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1991

Direct Immunofluorescence Detection of Giardiaspp. in Selected Mammals in Central Illinois, UsingAnti-Giardia Polyclonal and MonoclonalAntibodiesJeanette Aba Ackon BrownEastern Illinois UniversityThis research is a product of the graduate program in Zoology at Eastern Illinois University. Find out moreabout the program.

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Recommended CitationBrown, Jeanette Aba Ackon, "Direct Immunofluorescence Detection of Giardia spp. in Selected Mammals in Central Illinois, UsingAnti-Giardia Polyclonal and Monoclonal Antibodies" (1991). Masters Theses. 2246.https://thekeep.eiu.edu/theses/2246

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Date Author

Direct Imrnunof luorescence Detection of Giardia spp.

in Selected Mammals in Central Illinois, using

Anti-Giardia Polyclonal and Monoclonal Antibodies (TITLE)

BY

Jeanette Aba Ackon Brown

THESIS

SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF

Master of Science

IN THE GRADUATE SCHOOL, EASTERN ILLINOIS UNIVERSITY CHARLESTON, ILLINOIS

1 991 YEAR

I HEREBY RECOMMEND THIS THESIS BE ACCEPTED AS FULFILLING

THIS PART OF THE GRADUATE DEGREE CITED ABOVE

7 s .. n\<:, l'i <t \ DATE

Abstract: Giardia species occur in many kinds of mammals,

and some of these hosts have been postulated as potential

reservoirs for human infections. A study was initiated in

fall 1990 to determine the distribution and frequency of

giardiasis in populations of selected wild animals in 12

counties in Illinois. Fecal samples from 64 white-tailed

deer, 13 coyotes, 9 muskrats, 6 raccoons, and 5 badgers were

examined for the presence of Giardia spp. and specifically

for g. lamblia. Anti-Giardia lamblia cysts polyclonal (PAb)

and monoclonal (MAb), FITC-labeled antibody solutions were

used for the direct immunofluorescence detection of Giardia

cysts in the fecal samples. Fifty-five of the samples (37

white-tailed deer; 9 muskrats; 5 coyotes; 4 raccoons) reacted

to the PAb and were thus Giardia positive. one of five

positive coyote samples reacted with the g. lamblia specific

MAb indicating g. lamblia presence. The MAb result indicates

that coyotes can harbor g. lamblia and are possible

reservoirs. There was no indication that the other animal

species studied harbor g. lamblia.

i

Dedicated to my parents,

Abdul Karim and Khadija

"Surely every hour that follows is better for thee than

the one that precedes."

Qur'an 93 5

ii

Acknowledgments

I am indebted to my advisor, Dr. Bill T. Ridgeway, under

whose careful tutelage this work took shape. My deepest

gratitude goes to members of my committee: Dr. R. Andrews,

Dr. c. Costa and Dr. W. James), for their helpful discussion

and critical reading of this paper. My special thanks to

Dr. R. Andrews for his help in obtaining host specimens.

I wish to express my appreciation to Dr. John L. Riggs

(of BioVir Laboratories Incorporated, Benicia, California),

who provided the initial reagents for the study.

I am also grateful to my husband, Emmanuel for his moral

support.

Finally, I wish to thank my colleagues, who helped in

diverse ways; especially Eric Smith for trapping the

muskrats, Scott Kight for his technical help and c. Cox for

his assistance in collecting the coyotes.

This study was supported, in part, by the Zoology Gift

Fund, administered by the Department of Zoology.

iii

Table of Contents

Abstract •••

Dedication. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Acknowledgments.

Table of contents.

Introduction •••.••

. . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Materials and Methods. . . . . . . . . . . . . . . . . . . . . . . Results . ............. . . . . . . . . . . . . . . . . . . . . . . . Discussion/Conclusion •••••••••••••••••••••••

Figures/Tables . ............................ .

Literature Cited . .......................... .

iv

Page

i

ii

iii

iv

1

6

10

13

19

26

Introduction

The intestinal parasitic protozoan, Giardia spp. occur

in a variety of vertebrate animals including humans (Meyer,

1990, 1985; Ackers, 1980). Giardia is transmitted from host

to host by direct fecal-oral route (Bemrick, 1984; Shearer

and Lapham, 1984; Owen, 1984). It can be water-borne (Craun,

1990, 1979; Lippy and Logsdon, 1984), and outbreaks of food­

borne disease in humans have been reported by Barnard and

Jackson (1984).

According to Wright (1980), Giardia is endemic in the

tropic, sub-tropic and temperate climates. In the United

States, giardiasis is prevalent in the Northeastern, Pacific,

and Southwest states but has been reported in all regions of

the country (Lippy and Logsdon, 1984). Children are more

susceptible to infections with Giardia (Pickering and

Engelkirk, 1990). While often asymptomatic Giardia

infections in humans may cause diarrhea and fat

malabsorption; severe cases may lead to dehydration and

excessive weight loss (Beaver et al., 1984; Wright, 1980;

Wolfe, 1984, 1990).

Giardia exists in two distinct morphic forms (Feely et

al., 1990, 1984; Ackers, 1980; Levine, 1979), the motile

trophozoite (size: 9 um by 15 um) and the infective cyst

stage (size: 8 um by 11 um). It is the trophozoite (Fig. 1)

that invades the upper portion of the small intestine and

causes the symptoms of giardiasis (Knight, 1980;

Wolfe, 1990, 1984). The cyst is the transmission stage and

is egested in the feces. It can remain viable outside the

host for three months or more at optimum conditions (eg. cool

water temperature; Erlandsen and Bemrick, 1988).

Filice (1952), described three species groups of the

genus Giardia, (Q. muris group in rodents ; Q. agilis group

in amphibians; and Q. duodenalis group in various mammals),

based on the morphology of their trophozoites. The Giardia

duodenalis group can be divided into different races in

relation to host preference. Species of the Q. duodenalis

group have median bodies shaped like a claw hammer that lie

transversely across the body when viewed in stained specimens

of the trophozoite. The trophozoites are rounded anteriorly

and taper at the posterior end (Meyer, 1985). This group

contains Q. lamblia (=intestinalis) which is pathogenic to

humans, and other species including those living in domestic

animals such as dogs (Q. canis), cats (Q. catti) and bovines

(Q. bovis). The host-specificity of species of Giardia is

difficult to determine. Multiple Giardia spp. infections are

known to occur in some animals (Woo, 1984). Rats, for

example, are infected with both Q. muris and the mammalian

type Q. simoni. Q. lamblia (=intestinalis) is the only

species of Giardia that is infective to humans (Schmidt and

Roberts, 1989). However, Davies and Hibler, (1979), reported

infections in human volunteers, after being fed Giardia cysts

of beaver and mule deer source. The individual infected with

mule deer Giardia cysts experienced severe giardiasis.

2

Various studies have shown that Giardia lamblia can

infect other mammals. Davies and Hibler (1979), found

Q. lamblia cysts from humans to be infective to raccoons,

dogs, gerbils and beavers. Belosevic et al., (1983),

reported Q. lamblia infections in mongolian gerbils.

Strains of Giardia spp. with similar morphology to

Q. lamblia have been identified in beavers and muskrats

(Stibbs et al., 1988), suggesting that these animals, and

possibly others may serve as potential reservoirs for human

infection (Healy, 1990). Beavers, have been implicated as

the source of the infective cysts in some cases of waterborne

outbreaks of giardiasis (Stibbs et al., 1988; Erlandsen and

Bemrick, 1988).

The role of wild and domestic animals in the

transmission of Giardia to humans has been the focus of

several investigations. Buret et al., (1990), reported the

natural occurrence of Giardia spp. in domestic ruminants,

while the incidence of Giardia spp. in muskrats have been

reported by Kirkpatrick and Benson (1987), Pacha et al.,

(1987; 1985), and Erlandsen et al., (1990; 1988). Very few

studies have been conducted on white-tailed deer and coyote,

although Davies and Hibler (1979) reported the presence of

Giardia spp. in these animals and others. There is the need

to identify the potential sources and reservoirs for Giardia

infection (Buret et al., 1990). Animals other than beavers

and muskrats must be extensively studied, in order to obtain

the true prevalence of infections in animal communities.

3

Since ~- lall\blia is of importance to humans, it has been

necessary to investigate the relationship, if any, between

Giardia lall\blia and other members of the ~- duodenalis group.

This has been difficult with conventional light microscopy

due to morphological similarities in mammalian Giardia spp.

cysts and trophozoites (Bemrick, 1984). In the last decade

researchers have studied biochemical and immunologic

differences that may exist within the ~- duodenalis group

(Visvesvara and Healy, 1984; Engelkirk and Pickering, 1990).

Methods used to detect and identify Giardia cysts in

fecal samples have included immunodiagnostic tests, such as

Enzyme-Linked Immunosorbent Assay, ELISA, (Visvesvara and

Healy, 1984) and immunofluorescence (Visvesvara and Healy,

1979; Riggs et al., 1983, 1984; Sauch, 1985; Sterling et al.,

1988; Stibbs et al., 1988; Faulkner et al., 1989). The

traditional direct microscopic method of Giardia detection,

using flotation-sedimentation isolation techniques is time­

consuming and requires experience, skill and patience. In

some cases, detection of cysts in fecal samples proves

impossible due to low-level infections.

Recent studies show immunofluorescence methods to be

specific, reliable and rapid for detection of Giardia spp.

(Sterling et al., 1988). Indirect immunofluorescence assays

using polyclonal and monoclonal antibodies directed against

Giardia lamblia are currently being used for detection of

Giardia cysts from various animal and environmental sources,

with promising results.

4

Riggs et al., (1984), described a procedure for

detecting Giardia cysts by direct immunofluorescence antibody

testing. In this procedure the antibody is directly labeled

with the conjugate antibody and is later incubated with the

antigen and viewed by fluorescence microscopy.

A modification of Riggs' method was used in the present study

to examine Giardia spp. presence in fecal samples from white­

tailed deer (Odocoileus virginianus), muskrat (Ondatra

zibethicus), coyote (Canis latrans), raccoon (Procyon lotor),

and badger (Taxidea taxus).

This study was conducted: 1) to investigate the natural

occurrence of Giardia spp. in selected mammals in Central

Illinois, 2) to determine if the study animals harbor Giardia

lamblia, and 3) to determine any relationship between age,

and sex of animals and infection with Giardia.

5

Materials and Methods

All hosts used in this study were collected within a

twelve county area of Central Illinois (Figure 3).

Fecal pellets from white-tailed deer were obtained from

deer at check stations located in each of twelve counties in

Central Illinois during the 1990 hunting season (Figure 3).

Samples were placed in screw top vials, labeled with age,

sex, and county and returned to the laboratory.

Muskrats were kill-trapped from Lake Glenwood (a five­

acre water body in section 15 of Charleston Township, Coles

County) between November 21 and 27, 1990.

Raccoons and coyotes were trapped in Hutton Township,

Coles County. The colon and cecum were removed and placed in

Zip-loc freezer bags and transported to the laboratory. The

age and sex of each host was recorded.

Fresh fecal samples were collected between November 1990

and March 1991. Samples were stored in the refrigerator and

within 48 hours after collection examined using a modified

Riggs' Direct Immunofluorescence (DIF) method. Preserved

badger fecal samples were also examined.

Treatment Q.f Samples

Direct smears and concentrated cysts smears were made

for the test reactions. For the direct smear 2 mg of a fecal

sample was placed in a drop of water on a microscope slide,

and two small circular smears (about 1 cm in diameter) were

6

made and air-dried. Fecal samples were concentrated to

facilitate detection of cysts. For the concentrated cyst

smears two grams of feces was thoroughly comminuted in 15 ml

tap water in a paper cup, and strained through a metal

strainer (mesh size about 12 per cm) into a second paper cup.

The fine suspension was placed in 15 ml centrifuge tubes and

centrifuged at 500 x g for two minutes. The supernatant was

discarded, and using a wooden applicator stick, two spots of

sediment were applied to each slide and air dried. Both the

direct and concentrated cyst smear slides were labeled with

specific codes assigned to each fecal sample. The smears on

the slide were ringed with glass marker pencils and tested

for Giardia using antibody solutions.

Direct Immunofluorescence ~

Fecal samples were tested with both polyclonal and

monoclonal antibody solutions (John L. Riggs, BioVir

Laboratories Inc. Benicia, CA). One of the pair of smears

was treated with a drop of Fluorescein Isothiocyanate (FITC)

labeled, Goat anti-Giardia lamblia cyst polyclonal antibody

(PAb). The other smear on the slide was reacted with a drop

of FITC labeled anti-Giardia lamblia cyst monoclonal antibody

(MAb).

The slide was incubated at room temperature for 20

minutes in a covered petri dish lined with strips of damp

tissue paper, to provide a moist chamber. The slides were

removed and washed five times with 10 ml phosphate buffered

7

saline (PBS pH 7.2). The PBS washed slides were air dried

and then covered with a drop of buffered glycerol (90%

glycerol in PBS). A 22 mm x 40 mm coverglass was placed over

the preparation and sealed with clear nail lacquer. When dry

the slides were examined for Giardia cysts by fluorescence

microscopy, using American Optical light microscope (Model

2070) equipped with Vertical Illuminator for incident light

fluorescence microscopy with FOX 100-watts Halogen projector

lamp (USHIO, Tokyo, Japan) and a fluorescein-fluor cluster

(Exciter filter= 490 nm, Dichroic filter= 500 nm, and Barrier

filter= 515 nm).

Detection of Giardia Cysts

Slide preparations were scanned with low and medium

power objectives. A distinct apple-green fluorescence on the

surface of cysts (400x magnification), indicated a positive

reaction. Greatest length and greatest width of each cyst

was measured using a calibrated ocular micrometer.

Positive and Negative Controls

Positive and negative controls were important for the

proper interpretation of results. Giardia lamblia cyst

slides, were used for positive and negative controls. Each

slide had two wells with fixed Sh_ lamblia cysts (Riggs,

BioVir Lab. Inc., CA). A drop of anti-Giardia lamblia FITC

labeled monoclonal antibody solution was added to well one

for positive control, and PBS was placed in well number two

8

for negative control.

Statistical Analysis

Results of direct immunofluorescence test of fecal

samples from white-tailed deer were analyzed by Chi-square

(contingency tables). The relationship between age; sex and

infection with Giardia was considered significant if the

probability was less than five percent (P<0.05). Sample size

from muskrat, coyote and raccoon was too small to warrant

similar statistical analysis.

9

Results

Direct immunofluorescence antibody testing of fecal

samples indicated the presence of Giardia spp. in 57.8% of

white-tailed deer, 100% muskrat, 38.5% coyote, 66.8% raccoon.

None of five badger samples tested were positive for Giardia

spp. (Table 1).

Of 97 fecal samples examined, 56.7% reacted with

polyclonal antibody (PAb) solution indicating the presence of

Giardia spp. in the host animals; and of these, one (coyote­

ID # 13) reacted with the monoclonal antibody (MAb) solution

indicating ~. lamblia presence.

All Giardia cysts were measured at 400x magnification

and ranged in size from 10 um to 14 um long by 6 um to 9 um

wide (average; 12 um by 6 um). All cysts were elliptical in

shape, and no internal morphology was observed. Cysts

reacting with MAb solution fluoresced intensely (Figure 2)

compared to those reacting with the PAb, and did not fall

within the size ranges described above.

Odocoileus virginianus (white-tailed deer):

Samples from white-tailed deer from ten of twelve

counties surveyed were positive for Giardia spp. (Table 2).

Those from the other two counties were negative. Of the 64

white-tailed deer examined 57.8 % reacted with the PAb. All

were MAb negative.

Age distribution of white-tailed deer and Giardia

10

infection per age group are shown in figure 4. Infection on

the basis of sex is shown in figure 5.

Statistical analysis showed no relationship between age

and infection (X2=0.41; P>0.05), nor was there any

significant relationship between sex of white-tailed deer and

infection with Giardia spp. (X2=1.50; P>0.05).

Twenty-five Giardia cysts from white-tailed deer had

average dimensions (Table 1) of 10 um long (range 8-12) by 6

um wide (range 5-7).

Ondatra zibethicus (muskrat):

Eight males and one female muskrat were sampled; (3

adults, 4 juveniles and two of unknown age). All nine

samples were PAb positive. None were MAb positive. Thirty

Giardia cysts from muskrats (Table 1) had dimensions with

range of 12 um to 16 um long by 7 um to 11 um wide.

Canis latrans (coyote):

Five of thirteen (38.5%) adult coyotes were Giardia

spp. positive, and of these, one sample (a female, ID # 13)

reacted to the anti-Giardia lamblia cysts monoclonal antibody

solution indicating the presence of ~- lamblia. Three of the

five Giardia positive samples were from male coyotes and 2

from females (Table 3).

The size of 19 cysts measured from PAb positive coyotes

(Table 1) had average dimensions of 11 um long (range 13.5-8)

by 7 um wide (range 10-5). The mean size of 25 cysts from

11

the one monoclonal antibody positive sample was 13 um by 9

um, (range; 12 um to 15 um long by 8 um to 10 um wide).

Procyon lotor (raccoon):

Four of six fecal samples from adult raccoons reacted

with the PAb (Table 4). There was no MAb reaction. Three

male and one female raccoon were infected. Ten Giardia cysts

(Table 1) observed ranged from 13.5-10 um in length by 8.5-

7.0 um in width (average, 12 um by 8 um).

Taxidea taxus (badger):

Five badger samples, preserved in polyvinyl alcohol

since 1986, were tested for the presence of Giardia spp.

None of the samples reacted to either PAb or MAb.

One of three human stool samples examined for Giardia

presence, reacted to both PAb and MAb. The other two were

negative. Twenty-six ~- lamblia cysts had average dimensions

of 14 um long (range 16-12) by 8 um wide (range 10-7).

12

Discussion

The study confirms the presence of Giardia spp. in fecal

samples collected from white-tailed deer, coyote, muskrat and

raccoon, in Central Illinois.

Although a positive monoclonal antibody (MAb) reaction

was detected for one coyote sample, the lack of MAb reaction

among other tested hosts suggest that, the species of Giardia

found in host animals such as those studied here, are species

of the ~- duodenalis group but not ~- lamblia. Therefore,

with the exception of Canis latrans the animal species

studied probably do not constitute a potential source of

Giardia infection for humans in this area. A more intense

and prolonged study of the local coyote population seems to

be indicated in order to elucidate coyote-Giardia-human

relations.

The direct immunofluorescence antibody test is highly

specific and sensitive to Giardia spp. It detects the

surface of the cyst wall and not the actual internal

morphological characteristics, previously described by

earlier researchers such as Filice, (1952).

Data obtained on cyst measurements showed variable sizes

in length and width (Table 1). There was overlap in the

overall dimensions of cysts from the various animals. This

supports the long-standing view that, variations in Giardia

spp. cyst size are independent of host species (Bemrick,

1984; Filice, 1952; Meyer, 1985) and cannot be used

13

successfully to distinguish species within the G. duodenalis

group. However, in this study the ~- lamblia cysts from the

one coyote had dimensions (13 um by 9 um) which were greater

than the average range in size (12 um by 6 um) of cysts from

other wild animal hosts, but was similar in size to the

~- lamblia cysts (14 by 8 um) measured from the human

specimen.

The antibody testing method offered a rapid, direct

evidence of Giardia presence by the characteristic apple­

green fluorescence of cysts. There was no cross-reaction

with other organisms (for example: Chilomastix mesnili;

Enteromonas spp.) present in the fecal samples. studies by

Riggs et al. (1983; 1984); Erlandsen and Bemrick (1988), show

that the Riggs antisera does not cross-react with non-Giardia

organisms found in fecal samples.

The antibody solutions were tried on human stool samples

obtained from three individuals who had recently arrived in

the United States. Samples from a three year old male child

was Giardia positive, reacting with both polyclonal and

monoclonal antibody solutions. These individuals had

previous history of giardiasis and had been treated. It was

evident from the test results that the asymptomatic child was

still shedding Giardia lamblia cysts. This finding supports

the idea that the direct immunof luorescence antibody testing

enhances the detection of cysts in asymptomatic persons, and

that the monoclonal antibody solution used was indeed

sensitive to ~- lamblia in humans.

14

The MAb positive reaction of one fecal sample from an

adult female coyote is significant. The many cysts observed

fluorescing in the sample were Q. lamblia cysts, since the

monoclonal antibody recognizes antigenic determinants of

Q. lamblia origin. It is possible that this particular

coyote had come in contact with Giardia lamblia contaminated

feces and had subsequently become infected.

The presence of g. lamblia cysts (size 13 um long by 9

um wide) in the coyote could be incidental, since none of the

other coyote samples (all obtained within a one square mile

area in Hutton Township, Coles County, Illinois) reacted with

the g. lamblia specific monoclonal antibody. However, it is

an indication that coyotes can harbor g. lamblia and may be

potential reservoirs of human giardiasis.

Due to the short duration of this project it was not

possible to further investigate this interesting finding.

Further work therefore needs to be done before definite

conclusions can be drawn.

Ten of twelve Central Illinois counties surveyed for

white-tailed deer Giardia spp. were positive, although in

some cases only one fecal sample was received for the county.

This in itself is not a reliable indicator of the actual

prevalence of Giardia spp. in deer of the area. The Giardia

spp. occurring in white-tailed deer is not Q. lamblia since

fecal samples failed to react with the MAb solution.

The preferential shooting of male deer by hunters

accounts for the high percentage of male white-tailed deer

15

sampled for Giardia. This constitutes a biased sample,

however there was no statistically significant difference

between male and female white-tailed deer regarding Giardia

infection (p>0.05 by X2). Studies by Erlandsen et al.,

(1990), showed no difference in Giardia prevalence on the

basis of sex in beavers nor in muskrats.

Erlandsen et al., {1990), found the age of beavers to be

correlated to Giardia infection (juvenile 23.2%; adult 12.6%)

but found no correlation in muskrats (equally infected).

Buret et al., (1990), in their study, found high Giardia

infection rates in lambs (35.6%) and calves (22.7%).

According to Bemrick (1984) younger animals are more likely

to be infected than adults due to their low resistance. In

this study, however, there was no relationship between age of

white-tailed deer and infection with Giardia (Figure 3), as

determined by Chi-square analysis test (P>0.05). The

muskrat, coyote, and raccoon sample size was too small to

warrant any statistical evaluations.

Studies conducted on muskrats show high prevalence of

infection (82.5%, Pacha et al., 1987; 70% Kirkpatrick and

Benson, 1987). All the muskrats examined in this study were

infected. The muskrat Giardia cysts failed to react with the

monoclonal antibody solution even though studies indicate

that the cysts are morphologically similar to human Giardia

(Erlandsen and Bemrick, 1988). It is evident that the cyst

differ immunologically. Erlandsen and Bemrick (1988),

suggest that the muskrat (which has binary cysts unique to

16

microtine rodents) is infected by its own type of Giardia

spp. No evidence of Giardia presence was found in badger

fecal samples that had been preserved in polyvinyl alcohol

for six years. Very little background fluorescence was

observed and this negative reaction can be explained in

either of two ways. It is possible that these samples did

not have naturally occurring Giardia. On the other hand, it

is also possible that any Giardia cysts that were initially

present in the feces have been disintegrated by the long

preservation. The antibody solutions react poorly with

specimens that have been preserved in alcohol for long

periods of time (Stibbs et al., 1988). Faulkner et al.,

(1989), demonstrated the presence of Giardia spp. cysts in

ancient fecal samples of human origin (about 2500 years old),

using immunofluorescence assay. It appears then, that the

method works efficiently on unpreserved specimens.

The results obtained in the study indicate that,

muskrat, raccoon and white-tailed deer, may not play an

important role as potential reservoirs of ~. lamblia.

Coyotes on the other hand, are likely prospects because of

the positive monoclonal antibody reaction observed in this

study. The collection of more fecal samples from coyotes in

the area of Hutton Township and additional sites would be

necessary to determine the role coyotes could play in

harboring ~. lamblia.

In summary, the considerable potential of monoclonal

antibodies (MAb) in the study of Giardia spp. is of great

17

importance. It provides diagnostic advantage and it is

possible that, in the near future the MAb technique could be

used in differentiating strains or even species of Giardia

within the ~. duodenalis group.

18

disc

nucleus

median body

flagella -;/" . . . ~"

/ · :·. ~ ~ : ) --c y s t 1.-1 a 1 1 I . . . , I

(A) · Trophozoite

/. ·.j· w:-6. cytoplasm

I .. -. j \ '. . - .

Fig. 1.

Fig. 2.

(after Filice, 1952) s ize 14 um long by 10 um wide

\ . . ''" . , , __ -

(8) Cyst (from Meyer,1985) size 11 u m long

Structure of Giardia lamblia.

Photograph showing fluorescing ~. lamblia cysts (400x) from Canis latrans (arrows), stained with anti-Giardia lamblia FITC-labeled monoclonal antibody solution.

Fig. 3.

Counties

1. Edgar 2. Clark 3. Cumberland 4. Coles 5. Moultrie 6. Shelby 7. Fayette 8. Effingham 9. Clay

10. Richland 11. Edwards 12. Wayne

Map of Illinois showing region of white-tailed deer sample collection. * Muskrat, coyote and raccoon samples were collected in Coles County.

Table 1. Direct Immunofluorescence Giardia survey of fecal samples from various mammals of Central Illinois.

Animal Number Host tested *PAb *MAb

White-tailed 64 37 O deer

Coyote 13 5 1

Muskrat 9 9 o

Raccoon 6 4 o

Badger 5 o o

*PAb= polyclonal antibody *MAb= monoclonal antibody

% PAb positive

57.8

38.5

100

66.7

0

Cyst size (um)

10 x 6

11 x 7

14 x 9

12 x 8

0

Table 2. Giardia spp. in white-tailed deer from 10 Illinois counties based on *DIF analysis.

PAb County Samples tested *PAb positive

Edgar 10 7 70

Clay 10 6 60

Edward 10 6 60

Clark 9 6 67

Effingham 7 4 57

Fayette 4 4 100

Moultrie 3 1 33

Wayne 3 1 33

Richland 2 1 50

Coles 1 1 100

*DIF= Direct Immunofluorescence *PAb= pol¥clonal antibody All positive to PAb; no reaction with MAb

Central

35

30

25

... C1> 20 ~

~ 15 .c E :J z 10

5

0.5-1.5 2.0-3.0 3.5-4.5 unknown

Age Group (years)

Fig. 4. Age distribution of white-tailed deer and infections recorded for each group. Hatched bar indicates the number of individuals positive for Giardia spp.

.... Q) Q)

0 0

50

40

30

Ci> 20 .0 E ::> z

10

Male Female

Sex of Deer

Unknown

Fig. 5. Giardia infection in white-tailed deer on the basis of sex. Hatched bar indicates the number of positive indivi~uals.

Table 3a. Analysis of Giardia positive coyotes by sex

sex of Coyote

Male

Female

Table 3b.

Number tested

Number positive

6

7

PAb MAb

3 0

2 1

DIF reactions of fecal samples from 6 adult raccoons.

number positive Sex of raccoon PAb MAb

Male 3 0

Female 1 0

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