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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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