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Eastern Illinois UniversityThe Keep
Masters Theses Student Theses & Publications
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.
This is brought to you for free and open access by the Student Theses & Publications at The Keep. It has been accepted for inclusion in Masters Thesesby an authorized administrator of The Keep. For more information, please contact [email protected].
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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