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Effects of Age and Breed on the Prevalence of
Neospora caninum
in Commercial
Dairy Cattle from Pakistan
Author(s): Muhammad Mudasser Nazir , Azhar Maqbool , Muhammad Sarwar Khan , Afzal Sajjid , and
David S. Lindsay
Source: Journal of Parasitology, 99(2):368-370. 2013.
Published By: American Society of Parasitologists
DOI: http://dx.doi.org/10.1645/GE-3173.1
URL: http://www.bioone.org/doi/full/10.1645/GE-3173.1
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J. Parasitol., 99(2), 2013, pp. 368–370
Ó American Society of Parasitologists 2013
Effects of Age and Breed on the Prevalence of Neospor a caninum in Commercial Dairy
Cattle from Pakistan
Muhammad Mudasser Nazir, Azhar Maqbool, Muhammad Sarwar Khan*, Afzal Sajjid†, and David S. Lindsay‡, Department of Parasitology,
University of Veterinary and Animal Sciences, Lahore 54600, Pakistan; *Department of Clinical Medicine and Surgery, University of Veterinary and Animal
Sciences, Lahore 54600, Pakistan; †Veterinary Research Institute, Lahore 54810, Pakistan; ‡Department of Biomedical Sciences and Pathobiology, Virginia-
Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia 24061. Correspondence should be sent to: lindsayd@vt.edu
ABSTRACT: Neospora caninum is a major cause of bovine abortion
worldwide. A serological survey was carried out to determine the
seroprevalence of exposure to N. caninum in dairy cattle based on age
and breed from Punjab and Sindh provinces, Pakistan. Serum samples
from 641 animals from 12 herds from Punjab (n ¼ 7) and Sindh (n ¼ 5)
provinces were tested for antibodies against N. cani num using a
commercially available competitive enzyme-linked immunosorbent
assay. Positive reactions to N. caninum were seen in 277 (43%) of the
641 of the samples. Seropositive animals were present in all 12 herds.
Animals over 2 yr of age (47%) and crossbreds (55%) were more likely to
be seropositive than the other cattle examined. These results indicate that
N. caninum infection is widespread among dairy cattle in Pakistan.
Neospora caninum is an important cause of abortion in dairy cattle,
causing significant economic losses to the industry worldwide (Dubey and
Lindsay, 1996). The parasite was recognized in dogs in Norway in 1984 as
a cause of neuromuscular degeneration, leading to hind limb paralysis
(Bjerkas et al., 1984). Because of structural and antigenic differences from
Toxoplasma gondii, it was described as a new species in 1988 (Dubey,
Carpenter, et al., 1988) and isolated from naturally infected dogs that
same year (Dubey, Hattel, et al., 1988). In 1989 N. caninum was first
reported as a cause of bovine abortion (Thilsted and Dubey, 1989).
Domestic dogs are definitive hosts for N. caninum (McAllister et al., 1998;
Lindsay, Dubey, et al., 1999; Lindsay, Upton, et al., 1999). Horizontal
transmission of N. caninum can result in abortion storms (Thilsted and
Dubey, 1989), but vertical (transplacental) transmission of the N. caninum
is the most economically important outcome of infection (Bj
¨
orkman et al.,
1996; Par
´
e et al., 1996; Schares et al., 1998). The majority of congenital
infections result in the birth of apparently healthy calves. Cows may abort
repeatedly due to N. caninum or give birth to apparently healthy calves,
calves with subclinical N. caninum infection, or uninfected calves. Most
congenitally infected calves remain clinically asymptomatic. Most N.
caninum–induced abortions occur at 5–6 mo of gestation (Dubey, 1999).
Horizontal transmission occurs when cows ingest sporulated N.
caninum oocysts shed by dogs (McAllister et al., 1998). Domestic dogs
are natural definitive hosts for the parasite (McAllister et al., 1998;
Lindsay, Dubey, et al., 1999; Lindsay, Upton, et al., 1999). It has been
demonstrated that they can become infected by consuming N. caninum–
infected tissues (Gondim et al., 2002) or bovine placenta (Dijkstra et al.,
2001). Experimentally infected dogs excrete few oocysts (Lindsay et al.,
2001), and the role of dogs and N. caninum oocyst excretion is an area of
active research in the epizootiology of the disease (Bandini et al., 2011;
Cavalcante et al., 2011; King et al., 2012).
Dairy herd health is extremely important to the economy of Pakistan
because it is the third largest milk-producing country in the world (FAO,
2009). We have recently reported the seropositivity rate of N. caninum in
dairy herds with a history of high rates of abortion (Shabbir et al., 2011).
Brucella abortus was the most common agent found in aborting cattle
(77%), while N. caninum was present in 47% of aborting animals, and
dual infections with these agents were present in 28%of the animals.
However, N. caninum was found in 39% of nonaborting cattle, while B.
abortus was found in 27% of nonaborting cattle (Shabbir et al., 2011).
We became interested in determining if horizontal or vertical
transmission is more important in maintaining the parasite in the cattle
population in Pakistan. In the present study, we examined animals from 3
age groups to consider this question. Additionally, we examined the effects
of breed on prevalence to determine if genetic factors may play a role in
susceptibility to N. caninum. The present study was done in healthy dairy
cows from farms in Punjab and Sindh provinces, Pakistan.
Pakistan is an agricultural country, located in southern Asia with hot
dry summers, during which temperatures reach over 40 C, and mild
winters with average temperatures between 15 and 20 C. Pakistan is
located at latitude between 248 and 378 north and longitude between 618
and 768 east, and has 803,940 km
2
of land, a human population of 1.8
million, and a cattle population of 24.2 million. Between January 2009 and
August 2010, serum samples were taken from 641 clinically normal cattle
on 12 dairy farms, 7 from Punjab and 5 from Sindh provinces (Tables I–
III). Information regarding abortion history was available for only 2
farms (10 and 11). Farm 10 reported abortions in the last year, while farm
11 reported abortions over the last 4 yr. Only 6 cows aborted in herd 10 in
the previous year; in herd 11, 21 of the 37 cows had no history of abortion,
and the remaining 16 had aborted previously. Some of the cows may have
had repeated abortions, but no supporting data were available. Samples
were selected from clinically healthy cows of different breeds and different
age groups using animal numbers provided by the producers by an
individual who had no knowledge of animals status. We sampled 3 age
groups of cows, including calves (8 mo of age), heifers (.8moto2yr
old), and adult lactating cows (.2 yr old) to determine the variations of
serological prevalence against N. caninum between different age groups
(Table II). Cattle included crossbred (local 3 exotic), non-descript (no
cross-discernable based on animal appearance), and purebred (Table III).
Water and feed were available ad libitum.
Blood was collected from the jugular vein; serum was separated at the
Department of Parasitology, University of Veterinary and Animal
Sciences, Lahore, Pakistan, and stored at 20 C until assayed. Serum
reactivity to N. caninum was evaluated using a commercially available
competitive enzyme-linked immunosorbent assay kit (cELISA, VMRD,
Pullman, Washington). The procedures were performed according to the
manufacturer’s instructions, and results were expressed as percent
inhibition.
Pearson’s v
2
test was used to compare seropositive status with age and
breed using (SPSS for Windows, version 13.0; SPSS, Chicago, Illinois),
and P , 0.05 was considered significant.
The cELISA indicated that 277 (43%) of the 641 cows were seropositive
to N. caninum. A significant difference (P , 0.05) of seropositivity was
observed between different herds (Table I). The age of the cows were
categorized into 3 groups, below 8 mo of age, 8 mo to 2 yr of age, and
above 2 yr old. Neospora caninum seroprevalences were 39% (37/94), 36%
(59/162) and 47% (181/385), respectively. Age-wise prevalence, though
not statistically significant (P . 0.05), was greatest in animals older than 2
yr of age (Table II). The prevalence was significantly different (P , 0.05)
between purebred, crossbred, and non-descript animals. However, no
DOI: 10.1645/GE-3173.1
368
American Society of Parasitology. Muhammad Mudasser Nazir, Azhar Maqbool, Muhammad Sarwar Khan, Afzal Sajjid, and David S.
Lindsay (2013). "Effects of Age and Breed on the Prevalence of Neospora caninum in Commercial Dairy Cattle from Pakistan," Journal of
Parasitology, Vol. 99, No. 2, pp. 368-370. doi: http://dx.doi.org/10.1645/GE-3173.1
significant difference (P . 0.05) was found between sub-breeds, i.e., exotic
and local breeds.
The prevalence was not significantly different (P . 0.05) within the
exotic breeds, i.e., Jersey and Freisian, and local breeds, i.e., Sahiwal,
Rojan, Dajal, and Dhanni (Table III). Seroprevalence was highest in
crossbred animals (Table III).
The high prevalence of ex posure to N. caninum (43%) even in
apparently healthy animals indicates that N. caninum may be an important
risk factor of abortions in these provinces of Pakistan. The seroprevalence
of N. caninum varies within countries, between regions in countries, and
between beef and dairy cattle in areas examined (Dubey et al., 2007).
Seropr evalence depends largely on management and environmental
conditions of the country being studied (Dubey et al., 2007). The overall
high prevalence of N. caninum among dairy cattle herds corresponds to
previous observations made by Shabbir et al. (2011) who reported high
prevalence in aborting animals and in those that were at risk. The
prevalence of N. caninum in all the age groups strongly suggests
transplacental and horizontal transmission (Dubey et al., 2007), but the
rate of transmission may be different in different scenarios (Bartels et al.,
2007; Dijkstra et al., 2008; Williams et al., 2009).
Sadrebazzaza et al. (2004) concluded that there is no significant
relationship between N. caninum seropositivity and age. Howe ver,
Guimaraes et al. (2004) reported that N. caninum seropositivity was
higher in cattle above 2 yr of age and that horizontal transmission could
be responsible for this situation. In the present study, the highest
prevalence in age groups over 2 yr agrees with this study and observations
made by others that seroprevalence increases with animal age (Dubey et
al., 2007; Eiras et al., 2011).
The results of our study showed a significant difference in prevalence of
N. caninum among different breeds of cattle in our cattle populations and
that it was significantly (P , 0.05) higher in crossbreds. There are many
reports worldwide demonstrating differences in seropositivity of different
cattle breeds in N. caninum prevalence studies (Armengol et al., 2007;
Dubey et al., 2007; Duong et al., 2008; Munhoz et al., 2009). However,
these reports should be interpreted carefully because production systems
may be a possible explanation rather than breed-related susceptibility. The
extent of abortion and immune responses after N. caninum infection is
likely affected by the breed of cattle (Armengol et al., 2007; Almeria et al.,
2009; Romero-Salas et al., 2010; Santolaria et al., 2011). Experimental
studies need to be conducted with different bovine breeds before the
influence of breed can be accurately evaluated and its importance in the
epizootiology of bovine N. caninum infections can be determined.
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Number of
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Number of
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2 963 41 13 32
3 392 88 33 38
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Local Sahiwal 147 59 40
Rojan 24 7 29
Dajal 19 6 32
Dhanni 31 11 36
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