Isolation of Borrelia burgdorferi from Peromyscus leucopus in Oklahoma Author(s): Stanley W. Mukolwe, A. Alan Kocan, Robert W. Barker, and George L. Murphy Source: Journal of Wildlife Diseases, 28(2):281-283. Published By: Wildlife Disease Association DOI: http://dx.doi.org/10.7589/0090-3558-28.2.281 URL: http://www.bioone.org/doi/full/10.7589/0090-3558-28.2.281

BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.

BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.

Journal

Isolation

of Borrelia

burgdorferi

from

of Wildlife

Peromyscus

Diseases, 28(2), 1992, pp. 281-283 © Wildlife Disease Association 1992

leucopus

in Oklahoma Stanley W. Mukolwe,’ A. Alan Kocan,’ 4Robert W. Barker,2 and George L Murphy,3 ‘Department of Veterinary Parasitology, Microbiology, and Public Health, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma 74078, USA; 2Department of Entomology, Oklahoma State University, Stiliwater, Oklahoma 74078, USA; and 3Department of Veterinary Pathology, College of Veterinary Medicine, Oklahoma State University, Stiliwater, Oklahoma 74078, USA. Author to whom reprint requests should be addressed

Borrelia

ABSTRACT:

burgdorferi

isolated

was

a field-caught Peromyscus leucopus from Oklahoma (USA). The strain was idenas B. burgdorferi by reaction with mono-

from central

tified clonal

antibody

H5332

specific

for

the

the

known

Ixodes

vectors

dammini

outer

and

I.

pacificus.

Key

words: Borrelia burgdorferi, Peromyscus leucopus, Oklahoma.

Disease,

Lyme

disease

is

spirochetosis caused relia burgdorferi. ness

in the

ease

Control,

disease

vast

and for

been

Pacific

are

in

states,

case

definitions

occurred

since

midwestern

thus States Lyme

appear

increased the

using Traps

in

late

1980’s

transmission

and

of

transmission

in Oklahoma, B. burgdorferi from central

was conducted (1989 to 1991) Noble, central mice

double Trap

Wisconsin, USA) with a combination oatmeal.

exist

the

in the

states

occurrence

we

atfrom Okia-

over in five

a 20coun-

Osage, Pawnee, Oklahoma

and (USA)

All

laboratory

live and

No attempt was parasite infestations.

(An-

and

Sherman live-traps Inc., Tallahassee,

Tomahawk tional Live

of reports

southern

and

(35#{176}50’N to 36#{176}50’N, 97#{176}00’W to 98#{176}00’W).

Wild-caught

states,

to

numbers

and

ties (Lincoln, Payne) of

states (Centers Although vari-

1991).

the

The study month-period

illDis1988)

46

Isolation

report spirofrom non-en-

homa.

midwestern

northwestern

among

Bormost

recorded

upper

Control,

ations

lower

from

of cases

northeastern,

Disease

have

reported

majority

several

et al.,

only tick-borne spirochete in the eastern United et al., 1989). Although

has

the

Ciesielski

areas.

et al. (1991) arthropods

of B. burgdorferi tempted to isolate wild-caught rodents

tick-borne

human tick-borne States (Centers for

1991;

and is the far isolated (Anderson

zoonotic

demic

fluencing

Lyme

by the bacterium The disease is the

reported United

commonly

a

Teltow in several

the infective agent are essential for determining vectors and wildlife reservoirs in geographic locations outside of the highly endemic areas of the U.S. As pant of an ongoing evaluation of ecological factors in-

surface protein OspA of B. burgdorferi. This represents the first isolation of B. burgdorferi from a wild mouse outside of the normal range of

plished, chetes

rats

door Company, set of rodents killed made

were

(H. Florida,

collected

B. Sherman USA) and

live-traps (NaTomahawk,

weekly peanut

and baited butter and

were

returned

with

chloroform.

to determine Liven, spleen,

to

ectokidney,

derson et al., 1989; Miller et al., 1990). Confirmed human Lyme disease cases in Oklahoma between 1988 and 1990 are 8, 25, and 13 respectively (Reiner et al., 1991). Most information on the epidemiology,

heart and urinary bladder were removed aseptically under a sterile hood and placed separately into sterile mortars. Approximately 2 ml of sterile BSK II media (Banboun, 1984) was added and the tissue asep-

ecology,

tically ground with a pestle. A 100 l portion of media containing ground tissue was inoculated into tubes with 6.5 ml BSK

and

transmission

of

B.

burgdor-

feri in the U.S. comes from areas where I. dammini and/on I. pacificus are endemic. Information on vectors, transmission, and wildlife reservoirs from non-endemic locations of transmission

II media, and incubated Cultures were examined by dark-field microscopy of spirochetes. A 0.5 ml

the U.S. is scanty. Although studies have not been accom281

at 33 weekly for the portion of

to 34 C. for 6 wk presence cultures

282

JOURNAL

showing

OF WILDLIFE

spirochetes

cutaneously

was

into

oratory

each

after

BSK

6 wk media

II

VOL

28, NO

2, APRIL

inoculated

of

P.

neared

killed in

DISEASES.

six

sub-

tick-free

leucopus.

123

lab-

Mice

for spirochete following

1992

A

were

re-isolation same pro-

the

cedunes. Fifty-three

Sigmodon floridana)

rats

were one

were

County.

Based

tm

on

an

ranged

during

forms

(18

and

reared

3 wk

of culture;

were

observed

in

BSK

precedures

being

pus

Whole

were

in

solubulized

sod mm

teins

were

thick,

mm

electrophoresis) separating

rylamide

: bis)

(Laemmli,

transferred ifornia,

mm

gelatin

at

40V,

at

with

anti-OspA

antigen MAb M Ab-antigen with affinity kaline Inc.,

mm

monoclonal anti-41

polypeptides,

on

conjugate Maryland, the we

SDS-PAGE observed

dorferi-specific, blot reprobed MAb,

JDI,

after

and

recognizes

H5332,

MAb.

H9724,

the a 41

total

cellular

isolate

(PLOK),

B. herrnsii.

(3)

with

anti-OspA

which

of

mouse

probing

with

at

at

and

50V, with

1%

(MAb)

on the BorkDa flagellar

BRL,

analysis a major

protein

of

kDa

(data

burgdorferi-specific

Panel

kDa

Panel

A

B. burg-

the

B is the

same

genus-specific

Borrelia

flagellar

not

antigen.

Leptospira not shown)

shown).

MAb

in a Western of P10K and of similar size in B. burgdorferi cell lysates (Fig. 1) but did

The

H5332 blot with

(JDI)

not

re-

with the a protein whole

react

with

spp. whole cell antigens (data nor with B. hermsii whole cell

antigens (Fig. a similar sized B. burgdorferi

incubat-

(Gibco USA).

at 31

Cal-

mm

15

were

1983)

blot

acted strongly 31 kDa protein

H9724 (Barbour et al., 1986). interactions were detected purified, goat anti-mouse al-

phosphatase Gaithensbung,

Based

al.,

KDa

analysis

Oklahoma

B. burgdorferi

UV, ap-

antibody

et

(1)

B.

B. burgdorferi-specific,

the

H5332 (Barboun relia genus-specific

15V,

blots

Immunoblot

form

of P10K

Cali-

blocking

Inc.),

either

La Jolla,

at

20

After

75V.

(Bio-Rad

is the

vere

Richmond,

mm

15

(2)

Mini-Tnansblot

Inc.,

for

10% Pro-

(Duralose

the

-29

buffer

Proteins

Systems,

using

mm

(29:1,

1.

FIGURE

proteins

a 0.75 ac-

on

1970).

Cloning

USA) 15

in

buff-

a discontinuous

(Bio-Rad,

fornia, 30

using

USA)

paratus

mm,

(poly-

gel

to nitrocellulose

Stratagene

10 sample

SDS-PAGE

gel

KDa

are

blue).

10%

-43

spirochetes

(SDS)

by

w

P.

C for

bromophenol

system

25V,

100

separated

acrylamide

and

5% 2-mercaptoethanol,

0.1%

-

using

above,

sulfate

SDS,

(2.5%

glycerol,

II media

II media. isolated

at

dodecyl

B

laboratory-

laboratory-reared

in BSK of the

cells

longer

123

(P10K)

six

described

and

12

_______

maintained

leuco

to during

Spirochetes

P. leucopus

the

8.8

m)

all

KDa

micrometer,

first

from

29

1 Neo-

from

the

culture.

KDa

bladder of from Payne

ocular

in length

re-isolated

‘43 52

Spirochetes

to 28

subsequent

were

and

.

isolated from the urinary adult male P. leucopus

organisms

er

leucopus)

hispidus, trapped

(51

toma

ed

(P.

mice

1). MAb H9724 reacted antigen (41 K02) in (JDI) and B. hermsii

with P10K,

(Fig.

1). tion

This report is the of B. burgdorferi

a site

where

cificus mation

are does

both

first confirmed from P. leuco I. dammini

absent. Although not confirm the

and

isolapus at I. pa-

this inforrole of P. leu-

copus in the transmission to humans in Oklahoma,

of B. burgdorferi it does document

P. leuco

natural

pus

as a possible

reservoir

SHORT

for

this

organism

in this

efforts

try.

No

the

rodents

were were

rodents

from

region

made infested

the

of the

with

same

determinant

coun-

to determine areas

if

ticks, are

tick

infected

vectors

mice,

natural This

we

that

do

not

tick

vector

in

study

was

supported

could

disease

infested

on

a

University,

Medicine Technical

publication assistance

Chnistianne

Gray

in part

by

the

of

Veterinary

number 91-034. by Cindy McVey and is appreciated.

BRURE, PERNG,

cally

variable

Borrelia

cotton-tail

rabbits

and

urban

27:

13-22.

BARBOUR, 1983.

CITED

A.

areas.

G.,

Lyme

spirochetes

Journal

S. L. disease

share

Ixodes

isolated

dentatus

of Clinical

TESSIER,

spirochetes

a common

and

A Borrelia-specific

to a flagellar

epitope.

Infection

549-554.

DISEASE

A.

1991.

CONTROL.

Lyme

States

Mortality

Weekly

dis-

1989-1990. Report

C. A., L. E. MARKOWITZ, HIGHTOWER, H. RUSSELL,

W.

1988.

BROOME.

Lyme

disease

New

York

LAEMMLI,

U.

teins

40: 417-

T4.

1990.

in the

United

from

in rural

W.

AND

and

surface

J. ixodid

TODD.

The

disease

head

R. E. BAILEY,

States.

1987-1988.

HUYCKE,

AND

descriptive Journal

collected

Tropical

Medicine

for

dis-

Laboratory

J. B.

S.

epidemiology 84:

State

publication

AND

J.

A.

burgdorferi American

Hygiene

16

Lyme

Medical

503-509.

in Texas. and

MCNABB.

of

Oklahoma

G. J., P. U. FOURNIER, Isolation of Borrelia

thropods

Received

T.

AND

of Lyme

44:

tick

antigenic

pro-

of bacterio-

epidemiology

in Oklahoma.

Association TELTOW,

283-288.

285-289.

K. L., M. M.

1991.

1991.

Microbiology

21:

539:

of the

680-685.

CRAVEN,

The

V. of

Annals

of structural

of the

227:

C.

distribution

of Sciences

assembly

HORSELY, AND

States.

Cleavage

Nature

TSAI.

REINER,

United

1970.

G. L., R. B.

MILLER,

R.

geographic

Academy

during

phage

The in the

K.

Medicine

burgdorferi and

Lyme

Biology

M. E. SCHRUMFF,

1986.

52:

and

CIESIELSKI,

F.

J. F., L. A. MAGNARELLI, R. B. LEFET. G. ANDREADIS, J. B. MCANINCH, G. AND R. C. JOHNSON. 1989. Antigeni-

ANDERSON,

binds

Morbidity

of of

HEILAND,

serveillance-United

ease

LITERATURE

cultivation Journal

421.

Oklahoma.

College

R. A.

TESSIER.

FOR

ease

antibody.

795-804.

521-525.

Immunity

CENTERS

Oklahoma Center for the Advancement of Science and Technology (OCAST) Project No. HR9-012 and published a Oklahoma State

L.

antibody

on

and Yale

HAYES,

S.

283

monoclonal 41:

Isolation 57:

S. F. AND

and

by

Immunity

spirochetes.

Medicine

feed

speculate

and 1984.

but

with larval and nymphal Dermacentor variabilis, Ixodes scapularis, and larval Amblyomma americanum (A. A. Kocan, unpubl. data). Because of the wide range of potential

defined

Infection

COMMUNICA11ONS

August

1991.

RAWLINGS.

from

ar-

Journal

of

469-474.