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Seroprevalence of Neospora caninum in Cattle in Nigde Province, Turkey
Karatepe, B. and Karatepe, M.*
Nigde University, Bor Vocational School, Bor/Nigde-Turkey.
*Corresponding Author: Prof. Dr. Mustafa Karatepe, Nigde University, Bor Vocational School, 51700 Bor/Nigde-Turkey. Tel: +90 388 311 45 27,
Fax: +90 388 311 84 37. Email: mkaratepe@nigde.edu.tr
AB S T RAC T
This study was carried out in order to investigate the presence of Neospora caninum antibodies in local cattle
breeds in the Nigde province. The animals used in this study consisted of cattle of at least one year of age
slaughtered at a local abattoir in Nigde province of Turkey. Blood samples were obtained from 264 cattle (207
male, 57 female) and serum samples were tested for antibodies to N. caninum using commercial competitive
ELISA (c-ELISA) (VMRD, Pulman, WA, USA). According to the serological examination, 70 (26.51%) of
264 cattle were seropositive for N. caninum. Anti-N. caninum antibodies were detected in 63 (30.43%) of 207
male cattle and 7 (12.28%) of 57 female cattle. There was a statistically significant differences in seropositivity
between genders (p < 0.05). This is the first serologic survey for N. caninum antibodies performed on cattle
in Nigde province.
Keywords: Neospora caninum; Neosporosis; Enzyme-Linked Immunosorbent Assay (ELISA);
Seroprevalence; Cattle; Nigde; Turkey.
INTRODUCTION
Neospora caninum is an obligate intracellular protozoan
parasite, closely related to Toxoplasma gondii. Neosporosis
is a major cause of abortion in cattle in many countries
(1, 2). N. caninum has a heteroxenous life cycle. Dogs are
both intermediate and definitive host for N. caninum, and
intermediate hosts such as cattle, sheep, goats and horse
becomes contaminated with oocysts in infected dog feces
(3, 4). Dogs become infected with contaminated meat
containing tissue cysts of intermediate host animals and
at the same time they may infect other dogs with oocysts
in their feces (5).
There are three infectious stages of the parasite: tachyzoites, bradyzoites, and sporozoites. Tachyzoites and bradyzoites
occur in tissues of infected hosts (intermediate and definitive)
whereas sporozoites are present in oocysts that are excreted
in the feces of the definitive host. Neospora caninum can be
transmitted horizontally by ingestion of infected tissues or by
ingestion of food or drinking water contaminated oocysts, or
Israel Journal of Veterinary Medicine  Vol. 71 (1)  March 2016
March 2016.indb 39
it can be transmitted vertically from an infected dam to her
fetus during pregnancy (4, 5).
Neosporosis especially causes abortion and neonatal mortality in cattle. Most neosporosis-induced abortions occur at
5-6 month gestation. Fetuses may die in utero, be resorbed,
mummified, autolyzed, stillborn, born alive with clinical
signs, or born clinically normal but chronically infected.
Neosporosis-induced abortions occur all year-round (1-3, 6).
In Turkey the seroprevalence of N. caninum in cattle
have been reported in various studies (7, 8) and the prevalence of N. caninum was determined to be 2-37.7%. The
aim of the present study was to determine the prevalence of
antibodies to N. caninum in the cattle in the Nigde province
of Turkey.
MATERIALS AND METHODS
Study Area
This study was performed on local cattle breeds of Nigde
province, situated in central-Turkey (with an altitude of
N. caninum in Cattle in Nigde, Turkey
39
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1240 m, 37°58′ N longitude-34°41′ E latitude), where neosporosis has not previously recognized. The province has a
continental climate with warm and dry summers and cold
and snowy winters. Annual average precipitation is 348.8
mm, average temperature is 11.1°C and average relative
humidity is 55%.
Sampling of Cattle and Blood Collection
This study was carried out to detect the seroprevalence of
Neospora caninum on cattle of at least one year of age slaughtered at a local abattoir of Nigde. A total of 264 cattle which
consisted of 57 female and 207 male were selected randomly.
Blood samples were collected from the jugular vein of these
cattle into sterile tubes. Serum samples were obtained by
centrifugation at room temperature (25°C), at 4,000 rpm for
10 minutes and were stored at -20°C until analyzed.
Serologic Examination
The serum samples were tested for antibodies to N. caninum
using commercial competitive ELISA (c-ELISA) (VMRD,
Pulman, WA, USA). The test was performed according to
the procedure described by the manufacturer and the results
were assessed at a wavelength of 630nm using an ELISA
reader (spectrophotometer). The formula used for calculation
of percent inhibition:
% I = 100 - (Sample O.D. x 100) ÷ (Mean Negative
Control O.D.)
Where the test sample showed ≥ 30% inhibition, it was
considered as positive. Where a test sample showed < 30%
inhibition, it was considered as negative.
Statistical Analysis
In this study, chi-square test was used to compare the seropositivity rates in terms of gender. Statistical significance was
considered at p < 0.05.
RESULTS
According to the serological examination, 70 (26.51%) out of
264 cattle were seropositive for N. caninum (Table 1). Anti-N.
caninum antibodies were detected in 63 (30.43%) of the 207
male cattle and 7 (12.28%) of the 57 female cattle. There was
a statistically significant differences in seropositivity between
genders with a higher proportion of male cattle being infected compared to females (p < 0.05).
40
Karatepe, B.
March 2016.indb 40
Table 1: The seropositivity of Neospora caninum in cattle with ELISA
according to gender
Gender
Female
Male
TOTAL
Number of Sera
Examined
57
207
264
Number of
seropositivity sera
7
63
70
Seropositivity
(%)
12.28
30.43
26.51
DISCUSSION
Neospora caninum is one of the most important causes of
bovine abortion and neonatal mortalities (1, 2). Studies have
shown that seropositive animals are up to 3.5 times more
likely to abort than seronegative herd-mates (9). Economic
loss associated with N. caninum infection is not only due to
increased prevalence of abortions, but also to decreased milk
and meat production in infected cattle (10). Due to clinically
insufficient symptoms various serological tests (ELISA, IFAT
and DAT) have been used to detect antibodies to N. caninum
and identify infected animals (2, 3). Neospora caninum infections have been reported from most parts of the world and
the worldwide seroprevalences of N. caninum in cattle are
summarized in Table 2.
When the N. caninum seroprevalence results of the study
are compared with the results of the studies conducted
around the world, N. caninum prevalence rate of this study
was lower than Argentina (11), Spain in dairy cattle (12),
Portugal (13), Mexico (14), in Georgia (15) and Romania
(16). On the other hand, seroprevalence result of our study
was higher than the results obtained in Southern Vietnam
(17), in Wales and England (9), in Spain in beef cattle (12),
in Brazil (18), in Northern Alberta (19), in Korea (20), in
Texas (15), in Northwest Iran (21) and in South Australia
(22). The frequency differs depending on the geographical
location, breeding conditions and type of serological tests.
In Turkey, seropositivity has been reported with ELISA
as 9.2% in Sakarya (23), 7.01% in Eastern Anatolian region
(24), 13.96% in Central Anatolia (25), 8.02% in Thrace
(26), 2%, 7.2% in Kars (7, 27), 7.5% in Şanlıurfa (28), 7%
in Kayseri (29), 8.19% in Elazığ (30), 4.88% in Van (31),
10.65% in Eastern Turkey (32), 21.03% in Afyonkarahisar
(33), 37.7% in Kırıkkale, Aksaray, Burdur (8), 35.07% in
Central Veterinary Control and Research Institute (34),
10.77% in Kırıkkale, İzmir, Tokat (35) and 5.3% in Burdur
(36). Furthermore, seroprevalence of N. caninum in cattle
has been determined as 10% by using Immunocomb (IC)
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Table 2: Select reports of prevalence of Neospora caninum in cattle
Country
Argentina
Southern
Vietnam
Brazil
Spain
Wales and
England
Portugal
Northern
Alberta
Mexico
Korea
Georgia
and Texas
Romania
Northwest
Iran
South
Australia
Prevalence %
Test
Author
Reference
88.8
Campero et al.,
IFAT
11
(in aborting cattle)
1998
5.5
Huong et al.,
ELISA
17
(in dairy cattle)
1998
14.09
Gondim et al.,
IFAT
18
(in dairy cattle)
1999
17.9
Quintanilla(in beef cattle)
ELISA
Gozalo et al.,
12
36.8
1999
(in dairy cattle)
12.5
Davison et al.,
ELISA
9
(in dairy cattle)
1999
49
Thompson et al.,
ELISA
13
(in dairy cattle)
2001
9
Waldner et al.,
ELISA
19
(in beef cattle)
2001
59
García-Vázquez
ELISA
14
(in dairy cattle)
et al., 2002
4.1
IFAT
Kim et al., 2002
20
(in beef cattle)
32.1%
(in dairy cattle) WESTERN Ortega et al.,
15
10.3%
BLOT
2007
(in dairy cattle)
34.6
Gavrea et al.,
ELISA
16
(in dairy cattle)
2011
10.5
Nematollahi
21
ELISA
et al., 2011
(in dairy cattle)
2.5
(in beef cattle)
Nasir et al.,
ELISA
22
3.8
2012
(in dairy cattle)
assay in Ankara (37) and 19% by IFAT in Western Turkey
(38).
In the present study, seroprevalence of N. caninum was
found to be 26.51% in cattle from Nigde in Turkey. The rate
of seroprevalence of this study has been shown to resemble
results of research conducted by Celik et al. (33). However,
the seroprevalence results of our study are higher than the
results of studies carried out in cattle from different parts of
Turkey (7, 23-32, 35-38) while it is lower than the results
obtained by Öcal et al. (8) and Piskin and Utuk (34). The
different prevalence rates of disease may be associated with
the different geographical locations, management practices
and type of the serological tests used.
In this study, the cattle were classified according to
gender and N. caninum antibodies in the male cattle were
more prevalent than those in the female cattle. The differIsrael Journal of Veterinary Medicine  Vol. 71 (1)  March 2016
March 2016.indb 41
ences between the genders for the presence of antibody may
be due to factors such as age of the animals and different
breeding conditions.
From a prophylactic point of view, the removal of all
potentially infected tissues, such as aborted fetuses and placentas from the environment, that might serve as a source of
infection for susceptible hosts is advisable. In addition, fecal
contamination of feed and water sources by potential host
animals should be minimized (3-5).
This is the first serologic survey for N. caninum performed
on cattle in Nigde province which detected a seroprevalence
of N. caninum as 26.51%. Therefore, neosporosis should be
taken into consideration in abortion cases and neonatal
deaths. In addition, further investigations, including molecular techniques are required in order to determine the
importance of this parasite as a cause of clinical disease in
dogs which are definitive hosts and of abortion in cattle in
Nigde province.
ACKNOWLED GEMENTS
This study was presented at the XVIII. National Congress of
Parasitology (29 September to 5 October 2013, Denizli, Turkey).
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