SEROLOGIC SURVEY FOR BOVINE PATHOGENS IN FREE-RANGING EUROPEAN BISON FROM POLAND Author(s): Jerzy Kita and Krzysztof Anusz Source: Journal of Wildlife Diseases, 27(1):16-20. Published By: Wildlife Disease Association DOI: http://dx.doi.org/10.7589/0090-3558-27.1.16 URL: http://www.bioone.org/doi/full/10.7589/0090-3558-27.1.16

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Journal

SEROLOGIC

SURVEY

EUROPEAN Jerzy

Kita

BISON

of Infectious 272, Poland

From

ABSTRACT:

in Poland. serovars

virus,

bovine

bovine

leukemia

herpes

virus-i

in

28

bison,

infection

in

a

12-yr-old

to

isolate

Attempts cross

burnetii; foot Bison bonasus,

blood

was

taken

tested

for

the

presence

Chiamydia

spp., virus-i.

virus

and

bovine

from

the

prepuce

ranged

the

and

only

1967).

12th

last

and

Poland,

centuries

but they

forests

Caucasua bison

poachers

the

by

study

were

three

Primeval mountains. was

were

killed

purchased

in

1978;

has

the

are

bison

600

which

230

Forest Bieszczady

cows.

possibility

and

domestic

spp.,

Coxiella

European

bison,

Krasi#{241}ska

natural about

the bison have culling. These

and

allowed still

their some

carefully

et

in 1952.

the

pastures mentary receive

and the reMountains

adjacent bility

al.,

in

been animals

infectious

have

been

exposure

used

of wild

dis-

to cattle.

Se-

elsewhere

animals

to

to var-

with the cattle. single

occurrence During

bison

may

of diseases summer and wander

in au-

into

pas-

1987).

and farmed land. Winter supplefeeding places, where bison herds hay, beets and carrots from farms of

to the contact

forest with

are another possithese microorgan-

isms.

distribution hunting,

MATERIALS

but

monitored

in

During

culling

AND METHODS

of

herds

from

1980

to

1983

was collected from 60 bison before they were killed; 19 heifers (females that had calved and were 6 mo to 3 yr of age), 14 cows (2 to 17

blood

environment. 1971,

the

certain

be common

tures and farmed land adjacent to the forest. This creates a situation in which the bison may come into contact with organisms pathogenic for domestic cattle, or other animals, by means of naturally fertilized

European are

of may

tests

tumn

from

herd

over

of bison

in Poland

Since

of

Primeval are in the

number

their

are

Poland,

(Krasi#{241}ski,

The

as a free-living

there

which

ciation domestic

The

1919.

bison

forest

Bia}owieza mainder

the

virus-i,

virus

three

ious pathogens (Zarnke, 1983). The purposes of our study were to test bison for antibodies against bovine viral and bacteria! pathogens and evaluate their asso-

(Krysiak,

in Poland

in April

Presently, bison

suggest

Chiamydia

occurrence

rologic

a zoo in Sweden and introduced to an enclosed area in Bia}owieza. Successful captive breeding allowed bison to be released into

and bison

herpes

to isolate

chlamydial leukemia

bulls

results

14

disease

made

bovine

free-ranging

bovine

eases

bonasus)

Bia}owieza the

In 1929

was

five

interrogans, virus,

beginning of the 20th cenbison were present in only

free-living

by

These between

mouth

suggested

subclinical in

abortus,

and

attempt

tests

infection

successful.

the

of

larger

regions-the

Forest

heifer,

pathogens

leukemia

an

Warsaw

bonasus)

Brucella foot

Serological

Leptospira

bovine

(Bison

most

in the

At the European

tury, two

bison over

11th

found

addition,

(Bison

bison

against

burnetii;

03-849

serosurvey.

European

once

In

University,

European

Coxiella bulls.

not

bovine

abortus,

disease,

selected

virus-i

were

these

Agricultural

of antibodies

a 2-yr-old

herpes

virus-i of

Brucella

mouth

of

INTRODUCTION

The

16-20 1991

IN FREE-RANGING

Warsaw

60

of selected

bovine

herpes

from

herpes

Q-fever and

several

Serology,

Medicine,

1983,

bull of

and

of Veterinary

were

bovine

cattle in Poland. Key words:

Faculty

subclinical

transmission

pp.

1991,

Association

POLAND

interrogans,

infection

of

to

samples

Leptospira

of

PATHOGENS

Diseases, 27(1). © Wildlife Disease

Anusz

Diseases,

1980

Serum

BOVINE

FROM

and Krzysztof

Department Grochowska

FOR

of Wildlife

approximately

eliminated were

10%

of

yearly by used to assess

yr

of

animals 16

age)

and came

27

bulls

from

two

(6

mo

free-living

to

ii

yr). herds

These at

Bia-

KITA

AND

ANUSZ-SEROLOGIC

(52#{176}45’N, 23#{176}50’E)and Borki (53#{176}50’N, 21#{176}00’E) and two closed-herds at Komancza (49#{176}20’N, 22#{176}05’E) and Smardzewice (51#{176}30’N, 20#{176}00’E). Selection for culling was made during winter when animals gathered for supplementary feeding. Reasons for removal included poor condition; weak calves, especially those born after October; chronic reproductive problems, especially visible necrotic and/or purulent lesions of the prepuce and penis; advanced lameness in cows and bulls > 14-yr-old; tendency to remain near human settlements and attack humans or cause property damage; and presence of abnormal anatomical features. Serum was collected and frozen at -20 C until examination for antibodies against Brucelia abortus, 14 serovars of Leptospira interrogans, Chiamydia spp., Coxielia burnetii, foot and mouth diseases virus (FMD), bovine leukemia virus (BLV), bovine herpes virus 1 (BHV1). The serum agglutination test (SAT), salt-2mercaptoethanol tube agglutination test (SMTA) and the complement fixation test (CFT) described by Anczykowski (1960), Hoff and Hamblin (1974) and Alton et al. (1975), respectively, were used to detect antibodies against B. abortus. In SAT and SMTA the B. abortus agglutination suspension stained by TTC was prepared from strain S-19. In CFT unstained antigen prepared from strain S-19 of B. abortus was used (Kr#{243}lak and Biaszczyk, 1987). In both tests antigens were standardized using the second International Standard anti-Bruceila abortus serum (ISAbS-II). The CFT is considered to be more sensitive than SAT (Mathias and Pinto,

tibodies

1983).

serum

J.owieza

The scribed

microscopic

agglutination test (MAT) deby Cole et al. (1979) was used against 14 serovars of L.

sidered

CFT scribed Smertin applied mouth

A BLV,

to

Paris,

France)

was

used

tibodies against Chiamydia spp. higher were considered positive Truszczy#{241}ski, 1974; Truszczy#{241}ski

to detect anTiters 1:16 or (Sadowski and and Sadowski,

1978).

The described (1969), antigen

Krakow,

and microagglutmnation test (MA) by Anusz et al. (1986) and Fiset et al. respectively, using phase-Il Henzerling (Wytw#{243}rnia Surowic i Szczepionek,

Poland)

were

used

for detection

of an-

Enzygnost

Republic

detect

of Germany)

antibodies

against

bovine

An

attempt

was

made

to

BHV-1

isolate

from

the prepuce of bulls (Kita, 1978). Specimens were collected both from prepuces with visible lesions and without lesions, using sterile cotton swabs soaked in Hanks’ solution with added antibiotics. The material was delivered to the laboratory without cooling and stored at -20 C until examination. used

for

The cell culture was in Hanks’ solution with 10 to 15% added. The nutritional media

carcalf was

Fetal

bovine

isolation ried out

to a maintenance

the

media

infection

of

daily

Antibodies in

for

1:10

(20

18

samples;

12

samples

more to,

examined

lU/rn!

of 1:20

and

sensitive 1983),

ed.

was

tions. observed

1:40

(80 CFT,

than

SAT

Stryszak,

pres-

Titres

were

found

in

of

serum)

in

IU/rnl

of

of

serum)

considered all

in

there

previously

were

to be

(Mathias were

concluded

serologic

(Kr#{243}lak and

culture effects.

SAT.

IU/ml

The

nonspecific Such

serum) (40

Additionally,

dicated

in

negative

It was

serum

This

B. abortus

sera

sample.

without

the cells. cytopathologic

AND DISCUSSION

against

31

was

virus.

RESULTS

ent

cell culture

kidney

of the

changed

SMTA. CFT

immunosor-

virus (BLV). Serum samples were adquantitatively analyzed as positive or by the agar gel immunodiffusion test (Kita et al., 1988). The Bovine Leucosis Antigen (Behringwerke AG, Marburg, Federal Republic of Germany) was used. A commercial ELISA (Behringwerke Enzygnost IBR/IPV, Marburg, Germany) was applied to detect antibodies against bovine herpes virus 1 (BHV-i). Serum samples were diluted 1:20. Additionally the SN test described previously by Saxegard (1970) was used.

in one

Pasteur,

the

al., 1986;

(Behringwerke

Federal

applied

l’ornithose

Institute

to

et

leukemia ditionally negative (AGIDT)

observed

Favre,

(ELISA)

Marburg,

was

(Nicolass

(Behymer

enzyme-linked

assay

detect antibodies interrogans. The standard serovars were obtained from the WHO Reference Laboratory of the Royal Tropical Institute, Amsterdam, The Netherlands. It has been suggested that serologic reactions in the range of 1:100 in nonvaccinated individuals represent animals with either clinical disease or residual titers to a previous infection (Shotts et a!., 1970; Fournier et al., 1986). The CFT using Antigen pour le diagnostic de psittacose

17

According

burnetii.

commercial

bent

BISON

et a!., 1989) titres of 1:8 were conpositive. and seroneutralization test (SN) as deby Chemyaev and Sobko (1975) and by and Sviridov (1975), respectively, were to detect antibodies against foot and disease.

Ciecierski

was

OF EUROPEAN

investigations

to

to

C.

against

previous

prior

previously

SURVEY

that

and no

titers

in

SAT

in-

agglutination

reachave

in

and

1978).

test-

the

reactions cattle

Pin-

animals

been swine

18

JOURNAL

The

OF WILDLIFE

MAT

confirmed

against

tibodies

highest

titre

that

reacted against icterohaemorhagiae,

bataviae

and

the

L.

The vars

DISEASES,

L.

logic

believe that by serovars

duce ger,

cross-reactions 1963; Diaz

and

detectable titers of infection and of a few months. In seven sera,

or

of Ac28 posi-

new reactivaresidual titers

infections. Comare generally detime. Neuvonen

reached decline

antibodies

that

maximum

within 4 wk over a period against

C. bur-

were detected by CFT or MA. Of those, one 2-yr-old heifer which had an antibody titer 1:16 in the CFT showed a 1:8 titer in the MA. Based on the earlier netii

described infection agglutinating

criteria of the

our 2-yr-old antibodies

results indicate heifer. The that react

an MA with

phase II C. burnetii antigen are indicative of recent infection. ELISA confirmed the presence of antiBLV antibodies in the serum a 12-yr-old bull. This verified earlier AGIDT results. According to criteria (1988) in evaluation 12-yr-old AGIDT fected.

used of BLV

bull positive was considered This

case

is

by Kita infections, in ELISA subclinically

son

has

of bison with animal breeding

tures

forests

occupied

a!. the and in-

observed.

This

was

collected

prior

cation

of

BLV

1985 thors

(W. have

infection

KoJacz, discussed

9 yr) tested

bi-

to

this

eradiarea

in

Some auproblem

biting

and

(aged

and three positive

serum

the

pers. comm.). the potential

of transfer of BLV by insects (Ferrer, 1979). The sera of five bulls 4, 8 and yr-old)

positive

from

sucking

9 mo

and

2,

cows (13-, 17-, in the ELISA

18for

BHV-1. However the SN test did not confirm the results of any of the tested samples. All bulls that were serologically positive

to the

ELISA,

which

preferred (Forschner

is the

idently advanced necrotic sions of the prepuce and seropositive cows did not

and purulent lepenis. However, have any visible

lesions of the vulva. Attempts BHV-1 from the prepuce of the not successful. The BHV-1

possibility from local

of cattle

bison may

and 43% Though

to isolate bulls were

contracting be suggested

by the presence of antibodies virus in 38% of the local cattle in 1987, comm.).

method

in evaluation of BHV-1 et a!., 1986), had ev-

against the population

in 1988 (W. Ko1cz, we do not have

pers. infor-

mation about the prevalence of antibodies against BHV-1 during the 1980 to 1983 period when the samples were collected, the percent of infected animals probably was similar. The lesions of the prepuce may

be

due

possible bacterial

to

BHV-1

contribution agents cannot

infection, of

but

other viral be dismissed.

the and

ACKNOWLEDGMENTS

We wish to thank Lance Perryman, Patricia Mason (Washington State University, Pullman, Washington, USA), Stephen Lesniewski (Warsaw Agricultural University, Warsaw, Poland), Thomas Walton (USDA, ARS, ABADRL, Laramie, Wyoming, USA), and Marian Kr#{243}lak (Veterinary Institute, Puluwy, Poland) for helpful

discussions

regarding LITERATURE

areas of in pasby

been

sample

especially

interesting,

since the contact intensive domestic neighboring

et

1991

currently infection

detecttiters sera.

described earlier serologically

showed

are then

with pro-

and GodinFournier et were

with previous fixing antibodies only for a short 1981,

ti-

results of seroL. interrogans

samples with 1:32 in three

These may represent of old infections,

Pyorala,

these

reflect contact or ones which

to the criteria were considered

connected plement tectable

on

antibodies

ed by CFT in 28 1:16 in 25 sera and

tions

sero-

any of the bison of L. interro-

(Olitzki al., 1968;

et

1986). Anti-chlamydia

tive.

samples

hebdomadis,

B. abortus may microorganisms

cording bison

sera.

interrogans

Based

and these

a!.,

in six

Alternatively, these tests for serovars of

gans.

1, JANUARY

of an-

in 35

1:40

was

27, NO.

presence

interrogans

valbuzzi.

ters we do not were infected

VOL.

this

manuscript.

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