in Russia. (Rementsova,. 1964). In the United. States, a serologic sur- .... abortion or normal birth) from the cattle. Rectal swabs and blood samples were taken.
Journal
of Wildlife ©
INTERSPECIFIC
TRANSMISSION
EXPERIMENTALLY
INFECTED
TO PARTURIENT Donald
S. Davis,’
In
mascerated
Fred
four
C. Heck,2
separate
bovine
placed
were
COYOTES
24(3), 1988, pp. 533-537 Disease Association 1988
ABORTUS
(CANIS
FROM
LATRANS)
CATTLE
Department of Veterinary Pathology, College Station, Texas 77843, USA 2 Department of Veterinary Microbiology Texas A&M University, College Station,
ABSTRACT:
OF BRUCELLA
Diseases, Wildlife
D. Williams,2
and Parasitology, Texas 77843, USA
10 coyotes
tissue
isolation
T. R. Simpson,’
of Veterinary
trials,
placental
in a 1 ha
John College
Medicine, College
Texas of Veterinary
(Canis
latrans) inoculated six parturient, non-B.
with
L. Garry
Adams’
University, Medicine,
which had been individually with Brucella abortus strain abortus vaccinated, Brucella
experimentally
area
and
A&M
fed 2308 spp. spp.
of the heifers became Brucella tube, rivanol, complement fixation, exposure to the B. abortus infected of 10 coyotes in the second trial seroconverted to Brucella spp. positive by day 14 (PE) and by day 27 PE seven of the 10 coyotes were Brucella spp. reactive. The spp. seroreactive heifers subsequently aborted on days 35, 64 and 66 PE and B.
seronegative Hereford heifers. During the second trial,three seroreactive (as determined by the card, standard agglutination and enzyme labeled immunosorbent assay tests) 14 days after coyotes.
Five
postexposure three Brucella abort
us
2308
strain
isolated
was
from
vaginal
was no serologic or bacteriologic evidence susceptible
Key mestic
cattle
words: cattle,
recorded
Coyote, experimental
in the
other
swabs,
three
Canis latrans, study.
Brucella
abortus,
become
naturally
indicated
that
in Alaska
infected
with
me bruce!!osis (Nei!and, have been experimentally trapenitonea!
D.
has lupus)
inoculations
of
can
forming
units
foxes (Vulpes and Zabrodin, 1975).
(CFU)
1 x
vulpes) 1970)
Brucella
naturally
and
suis
in red
Russia Alaska
was
foxes
10
found
to
co!-
et a!., 1960) and in Russia (Rementsova, 1964). In the United States, a serologic survey for Brucella canis conducted on seven
in
and
prevalence determined
from
1974).
test. In a serologic for B. abortus
evidence been Ireland of wild
of Brucella reported in (McCaughey, foxes
a!., abortus
red
(Dusicyon
foxes 1968).
Serologic
infections
has
in Northern Two species
gymnocercus
and
B.
by
shown
methods B. abortus
to be biovar
1966).
Se-
was
re-
abortus
(Canis cropieScangray
spp.
as et
antibody
test (Rahdhawa
a!., 1977). Hoq (1978) in a serologic survey of Brucella spp. agg!utinins in wildlife and sheep of the
et
were
Gonzales-Tome, of
do-
serology,
Argentina
of Brucella by the card
from five states, in a red fox from (Canis latrans)
(Hoff
There
(Urocyon cinereoargen teus) with 10 CFU of B. abortus strain 2308. from coyotes collected in southern were found to have a 6% (11 of 198)
species of wild carnivores detected positive reactions New York and two coyotes Texas
tissues.
transmission,
evidence
foxes 4.4 x Sera Texas
occur
(Pavlov
fetal
infected coyotes to the
ported in the black-backed jackal mesomelas), spotted hyena (Crocuta cuta) and wild hunting dog (Lycaon tus) in Tanzania (Sachs et a!., 1968). lan et al. (1984) successfully infected
(Pinigin (Neiland,
in Bulgaria
and
and bacteriologic infected with
no!ogic
of
in
griseus)
1 (Szyfres
wolves by in-
Brucella suis biovar 4 (Neiland and Miller, 1981). Rangifenine bnuce!losis also has been reported in arctic foxes (Alopex lago pus) and red ony
placental
interspecific
serologic naturally
rangifer-
1970) and infected
and
trials.
INTRODUCTION
Serologic evidence timber wolves (Canis
milk,
of transmission from B. abortus
by
the
in California coyotes tested standard
rapid
533
nivanol,
3% (five a titer of plate
standard
of 148) >1:100
agglutination
and bacteriologic in coyotes collected
eastern Texas, 18% no!ogica!!y positive card,
found had
(nine of 51) as determined agglutination
survey from were by
sethe tube,
534
JOURNAL
OF WILDLIFE
DISEASES,
and cold complement abortus biovar 1 was tissues genital
from seven transmission
(Davis
et a!.,
VOL. 24, NO. 3, JULY
fixation tests; B. isolated from various of 43 was
1979).
The
conducted to evaluate ious serologic techniques
coyotes and conalso documented present
the
study
was
efficacy of varto diagnose B.
abortus infections in coyotes and to determine the transmission potential of B. abortus from experimentally infected coyotes to susceptible pantunient domestic cattle under controlled conditions. MATERIALS
AND METHODS
Coyotes were trapped eastern Texas (USA) by
from personnel
24
counties in of the Texas
Anima!DamageControlService(USDA/APHISADC, P.O. Box 604, Bryan, Texas 77806,
USA) and transported to the Veterinary Research Park (College of Veterinary Medicine, Texas A&M University, College Station, Texas 77843, USA). The coyotes were tested for Brucella spp. antibodies by the card, rivano! precipitation (RIV), standard agglutination tube (SAT) (National Animal Disease Laboratory, Diagnostic Services, Ames, Iowa 50010, USA) cold complement fixation test (CCFT) (Jones et al., 1963), enzyme labeled immunosorbent assay (ELISA) (Heck et a!., 1980, 1982; Nielsen et a!., 1983), and hemolysis in gel (HIG) techniques (Nielsen et a!., 1983). Only those coyotes shown to be serologically negative for Brucella spp. for 30 days postcapture were utilized in the study. In each of the four repetitions of the experiment, 10 Brucella spp. sero!ogica!ly negative coyotes were individually fed for three consecutive days approximately 100 g daily of mascerated bovine placenta! and fetal tissue which had been inoculated with 1 x 10” to 1 x 108 CFU B. abortus strain 2308 per g of tissue (Table 1). Prior to, and after, being fed the B. abort us inoculated bovine tissue, coyotes were maintained on an ad libitum diet of a commercial dog food ration (Purina Hi Pro, Ralston Purina Company, St. Louis, Missouri 63164, USA). After consumption of the B. abortus inoculated bovine tissue, the 10 coyotes were placed in a 1 ha pasture (which had been modified with high fences, buried aprons and electrified fences to provide suitable isolation and security) with six B. abortus susceptible parturient heifers. During each of four trials, the coyotes remained in the pasture with the susceptible cattle until the termination of all cattle pregnancies for that repetition of the study. The 24 parturient heifers used
in the
investigation
were
registered
Here-
1988
fords.
Brucella
They spp.
were B. abortus non-vaccinated, seronegative and from a single B. abortus-free herd. At the time the heifers were exposed to the B. abortus infected coyotes the cattle were in the second trimester of gestation.
Blood
samples
were
collected
from
all
of the
animals weekly posure mination
(10 coyotes and six heifers) for each repetition from 3 wk prior to the exof the cattle to the coyotes until the terof pregnancy in all of the cattle. Coyotes were bled through the cepha!ic vein and the cattle were bled through jugular venipunctune. The blood samples were collected by sterile needle and syringes, transferred to sterile vacuum blood tubes, refrigerated, allowed to clot for 4 to 6 hr, and centrifuged at 600 g for 10 mm. The resulting sera were decanted and stored at -20 C. The sera were evaluated for Brucella spp. antibodies by the card, RIV, SAT, CCFT, ELISA, and HIG tests. Heifers that became Brucelia spp. seroreactive were separated immediately prior to parturition from the rest of the cattle so that cross transmission of B. abortus
between Media!
cattle would retropharyngeal
not occur. lymph
nodes,
pala-
tine tonsils, spleen, superficial inguina! lymph nodes and the testes or uterus were collected aseptically from the coyotes at necropsy and stored at -20 C. Placenta! tissue, vagina! swabs and quarter milk samples from the four mammary glands were collected at the termination of pregnancy (either abortion or normal birth) from the cattle. Rectal swabs and blood samples were taken from a!! live-born calves. Aborted fetuses were necropsied and lung, abomasum, mediastinal lymph node and stomach swabs were collected. The tissue samples and swabs from the cattle were also stored at -20 C. For bactenio!ogic culture, the tissue or swab was thawed and streaked on agar media (Bacto Brucel!a Agar, Difco, Detroit, Michigan 48323, USA; BB1 Formu!a Agar, National Veterinary Services Laboratory, Ames, Iowa 50010, USA) and Farrell’s Media (Farrell, 1974). Inoculated plates were incubated at 37 C in 10% C02, and after 3 days the cultures were examined. Bacteria from Brucelia spp. -like colonies were characterized by a
rapid slide agglutination test (National Animal Disease
Laboratory).
Those
reacting
to the
slide
agglutination test were inoculated onto agar slants and B. abortus isolates were identified by standard serologic and biochemical criteria ton et a!., 1975). Subcultures were submitted the National Veterinary Services Laboratory
(A!to for
confirmation. RESULTS
In the
all repetitions coyotes became
AND DISCUSSION
of the study, infected with
some of B. abor-
DAVIS ET AL-TRANSMISSION
OF BRUCELLA
FROM COYOTES
ABORTUS
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