Evaluation of Coprological and Serological Techniques For Diagnosis of Bovine Fasciolosis

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Israel Journal of Veterinary Medicine  Vol. 69 (4)  December 2014 203 Diagnostic Tests for Bovine Fasciolosis
INTRODUCTION
Fasciolosis caused mainly by Fasciola hepatica as well as F.
gigantica is a widespread parasitic disease of ruminants. In
adult cattle, the infection usually follows a chronic course,
with no obvious clinical signs. Even when asymptomatic,
fasciolosis may cause economic losses in the cattle industry
(1) due to compromised weight gain (2), milk yield, and fer-
tility (3). In endemic areas grazing animals have the same
susceptibility of fasciolosis as confned animals, only the risk
for infection is lower (4).
Te diagnosis of fasciolosis is usually based on the de-
tection of F. hepatica eggs in feces or F. hepatica specifc
antibodies in serum or milk. Te enzyme-linked immuno-
sorbent assay (ELISA) often relies on excretory-secretory
(ES) products from the liver fuke (3, 5). Recently, a method
based on detection of a F. hepatica specifc coproantigen has
been developed and commercialized (4). Te ELISA method
developed for determination of Fasciola coproantigens in fe-
ces appears to be an alternative to coprological examination
(6, 7).
Evaluation of Coprological and Serological Techniques For Diagnosis
of Bovine Fasciolosis
Avcioglu, H.
1
*, Guven, E.
1
, Balkaya, I.
1
, Kaynar, O.
2
and Hayirli, A.
3
1
Ataturk University, Faculty of Veterinary Medicine, Department of Parasitology, Erzurum, Turkey.
2
Ataturk University, Faculty of Veterinary Medicine, Department of Biochemistry, Erzurum, Turkey.
3
Ataturk University, Faculty of Veterinary Medicine, Department of Animal Nutrition & Nutritional Disorders, Erzurum, Turkey.
*
Corresponding Author: Assoc. Prof. Hamza Avcioglu, Ataturk University, Faculty of Veterinary Medicine, Department of Parasitology, Erzurum, Turkey.
Tel: +90-442-2315536, Email: hamzaavcioglu@yahoo.com
ABSTRACT
Tis study was carried out to determine the prevalence bovine fasciolosis in Erzurum Province in Turkey
and to evaluate sensitivity and specifcity of the coproscopy by sedimentation and antibody-ELISA
tests considering the copro-ELISA test as the gold standard. A total of 282 cattle (230 female and 52
male; Holstein, n=6; Simmental, n=6; Brown Swiss, n=159; and crossbreed cattle, n=111), at an average
of 3.55±2.49 [mean±SD, (range 1.00-15.00)] years old, from local farms were monitored for fasciolosis
between April 2011 and June 2011. Animals were grazed and watered on communal areas during days and
housed in barns at nights. Blood and fecal samples were collected for coproscopy and serology for F. hepatica.
Data were subjected to Chi-square analysis, analysis of variance, and receiver operating characteristics
curve development. Te prevalence rate was 35.46, 61.70, and 34.04% when assessed by the copro-ELISA,
antibody-ELISA, and sedimentation tests, respectively (P < 0.0001). Cattle that were female, purebred, and
in advanced ages (≥6 years) had greater fasciolosis prevalence than those were male, crossbred, and in younger
ages (2≤ years) (40.4 vs. 3.6%, P < 0.0002 for sex; 22.8 vs. 55.0%, P < 0.0001 for breed; and 57.8 vs. 20.0%,
P < 0.0001 for age) as attained by the copro-ELISA test. Sensitivity and specifcity were 100.0% and 59.3%
for the antibody-ELISA technique and 96.0% and 100% for the sedimentation technique, considering
copro-ELISA technique gold standard. Tese data suggest that fasciolosis prevalence can greatly vary by
the diagnostic methods and should be cautiously interpreted as they refect disease status at diferent stages.
Keywords: Diagnostic method; Fasciolosis; Prevalence; Sensitivity; Specifcity.
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Nevertheless, the diagnosis of fasciolosis is complicated
due to the liver fuke’s biological cycle within the defni-
tive host. After ingestion of metacercariae, juvenile worms
migrate through the intestinal wall to the peritoneal cavity,
penetrate the liver parenchyma through which they migrate,
and pass into the biliary tract, where they ultimately reach
maturity and start oviposition. Tat is, eggs become present
in feces several weeks after the ingestion of metacercariae (8).
Moreover, ingested parasites, depending on immune status
of the host, may not always maturate in the liver. In less
severe infestation, eggs can be observed in only a serial fecal
collection because egg counts exhibit diurnal variation. In
contrast to Charlier et al. (9) who found that egg enumera-
tion, depending on the method used, may be used to identify
the heavily infected animals, and thus can guide treatment
decisions (4, 5).
Alternative immuno-serological methods have been de-
veloped for early diagnosis of fasciolosis. Te ELISA test is
easy to perform for herd monitoring at early stage of fas-
ciolosis (5, 10). Especially, the copro-ELISA test, based on
determination of ES antigens in feces, has high specifcity
as confrmed by necropsy and high sensitivity as confrmed
by lacking cross-reaction with antigens from other helmin-
thes. It also shows positivity during the frst 1-5 weeks after
parasites reaching the biliary tracts (8). Te objectives of this
study were to determine prevalence of bovine fasciolosis,
with associated risk factors in Erzurum Province in Turkey
and to compare sensitivity and specifcity of the coprology
by sedimentation and antibody-ELISA tests considering
the copro-ELISA test as the gold standard in diagnosis of
fasciolosis.
MATERIALS AND METHODS
Study area and animals
Te study was conducted in Erzurum (39°52’ N, 41° 17′ E,
1853 m above sea level) province, located in the eastern part
of Turkey. Te region receives 453 mm rainfall annually with
a temperature ranging from -35 to 35°C. Te animal produc-
tion is mainly based on intensive grazing.
Te requied sample size was estimated to be 323, using
the following formula: n = (Z
1-a
2
x [P x (1-P) / D
2
], where
P = prevalence, Z = confdence interval at 95%, P = absolute
precision, % (11). A total of 282 cattle (230 female and 52
male in terms of sex / Holstein, n=6; Simmental, n=6; Brown
Swiss, n=159; and crossbreed cattle, n=111 in terms of breed),
an average of 3.55±2.49 (mean±SD, range 1.00-15.00) years
old, from local farms were monitored for fasciolosis between
April 2011 and June 2011. Animals were grazed and wa-
tered on communal areas during days and housed in barns
at nights.
Blood and fecal samples
Ten milliliters of whole blood was drawn from the jugular
vein into additive-free vacutainers. Sera were harvested fol-
lowing centrifugation of clotted blood at 1,500 rpm at 20°C
and aliquots were stored at -20°C until ELISA analysis. Fecal
samples (50-100 g) were collected per rectum using gloved
fngers for coprological tests.
Te study protocol was approved by the Animal Care
and Use Committee at Atatürk University (20.10.2009-
2009/105 decision number).
Coprological examination
Coproscopy was performed by Benedect sedimentation.
Briefy, 6 g fecal samples were suspended in tap water, sieved
through a grid (mesh size 250 µm) into a beaker in a vol-
ume of 250 ml. After 3 min the solution was decanted and
refilled with water. Tis process was repeated twice. Ten,
the sediment was stained with 2 drops of methylene blue,
decanted into a Petri dish and scanned for Fasciola eggs using
a binocular with 100 X magnifcation (12).
Fecal samples were also subjected to determination of
F. hepatica ES antibody using the ELISA test (BIO K 201,
Fasciola hepatica Antigenic ELISA Kit, BIO-X Diagnostics,
Jemelle, Belgium). Te optical density (OD) was measured
at 450 UV wavelength by a fully automatic ELISA reader
(µQuant, Bio-Tek Instruments, Inc., Winooski, VT, United
States) equipped with a fully automatic microplate wash-
er (ELx50 microplate washer, Bio-Tek Instruments, Inc.,
Winooski, VT, United States). Blank reading was subtracted
from each sample reading. As recommended by the manu-
facturer, the cut-of OD value was > 0.150 and the positive
control OD value was > 1.533.
Serological examination
Te odd colums (1,3,5,7,9,11) of microplates were coated
with Fasciola hepatica ES antigen captured by the monoclonal
antibody (BIO K 211, Bio-X Diagnostics, Jemelle, Belgium).
Whereas, the even columns (2,4,6,8,10,12) contained only
Research Articles
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Israel Journal of Veterinary Medicine  Vol. 69 (4)  December 2014 205 Diagnostic Tests for Bovine Fasciolosis
the monoclonal antibody (BIO K 211, Bio-X Diagnostics,
Jemelle, Belgium) serving as a negative controls to distin-
guish specifc anti F. hepatica antibodies from non-specifc
ones.
Te test blood sera were diluted 1:100 in the dilution
bufer and each serum sample was applied to a coated cell
and an uncoated well. After washing at the end of incubation
period plates were added with the conjugate (a peroxidase-
labelled anti-bovine IgG1 monoclonal antibody (Bio-X
Diagnostics, Jemelle, Belgium)). Te plates were incubated
at room temperature and washed again. Ten, the enzyme’s
substrate (hydrogen peroxide) and the chromogen tetra-
methylbenzidine were added. Te intensity of the resulting
blue color was proportional to the titer of specifc antibody
in the sample. Te OD’s in the microwells were evaluated
spectrophotometrically (µQuant, Bio-Tek Instruments, Inc.,
Winooski, VT, United States) using a 450 nm flter and the
absorbance of the uncoated well was subtracted from the ab-
sorbance of the coated well. Te corrected absorbance values
were divided by the corresponding positive control serum
OD value. Te results were categorized as negative and three
positive antibody levels; -, +, ++, and +++, respectively . For
titer levels the results were characterized as < 15, 15-45, 45-
75, and > 75, respectively.
Statistical analysis
Due to small sample size for Holsteins and Simmentals,
animals were categorized as purebred and crossbred.
Animals were also categorized by age (< 2, 2-3, and > 6
years). Cross-tables were generated using the PROC
FREQ procedure to determine association of animal
factors (sex, age, and breed) with fasciolosis in the Chi-
square analysis (29). Antibody titers were analyzed using
the PROC MIXED procedure and mean diferences by
the degree of antibody-ELISA score were assessed using
the PDIFF option.
Te receiver operating characteristics (ROC) curves
were developed to compare sensitivity (ability to detect fas-
ciolosis), specifcity (ability to avoid misclassifying healthy
animals as animals with fasciolosis), positive likelihood ratio
(low specifcity or how much the odds of the disease in-
creases when a test is positive), and negative likelihood ratio
(low sensitivity or how much the odds of the disease de-
creases when a test is negative) for the antibody-ELISA and
sedimentation methods under consideration that the copro-
ELISA method is a gold standard (MedCalc version 9.6.2.0,
MedCalc Software, Mariakerke, Belgium). Tese tests were
compared based on their under areas of curves using z-test.
Statistical signifcance was declared at P < 0.05.
RESULTS
Prevalence and risk factors
Te overall prevalence of fasciolosis was 35.46, 61.70, and
34.04% as assessed by the copro-ELISA, antibody-ELISA,
and sedimentation methods, respectively (Table 1). Females
Table 1: Risk factors for bovine fascioliasis
Diagnostic Test
Copro-ELISA Antibody-ELISA Coproscopy by Sedimentation
Risk Factor - (n=182; 64.54%) + (n=100; 35.46%) - (n=108; 38.30%) + (n=174; 61.70%) - (n=186; 65.96%) + (n=96; 34.04%)
Sex
Female (n=230; 81.56%) 137 93 73 157 140 90
Male (n=52; 18.44%) 45 7 35 17 46 6
X
2
= 13.48; P < 0.0002 X
2
= 22.71; P < 0.0001 X
2
= 14.38; P < 0.0001
Age (yr)
2≤ (n=130; 46.10%) 104 26 72 58 107 23
3-5 (n=107; 37.94%) 59 48 25 82 60 47
≥6 (n=45; 15.96) 19 26 11 34 19 26
X
2
= 27.50; P < 0.0001 X
2
= 29.81; P < 0.0001 X
2
= 31.42; P < 0.0001
Breed
Purebred (n=171; 60.64%) 132 39 87 84 134 37
Crossbred (n=111; 39.36%) 50 61 21 90 52 59
X
2
= 30.40; P <.0001 X
2
= 29.09; P < 0.0001 X
2
= 29.77; P < 0.0001
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Israel Journal of Veterinary Medicine  Vol. 69 (4)  December 2014 Avcioglu, H. 206
were afected by fasciolosis at a greater incidence than
males as diagnosed by the copro-ELISA (40.4 vs. 3.6%; P <
0.0002), antibody-ELISA (68.3 vs. 8.8%; P < 0.0001), and
sedimentation (39.1 vs. 2.6%; P < 0.0001) methods (Table 1).
Bovine fasciolosis was more common in advanced ages
when determined by the copro-ELISA (57.8%) and sedimen-
tation (57.8%) techniques (Table 1). Fasciolosis was the most
frequent in mid-age group when the assessment was made
by the antibody-ELISA method (76.6%). Te frequency
of afected animal younger than 2 years old was 20.0, 44.6,
and 17.7% when diagnosis was made by the copro-ELISA,
antibody-ELISA, and sedimentation techniques, respectively.
Te frequency of crossbred animals with fasciolosis was
1.36, 1.65, and 2.46-fold greater than purebreds when diag-
nosis was made by the copro-ELISA, antibody-ELISA, and
sedimentation methods (P < 0.0001 for all; Table 1).
Seropositivity assessed by the antibody-ELISA tech-
nique increased in females, advanced ages, and purebreds,
whereas it decreased in males, young animals, and hybrids
(P < 0.0001 for all; Table 2). Te frequency of animals with
fasciolosis detected by the copro-ELISA and sedimentation
techniques increased with the degree of seropositivity ELISA
score (P < 0.0001 for both; Table 2).
ROC curves for diagnostic methods
Te antibody-ELISA method had high sensitivity (100.0%)
and low specifcity (59.3%), whereas the coproscopy by sedi-
mentation method had both high sensitivity (96.0%) and
specifcity (100.0%) when the copro-ELISA technique was
considered gold standard (Table 3; Figure 1). Te ROC
curves for the antibody-ELISA and sedimentation methods
were diferent (P < 0.001; Table 3; Figure 1). Te results of
diagnostic techniques varied by the method principles that
are related to life cycle of F. hepatica and refect pathobiology
of fasciolosis (Figure 2).
Figure 1: Comparison of specifcity and sensitivity of the antibody-
ELISA and sedimentation techniques when the copro-ELISA
technique was considered gold standard.
Table 2: Distributions of cattle and diagnostic test results by antibody levels for F. hepatica infection
Degree and Titer
1
- (n=108; 38.30%) + (n=34; 12.06%) ++ (n=44; 15.60%) +++ (n=99; 34.04%) Signifcance
2.6±0.3
d
27.6±1.4
c
58.4±1.5
b
122.1±3.4
a
P < 0.0001
A
n
i
m
a
l
F
a
c
t
o
r
s
Sex
Female (n=230) 73 32 38 87 X
2
= 23.49;
P < 0.0001 Male (n=52) 35 2 6 9
Age (yr)
2≤ (n=130) 72 15 9 34
X
2
= 42.47;
P < 0.0001
3-5 (n=107) 25 17 20 45
≥6 (n=45) 11 2 15 17
Breed
Purebred (n=171) 87 19 24 41 X
2
= 31.89;
P < 0.0001 Crossbred (n=111) 21 15 20 55
D
i
a
g
n
o
s
t
i
c
M
e
t
h
o
dCopro-ELISA
- (n=182) 108 29 19 26 X
2
= 133.36;
P < 0.0001 + (n=100) 0 5 25 70
Sedimentation
- (n=186) 108 30 20 28 X
2
= 129.37;
P < 0.0001 + (n=96) 0 4 24 68
1
Data are presented as LS means ± SE. Diferent superscripts among the columns difer (P < 0.05)
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Israel Journal of Veterinary Medicine  Vol. 69 (4)  December 2014 207 Diagnostic Tests for Bovine Fasciolosis
DISCUSSION
Te coproscopy by sedimentation, antibody-ELISA, and
copro-ELISA (gold standard) techniques were utilized to at-
tain the prevalence of bovine fasciolosis in Erzurum Province,
considering associated risk factors as well as their diagnostic
sensitivity and specifcity. In Turkey, epidemiology studies on
fasciolosis are mostly based on fecal examination or inspec-
tion at slaughterhouse, and few researchers have employed
immuno-serological methods. Recent surveys employing the
antibody-ELISA technique by Yildirim et al. (13) and the
copro-ELISA technique by Sen et al. (14) reported that
prevalence of bovine fasciolosis in the Cappadocia region
was 65.2 and 3.03%, respectively. In the present study,
bovine fasciolosis prevalence in the Erzurum Province
was 34.04, 35.46, and 61.70% as determined by the co-
proscopy by sedimentation, copro-ELISA, and antibody-
ELISA techniques, respectively (Table 1). Prevalence re-
ports based on the anitibody-ELISA test were greater
than those based on the copro-ELISA test. Tis could be
related to development of antibodies much earlier than
presence of eggs in feces during the course of fasciolosis
(10, 15, 16).
Te literature coping with gender association with
fasciolosis prevalence is inconsistent. Studies reporting
no sex efect on fasciolosis are available (17, 18). In agree-
ment with the present study (Table 1), dairy cattle were
shown to be more vulnerable to fasciolosis than beef cattle
(13, 19). It appears that this is not a direct efect of sex,
but of the animal production system (19). Te facts in the
region are that males have shorter life-spans than females
and that females are grazed whereas males are confned.
Tese (age and grazing vs. confned) increase predisposi-
tion to fasciolosis.
Cattle in advanced ages (> 2-3 years) are more prone to
fasciolosis than those in younger ages (< 2 years) (13, 17, 19,
20, 21). In this study, cattle older than 2 years were afected by
fasciolosis in all techniques at a greater frequency than those
younger than 2 years (P < 0.0001; Table 1). Tis could be a
result of decreased immune-potency as age advances (17).
It could also be probable that older cattle have prolonged
host-parasite association and they are exposed to intermedi-
ate hosts in longer periods (21).
Table 3: Sensitivity and specifcity of diagnostic tests for fascioliasis*
Sensitivity Specifcity
Likelihood
Ratio (LR)
Area Under Curve (AUC) Statistics
Test Mean 95% CI Mean 95% CI +LR -LR Mean SE 95% CI z P <
Antibody-ELISA 100.0 96.4-100.0 59.34 51.8-66.5 2.46 0 0.797 0.0183 0.745-0.842 16.253 0.0001
Coproscopy by
Sedimentation
96.0 90.1-98.9 100.00 98.0-100 - 0.04 0.980 0.00985 0.956-0.993 48.744 0.0001
Antibody-ELISA vs. Coproscopy by Sedimentation
Diference SE 95% CI z P <
0183 0.0207 0.143-0.224 8.837 0.001
*
Te copro-ELISA test considered golden standard.
Figure 2: Interpretation of the fasciolosis detection rate by diagnostic tests.
a: Surely healthy animals based on all tests; b: Animals with seropositivity
based on the antibody-ELISA technique, refecting animals with current
fasciolosis, infected with other trematodes, recovered from past infection,
or all; c: Animals with positivity based on the copro-ELISA technique,
refecting a patent infection. d: Animals with positivity based on the
coproscopy by sedimentation, indicating a chronic infection; e: Animals
with positivity based on the copro-ELISA technique and negativity
based on the coproscopy by sedimentation method, indicating a prepatent
infection.
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In disagreement with the present study, several research-
ers reported lacking breed efect on fasciolosis (13, 14, 21).
In all three methods, fasciolosis was more common in cross-
bred cattle than purebred cattle (1.36-2.46 folds; P < 0.0001;
Table 1). Tis is opposed to expectations that local breeds and
crossbreeds are more resistant to bacterial and parasitic infec-
tions than purebred breeds due to their adaptation to habitat.
Tus, this fnding could also be a consequence of production
system in the region. Te fact is that purebred breeds are
mostly raised in confned system or fenced grasslands, where-
as local breeds and crossbreeds are grazed on communal areas
with poor vegetation. Indeed, metacercariae (infective form
of the parasite) are colonized in parts of grasses close to soil
and/or water (22), which could contribute greater prevalence
of cattle grazing at a late stage of the vegetation period.
Te diagnosis of fasciolosis is confrmed by the observa-
tion of parasite eggs in the feces of infected animals, but due
to the long pre-patent period in cattle, coprological meth-
ods are only sensitive 8-9 weeks after infection (23). Tus,
more accurate diagnostic methods for the early detection of
fasciolosis are invaluable (24). Because antibodies are pres-
ent approximately 1-5 weeks before eggs evident in feces or
infection matures, the antibody-ELISA technique is more
sensitive than the coproscopy by sedimentation technique
(10, 15, 16). Tis suggests that cattle with mature infections
may not excrete detectable numbers of eggs in feces. Indeed,
4 and 78 cattle with negativity based on the coproscopy by
sedimentation technique yielded positivity based on the
copro- and antibody-ELISA techniques, respectively (Table
1). One disadvantage of the antibody-ELISA is that posi-
tive results only imply exposure to the trematode at some
time, but not necessarily current infection (25). In addition
antibody levels in most animals persist above the positive
threshold of the antibody-ELISA for about 12 weeks after
treatment (26), suggesting that prevalence data based on the
antibody-ELISA may not be reliable about current infection
status. Cattle with increased antibody levels refected positiv-
ity at a higher percentage in both the coproscopy by sedimen-
tation and copro-ELISA techniques (Table 2), which could
be related to the fact that currently infected animals may
have higher level of antibodies than those recovered from
the past infection(s).
Te copro-ELISA method was developed for determina-
tion of F. hepatica and F. gigantica coproantigens in feces (6,
7). Te copro-ELISA technique is considered gold standard
to diagnose fasciolosis in the prepatent phase of infection
when fecal examination remains negative (8, 27, 28). Tis
test is highly sensitive (confrmed by necropsy) and specifc
(no cross reaction was observed with antigens from other
helminthes), and enables detection of Fasciola infections 1-5
weeks before patency (27).
Te copro-ELISA technique is reported to have high
sensitivity (94%) and specifcity (100%) (27). Coproantigen
detection was carried out using copro-ELISA test (Bio-X
Diagnostics, Jemelle, Belgium), with a specifcity of 100%
(24). Salimi-Bejestani et al. (5) reported 98% sensitivity and
96% specifcity for the antibody-ELISA technique. Similar
to the present survey, Charlier et al. (9) conducted an experi-
ment under feld conditions to compare sensitivity and speci-
fcity of diagnostic tests. Tey (9) reported that overall, sensi-
tivity and specifcity of, 64% (53-74%) and 93% (87-97%) for
the coproscopy by sedimentation (10 g), 87% (78-93%) and
90% (83-95%) for the antibody-ELISA, and 94% (87-98%)
and 93% (86-97%) for the copro-ELISA, respectively (9).
In this feld survey, the antibody-ELISA technique had high
sensitivity (100%) and low specifcity (59.3%), whereas the
coproscopy by sedimentation method had both high sensitiv-
ity (96%) and specifcity (100%) (Table 3; Figure 1).
Interpretation of prevalence rate attained by diferent
techniques requires caution. For instance, the antibody-
ELISA test result (61.70%) may refect antibodies developed
from past fasciolosis despite being treated, cross-reactions
among other trematodes, current fasciolosis, or all these fac-
tors together (Figure 2, arrow b). Diference in prevalence
rates between the coproscopy by sedimentation (34.04%)
and copro-ELISA (35.46%) techniques was 1.42%. Te co-
proscopy by sedimentation method is based on enumeration
of eggs in feces that are produced by mature parasite in the
host (Figure 2 arrow d), whereas the copro-ELISA method is
based on determination of ES-antigens of immature/mature
parasites residing in biliary tract, (Figure 2 arrow c). Tus,
1.42% diference may refect parasites not mature enough to
produce eggs that could be determined in feces and indicates
prepatent infections (Figure 2 arrow e).
In conclusion, in this study a greater predisposition of
females and crossbreds to fasciolosis than males and pure
breeds could be artifacts, probably resulting from the produc-
tion system practice in this region, and as a consequence the
results cannot be generalized. Older cattle (> 2 years) were
more prone to fasciolosis than younger cattle (< 2 years).
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Israel Journal of Veterinary Medicine  Vol. 69 (4)  December 2014 209 Diagnostic Tests for Bovine Fasciolosis
Diferent bovine fasciolosis prevalence rates obtained from
the antibody-ELISA (61.70%), copro-ELISA (35.46%), and
coproscopy by sedimentation (34.04%) methods could be re-
lated to principles of methods (determination of antibody in
serum, ES-antibody in feces, and egg enumeration in feces,
respectively), in association with diferent course of fascio-
losis. Considering the copro-ELISA gold standard, sensitiv-
ity and specifcity were 100 and 59.3% for antibody-ELISA
technique and 96.0 and 100% for sedimentation technique.
It is recommended that to determine active infection, the
coproscopy by sedimentation method should be performed
simultaneously with the copro-ELISA technique.
ACKNOWLEDGMENTS
Tis study was supported by Ataturk University Scientifc
Researches Projects (Project number: BAP-2010-121).
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