ANTIBODIES TO BORRELIA BURGDORFERI IN DEER AND RACCOONS Author(s): Louis A. Magnarelli, James H. Oliver, Jr., H. Joel Hutcheson, and John F. Anderson Source: Journal of Wildlife Diseases, 27(4):562-568. Published By: Wildlife Disease Association DOI: http://dx.doi.org/10.7589/0090-3558-27.4.562 URL: http://www.bioone.org/doi/full/10.7589/0090-3558-27.4.562

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

of Wildlife

Diseases,

© Wildlife

ANTIBODIES

TO

BURGDORFERI

BORRELIA

Louis A. Magnarelli,’ James and John F. Anderson’

H. Oliver, Jr,2 H. Joel

Department of Entomology, The Connecticut New Haven, Connecticut 06504, USA 2 Institute of Arthropodology and Parasitology, Statesboro, Georgia 30460, USA An

ABSTRACT:

antibodies

enzyme-linked

to Borrella

virginlanus)

and

Connecticut,

raccoons

munoglobulmns detected was more sensitive than

353

Carolina,

Experiment

sera)

and

University,

assay

(ELISA)

had

Association

1991

RACCOONS

was

developed

to

detect

agent of Lyme borreliosis, in deer samples were collected from these and

Maryland also (79%

Disease

562-568

pp.

Station,

Southern

Georgia

and Florida in Maryland

AND

1991,

Hutcheson,2

the causative lotor). Blood

(Procyon North

in Connecticut (56% of Maryland, North Carolina, positive sera was highest

Georgia

immunosorbent

burgdorferi,

Maryland,

Agricultural

IN DEER

27(4),

Florida.

Seropositivity

for

serum

(Odocolleus mammals

deer

was

in

highest

(51% of 35 sera). Raccoons in Connecticut, antibodies to B. burgdorferi, but prevalence of 14 samples). Based on adsorption tests, the im-

of in these mammals were probably an indirect fluorescent antibody

analyzing large numbers of serum samples. words: Borrella burgdorferi, Lyme borreliosis, assay, deer Odocolleus virginianus, raccoons Procyon

specific staining

to B. burgdorferi. method and was

The ELISA more suitable

for

Key

INTRODUCTION

Ticks

in the

Ixodes

ricinus

complex

asitize and transmit Borrelia to human beings, domestic wildlife (Carey et al., 1980; and

Keirans,

derson, narelli terium

1983;

par-

1989;

develop

An-

been verified by isolation and immunochemical characterization procedures (Burgdorfer et al., 1982; Burgdorfer, 1989; et al., 1990). In addition, to B. burgdorferi have

and Anderson, Godsey et al.,

latter

is often

method

of automation,

ease

preferred

of standardization

by

al., 1984a, lected from

Ixodes dammini to B. burgdorferi 1986). Since deer during

MagThe

because and

and develop (Magnarelli et

sera can the hunting

ELISA

for

deer in Anthe in

AND METHODS

Blood samples were collected from whitetailed deer, Sika deer (Cervus nippon) and raccoons captured in tick-infested areas of Connecticut, Maryland, North Carolina, Georgia and Florida (USA) in 1984, 1987 to 1989 (deer) or during 1977 to 1984 (raccoons). Examination of these mammals for I. dammini, I. scapularis and other ticks and prevalence of tick parasitism will be reported separately. Blood specimens were collected from deer that had been killed during fall hunting seasons. In Connecticut, samples were obtained at state check stations located in the towns of Barkhamsted, Bethel, East Haddam, East Lyme, Haddam, Marlborough, West Stafford, and Voluntown. Ixodes dammini is common in many of these towns. Additional deer sera (normal specimens) were collected in northwestern Connecticut (check stations at Sharon, Litchfield, and Middlebury), where I. dammini and Lyme borreliosis are rare. Other deer sera were collected in Maryland and

more objective test results that can be statistically analyzed. White-tailed deer (Odocoileus virginianus) and raccoons (Proc yon lotor) are parasitized antibodies

an

Study sites and sampling

serum been de-

1989; 1987).

that are easily capare particularly suitsurveillance programs. of this study was to

evaluate

MATERIALS

tected by indirect fluorescent antibody (IFA) staining procedures or enzymelinked immunosorbent assays (ELISA) (Magnarelli et al., 1984a, b, 1986, 1990a, b; Magnarelli narelli, 1989;

and

immunosorbent

and raccoons to facilitate seroanalyses the surveillance of Lyme borreliosis. other goal was to further determine geographic distribution of this disease the eastern United States.

1989; Anderson et al., 1990; Maget al., 1990a). Presence of this bacin ticks, mammals and birds has

Anderson antibodies

enzyme-linked

and from raccoons tured, these animals able for testing and The main objective

burgdorferi animals and Burgdorfer

Burgdorfer,

antibodies, lotor.

be colseason 562

MAGNARELLI ET AL-ANTiBODIES

Georgia, scapularis,

states

with records of I. dammini or I. respectively. Positive and negative serum controls were available from previous work (Magnarelli et al., 1986). Raccoons were captured alive in Tomahawk traps during summers from sites in southern Connecticut and in Maryland, North Carolina, and Florida. Animals were anesthetized as previously described (Magnarelli et al., 1984a) before blood samples were collected by cardiac puncture. Following a recovery period, animals were released unharmed into their natural habitats. Sera collected during 1980 in Newtown, Connecticut, a town where I. dammini and Lyme borreliosis was uncommon, served as negative controls. For positive serum controls, blood samples were obtained from two raccoons 6 to 8 wk after they were each inoculated four times over 19 days intraperitoneally or in the region of the left axillary lymph node with living B. burgdorferi (CT strain 2591). Numbers of spirochetes in inocula (0.5 ml of BSK II medium) ranged from 2.0 to 7.0 x 10. Additional positive serum samples used previously (Magnarelli et al., 1984a) also were included. All serum samples were stored at -60 C until analyses could be performed. Serologic

tests

Indirect fluorescent antibody staining methods and ELISA were used to detect antibodies to B. burgdorferl (Magnarelli et al., 1984a, b, 1986). Procedures for ELISA were similar to those described for analyses of human and dog sera (Magnarelli et al., 1984b). Washed whole cells of B. burgdorferi (5 to 15 g protein/ml of CT. 2591) were coated to the solid phase at

37 C for blocking diluted

18 to 20 hr.

solutions to 1:160,

and 1:320,

Plates

were

filled

with

washed. Test sera were and 1:640 in phosphate

buffer saline (PBS) solution containing 0.05% Tween 20, 5.0% horse serum, and 50 sg of dextran sulfate/ml. Plates with diluted sera were incubated at 37 C for 1 hr and washed to remove unbound antibodies. Horseradish peroxidase-labeled rabbit anti-deer and goat anti-raccoon immunoglobulin

Perry

G

Laboratories, USA) were and 1:800,

antibodies

(Kirkegaard

Gaithersburg, diluted in PBS

and

Maryland solution to

20879, 1:4,000 respectively. Plates containing these polyvalent conjugates were incubated and washed as in previous steps. Following the addition of 2,2’-azino-di-(3-ethyl-benzthiazoline sulfonate) substrate (Kirkegaard and Perry Laboratories) to each well and a 45-mm incubation period, absorbance values (optical densities) were recorded at 414 nm by using a microtiter spectrophotometer. Net absorbance values were calculated by subtracting readings for PBS controls (i.e., wells without antigen) and

TO BORRELIA

BURGDORFERI

563

compared to values determined for negative Serum controls. Critical regions for positivity were determined by statistical analyses (three standard deviations + mean) of net absorbance values for 30 normal deer sera or 30 normal raccoon sera. Readings for deer sera were considered positive if net optical densities exceeded 0.10, 0.08, and 0.07 for the respective serum dilutions of 1:160, 1:320, and 1:640. Although antise-

rum to white-tailed deer was used to prepare the conjugated reagent, the polyvalent, peroxidase-labeled goat anti-deer antibodies reacted similarly in assays for both genera of deer. Therefore, the same cut-off optical density values were used to determine positive results. A net optical density of >0.08 was considered positive for diluted raccoon sera. All analyses were standardized by using the same negative and positive control sera. Newly purchased enzymelabeled antibodies were tested to determine optimal ibility

working dilutions, of results were

standardization

had

and tests conducted

been

on reproducto verify that

achieved.

Specificity

To determine tained antibodies

if deer and raccoon sera conto Leptospira sp. or Treponerna sp. spirochetes, unadsorbed and adsorbed samples were screened by an IFA staining method or ELISA. In tests for Leptospira sp. antibodies, Leptospira Interrogans serovars canicola (strain Moulton) or pomona (strain MLS) were used as antigens in an IFA assay. Details on the sources and preparation of these and other reagents and on adsorption procedures have been described (Magnarelli et al., 1986, 1990b). The following were used in adsorption experiments: a 1:2 or 1:5 dilution of commercially prepared sorbent (Treponema phagedenis biotype Reiter) or washed cells of B. burg-

dorferi

or L. Interrogans serovars canicola Sorbents were added to test sera wells of microtiter plates prior to analyses remove heterologous or homologous antibodies. Protein contents of washed L. Interrogans or

pomona.

Borrella sp. cells ing a commercially

were

determined available

by assay

or in to of

perform(Bio-Rad,

Richmond, California 98404, USA) and were standardized to 30 g protein/mi after preliminary testing of a range of concentrations (20 to 60 g protein/mi). Each sorbent was mixed in equal volumes (1:1) with test sera. Adsorbed and untreated (control) sera were held at room temperature (23 ± 3 C) for 90 mm and subsequently tested against B. burgdorferl in antibody assays. RESULTS

and

Deer sera Georgia

from Connecticut, were tested

for

Maryland antibodies

564

JOURNAL OF WILDLIFE DISEASES, VOL. 27, NO.

1. tailed deer) ELISA.

Number of serum in eastern United

TABLE

samples States

4, OCTOBER

from Cervus with antibodies

1991

nippon (Sika deer) or Odocolleus to Borrella burgdorfer; by IFA

virginlanus

IFA

Total sera Sites

Years

Connecticut1’

1984,

Cumberland

Is.,

Assateague

Is.,

1989

1987

Georgia1’ Marylan&

1987,

IFA

positive

1:64;

=

ELISA

deer;

IFA

are

here

for

included

‘Sika

to

positive

results

for

Number positive’

(%)

197 (56)

83 (22) 2 (2)

42

3 (7)

13 (34)

35

18 (51)

38

353

98(18)

430

218

(51)

sera

obtained

in Connecticut

have

reported

been

(Magnarelli

earlier

et al., 1986)

and

comparison.

deer.

B.

burgdorferi.

specimens

alence

There

each of seropositive

were

state (Table specimens

from

positive

trations

1). Prevwas high-

method. ples from

For example, Connecticut

method. Seropositivity more than 2-fold tained endpoints

by

more were greater

than tested

in ELISA than

were

IFA staining for high-titered

antibody concentraIFA analyses were

1:10,240

and

respectively.

termine reproducibility, negative serum samples in ELISA. In the second imens remained positive, all 15 negative samples A single dilution negative the

serum

sample

changed

antibody

(n

9) or

=

4 fold

Serum

(n

positive

at

Seropositivity from 15%

79% (Maryland). ranged between majority were

were

by

2 fold

un(n

Although 1:160

relatively

Connecticut,

=

captured

North antibodies

Carolina, to

in ELISA (Connecticut)

in comparative

=

sera

positive

by this

and ELISA. For samples, ELISA seven specimens

method

and

by IFA staining. There itive by immunofluorescence

were

non-reactive

Maximal

in ELISA.

reproducibility, reanalyzed

of

results for 155 11) or negative

negative

four sera staining

posand

antibody than verify

40 serum samples by an ELISA on different

Of these, 24 of 25 positive mained reactive in the second tion end points were identical

were days.

samples retrial. Titra(n = 8) or

differed 3). One

by 2 fold (n sample was

160) and retesting remained

non-reactive subsequently. In the of 15 negative serum samples, 14 negative in the second trial. One was

negative

13) or 4 fold positive initially

=

in

the

in

positive (1:160) after titers for inoculated

reanalysis. raccoons

B.

and

staining

(Tato

1:256

1:5,120 Deer

by

an

IFA

by ELISA. and raccoon

sera

antibody titers and 1:40,968, the low (1:160-i:

antibodies to L. Interrogans an IFA staining method. tested from deer killed

antibody

Maryland

concen-

for

analyses

144) by IFA staining remaining 51 serum more sensitive. Forty

sample

of raccoons

Maryland, contained

burgdorferi. ble 2) varied

In

varied

titers

a

was in

1).

=

samples

Connecticut, and Florida

1,280).

25 positive and 15 were reanalyzed test, 24 of 25 specwhile results for were unchanged. was

1:5,120)

titers were higher in ELISA (1:40,960) by the IFA method (1:2,048). To

de-

of 1:160 in the first trial and in the second test. Differences

remaining or

To

(

procedures. Of these, were positive (n =

were

(22%). Titration samples were

maximal ELISA and 1:4,096,

(n the was

was ob-

comparable; tions in

highest

included

testing samples

350 samby each (56%) results

were

collected during June through August. Serum samples from all four states (n = 206)

est in Connecticut and Maryland, regardless of the assay method used; the ELISA appeared to be more sensitive than the IFA

14)

Total sera tested

1:160.

=

some

(%)

124 549

White-tailed

or

387

1988

Totals

methods

ELISA N umber positive’

tested

(white-

staining

and

Georgia,

first

test

(n

=

(1:

and

Antibody were 1:128 method

were

and

tested

for

spirochetes by Of the 47 samples in Connecticut, none

had

serologic

MAGNARELLI ET AL-ANTiBODIES 2.

TABLE

Number

burgdorferl

of

serum

samples

from

raccoons

Total sera tested

Sites Connecticut

evidence rogans corded collected

recorded

14

40 12 370

The

acted and

to L. pomona

two

interrogans at dilution

specimens

serovars of 1:64,

tibody titers to B. burgdorferi siderably higher (1:20,480 and Adsorption procedures were termine whether with B. burgdorferi

antibodies were

Following

B. burgdorferi in

corded

for

periments as sorbent, identical

two

washed

sorption

IFA

titers

was

In

4 fold

and

N umber

at reciprocal

samples

tit ers

2,560-5,120

640-1,280

10,240

45(15)

17

19

7

2

11(79)

5

5

1

0

6 1 29

7 3 34

0

2’

0 8

0 4

Five

of 1:512 chosen

Following

beyond remaining

increased

cells

or without of deer

to sera,

the use of sorbents. and raccoon sera,

concentrations

sorbent tibody

of T. phagedenis

(1:2 dilution) concentrations.

did

not

reduce

an-

DISCUSSION

Indirect methods

fluorescent are suitable

reliosis

were

and

sera.

derson, 1990a)

normal, titers

the

1987;

animals

geographic

seemed

to be

presence (Steere, 1989).

treatment

with

are

of ticks 1989; Attack

relatively

eastern Steere, Numerous

and

(Magnarelli et present study,

more

sensitive.

distribution correlated

of Lyme with the

in the

I. ricinus

complex

1989; human

Anderson, infections

in

b, on

of B. burgdor-

Burgdorfer, rates for high

An-

et al., 1984a, information

distribution

The geographic borreliosis is strongly

1989; 1989)

(Magnarelli

in wildlife populations 1984a, 1986). In the

ELISA

ence mals

(Steere,

Magnarelli,

1989; Magnarelli and have provided

len al.,

ELISA serum

Both assays of Lyme bor-

in humans

et al.,

domestic

sera,

changes in titration end points were 4 fold or less by IFA staining. There were no changes in antibody titers when sera from inoculated raccoons were tested. However,

antibody and for detecting

burgdorferi. diagnoses

infections

to B. ad-

dilution) serovars,

to B. clinical

Magnarelli

to 1:2,048 for similar

either T. phagedenis sorbent (1:5 or washed cells of L. interrogans

of B. burgdorferi

dorferi with In reanalyses

ex-

speciused.

raccoon

addition

including the positive controls, in duplicate trials resulted in an 8-fold decline in antibody concentrations. Negative serum samples (controls) for deer and raccoons remained non-reactive in tests for B. burg-

antibodies have aided

whole cells sera in duor greater for 13 method

the

recells

unadsorbed

was The

or less.

titers were

separate

to B. burgdorferi

in antibody titers regardless of the assay

tests.

A 4-fold

L. interrogans

for adsorbed

by

to Borrelia

posi-

of T. there titers to

specimens.

samples.

with

having IFA burgdorferi,

23

concentration

This change in results two-fold variability. varied

reacted or cross-

of

However, application of washed of B. burgdorferi to positive plicate trials resulted in 8-fold declines mens,

were con1:40,960). used to de-

a 1:5 dilution Reiter sorbent, in antibody

in 21

antibody

re-

canicola but an-

that specific

treatment

tive deer sera with phagedenis biotype was a 2-fold decline drop

antibodies

(1:40,960).

remaining

reactive.

565

BURGOORFERI

with

States

160-320

15 (38) 4 (33) 75 (20)

of previous or current L. interinfections. Similar results were refor 55 of 57 raccoon serum samples in Connecticut and North Car-

olina.

United

Number (%) positive

304

Island, Maryland Cape Hatteras, North Carolina Merritt Island, Florida Totals Assateague

titer

eastern

BORRELIA

ELISA.

in

Maximum

in

TO

parts

of

United States (Petersen 1989) where I. dammini studies have verified

the

north-

et al., 1989; abounds. the pres-

of B. burgdorferi in ticks and mam(including raccoons) collected in

Connecticut,

New

York

State

and

Rhode

566

JOURNAL OF WLDLIFE DISEASES, VOL. 27, NO.4,

Island

(Anderson,

1989).

Based

on

OCTOBER

our

se-

1991

Treponemal

antibody in serologic

reactivity assays for

rologic results for deer and raccoons and on reports of human infections in tick-infested areas of Maryland, North Carolina,

burgdorferi

Georgia appears

due to Leptospird sp. antibody In addition to flagellin, there

and that

Florida Lyme along

(Tsai et al., 1989), borreliosis occurs

many

sites

United alence pared

States. The relatively lower prevof seropositive raccoons, as comto deer, in Connecticut might be due

to differences infected ticks, B. bungdorfeni

in the host and/or

tibodies. Aside from I. dammini, zootiology

eastern

However,

borreliosis

is unclear.

B. by

burgdorferi,

dark-field

blood (Bosler

samples et al.,

organisms

microstaken from 1983, 1984).

were

antibody

not

sub-

with

memu-

(H5332).

The

of

these organisms is In other studies (Telauthors have concluded

white-tailed

deer

are

In southeastern

reservoir

states,

incom-

I. scapularis

vector of B. burgdorfeni, yet to be isolated from

but these

ticks or from lone-star ticks (Amblyomrna amenicanum) there. Accordingly, further studies of arthropod vectors and vertebrate reservoirs are needed to support serologic test findings and clinical reports that indicate the occurrence of Lyme borreliosis United

There sults when

chetes Zander 1989).

for

in serologic tests whole cell B. the and

false

and

Treponema

re-

borreliosis is

flagellar components Treponema sp.

share common and Lukehart, Consequently,

sp.

positive

for Lyme bungdonfeni

used.

of spiro-

epitopes (Baker1984; Luft et al., immunoglobulins

directed to these polypeptides in heterologous antibody tests

relia

1988). Homologous antibodies bacteria may also contribute to cific reactivity in tests for Lyme

cross-react against Borsp.

antigens.

of

those

to these non-speborrelio-

deer

or

phageof L. on se-

raccoon

Conversely,

the

sera use

of

B. burgdorferi cells as sorbent effectively removed antibodies from sera. Since cutoff levels for positive results were appropriately set to minimize or eliminate cross-reactivity of low concentrations of nonspecific antibodies, the immunoglobulins detected in the present study were probably specific to B. burgdorferi. The high-titered samples collected in Connecticut, where I. dammini and B. burgdorferi are widely distributed, are particularly idase-labeled

convincing. antisera

are commercially analyses can be body production

other of B.

less, isolation mately required

of

in

Now that peroxto deer and raccoons

available, conducted to specific

protein A or polypeptides

present

States.

is potential

For example, Borrelia sp.

as

of about 60,000 to Escherichia (Hansen et al.,

reactivity

monoclonal

in southern

such

having a molecular mass daltons, that are common coil and other bacteria

burgdorferi.

and deer

is a suspected this agent has

is minimal. are surface

burgdorfeni,

B.

al., 1987, reactivity

to B.

these

petent.

of

et

rologic

specific identity therefore unknown. ford et al., 1988), that

proteins

(Magnarelli false-positive

Lyme

cultured in Barbour-Stoenner-Kelly dium or tested by IFA staining rine

the

is reported whereas

sis. However, adsorption with T. denis sorbent or washed cells interrogans had little or no effect

observed

copy in urine white-tailed

of

degree of exposure to immune responses to in persistence of an-

presumably

been

coast

i990b),

being chief hosts for adult the role of deer in the epi-

of

Spirochetes, have

the

it in

sera

to B. human

key

immunodominant Nonethe-

burgdonferi. B. burgdonferi

to confirm deer

Western blot to verify antiouter surface

and

that

raccoons

is ultiantibodies in

more

southern states are indeed specific to this bacterium. Serologic analyses can provide important information in surveillance programs. Detection of antibodies to B. burgdorferi in dogs has been especially useful in identifying foci of Lyme borreliosis (Magnarelli et al., 1985; Greene et al., 1988). These analyses have supplemented tick surveillance studies and evaluations of human case

reports

and

can

be

particularly

help-

MAGNARELLI ET AL-ANTiBODIES

ful

in areas

where

tick

less

prevalent.

dorfeni

are

recent

introduction

vaccine icut

for and

canine

an

in

may

serologic

test

Connectinter-

results

for

dogs

in the future. If this vaccine receives spread acceptance and use, serologic for antibodies in deer, raccoons, footed

mice

leuco

(Peromyscus

other wildlife important in

species national

wideassays white-

pus)

1317-1319.

more pro-

ACKNOWLEDGMENTS

is expressed

to the National

Park

and Fish and Wildlife Services, U.S. Department of Interior, for cooperation at Assateague, Cumberland and Merritt Islands, and Cape Hatteras. We also thank the Georgia Southern Acarology Group, especially Ellen Dotson, Jordi Galbe, Angela James, Sturgis McKeever and Greg Vogel, for help in sera collection and are grateful to the following persons at The Connecticut Agricultural Experiment Station who gave technical assistance: Niel Infante, Harvey

Levine,

Bonnie

research Institute AI-24899

Hamid,

and Carol

Lemmon.

was supported in part of Allergy and Infectious to J. H. Oliver, Jr.

by

A.

Ixodes ixodid Journal

grams.

Appreciation

B., W. L. KRINSKY, AND A. J. MAIN. 1980. dammini (Acari: Ixodidae) and associated ticks in southcentral Connecticut USA. of Medical Entomology 17: 89-99. GODSEY, M. S., T. E. AMUNDSON, E. C. BURGESS, W. SCHELL, J. P. DAVIS, R. KASLOW, AND R. EDELMAN. 1987. Lyme disease ecology in Wisconsin: Distribution and host preferences of Ixodes dammini, and prevalence of antibody to Borrella burgdorferi in small mammals. American Journal of Tropical Medicine and Hygiene 37: 180CAREY,

and

may become surveillance

567

Rheumatic Disease Clinics of North 15: 775-787. , AND J. E. KEIRANS. 1983. Ticks and Lyme disease in the United States. Annals of Internal Medicine 99: 121. A. C. BARBOUR, S. F. HAYES, J. L. BENACH, E. GRUNWALDT, AND J. P. DAVIS. 1982. Lyme disease-A tick borne spirochetosis? Science 216:

the

confuse

BURGOORFERI

dorferi. America

B. burg-

experimental

borreliosis states

of

and However,

of

other

pretations

vectors

TO BORRELIA

187. R. T.,

GREENE,

J.

F. LEVINE,

relia burgdorferl American Journal 476. HANSEN, K., J. M.

of

the

gene

nodominant

wide 56:

to BorCarolina.

Antibodies

dogs in North of Veterinary Research in

49:473-

BANGSBORG,

AND

munochemical

the National Disease grant

1988.

H. FJORDVANG, N. P. HINDERSON. 1988. Imcharacterization of and isolation for a Borrelia burgdorferl immu-

S. PEDERSEN,

This

E. B. BREITSCHWERDT,

H. A. BERKHOFF.

AND

60-kilodalton

range

antigen

of bacteria.

common

Infection

and

to a

Immunity

2047-2053.

B. J., W. JIANG, P. MUNOZ, R. J. DATTWYLER, AND P. D. GOREVIC. 1989. Biochemical and immunological characterization of the surface proteins of Borrella burgdorferl. Infection and Immunity 57: 3637-3645. MAGNARELLI, L. A., J. F. ANDERSON, H. R. LEVINE, AND S. A. LEVY. 1990a. Tick parasitism and antibodies to Borrella burgdorferl in cats. Journal of the American Veterinary Medical Association LUFT,

LITERATURE

J.

ANDERSON,

Ixodes of

F.

tick

1989.

L. III.

Epizootiology

vectors

Infectious

and

Diseases,

A.

CITED

1990.

hosts.

11 Suppl.

MAGNARELLI,

ticks

C.

Borrelia

burgdorferi

that

hamsters.

Journal

of Wildlife

Diseases

S. A.,

Antigenic family 46:

and

infect

between

other

Syrian 26:

1-10.

S. A. LUKEHART.

AND

cross-reactivity

pallidum

STAFFORD

transstadially

transmit BAKER-ZANDER,

Reviews

6: S1451-1459. K.

AND

Bird-feeding

in

of Borrella

reservoir

pathogenic

Spirochaetaceae.

1984.

197:

members

Infection

and

of

the

Immunity

116-121. E.

BOSLER,

M.,

B. G.

J.

ORMI5T0N,

J.

L.

J.

COLEMAN,

Prevalence of the Lyme disease spirochete in populations of white-tailed deer and white-footed mice. Yale Journal of Biology and Medicine 57: 651P. HANRAHAN,

AND

1984.

L. BENACH.

659.

J. L.

J. P.

BARBOUR.

Ixodes 322. the

W.

BENACH,

D. A.

BURGDORFER,

Natural

1983.

dammini

spirqchete.

MASSEY,

AND

distribution Science

A.

of 220:

J. F. ANDERSON, AND C. R. 1990b. Cross reactivity of non-specific treponemal antibody in serologic tests for Lyme disease. Journal of Clinical Microbiology 28: 12761279. 1989. Laboratory diagnosis of Lyme disease. Rheumatic Disease Clinics of North America 15: 735-745. AND J. F. ANDERSON. 1989. Class-specific and polyvalent enzyme-linked immunosorbent assays

G. the

W.

Lyme

1989. disease

Vector/host

for

burgdorferl Veterinary

321-

spirochete,

and

relationships

Borrella

burg-

MILLER,

RIvIERE.

reactivity

BURGDORFER, of

J. L.

COLEMAN,

HANRAHAN,

63-66.

J. N.

Treponema

fectious

detection

in

other

of

antibodies

in equids. Journal Medical Association AND R. C. JOHNSON. serological

spirochetal Diseases

tests

infections. 156:

183-188.

to

Borrella

of the American 195: 1365-1368. 1987. Crossfor

Lyme

Journal

disease of

In-

568

JOURNAL

OF WILDLIFE

C. S. JOHNSON,

AND

rochetes dorferl New

in ticks in York

VOL. 27, NO. 4, OCTOBER

D.

APPERSON,

W.

A.

and and

North

FISH,

1986.

CHAPPELL.

antibodies deer

white-tailed State,

DISEASES,

R. C. Spi-

to Borrella burgfrom Connecticut, Carolina.

Journal

of

Wildlife

Diseases 22: 178-188. A. F. KAUFMANN, L. L. LIEBERMAN, AND C. D. WHITNEY. 1985. Borreliosis in dogs from southern Connecticut. Journal of the American Veterinary Medical Association 186: 955959. W. BURGDORFER, AND W. A. CHAP1984a. Parasitism by Ixodes dammini (Acari: Ixodidae) and antibodies to spirochetes in mammals at Lyme disease foci in Connecticut, USA. Journal of Medical Entomology 21: 52-57. J. M. MEEGAN, J. F. ANDERSON, AND PELL.

1984b. orescent-antibody immunosorbent

Comparison test assay

with for

of an an serological

indirect

flu-

enzyme-linked

studies

of

1991

Lyme disease. Journal of Clinical Microbiology 20: 181-184. PETERSEN, L. R., A. H. SWEENEY, P. J. CHECKO, L. A. MAGNARELLI, P. A. MSHAR, R. A. GUNN, AND J. L. HADLER. 1989. Epidemiological and clinical features of 1,149 persons with Lyme disease identified by laboratory-based surveillance in Connecticut. Yale Journal of Biology and Medicine 62: 253-262. STEERE, A. C. 1989. Lyme disease. New England Journal of Medicine 321: 586-596. TELFORD, III, S. R., T. N. MATHER, S. I. MOORE, M. L. WILSON, AND A. SPIELMAN. 1988. Incompetence of deer as reservoirs of the Lyme disease spirochete. American Journal of Tropical Medicine and Hygiene 39: 105-109. TSAI, T. F., R. E. BAILEY, AND P. S. MOORE. 1989. National surveillance of Lyme disease, 1987-1988. Connecticut Medicine 53: 324-325. Received

for

publIcation

13 February

1991.