seroepidemiology of leptospirosis in minnesota wolves - BioOne

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Wildlife. Diseases,. 27(2),. 1991, pp. 248-253. U) Wildlife. Disease. Association. 1991. SEROEPIDEMIOLOGY. OF LEPTOSPIROSIS. IN MINNESOTA. WOLVES.
Journal

SEROEPIDEMIOLOGY Muhammad

A. Khan,

Department

of Veterinary

University

of Minnesota,

(Mailing

address:

Sagar

North

Central

collected

Forest

tumnalls,

ballum, and

of L. grippotyphosa, penhageni. No indicate that L. Key words: serovars

miology,

zoonosis,

1992

=

1986

the

most

world

and

et at.,

infected vironment McCulloch,

1989).

become infected urine or indirectly contaminated

1970;

moist

stow-moving can survive

by

the

(Diesch hosts contact with soil,

and can

horses;

and headache, failure

pathy,

1981;

tospirosis

1983a).

interrogans

Adolf

acute

is

febrite

Fame,

Weit

first

1986).

The

disease

in Germany

with

pigs; in my-

occasional and Tn-

In humans,

reported

in

(Khan

wild

species

can

act

interrogans

USA Minnesota

and

for

tep-

1886

other

(Odocoileous (Ingebrigtsen

virginianus) et a!., 1986).

of the

study

present

55108,

USA)

wild

or

do-

was

of Minnesota The objective to determine

the

of 12 selected serovars of in wolves (Canis lupus) (USA).

MATERIALS

by

Diesch,

hageni,

1987).

Many

St. Paul,

AND

METHODS

Four hundred fifty seven serum samples were collected from wolves in northern Minnesota that were live-trapped and anesthetized. Blood was collected in glass tubes by saphenous or cephalic venipuncture. Serum was separated by centrifugation and stored at -20 C until tested. Most samples were collected during June through October 1972 to 1986 and were tested in 1987. These sera have been used in previous surveys for canine pathogens (Mech et al., 1986; Goyat et a!., 1986). The following L. Interrogans serovars were included in the study: australls, autumnalla, ballum, batavlae, bratislava, canicola, copen-

characterized

and (Hanson

L.

stagnant

illness

meningitis in humans

was

20708,

Maryland

Avenue,

seroprevalence L. interrogans from Minnesota

with urine-

by abortion and stillbirth in cattle and periodic ophthalmia (moon blindness) a!gia, kidney

en-

streams where L. for weeks (Roth,

et a!.,

L.

Laurel, FoIwell

reported in foxes ( Vulpes sp.) (Clark et a!., 1961), coyotes (Canis latrans) (Cirone et a!., 1978; Drewek et a!., 1981), and jackals (Canis sp.) (van der Hoeden, 1955). Serological evidence of !eptospirosis has recently been reported in white-tailed deer

of the

into

directly by by contact

Hathaway

a

interro-

kidneys

their urine Susceptible

water,

ponds and interrogans caused

Leptospira

is excreted

through 1966).

by

the genus LepAlt pathogenic

in the

and

zoo-

under one species, contains more than into 19 serogroups

occurs

host

H. Fritts2

mestic animals and even for humans (Hathaway et al., 1981; Hathaway and Blackmore, 1981). Leptospirosis has been

mammals is caused

of 1984).

leptospires are classified L. interrogans, which 170 serovars organized usually

Steven

from and were test. Twelve

distributed

affects

pathogenic spirochete tospira (Thiermann,

(Nielsen

and

Medicine,

457)

of

is a widely

which

gans

Mech,2

WOLVES

survey.

Leptospirosis throughout

L David

of Veterinary Center,

Station,

INTRODUCTION

nosis

IN MINNESOTA

L Diesch,

College

Experiment

(n

samples

from 1972 through using a microtiter agglutination

pyrogenes,

Stanley

Investigation,

27(2), 1991, pp. 248-253 Disease Association 1991

Diseases, U) Wildlife

wolves (Canis lupus) in northern Minnesota were tested for antibodies against Leptospira interrogans serovars included in the study were: australls, aubataviae, bratislava, canicola, copenhageni, grippotyphosa, hard Jo, pomona, tarassovi. Fifty-two (11%) sera had antibody titers of 1:50 against one or more Interrogans. The seroprevatence of different serovars in decreasing order was: bratislava, autumnalis, canicola, pomona, ballum, pyrogenes, hard Jo, and coantibodies were found against australia, batavlae, and tarassovi. These results interrogans infection may occur in wolves of Minnesota. Leptospirosis, Leptospira interrogans, serovars, wolves, Canis lupus, seroepide-

Serum

ABSTRACT:

M. Goyal,1

Diagnostic

St. Paul, Minnesota 55108, USA Service, Patuxent Wildlife Research

Fish and Wildlife

2

OF LEPTOSPIROSIS

Wildlife

of

grippotyphosa, tarassovi.

genes, and incriminated

as reservoirs 248

hardJo,

pomona,

These serovars as a cause of disease

pyro-

have been in different

KHAN

TABLE

1.

Titers

of Leptospira

interrogans

Number (%) at

Serovar

Autumnalis

positive 1:50

antibodies

in sera

of

wolves

50

in

of sera

Number

.

ET AL-LEPTOSPIROSIS

100

IN WOLVES

249

Minnesota.

sbowing

titerb

200

of

400

800

>1,600

15 (3.3)

5

1

4

1

2

2

3 (0.7)

2

1

0

0

0

0

Bratislava

18 (3.9)

14

1

2

1

0

0

Canicola

13(2.8)

5

3

3

0

0

3 (0.7)

3

0

0

2 0

0

0

24 (5.3)

9

4

6

3

1

1’

Hardjo

2(0.4)

1

1

0

0

0

0

Pomona

7(1.5)

2

1

3

1

0

0

Pyrogenes

7 (1.5)

1

1

3

1

1

0

9

4

3

Ballum

Copenhageni Grippotyphosa

92 No

serum

was

positive

for

Reciprocal

of highest

One

serum

was

positive

This

serum

had

a titer

‘Of

sera

457

of positive

species

examined, sera

of

tional States 50010,

australi.s,

dilution 1:1600

of

1:1600.

52

were

appears

hosts

(Fame, 1982). were obtained

serovars

serum at

42

(20)’

the

positive

and

batavlae,

showing and

13

50% other

at

at

1:6400.

1:50.

Because

the

world

some

in

various

parts

in semi-solid

of

had

antibodies

to more

than

one

serovar,

the

number

RESULTS

these bacteria media from the Na-

cultures

of

Veterinary Services Laboratory (United Department of Agriculture, Ames, Iowa USA). These organisms were sub-cul-

test.

The microscopic agglutination at., 1979) was performed in 96-well,

test

microtiter McLean,

Laboratories,

(Linbro; 22102,

Flow USA).

(Cole

et

flat-bottom Briefly,

a

Of

457

sena

examined,

52

positive at titers of were positive at titers

laboratory. The stock cultures were maintained in semi-solid media and liquid cultures were used as sources of antigen for the microagglu-

1:25

dilution of test serum was made in phosphate buffered saline, pH 7.2. To 50 j.il of this diluted serum was added 50 tl of live antigen at 25 nephetometenic turbidity units (NTU) making a final serum dilution of 1:50. The plates were incubated at 29 C for 120 ± 30 mm and examined for agglutination under a dark-field microscope. The samples found positive on screening at a 1:50 dilution were further titrated using serial two-fold dilutions of up to 1:6,400. We considered microagglutmnation titers of 1:50 as positive. Data were analyzed via a Chi square test with Statistix software (N.H. Analytical Software, St. Paul, Minnesota 55104, USA, 1985) and an IBM personal computer (IBM, PC, International Business Machines, Rochester, Minnesota 55901,

USA).

sera

to be 92.

Stock

plates Virginia

tarassovi.

agglutination.

tured in both semi-solid and liquid (bovine atbumin-polysorbate 80, Intergen Company, Purchase, New York 10577, USA) media in our

tination

21

bodies against interrogans. itive

for

ben

of

one Because

more

in Table

to more 2 serum

,

more some

one

senovar sera were

52 positive

of L. pos-

the

is shown

than one samples

were

31 (6.8%) for anti-

senovan,

samples

1. Of the

positive 1 1 and

1:50, and of 1:100 or

than

positive

(1 1 .4%)

num-

to be

92

sena,

25 were

serovar. contained

Twelve, anti-

bodies against two, three, and four serovans, respectively. At titers of 1:50, the prevalence of antibodies to serovans in decreasing

order

bratislava nicola

was:

(3.9%), (2.8%),

(1.5%),

ballum

and was

hardjo positive

viae,

and

grippotyphosa (1.5%),

(0.7%),

copenhageni

tarassovi. to

(3.3%),

pomona (0.4%). None for senovans

antibodies

The

different

1:50-400

for bratislava, and

hard

Jo. Antibody

penhageni Of

wolves

range

in titers was:

1:50-1,600 for pyrogenes;

sex

at was

and

ballum

against

positive

whose

samples bata-

senovars

for

titers

were

(0.7%),

the australis,

canicola

1:50-100

Ca-

pyrogenes

of

50-6,400 for autumnalis; grippotyphosa; 1:50-800 mona;

(5.3%),

autumnalis

serovar 1:50

of 1: for Poand co-

only.

known,

212

250

JOURNAL

TABLE

2.

OF WiLDLIFE

Distribution

DISEASES,

VOL. 27, NO. 2, APRIL

of Leptospira

1991

antibodies

interrogans

in wolves

Number Number County

Number

tested

Lake

positive

181

Bal-

Brati-

nalis

lum

slava

3

0

4

1 1 1 1

0 0 2 0

0 2 4 1

15(8.3)

Minnesota

a t iter

sh owing

Autum(%)

from

of

according

>1 :50

to counties.

against

Canic-

Pomo-

ola

Pyro-

Grippo

Hardjo

na

genes

8

2

0

0

1

0 1 0 1

1 4 7 1

0 0 0 1

0 1 1 1

0 1 0 0

Koochiching St. Louis Kittson

9 22 20 62

1(11.1) 5 (22.7) 7 (35.0) 3(4.8)

Itasca

11

3(27.3)

1

0

1

0

2

0

1

0

2

0(0.0)

0

0

0

0

0

0

0

0

8

2

11

10

17

1

4

2

Beltrami

Roseau Total Some

307b

of the animals

Excludes

were

samples

mates

showed

titers

for which

county

and

of seropositivity for females statistically in the rates

34 (11.2)

198 for

to two

were

of origin

was

females.

mates

was

serovars. unknown.

The 11.6%

rate

found

and

ka

it was 12.3%. There was no significant difference (F> 0.05) of seropositivity between males

and

females. Wolves were

ties

in northern

collected

from

Minnesota.

seven

of seropositive wolves near (20.1%) was 2.6 times greater wolves living in wilderness (7.7%)

(Table

coun-

Prevalence

antibody positive wolves from ties is shown in Table 2. The

farms

or more

of

these counprevalence

farming than away

areas that of from

3).

DISCUSSION

The 11.4% seroprevatence study is lower than

in wolves observed

in by

only

1 of 80 wolf

positive

Presence

for of

L.

samples

antibodies

in

dicate previous or current may have resulted either tact with urine of other eating have

infected indicated

may

Alas-

antibodies. wolves

may

in-

infection which from direct conwolves or from

prey. Reilly et that oral infection

idae and other carnivores rogans is possible. They interrogans

from

interrogans

a!.

with observed

be protected

L.

(1970) of Caninterthat L.

from

gastric

acidity because the gulped bolus passes partially digested into the alkaline duodenum. Once infected, wolves may act as maintenance hosts for L. interrogans on account of their highly social behavior.

Everard animals examined

et a!. (1983) in 31 species of wild from Trinidad and Grenada. They 894 animals and found 198

Leptospirosis is endemic in bovine, percine and equine populations of Minnesota. Of 17,014 animal sera examined from 1984 to 1986 at the Minnesota Veterinary Diagnostic Laboratory (St. Paul, Minnesota

(22.1%) results,

positive however,

Our to

55108, against

who

rogans at titers of of antibodies against

our

those

of

Zarnke

that

at titers are in and

of 1:100. sharp contrast

Ballard

(1987)

TABLE 3. Relative prevalence of antibodies against Leptospira interrogans in wolves depending upon their proximity to dairy and beef cattle farms.

Wolf population

Seropositive

at 1:50

Preva-

Sero-

negative at 1:50

lence Total

(%)

27

107

134

20.1

related

25

298

323

7.7

Total

52

405

457

Farm

related

Non-farm

USA), 3,040 (17.9%) had antibodies one or more serovars of L. Inter1 :50. The different

prevalence serovars

in

decreasing order was: icterohaemorrhaglae (24.9%), hardjo (2 1 .4%), canicola (20. 1 %), pomona (18.5%), and grippotyphosa (15.1%). It is possible for a particular serovar to be shared by many maintenance hosts or to be specific to a single host (Hathaway highest for study

et a!., 1983b). seroprevalence

serovar

In our study, (5.3%) in wolves

grippotyphosa.

of moose

in Minnesota,

In

the was

a previous Diesch

et al.

KHAN ET AL.-LEPTOSPIROSIS

(1972) found that var grippotyphosa 328 (27.1%), white ovar pomona were

the prevalence of serowas the highest, 89 of antibodies against serthe least prevalent (18

of 328) (5.5%). Serovar grippotyphosa many countries including China mitsu

(Tonkonozhenko et at., 1987)

from dog, zibethicus),

cattle, swine, squirrels

striped

red fox ( Vu! pes cinereoargenteus), us)

and

opossum

(Hanson,

1982;

species of interrogans

(Ondatra niger,

(Mephitis

vu! pes) hare

,

S.

mephitis),

(Diadeiphus et at.,

may organisms

be

36 (31%)

were (47%)

53

gray

had titers

foxes

Two animals hard Jo albeit 1:100. Collins

most foxes

having

prevalent and 11 of

titers.

Juvenile

higher

geometric

were positive to serovar at tow titers e.g., 1:50 and et at. (1981) have reported

the presence of antibodies Jo in pronghorns (Antilocapra

against

may

indicate

sometime subsequently in L. titers

existence

earlier

the

and decreased.

interrogans may mean

cination residual mona infection

that

persist or longer of

that

were their This

infected

titers have is common

infections where high active infection or vac-

and a fall infection. can

wolves

in titer may indicate Titers to serovar Poin deer (Ferris

antibodies

for 3 mo et a!. 1960). to

multiple

after The sero-

serovars. in wolves was 20.1%

to 7.7% prevalence in wilderis not surprising because L. in(especially serovar pomona)

has

isolated

been

such farm

from

as in recreational waters (Diesch

where

it has

long time. vived for taminated

been

found

of infection

to

survive

and 1966) for

to wild

animals.

It was

disinfection of break the chain to wild animals,

livestock of infecdomestic

and

rodent populations (Will and Contrary to this opinion, it postulated that wild animals may

demiology need for mine the

of L. interrogans infection cattle. However, conclusive area of L. interrogans epi-

is not available indicating more detailed studies to actual rote of wolves and in spreading

and

tospirosis in domestic and elsewhere.

the deterother

maintaining

animals

lep-

in Minnesota

ACKNOWLEDGMENTS We tance,

thank Mary W. K. GomI,

Thurn for secretarial M. E. Nelson, W.

assis-

J. Paul, and many volunteer technicians for assisting with blood-sampling, and U.S. Seal for serum storage. This study was funded by the U.S. Fish and Wildlife Service and the U.S.D.A. North Central Forest Experiment Station. LITERATURE T. C.,

AMUNDSON,

La

AND

virus

Crosse

CITED

T. M.

infection

YUILL. in the

1981. red

fox

Natural (Vulpes

fulva), raccoon

gray fox (Urocyon cinereoargenteus), (Procyon lot or), and opossum (Dideiphis virginians). American Journal of Tropical Medicine

ARMITSU,

a

surconwhich

1972).

act as sources for deer and proof in this

wildlife

environment

(1971) to believe that manure farm area was a potential pubproblem and could also be a

suggested that wastes may help tion from farms animals, Diesch, has been

the

waters, ponds, and McCultoch,

For example, teptospires 61 days in experimentally oxidation ditch manure

L. hardamericana)

of Colorado. Antibodies against serovar pomona, which is a common serovar in domestic animals, were also tow in our study. The tow antibody titers in wolves

with

compared ness. This terrogans

source

Many with L. 1981).

in red and gray of southwestagainst serovar

significantly than adults.

various

251

especially cross-reactivity

The prevalence of antibodies mnhabitating farming areas

virginianus) infected (Shotts,

the red

among

1971).

and Yuill (1981) examined the of antibodies to five different

grippotyphosa with 25 of

of L. interrogans, titers, may indicate

led Diesch in the open lic health

gray fox (Urocyon (Lepus american-

serovars of L. interrogans foxes (U. cinereoargenteus) ern Wisconsin. Antibodies

foxes mean

in and

(Lynx rufus), cottonfloridanus), swamp raccoon (Procyon lo-

Shotts

foxes

Amundson prevalence

muskrat (Sciurus

,

skunk

prevalent U.S.S.R.

et at., 1965; Arhas been isolated

and

carolinensis), bobcat tail rabbit (Sylvilagus rabbit (S. aquaricus) tor),

is the

types tow

IN WOLVES

and Y.,

Hygiene T.

30:

MATUHASI,

706-714.

S. KOBAYASHI,

T. SATO,

252

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gie

und

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J.

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(Canicola)

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AND

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health Animal

for

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in Alaska, 23:

B.

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