received from
November through February. They were submitted
from 56 different herds, 54 of which were from dairies.
There were multiple abortions reported in 36 herds.
Twelve of these herds had more than one fetus (36
fetuses from 12 herds) included in this study group.
There were no reports of clinical signs in the cows that
aborted.
At necropsy, most fetuses were autolyzed to some
degree, with red-tinged fluid in body cavities and subcutaneously, and had no significant gross pathologic
findings. Pale-tan streaks were found in the muscles of
three fetuses, mildly enlarged lymph nodes were found
in seven fetuses, and fibrin strands were found in the
abdomen of one fetus. Figure 1 summarizes the gestational age of the fetuses. Fetuses were between 3 and
9 months gestation, although at least 75% of the fetuses
submitted were between 5 and 7 months gestation.
Serologic results

Fifty-two of the 69 fetal fluids were negative for all

tests performed. Total fetal immunoglobulin G levels
were greater than 20 mg/dl in 12 fetuses (fetus Nos. 2,
3,6, 12, 16, 18,25, 30,45,48,70, 80). Antibody titers
were detected in fetal fluids to BVD in three fetuses
(fetus Nos. 36,48,49); IBR in three fetuses (fetus Nos.
9, 19, 36); Parainfluenza 3 virus in two fetuses (fetus
Nos. 13, 48); and T. gondii in one fetus (fetus No. 80
with a titer of 1 : 60).

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356

Barr et al.

Table 2. The size and location of protozoa observed in

aborted fetuses.
NumSize of Zoite
Cluster (pm)
ters

Gestational Age (Months)

Fig. 1. Gestational age, in months, of all bovine fetuses

diagnosed with nonsuppurative encephalitis and myocarditis, and the number of fetuses where protozoa were found
in the tissues.

A total of 39 sera were submitted from cows that

aborted. Antibody titers to IBR, BVD, and Leptospira
spp. were within normal or acceptable ranges. In eight
of these samples, antibody titers to T. gondii were determined. Two of these cows had antibody titers of 1 :
64 and 1 : 128 by latex agglutination.
Multiple paired cow sera were also obtained from
cows in two herds diagnosed with this particular abortion problem. In the first herd, blood that was collected
from 13 cows had no seroconversions or elevated titers
to IBR, Leptospira spp., B. abortus, or T. gondii. In
Table 1. The frequency, distribution, and nature of tissue
inflammation in 82 aborted fetuses with encephalitis and
myocarditis associated with protozoal infection.

Tissue
Inflammation

Number
Affected/ PercentNumber age AfExam- fected
ined

Encephalitis

82/82

100

Myocarditis
Adrenalitis
Myositis
Nephritis
Hepatitis

82/82
43/54
46/64
52/79
48/77

100
80
72
66
62

Peritonitid
steatitis
Placentitis

25/46

54

10/19

53

Pneumonia

35/80

44

* Nonsuppurative = mononuclear

Histologic Findings

Focal nonsuppurative,*
focal necrosis
Nonsuppurative
Focal nonsuppurative
Focal nonsuppurative
Focal nonsuppurative
Portal nonsuppurative,
focal necrosis
Focal nonsuppurative
Focal nonsuppurative,
focal necrosis
Focal nonsuppurative,
focal necrosis
inflammatory cell infiltrates.

32

2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17

15
1
1
3
6
5
6
1
1
1
1
1
2
5
1
11

Brain, lymph node,

abomasum, thymus, adrenal
Brain, lung, omasum
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Brain
Heart
Brain
Brain, placenta

13-35 x 16-35
12-28 x 18-36
14 x 19
Free zoites
1-14 x 13-14
0-16 x 10-16
4-20 x 14-23
6-18 x 16-18
14 x 15
15 x 19
11 x 14
13 x 13
Free zoites
Free zoites
10-12 x 10-20
13 x 20
12-24 x 12-36

the second herd blood was collected from 41 cows.

Only elevated antibody titers to BVD were found. The
owner indicated the herd had not been vaccinated for
BVD for at least 2 years. Seven cows in this herd maintained persistent titers to BVD (greater than or equal
to 1 : 5 12), two cows seroconverted to BVD, while ten
cows had no titer to BVD. This herd had seven recent
abortions. Two of the aborted cows had no antibody
titer to BVD and one aborted cow had a persistent
antibody titer of 1 : 5 12. Three fetuses were submitted
to the laboratory and met the histologic criteria for this
study (Nos. 5, 80, 82). Protozoa were found in one
fetus (No. 5).
Histologic results

Protozoa were found microscopically in 17 fetuses

that were submitted from 17 of the 56 herds in this
study. They were seen in fetuses ranging from 3 to 9
months gestation, although protozoa were found more
frequently in fetuses between 3 and 5 months gestation
(Fig. 1). Histologically, focal, nonsuppurative infiltrates could be found in several organs (Table 1). Of
these affected organs, the pattern and degree of inflammation in the brain, striated muscle, and liver were
the most significant. In the brain, there were random
small, scattered foci of microglia and necrosis. The
focal gliosis was often adjacent to capillaries that had
hyperplastic endothelium, with perivascular collec-

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Bovine Fetal Encephalitis and Myocarditis

351

Fig. 2. Brain; fetus No. 7. There are foci of microglia in the parenchyma adjacent to a group of capillaries (arrows).
The capillaries have thickened walls and are ensheathed by mononuclear cells. Four protozoal zoite clusters (arrowheads
show two) lie adjacent to the microglial foci. Bar = 100 pm.
Fig. 3. Brain; fetus No. 36 with focal necrosis. Central necrotic debris is surrounded by a thin band of glial cells.
Bar = 100 pm.
Fig. 4. Brain; fetus No. 7. Higher magnification of the four protozoal clusters in Fig. 1. The nuclei of individual zoites
can be seen in these clusters, and a cyst-like membrane or wall is clearly visible around one group (arrow). An unidentified
cell nucleus is adjacent to another cluster (arrowhead). Bar = 20 pm.
Fig. 5. Liver; fetus No. 23. There is a dense portal infiltrate of small mononuclear cells (arrow) and a large focus of
hepatocellular and inflammatory cell necrosis in the adjacent parenchyma (arrowheads). Bar = 100 pm.

tions of mononuclear cells (Fig. 2). Additionally, there

were random foci of necrosis, often in white matter or
brain stem, that vaned from focal, swollen, eosinophilic axons to foci of necrosis, surrounded by rims of
glial cells (Fig. 3). Protozoal clusters could be seen
sometimes in association with these foci of inflam-

mation (Figs. 2, 4), although they were usually randomly located.

In the heart, there was severe autolysis, that often
exceeded the degree of autolysis in other organs. There
were varying numbers of mixed mononuclear cell infiltrates in the epicardium, myocardium, and endo-

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358

Barr et al.

Fig. 6. Brain; fetus No. 1. A cluster of zoites lies within an endothelial cell of a small vessel. The individual zoites are
arranged in a rosette-like pattern around a more lucent central region. There is an endothelial cell nucleus at one side of
the cluster of zoites. Bar = 10 pm.
Fig. 7. Brain; fetus No. 4. A cluster of zoites lies within a capillary. Bar = 10 pm.
Fig. 8. Brain; fetus No. 6. There is a dense cluster of zoites lying directly within the brain parenchyma. The zoites are
randomly arranged and surrounded by a peripheral cyst-like wall or membrane. Bar = 10 pm.
Fig. 9. Brain; fetus No. 14. A free cluster of zoites lying within the brain parenchyma. There is no obvious wall or
membrane around the zoites and they are dispersed. Bar = 10 pm.

cardium. Infrequent, focal, interstitial infiltrates of

similar mixed mononuclear cells were also found in
skeletal muscle.
In the liver two patterns of inflammation were encountered. Mononuclear inflammatory infiltrates were
present in portal areas in 43/77 fetuses (56%), and
scattered foci of either hepatocellular necrosis, or sinusoids filled with fibrin thrombi were present in 27/77
fetuses (35%). In a few cases, both types of lesions were
present (Fig. 5).
Random scattered foci of mononuclear inflammatory cells were found in several other organs (Table 1).
Occasional, small foci of necrosis were also found in
both the interstitium of the lung, and along the surface
of cotyledonary villi in the placenta.
Protozoa found in tissues of 17 fetuses were generally
round or ovoid, 1.5-2 x 2-6 pm, with prominent nuclei and, in most cases, a distinct plasmalemma. Parasites were often in clusters surrounded by a limiting

membrane or wall. These parasites will be referred to

as zoites throughout this paper, because a distinction
as to whether they were merozoites budding from schizonts or tachyzoites/bradyzoiteswithin cysts could not
be made by light microscopy. The location and size of
the parasite clusters are summarized in Table 2.
In four fetuses (Nos. 1-4), clusters of zoites were
primarily or exclusively in endothelial cells (Table 2).
In two of these fetuses (Nos. 1 and 2), multiple clusters
of organisms were found within endothelial cells in
several tissues (Table 2), and in several of these clusters, the zoites had a rosette formation (Fig. 6). In fetus
Nos. 3 and 4, only one endothelial cluster of zoites was
found in the brain, and there was no rosette formation
(Fig. 7).
Eleven fetuses (Nos. 5-1 5 ) had clusters of zoites either
in neural cells in the brain or in cardiac myofibers.
Most of the clusters had the appearance of cysts, as
shown in Figs. 4 and 8, with a distinct peripheral wall

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Bovine Fetal Encephalitis and Myocarditis

which ranged in thickness from 0.2-2 pm. Zoites were

observed in random arrangements, usually close together, with no rosette formation. In fetus Nos. 13 and
14 there were clusters of zoites with no limiting membrane or wall, which were located directly in the brain
parenchyma (Fig. 9).
In the remaining two fetuses, Nos. 16 and 17, some
of the zoite clusters were closely associated with endothelial cells, but whether they were within these cells
could not be determined. Fetus No. 16 had one cluster
of 28 zoites adjacent to the nucleus of a capillary endothelial cell in the brain. In fetus No. 17, eleven zoite
clusters were seen in the brain and placenta. Two of
the six clusters in the brain of fetus No. 17 were associated with endothelialcells. The clusters in the brain,
as well as three clusters in the placenta, were similar
to those shown in Figs. 4 and 8. In the remaining two
placental clusters, the zoites were arranged in rosettes
and surrounded by a membrane or wall that was 1-2
pm in thickness (Fig. 10).

359

Fig. 10. Placenta; fetus No. 17. There is a dense interstitial cluster of zoites. They are arranged in a rosette-like
pattern and are surrounded by a cyst-like wall or membrane
with an adjacent unidentified cell nucleus. A large vessel
containing red cells lies above the cluster. Bar = 10 km.

tropism, and the presence of rosetting arrays of organDiscussion

isms reflecting its form of asexual reproduction, termed
Eighteen percent (82/445) of the total bovine fetal e n d ~ p o l y g o n y . ' ~ J A
caninum-like or~ , ~Neospora
',~~
submissions to the diagnostic laboratory during a 2-year ganism has also been identified in aborted bovine feperiod were found to have a consistent pattern of mul- tuses. It is very similar in appearance to Toxoplasma
tifocal nonsuppurative encephalitis and myocarditis, and has only been differentiated in fetuses by immusuggesting a common infectious disease process. Mi- noperoxidase using specific antisera. 1,20,29
crobiologic, serologic, and histopathologic examinaThe identity of the protozoa in most of these 17
tions did not reveal viral, or bacterial pathogens. The fetuses is unknown. Sarcocystis was diagnosed in only
fetuses shared a similar pattern of encephalitis, with two fetuses (Nos. 1 and 2) and was based on the presdistinctive foci of necrosis (Fig. 3) or focal vascular ence of protozoal clusters in endothelial cells of muloriented gliosis (Fig. 2). This inflammatory pattern is tiple tissues, and the rosette arrangement of
similar to the encephalitis induced by Toxoplasma Z O ~ ~ ~ S '(Fig.
~ J 6).
~ , In
~ ten
~ - of
~ the
~
17 fetuses (Nos. 5gondii, Sarcocystis species, and Neospora caninum in- 14), however, Sarcocystis could be ruled out since the
fections in ruminant ~ ~ ~ u s ~ s Nonsuppurative
. ~ J ~ J ~ , ~ protozoa
~ , ~ ~ in these cases were located directly within
myocarditis, myositis, hepatitis, nephritis, and focal brain parenchyma (Figs. 8 and 9). Additionally, of the
placentitis found in a large percentage of the fetuses 42 fetuses examined for antibody titers to T. gondii,
(Table 1) have also been reported in protozoal abor- only one fetus (No. 80) was positive, and there were
t i o n ~ . ~ , ' Protozoa
~,~~,~
were
~,~
found
~ in 17 of the 82 no significant titers to T. gondii among several cows
fetuses with nonsuppurative encephalitis and myocar- from the two affected herds examined. Collectively,
ditis. We conclude that protozoal infections are the these data would suggest that Sarcocystis is the etiocause of these abortions.
logic agent in a few cases, and, Toxoplasma may have
Sarcocystis species, Neospora caninum and Toxo- been present in at least one fetus (No. SO), but the
plasma gondii, have all been documented to cause etiology in the vast majority of cases is unknown.
abortion or congenital infection in r u m i n a n t ~ , ~ , ~ JReports
~
of multiple protozoal abortions in the cow
15,L7,22,24,29,31,32although there is a controversy whether
are infrequent. Thilsted et al.32reported recently on
Toxoplasma is a significant cause of bovine abortion. multiple protozoal abortions in dairy cows caused by
Attempts to induce experimentally bovine fetal Tox- a Neospora caninum-like organism.32McCausland et
oplasma infection and abortion have been al. reported multiple protozoal abortions in cows from
unsuccessfu16~23~30~3
Dubey has reviewed reports of bo- several dairies, and the authors speculated that they
vine Toxoplasma abortion, and concluded that Tox- were due to Sarcocystis infections.22Multiple abortions
oplasma infection is probably not a significant cause were also seen in one dairy, associated with adult boof bovine a b ~ r t i o n Sarcocystis
.~
has been well docu- vine sarcocystosis, although fetal infections were not
mented as a cause of sporadic abortion in the cow. The demonstrated.2 In our study, multiple abortions were
organism is identified by its specific endothelial cell seen in 36 herds. In 12 of these herds, presumptive or

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360

Barr et al.

confirmed protozoal fetal infection was diagnosed in

multiple fetuses. No clinical signs were reported in the
aborting cows. These clinical and pathologic findings
closely resemble the reports by Thilsted et al.32and
McCausland et a1.,22although our data demonstrate a
markedly higher incidence of protozoal infections.
Protozoa were found most frequently in the youngest
of the submitted fetuses (Fig. 1). Microscopically, in
the majority of cases where multiple protozoa were
found, the degree of inflammation in the brain was
often mild. Vascular oriented, focal gliosis was the
dominant inflammatory change, and necrotic foci were
infrequent in these cases. This association between
younger fetuses, more visible organisms, and less severe tissue inflammation may reflect a more poorly
developed inflammatory or immune system in these
younger fetuses.
The serologic results, from one of the two herds
where multiple paired dam sera were examined, indicate that several cows in this herd were exposed to
BVD virus. Two fetuses from this herd were diagnosed
with presumptive protozoal infection and protozoa
were found in a third fetus (Table 2, fetus No. 5). BVD
was not isolated or demonstrated by fluorescent antibody tests in any of these fetuses, but given the serological results, a possible interaction between BVD and
fetal protozoal infections cannot be ruled-out in this
particular herd. In conjunction with these findings, it
is interesting to note that noncytopathogenic BVD has
been isolated from an aborted calf infected with a Neospora caninurn-like organism."
The results of this retrospective study suggest that
protozoal infections are a significant cause of abortion
in California dairies. Further studies are necessary to
define the etiology. Isolation of an organism is perhaps
the first step in precisely defining the agent, its life cycle,
and pathogenesis of the disease process.

4
5
6
7

8
9
10
11
12
13
14
15

16
17

Acknowledgements
The authors thank Dr. S. Hietala, M. Stillian, J. Goodbrod,
and P. Chiu for their technical assistance and S. Arnold for
assistance in preparing this manuscript. Supported in part
with funding provided through Livestock Diseases Research
Laboratory, School of Veterinary Medicine, University of
California, Davis, CA.

20

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21

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