Accepted Manuscript Neospora caninum seropositivity and reproductive risk factors in dogs Domenico Robbe, Alessandra Passarelli, Alessia Gloria, Angela Di Cesare, Gioia Capelli, Raffaella Iorio, Donato Traversa PII:
S0014-4894(16)30018-2
DOI:
10.1016/j.exppara.2016.02.003
Reference:
YEXPR 7189
To appear in:
Experimental Parasitology
Received Date: 13 April 2015 Revised Date:
4 February 2016
Accepted Date: 5 February 2016
Please cite this article as: Robbe, D., Passarelli, A., Gloria, A., Di Cesare, A., Capelli, G., Iorio, R., Traversa, D., Neospora caninum seropositivity and reproductive risk factors in dogs, Experimental Parasitology (2016), doi: 10.1016/j.exppara.2016.02.003. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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ACCEPTED MANUSCRIPT Neospora caninum seropositivity and reproductive risk factors in dogs Domenico Robbe1, Alessandra Passarelli1, Alessia Gloria1*, Angela Di Cesare1, Gioia Capelli2, Raffaella Iorio1, Donato Traversa1 1
Faculty of Veterinary Medicine, University of Teramo, Piazza Aldo Moro 45, 64100, Teramo, Italy Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10-35020 Legnaro (PD), Italy.
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*Corresponding author. Tel.: +390861266995 E-mail address: [email protected] (Alessia Gloria).
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ACCEPTED MANUSCRIPT Abstract
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Despite the importance of Neospora caninum in veterinary medicine, knowledge of distribution of
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neosporosis in dog populations in some countries is still poor. The aims of the present study were to
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determine the occurrence of anti-N. caninum antibodies in one-hundred dogs living in cattle farms
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or dog breedings in central Italy and to evaluate the risk factors associated with seropositivity. The
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incidence of reproductive system disorders (e.g. infertility after first pregnancy) was also evaluated.
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Serum from breeding and farm dogs was tested to an indirect immunofluorescent antibody test
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(IFAT) to assess the occurrence of seropositivity. Management and individual data were collected
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and analysed both by linear and logistic multiple-regression models to find reliable predictors of
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seroprevalence and anti-N. caninum antibody level. The seropositivity for N. caninum was 32%.
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Dogs reared for breeding and presence of cattle on the farm were associated with seropositivity for
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N. caninum. Dogs living in the cattle farms showed a higher seropositivity for N. caninum (46%)
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compared with those living in dogs breeding (18%) (P or < 3 years), sex, and breed (mongrel/pure breed) were offered to binary
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logistic models to evaluate possible risk factors for N. caninum-seropositivity. Breed was
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introduced in the models as interaction with the provenance. The differences were considered
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significant with P0.05).
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3.2 Group 2
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Out of the 50 dogs belonging to Group 2, nine (18%) were positive for anti-N. caninum antibodies.
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None of the nine seropositive animals, which belonged to four different breeds, showed past or
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present reproductive problems (Table 2). The presence of N. caninum antibodies was detected in
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four of the six facilities examined. However, most of the infected animals (60%) were hunting dogs,
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and the differences between this typology of dogs and the other breeds included in the study were
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not statistically significant (P>0.05) (Table 2). All variables were not statistically related to
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seropositivity (P>0.05).
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3.3 Risk factors
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The model which best fitted the data is shown in Table 3. The only significant predictor of
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Neospora-seropositity was the interaction provenance-breed, i.e. pure breed dogs living in a cattle
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farm had 2.7 more probability to be Neospora seropositive than the other animals.
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4. Discussion
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ACCEPTED MANUSCRIPT The present study showed for the first time the presence of anti-N. caninum specific antibodies in
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dogs living in this area of Italy. Despite the examined groups of dogs do not represent the general
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canine population of Italy, the results demonstrate that the pathogen may circulate in selected
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populations of dogs in particular geographic areas that have been never investigated for
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neosporosis. Hence, the achievements are of particular relevance in those settings where zootechnic
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activities are of economic importance.
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Despite dogs play a major role in the epidemiology of the infection, studies on the occurrence of N.
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caninum and the risk factors for neosporosis in dogs are still scant in several countries, including
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Italy. The prevalence recorded in the present study (i.e. 32%) is similar to data (36.4%) reported in
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north-western Italy in dogs living in rural habitats (Ferroglio et al., 2007), and higher than data
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reported (10.9%) in kenneled and owned dogs living in north-eastern Italy (Capelli et al., 2004), and
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in kenneled and farm dogs from the south of the Country (14.6% and 26.5% respectively) (Paradies
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et al., 2007).
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Prevalence recorded in other European countries, i.e. Austria (5.3% in dogs from rural areas and
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2.1% from the urban area of Vienna) (Wanha et al., 2005), Czech Republic (4.7% in dogs at the
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service of the Czech Army and 2.6% in pet dogs) (Václavek et al., 2007), Serbia (17.2% in hunting,
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farm and stray dogs) (Kuruca et al., 2013), were lower than data showed in the present survey. A
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recent study carried out in Romania reported a similar rate of infection (i.e. total rate of infection of
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32.7%) (Gavrea et al., 2012). In this latter study dogs were divided in different categories showing
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the following results: 54.5% of hunting, 50% of farm, 29.6% of guard, 31.2% of shelter, 34.8 % of
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pet and 0% of kennel dogs were positive. Moreover, 34.5% of dogs from rural and 31.4% from
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urban areas presented anti-N. caninum specific antibodies (Gavrea et al., 2012).
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In the present study, dogs living in cattle farms showed a highest percentage of seropositivity
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infection (46%) compared to dogs in breeding facilities (18%). This difference could be explained
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by wrong feeding habits. In fact, dogs living outdoor in livestock premises have more chances to
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become in contact with bovine infected tissues (e.g. fetuses/placentas) or with other intermediate
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ACCEPTED MANUSCRIPT hosts of N. caninum (e.g. rodents, birds). However, a history of ingestion of bovine tissues resulted
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equivalent in both dogs which tested positive or negative for N. caninum.
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Although owners of dogs belonging to both groups declared to feed them with commercial food, the
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administration of raw or undercooked meat cannot be ruled out. Also, dogs living in rural farms are
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more likely to be in contact with wildlife and other potential ruminant hosts and more likely to
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show their carnivorous and scavenging behavior by eating small mammals and birds which possibly
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contain N. caninum tissue cysts (Wouda et al., 1999). These features could explain the prevalence
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differences between the two study groups.
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Despite no specific risk factors were evidenced for dogs living in breeding facilities, a relatively
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high value of seropositivity was recorded. This result may be due to the presence, in the study
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kennels, of open fences and outdoor areas which allow easy access to rodents or birds, as previously
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described (Fernandes et al., 2004).
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The higher seropositivity in pure breed animals than in mixed breed dogs fits with the results of
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previous studies, which suggested a genetic factor predisposing to infection, or a more efficient
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vertical transmission (Capelli et al., 2004). However, other works showed a higher prevalence in
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mixed breed dogs (Fernandes et al., 2004; Collantes-Fernandez et al., 2008), thus the role of breeds
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in the epidemiology of canine neosporosis is not well established.
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In any case, it can be argued that hunting dogs have a high chance of being fed with hunted animals
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that can act as intermediate hosts for the parasite (Gavrea et al., 2012). Furthermore, a vertical
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transmission is also possible in breeding facilities due to the introduction of infected animals in the
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farms or to a relapse of congenital infection during pregnancy (Rasmussen and Jensen, 1996;
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Wouda et al., 1999).
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Other factors, i.e. sex, age, reproductive status and/or reproductive disorders, were not significant.
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Analogously, other studies have indicated that sex does not influence the occurrence of N. caninum
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(Cheadle et al., 1999; Ferroglio et al., 2007; Collantes-Fernández et al., 2008). Indeed, dogs of any
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age can acquire the infection by N. caninum. Some studies have shown that older animals may have
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ACCEPTED MANUSCRIPT a higher risk to be infected due to the chronic pattern of the disease (Wouda et al., 1999; Fernandes
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et al., 2004; Ferroglio et al., 2007).
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Although provenance and breed resulted predictors of seropositivity further studies are warranted to
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eventually support these results in other categories of dogs, e.g. non-breeding animals or dogs
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Among the risk factors evaluated in the present study, the positive percentage was significantly
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higher in farm dogs who had a full-term pregnancy. Although dog neosporosis is a cause of
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reproductive disorders in dogs (Barber and Tress, 1998), information on abortion rates, infertility or
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other reproductive disorders were not well defined. Further studies are warranted to understand the
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actual impact of neosporosis in causing reproductive infertility, abortion and stillbirths in bitches.
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On the whole, the data showed in the present study provide new information of the prevalence of N.
198
caninum in dogs and contribute to the knowledge of the role of the parasite in reproductive
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disorders in dogs. The high seropositivity in dogs living with livestock confirms that habitats, such
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as rural areas, and the close contact with ruminants may indeed represent risk factors for the
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occurrence of canine neosporosis. Moreover, an association between the presence of dogs and the
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prevalence of anti-N. caninum antibodies in cattle was observed in previous studies (Wouda et al.,
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1999; Dijkstra et al., 2002), thus suggesting a potential transmission of the parasite between dog
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and cattle according to the an higher exposure to environmental sources of infection (Regidor-
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Cerrillo et al., 2010). Therefore, dogs should be not allowed to have access to infected materials
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such as bovine carcasses, aborted fetuses, placenta and uterine discharge, in order to limit the
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prevalence of the infection in both dogs and cattle. In fact, N. caninum is a major cause of abortion
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in cattle all over the world. Also, knowledge of the occurrence of the infection in dog breeding
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facilities is of particular importance to understand the correlation between seropositive dogs and the
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effects on their reproductive performances. While the transplacental transmission has been
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demonstrated in experimental infections, the frequency of vertical transmission in naturally infected
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dogs is low (Barber and Trees, 1998; Cavalcante et al., 2012). Nonetheless, it should be borne in
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mind that clinical neosporosis in adult dogs may result either from a recent infection or to the
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ACCEPTED MANUSCRIPT recrudescence of an existing infection as a result of pregnancy or other immune-compromising
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conditions. Stressing and immune-compromising drivers can cause bradyzoites to transform back
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into tachyzoites, resulting in reproductive disorders (e.g. abortion) and infection of fetuses in
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pregnant animals (e.g. stillbirths) (Barber and Tress, 1998; Lyndsay et al., 1999; Dubey et al., 2007,
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2011).
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5. Conclusions
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Serologic testing of bitches can be used as a method to determine whether neosporosis is a potential
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or evident reproductive problem in dog breeding. This strategy can be helpful when investigating
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dog farms with cases of abortion, stillbirths or litter with neuro-muscular disorders. In these cases,
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seropositive animals should be excluded from breeding. Although the number of positive dogs in
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the study area is not particularly high, the pathogenic potential of this parasite should be always
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taken into appropriate account for the reproductive disorders they may cause.
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On the contrary, the high level of seropositivity in dogs living in cattle farms implies that more
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attention is warranted in the management (i.e. feeding habits) of dogs living with livestock and/or in
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rural conditions, and in the introduction of new dogs in a farm, in order to limit the spreading of
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neosporosis in both dogs and cattle.
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Conflict of interest: The authors declare that they have no conflict of interest.
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References
235
Aguiar, D.M., Cavalcante, G.T., Rodrigues, A.A.R., Labruna, M.B., Camargo, L.M.A., Camargo,
237
E.P., Gennari, .S.M., 2006. Prevalence of anti-Neospora caninum antibodies in cattle and dogs from
238
Western Amazon, Brazil, in association with some possible risk factors. Vet. Parasitol. 142, 71-77.
RI PT
236
239
Barber, J.S., Trees, A.J., 1998. Naturally occurring vertical transmission of Neospora caninum in
241
dogs. Int. J. Parasitol. 28, 57-64.
SC
240
242
Björkman, C., Jakubek, E.B., Arnemo, J.M., Malmsten, J., 2010. Seroprevalence of Neospora
244
caninum in gray wolves in Scandinavia. Vet. Parasitol. 173, 139-142.
M AN U
243
245
Björkman, C., Lundën, A., Uggla, A., 1994. Prevalence of antibodies to Neospora caninum and
247
Toxoplasma gondii in Swedish dogs. Acta Vet. Scand. 35, 445-447.
248
TE D
246
Capelli, G., Nardelli, S., di Regalbono, A.F., Scala, A., Pietrobelli, M., 2004. Sero-epidemiological
250
survey of Neospora caninum infection in dogs in north-eastern Italy. Vet. Parasitol. 123, 143-148.
251
EP
249
Cavalcante, G.T., Soares, R.M., Nishi, S.M., Hagen, S.C., Vannucchi, C.I., Maiorka, P.C., Paixão,
253
A.S., Gennari, S.M., 2012. Experimental infection with Neospora caninum in pregnant bitches.
254
Rev. Bras. Parasitol. Vet. 21, 232-236.
AC C
252
255 256
Cheadle, M.A., Lindsay, D.S., Blagburn, B.L. 1999. Prevalence of antibodies to Neospora caninum
257
in dogs. Vet. Parasitol. 85, 325-330.
258
11
ACCEPTED MANUSCRIPT 259
Collantes-Fernández, E., Gómez-Bautista, M., Miró, G., Alvarez-García, G., Pereira-Bueno, J.,
260
Frisuelos, C., Ortega-Mora, L.M., 2008. Seroprevalence and risk factors associated with Neospora
261
caninum infection in different dog populations in Spain. Vet. Parasitol. 152, 148-151.
262
Costa, K.S., Santos, S.L., Uzêda, R.S., Pinheiro, A.M., Almeida, M.A.O., Araújo, F.R., McAllister,
264
M.M., Gondim, L.F., 2008. Chickens (Gallus domesticus) are natural intermediate hosts of
265
Neospora caninum. Int. J. Parasitol. 38, 157-159.
SC
266
RI PT
263
Costa-Júnior, L.M., Rembeck, K., Ribeiro, M.F.B., Beelitz, P., Pfister, K., Passos, L.M.F., 2007.
268
Sero-prevalence and risk indicators for canine ehrlichiosis in three rural areas of Brazil. Vet. J. 174,
269
673-676.
M AN U
267
270
Costa-Júnior, L.M., Ribeiro, M.F.B., Rembeck, K., Rabelo, E.M.L., Zahler-Rinder, M., Hirzmann,
272
J., Pfister, K., Passos, L.M., 2009. Canine babesiosis caused by Babesia canis vogeli in rural areas
273
of the State of Minas Gerais, Brazil and factors associated with its seroprevalence. Res. Vet. Sci. 86,
274
257-260.
EP
275
TE D
271
Crookshanks, J.L., Taylor, S.M., Haines, D.M., Shelton, G.D., 2007. Treatment of canine pediatric
277
Neospora caninum myositis following immunohistochemical identification of tachyzoites in muscle
278
biopsies. Can. Vet. J. 48, 506-508.
279
AC C
276
280
Cunha Filho, N.A., Lucas, A.S., Pappen, F.G., Ragozo, A.M.A., Gennari, S.M., Júnior, T.L., Farias,
281
N.A., 2008. Fatores de risco e prevalência de anticorpos anti-Neospora caninum em cães urbanos e
282
rurais do Rio Grande do Sul, Brasil. Rev. Bras. Parasitol. Vet. 17, 301-306.
283
12
ACCEPTED MANUSCRIPT 284
Dijkstra, T., Barkema, H.W., Eysker, M., Hesselink, J.W., Wouda, W., 2002. Natural transmission
285
routes of Neospora caninum between farm dogs and cattle. Vet. Parasitol. 105, 99-104.
286
Dubey, J.P., Carpenter, J.L., Speer, C.A., Tooper, M.J., Uggla, A., 1988. Newly recognized
288
protozoan disease of dogs. J. Am. Vet. Med. Asso. 192, 1269-1285.
289
RI PT
287
Dubey, J.P., Lindsay, D.S., 1996. A review of Neospora caninum and neosporosis. Vet. Parasitol.
291
67, 1-59.
SC
290
292
Dubey, J.P., Vianna, M.C., Kwok, O.C., Hill, D.E., Miska, K.B., Tuo, W., Velmurugan, G.V.,
294
Conors, M., Jenkins, M.C., 2007. Neosporosis in Beagle dogs: Clinical signs, diagnosis, treatment,
295
isolation and genetic characterization of Neospora caninum. Vet. Parasitol. 149, 158-166.
M AN U
293
296
Dubey, J.P., Jenkins, M.C., Rajendran, C., Miska, K., Ferreira, L.R., Martins, J., Kwok, O.C.,
298
Choudhary, S., 2011. Gray wolf (Canis lupus) is a natural definitive host for Neospora caninum.
299
Vet. Parasitol. 181, 382-387.
EP
300
TE D
297
Dubey, J.P., Schares, G., 2011. Neosporosis in animals-the last five years. Vet. Parasitol. 180, 90-
302
108.
303
AC C
301
304
Fernandes, B.C.T.M., Gennari, S.M., Souza, S.L.P., Carvalho, J.M., Oliveira, W.G., Cury, M.C.,
305
2004. Prevalence of anti-Neospora caninum antibodies in dogs from urban, periurban and rural
306
areas of the city of Uberlândia, Minas Gerais-Brazil. Vet. Parasitol. 123, 33-40.
307
13
ACCEPTED MANUSCRIPT 308
Ferroglio, E., Pasino, M., Ronco, F., Benà, A., Trisciuoglio, A., 2007. Seroprevalence of antibodies
309
to Neospora caninum in urban and rural dogs in north-west Italy. Zoonoses Public Health 54, 135-
310
139.
311
Gavrea, R., Mircean, V., Pastiu, A., Cozma, V., 2012. Epidemiological survey of Neospora
313
caninum infection in dogs from Romania. Vet. Parasitol. 188, 382-385.
RI PT
312
314
Gennari, S.M., Yai, L.E., D'Auria, S.N., Cardoso, S.M., Kwok, O.C., Jenkins, M.C., Dubey, J.P.,
316
2002. Occurrence of Neospora caninum antibodies in sera from dogs of the city of São Paulo,
317
Brazil. Vet. Parasitol. 106, 177-179.
M AN U
318
SC
315
319
Gondim, L.F.P., McAllister, M.M., Pitt, W.C., Zemlicka, D.E., 2004. Coyotes (Canislatrans) are
320
definitive hosts of Neospora caninum. Int. J. Parasitol. 34, 159-161.
TE D
321
Guedes, M.H.P., Guimarães, A.M., Rocha, C.M.B.M., Hirsch, C., 2008. Frequência de anticorpos
323
anti-Neospora caninum em vacas e fetos provenientes de municípios do sul de Minas Gerais. Rev.
324
Brazil. Parasitol. Vet. 17, 189-194.
EP
322
325
Guimarães, A.M., Rocha, C.M.B.M., Oliveira, T.M.F.S., Rosado, I.R., Morais, L.G., Santos,
327
R.R.D., 2009. Fatores associados à soropositividade para Babesia, Toxoplasma, Neospora e
328
Leishmania em cães atendidos em nove clínicas veterinárias do município de Lavras, MG. Rev.
329
Brazil. Parasitol. Vet. 18, 49-53.
AC C
326
330 331
Hemphill, J., Gottstein, B., 2000. A European perspective on Neospora caninum. Int. J. Parasitol.
332
30, 877-924.
333 14
ACCEPTED MANUSCRIPT 334
Kuruca, L., Spasojević-Kosić, L., Simin, S., Savović, M., Lauš, S., Lalošević, V., 2013. Neospora
335
caninum antibodies in dairy cows and domestic dogs from Vojvodina, Serbia. Parasite 20, 40.
336
Lindsay, D.S., Dubey, J.P., Duncan, R.B., 1999. Confirmation that the dog is a definitive host for
338
Neospora caninum. Vet. Parasitol. 82, 327-333.
RI PT
337
339
Martins, J., Kwok, O.C.H., Dubey, J.P., 2011. Seroprevalence of Neospora caninum in free-range
341
chickens (Gallus domesticus) from the Americas. Vet. Parasitol. 182, 349-351.
SC
340
342
McAllister, M.M., Dubey, J.P., Lindsay, D.S., Jolley, W.R., Wills, R.A., McGuire, A.M., 1998.
344
Dogs are definitive hosts of Neospora caninum. International J Parasitol. 28, 1473-1478.
M AN U
343
345
Packham, A.E., Sverlow, K.W., Conrad, P.A., Loomis, E.F., Rowe, J.D., Anderson, M.L., Marsh,
347
A.E., Cray, C., Barr, B.C., 1998. A Modified Agglutination Test for Neospora caninum:
348
Development, Optimization, and Comparison to the Indirect Fluorescent-Antibody Test and
349
Enzyme-Linked Immunosorbent Assay. Clin. Diagn. Lab. Immunol. 5, 467-473.
EP
350
TE D
346
Paradies, P., Capelli, G., Testini, G., Cantacessi, C., Trees, A.J., Otranto, D., 2007. Risk factors for
352
canine neosporosis in farm and kennel dogs in southern Italy. Vet. Parasitol. 145, 240-244.
353
AC C
351
354
Poglayen, G., Giannetto, S., Foti, S., Foti, M., Finocchiaro, B., Genchi, C., 2000. Serological survey
355
of Neospora caninum in dogs from Sicily. Parassitol. 42, 186.
356 357
Rasmussen, K., Jensen, A.L., 1996. Some epidemiologic features of canine neosporosis in
358
Denmark. Vet. Parasitol. 62, 345-349.
359 15
ACCEPTED MANUSCRIPT 360
Romanelli, P.R., Freire, R.L., Vidotto, O., Marana, E.R.M., Ogawa, L., De Paula, V.S.O., Garcia,
361
J.L., Navarro, I.T., 2007. Prevalence of Neospora caninum and Toxoplasma gondii in sheep and
362
dogs from Guarapuava farms, Paraná State. Brazil. Res. Vet. Sci. 82, 202-207.
363
Václavek, P., Sedlák, K., Hůrková, L., Vodrázka, P., Sebesta, R., Koudela, B., 2007. Serological
365
survey of Neospora caninum in dogs in the Czech Republic and a long-term study of dynamics of
366
antibodies. Vet. Parasitol. 143, 35-41.
RI PT
364
SC
367
Wanha, K., Edelhofer, R., Gabler-Eduardo, C., Prosl, H., 2005. Prevalence of antibodies against
369
Neospora caninum and Toxoplasma gondii in dogs and foxes in Austria. Vet. Parasitol. 128, 189-
370
193.
371
Wouda,
Dijkstra,
T.,
Kramer,
A.M.,
373
Seroepidemiological evidence for a relationship between Neospora caninum infections in dogs and
374
cattle. Inter. J. Parasitol. 29, 1677-1682.
AC C
EP
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375
W.,
M AN U
368
16
van
Maanen,
C.,
Brinkhof,
J.M.,
1999.
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Table 1.
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Prevalence of anti-Neospora caninum antibodies in farm dogs according to the variables age, sex, breed, reproductive
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status, feeding habits, pregnancy, reproductive disorders. Variables “Age and Feeding habits” were recorded for each
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animal according to the owners knowledge.
380 381 % positivity 44.8% 382 42.1%
13 14
11 12
45.8% 46.2%
Pure breed Mixed breed
Reproductive status
Intact Neutered
Feeding habits (fetuses/placenta)
Yes No
Pregnancy
Yes No
Reproductive disorders
Yes No
10 13
76.9% 35.1%
21 6
16 7
43.2% 53.8%
24 3
18 4
42.9% 57.1%
4 23
8 15
66.7% 39.5%
0 27
2 21
100% 43.8%
3 24
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Breed
Male Female
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Sex
3 years
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IFAT + 13 8
IFAT -
Variables Age
383 384 385 386 387 388 389
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Pure breed: Malamute, Newfoundland, Boxer, German Shepherd, Bull Terrier.
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ACCEPTED MANUSCRIPT 394 Table 2. 395 Prevalence of anti-Neospora caninum antibodies in dogs breeding according to the variables age, sex, breed, reproductive 396 status, pregnancy, reproductive disorders. Variables “Reproductive status, Pregnancy and Reproductive disorders” have 397 been evaluated only for female dogs.
IFAT -
400 401 402 403
Sex
Male Female
14 26
3 6
Breed
Malamute Newfoundland Boxer German Shepherd Hunting dogs Bull Terrier
13 10 4 7 2 5
Reproductive status
Intact Neutered
Pregnancy
None Normal Patological
Reproductive disorders
Yes No
17.6% 18.8%
4 1 0 1 3 0
23.5% 9.1% 0% 12.5% 60.0% 0%
39 1
9 0
18.8% 0%
15 7 3
3 1 0
16.7% 12.5% 0%
1 25
0 6
0% 19.4%
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% positivity 16.7% 19.2%
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IFAT + 4 5
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3 years
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Variables Age
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404 405 406 407 408 18
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Table 3.
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Results of the multivariate analysis of potential risk factors.
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Provenance Breed by provenance Age Sex Constant
B
S.E.
Wald
df
Sig.
Exp(B)
0.365 0.999 0.012 -0.068 -0.151
0.264 0.377 0.251 0.255 0.379
1.918 7.002 0.002 0.072 0.158
1 1 1 1 1
0.166 0.008 0.960 0.788 0.691
1.441 2.715 1.013 0.934 0.860
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95.0% C.I.for EXP(B) Lower Upper 0.859 1.296 0.620 0.567
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Independent variables
2.415 5.690 1.655 1.538
ACCEPTED MANUSCRIPT Highlights • The seropositivity for Neospora caninum in breeding and farm dogs was determine. • Seropositivity was performed by indirect immunofluorescent antibody test (IFAT). • Dogs living in the cattle farms showed a higher seropositivity for N. caninum.
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• The potential risk of horizontal transmission of N. caninum between dogs and cattles was confirmed.
S0014-4894(16)30018-2
DOI:
10.1016/j.exppara.2016.02.003
Reference:
YEXPR 7189
To appear in:
Experimental Parasitology
Received Date: 13 April 2015 Revised Date:
4 February 2016
Accepted Date: 5 February 2016
Please cite this article as: Robbe, D., Passarelli, A., Gloria, A., Di Cesare, A., Capelli, G., Iorio, R., Traversa, D., Neospora caninum seropositivity and reproductive risk factors in dogs, Experimental Parasitology (2016), doi: 10.1016/j.exppara.2016.02.003. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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ACCEPTED MANUSCRIPT Neospora caninum seropositivity and reproductive risk factors in dogs Domenico Robbe1, Alessandra Passarelli1, Alessia Gloria1*, Angela Di Cesare1, Gioia Capelli2, Raffaella Iorio1, Donato Traversa1 1
Faculty of Veterinary Medicine, University of Teramo, Piazza Aldo Moro 45, 64100, Teramo, Italy Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università, 10-35020 Legnaro (PD), Italy.
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*Corresponding author. Tel.: +390861266995 E-mail address: [email protected] (Alessia Gloria).
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ACCEPTED MANUSCRIPT Abstract
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Despite the importance of Neospora caninum in veterinary medicine, knowledge of distribution of
15
neosporosis in dog populations in some countries is still poor. The aims of the present study were to
16
determine the occurrence of anti-N. caninum antibodies in one-hundred dogs living in cattle farms
17
or dog breedings in central Italy and to evaluate the risk factors associated with seropositivity. The
18
incidence of reproductive system disorders (e.g. infertility after first pregnancy) was also evaluated.
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Serum from breeding and farm dogs was tested to an indirect immunofluorescent antibody test
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(IFAT) to assess the occurrence of seropositivity. Management and individual data were collected
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and analysed both by linear and logistic multiple-regression models to find reliable predictors of
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seroprevalence and anti-N. caninum antibody level. The seropositivity for N. caninum was 32%.
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Dogs reared for breeding and presence of cattle on the farm were associated with seropositivity for
24
N. caninum. Dogs living in the cattle farms showed a higher seropositivity for N. caninum (46%)
25
compared with those living in dogs breeding (18%) (P or < 3 years), sex, and breed (mongrel/pure breed) were offered to binary
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logistic models to evaluate possible risk factors for N. caninum-seropositivity. Breed was
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introduced in the models as interaction with the provenance. The differences were considered
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significant with P0.05).
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3.2 Group 2
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Out of the 50 dogs belonging to Group 2, nine (18%) were positive for anti-N. caninum antibodies.
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None of the nine seropositive animals, which belonged to four different breeds, showed past or
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present reproductive problems (Table 2). The presence of N. caninum antibodies was detected in
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four of the six facilities examined. However, most of the infected animals (60%) were hunting dogs,
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and the differences between this typology of dogs and the other breeds included in the study were
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not statistically significant (P>0.05) (Table 2). All variables were not statistically related to
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seropositivity (P>0.05).
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3.3 Risk factors
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The model which best fitted the data is shown in Table 3. The only significant predictor of
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Neospora-seropositity was the interaction provenance-breed, i.e. pure breed dogs living in a cattle
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farm had 2.7 more probability to be Neospora seropositive than the other animals.
134 135
4. Discussion
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ACCEPTED MANUSCRIPT The present study showed for the first time the presence of anti-N. caninum specific antibodies in
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dogs living in this area of Italy. Despite the examined groups of dogs do not represent the general
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canine population of Italy, the results demonstrate that the pathogen may circulate in selected
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populations of dogs in particular geographic areas that have been never investigated for
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neosporosis. Hence, the achievements are of particular relevance in those settings where zootechnic
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activities are of economic importance.
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Despite dogs play a major role in the epidemiology of the infection, studies on the occurrence of N.
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caninum and the risk factors for neosporosis in dogs are still scant in several countries, including
144
Italy. The prevalence recorded in the present study (i.e. 32%) is similar to data (36.4%) reported in
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north-western Italy in dogs living in rural habitats (Ferroglio et al., 2007), and higher than data
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reported (10.9%) in kenneled and owned dogs living in north-eastern Italy (Capelli et al., 2004), and
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in kenneled and farm dogs from the south of the Country (14.6% and 26.5% respectively) (Paradies
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et al., 2007).
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Prevalence recorded in other European countries, i.e. Austria (5.3% in dogs from rural areas and
150
2.1% from the urban area of Vienna) (Wanha et al., 2005), Czech Republic (4.7% in dogs at the
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service of the Czech Army and 2.6% in pet dogs) (Václavek et al., 2007), Serbia (17.2% in hunting,
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farm and stray dogs) (Kuruca et al., 2013), were lower than data showed in the present survey. A
153
recent study carried out in Romania reported a similar rate of infection (i.e. total rate of infection of
154
32.7%) (Gavrea et al., 2012). In this latter study dogs were divided in different categories showing
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the following results: 54.5% of hunting, 50% of farm, 29.6% of guard, 31.2% of shelter, 34.8 % of
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pet and 0% of kennel dogs were positive. Moreover, 34.5% of dogs from rural and 31.4% from
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urban areas presented anti-N. caninum specific antibodies (Gavrea et al., 2012).
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In the present study, dogs living in cattle farms showed a highest percentage of seropositivity
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infection (46%) compared to dogs in breeding facilities (18%). This difference could be explained
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by wrong feeding habits. In fact, dogs living outdoor in livestock premises have more chances to
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become in contact with bovine infected tissues (e.g. fetuses/placentas) or with other intermediate
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ACCEPTED MANUSCRIPT hosts of N. caninum (e.g. rodents, birds). However, a history of ingestion of bovine tissues resulted
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equivalent in both dogs which tested positive or negative for N. caninum.
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Although owners of dogs belonging to both groups declared to feed them with commercial food, the
165
administration of raw or undercooked meat cannot be ruled out. Also, dogs living in rural farms are
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more likely to be in contact with wildlife and other potential ruminant hosts and more likely to
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show their carnivorous and scavenging behavior by eating small mammals and birds which possibly
168
contain N. caninum tissue cysts (Wouda et al., 1999). These features could explain the prevalence
169
differences between the two study groups.
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Despite no specific risk factors were evidenced for dogs living in breeding facilities, a relatively
171
high value of seropositivity was recorded. This result may be due to the presence, in the study
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kennels, of open fences and outdoor areas which allow easy access to rodents or birds, as previously
173
described (Fernandes et al., 2004).
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The higher seropositivity in pure breed animals than in mixed breed dogs fits with the results of
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previous studies, which suggested a genetic factor predisposing to infection, or a more efficient
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vertical transmission (Capelli et al., 2004). However, other works showed a higher prevalence in
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mixed breed dogs (Fernandes et al., 2004; Collantes-Fernandez et al., 2008), thus the role of breeds
178
in the epidemiology of canine neosporosis is not well established.
179
In any case, it can be argued that hunting dogs have a high chance of being fed with hunted animals
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that can act as intermediate hosts for the parasite (Gavrea et al., 2012). Furthermore, a vertical
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transmission is also possible in breeding facilities due to the introduction of infected animals in the
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farms or to a relapse of congenital infection during pregnancy (Rasmussen and Jensen, 1996;
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Wouda et al., 1999).
184
Other factors, i.e. sex, age, reproductive status and/or reproductive disorders, were not significant.
185
Analogously, other studies have indicated that sex does not influence the occurrence of N. caninum
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(Cheadle et al., 1999; Ferroglio et al., 2007; Collantes-Fernández et al., 2008). Indeed, dogs of any
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age can acquire the infection by N. caninum. Some studies have shown that older animals may have
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ACCEPTED MANUSCRIPT a higher risk to be infected due to the chronic pattern of the disease (Wouda et al., 1999; Fernandes
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et al., 2004; Ferroglio et al., 2007).
190
Although provenance and breed resulted predictors of seropositivity further studies are warranted to
191
eventually support these results in other categories of dogs, e.g. non-breeding animals or dogs
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Among the risk factors evaluated in the present study, the positive percentage was significantly
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higher in farm dogs who had a full-term pregnancy. Although dog neosporosis is a cause of
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reproductive disorders in dogs (Barber and Tress, 1998), information on abortion rates, infertility or
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other reproductive disorders were not well defined. Further studies are warranted to understand the
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actual impact of neosporosis in causing reproductive infertility, abortion and stillbirths in bitches.
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On the whole, the data showed in the present study provide new information of the prevalence of N.
198
caninum in dogs and contribute to the knowledge of the role of the parasite in reproductive
199
disorders in dogs. The high seropositivity in dogs living with livestock confirms that habitats, such
200
as rural areas, and the close contact with ruminants may indeed represent risk factors for the
201
occurrence of canine neosporosis. Moreover, an association between the presence of dogs and the
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prevalence of anti-N. caninum antibodies in cattle was observed in previous studies (Wouda et al.,
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1999; Dijkstra et al., 2002), thus suggesting a potential transmission of the parasite between dog
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and cattle according to the an higher exposure to environmental sources of infection (Regidor-
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Cerrillo et al., 2010). Therefore, dogs should be not allowed to have access to infected materials
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such as bovine carcasses, aborted fetuses, placenta and uterine discharge, in order to limit the
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prevalence of the infection in both dogs and cattle. In fact, N. caninum is a major cause of abortion
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in cattle all over the world. Also, knowledge of the occurrence of the infection in dog breeding
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facilities is of particular importance to understand the correlation between seropositive dogs and the
210
effects on their reproductive performances. While the transplacental transmission has been
211
demonstrated in experimental infections, the frequency of vertical transmission in naturally infected
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dogs is low (Barber and Trees, 1998; Cavalcante et al., 2012). Nonetheless, it should be borne in
213
mind that clinical neosporosis in adult dogs may result either from a recent infection or to the
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ACCEPTED MANUSCRIPT recrudescence of an existing infection as a result of pregnancy or other immune-compromising
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conditions. Stressing and immune-compromising drivers can cause bradyzoites to transform back
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into tachyzoites, resulting in reproductive disorders (e.g. abortion) and infection of fetuses in
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pregnant animals (e.g. stillbirths) (Barber and Tress, 1998; Lyndsay et al., 1999; Dubey et al., 2007,
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2011).
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5. Conclusions
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Serologic testing of bitches can be used as a method to determine whether neosporosis is a potential
222
or evident reproductive problem in dog breeding. This strategy can be helpful when investigating
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dog farms with cases of abortion, stillbirths or litter with neuro-muscular disorders. In these cases,
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seropositive animals should be excluded from breeding. Although the number of positive dogs in
225
the study area is not particularly high, the pathogenic potential of this parasite should be always
226
taken into appropriate account for the reproductive disorders they may cause.
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On the contrary, the high level of seropositivity in dogs living in cattle farms implies that more
228
attention is warranted in the management (i.e. feeding habits) of dogs living with livestock and/or in
229
rural conditions, and in the introduction of new dogs in a farm, in order to limit the spreading of
230
neosporosis in both dogs and cattle.
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References
235
Aguiar, D.M., Cavalcante, G.T., Rodrigues, A.A.R., Labruna, M.B., Camargo, L.M.A., Camargo,
237
E.P., Gennari, .S.M., 2006. Prevalence of anti-Neospora caninum antibodies in cattle and dogs from
238
Western Amazon, Brazil, in association with some possible risk factors. Vet. Parasitol. 142, 71-77.
RI PT
236
239
Barber, J.S., Trees, A.J., 1998. Naturally occurring vertical transmission of Neospora caninum in
241
dogs. Int. J. Parasitol. 28, 57-64.
SC
240
242
Björkman, C., Jakubek, E.B., Arnemo, J.M., Malmsten, J., 2010. Seroprevalence of Neospora
244
caninum in gray wolves in Scandinavia. Vet. Parasitol. 173, 139-142.
M AN U
243
245
Björkman, C., Lundën, A., Uggla, A., 1994. Prevalence of antibodies to Neospora caninum and
247
Toxoplasma gondii in Swedish dogs. Acta Vet. Scand. 35, 445-447.
248
TE D
246
Capelli, G., Nardelli, S., di Regalbono, A.F., Scala, A., Pietrobelli, M., 2004. Sero-epidemiological
250
survey of Neospora caninum infection in dogs in north-eastern Italy. Vet. Parasitol. 123, 143-148.
251
EP
249
Cavalcante, G.T., Soares, R.M., Nishi, S.M., Hagen, S.C., Vannucchi, C.I., Maiorka, P.C., Paixão,
253
A.S., Gennari, S.M., 2012. Experimental infection with Neospora caninum in pregnant bitches.
254
Rev. Bras. Parasitol. Vet. 21, 232-236.
AC C
252
255 256
Cheadle, M.A., Lindsay, D.S., Blagburn, B.L. 1999. Prevalence of antibodies to Neospora caninum
257
in dogs. Vet. Parasitol. 85, 325-330.
258
11
ACCEPTED MANUSCRIPT 259
Collantes-Fernández, E., Gómez-Bautista, M., Miró, G., Alvarez-García, G., Pereira-Bueno, J.,
260
Frisuelos, C., Ortega-Mora, L.M., 2008. Seroprevalence and risk factors associated with Neospora
261
caninum infection in different dog populations in Spain. Vet. Parasitol. 152, 148-151.
262
Costa, K.S., Santos, S.L., Uzêda, R.S., Pinheiro, A.M., Almeida, M.A.O., Araújo, F.R., McAllister,
264
M.M., Gondim, L.F., 2008. Chickens (Gallus domesticus) are natural intermediate hosts of
265
Neospora caninum. Int. J. Parasitol. 38, 157-159.
SC
266
RI PT
263
Costa-Júnior, L.M., Rembeck, K., Ribeiro, M.F.B., Beelitz, P., Pfister, K., Passos, L.M.F., 2007.
268
Sero-prevalence and risk indicators for canine ehrlichiosis in three rural areas of Brazil. Vet. J. 174,
269
673-676.
M AN U
267
270
Costa-Júnior, L.M., Ribeiro, M.F.B., Rembeck, K., Rabelo, E.M.L., Zahler-Rinder, M., Hirzmann,
272
J., Pfister, K., Passos, L.M., 2009. Canine babesiosis caused by Babesia canis vogeli in rural areas
273
of the State of Minas Gerais, Brazil and factors associated with its seroprevalence. Res. Vet. Sci. 86,
274
257-260.
EP
275
TE D
271
Crookshanks, J.L., Taylor, S.M., Haines, D.M., Shelton, G.D., 2007. Treatment of canine pediatric
277
Neospora caninum myositis following immunohistochemical identification of tachyzoites in muscle
278
biopsies. Can. Vet. J. 48, 506-508.
279
AC C
276
280
Cunha Filho, N.A., Lucas, A.S., Pappen, F.G., Ragozo, A.M.A., Gennari, S.M., Júnior, T.L., Farias,
281
N.A., 2008. Fatores de risco e prevalência de anticorpos anti-Neospora caninum em cães urbanos e
282
rurais do Rio Grande do Sul, Brasil. Rev. Bras. Parasitol. Vet. 17, 301-306.
283
12
ACCEPTED MANUSCRIPT 284
Dijkstra, T., Barkema, H.W., Eysker, M., Hesselink, J.W., Wouda, W., 2002. Natural transmission
285
routes of Neospora caninum between farm dogs and cattle. Vet. Parasitol. 105, 99-104.
286
Dubey, J.P., Carpenter, J.L., Speer, C.A., Tooper, M.J., Uggla, A., 1988. Newly recognized
288
protozoan disease of dogs. J. Am. Vet. Med. Asso. 192, 1269-1285.
289
RI PT
287
Dubey, J.P., Lindsay, D.S., 1996. A review of Neospora caninum and neosporosis. Vet. Parasitol.
291
67, 1-59.
SC
290
292
Dubey, J.P., Vianna, M.C., Kwok, O.C., Hill, D.E., Miska, K.B., Tuo, W., Velmurugan, G.V.,
294
Conors, M., Jenkins, M.C., 2007. Neosporosis in Beagle dogs: Clinical signs, diagnosis, treatment,
295
isolation and genetic characterization of Neospora caninum. Vet. Parasitol. 149, 158-166.
M AN U
293
296
Dubey, J.P., Jenkins, M.C., Rajendran, C., Miska, K., Ferreira, L.R., Martins, J., Kwok, O.C.,
298
Choudhary, S., 2011. Gray wolf (Canis lupus) is a natural definitive host for Neospora caninum.
299
Vet. Parasitol. 181, 382-387.
EP
300
TE D
297
Dubey, J.P., Schares, G., 2011. Neosporosis in animals-the last five years. Vet. Parasitol. 180, 90-
302
108.
303
AC C
301
304
Fernandes, B.C.T.M., Gennari, S.M., Souza, S.L.P., Carvalho, J.M., Oliveira, W.G., Cury, M.C.,
305
2004. Prevalence of anti-Neospora caninum antibodies in dogs from urban, periurban and rural
306
areas of the city of Uberlândia, Minas Gerais-Brazil. Vet. Parasitol. 123, 33-40.
307
13
ACCEPTED MANUSCRIPT 308
Ferroglio, E., Pasino, M., Ronco, F., Benà, A., Trisciuoglio, A., 2007. Seroprevalence of antibodies
309
to Neospora caninum in urban and rural dogs in north-west Italy. Zoonoses Public Health 54, 135-
310
139.
311
Gavrea, R., Mircean, V., Pastiu, A., Cozma, V., 2012. Epidemiological survey of Neospora
313
caninum infection in dogs from Romania. Vet. Parasitol. 188, 382-385.
RI PT
312
314
Gennari, S.M., Yai, L.E., D'Auria, S.N., Cardoso, S.M., Kwok, O.C., Jenkins, M.C., Dubey, J.P.,
316
2002. Occurrence of Neospora caninum antibodies in sera from dogs of the city of São Paulo,
317
Brazil. Vet. Parasitol. 106, 177-179.
M AN U
318
SC
315
319
Gondim, L.F.P., McAllister, M.M., Pitt, W.C., Zemlicka, D.E., 2004. Coyotes (Canislatrans) are
320
definitive hosts of Neospora caninum. Int. J. Parasitol. 34, 159-161.
TE D
321
Guedes, M.H.P., Guimarães, A.M., Rocha, C.M.B.M., Hirsch, C., 2008. Frequência de anticorpos
323
anti-Neospora caninum em vacas e fetos provenientes de municípios do sul de Minas Gerais. Rev.
324
Brazil. Parasitol. Vet. 17, 189-194.
EP
322
325
Guimarães, A.M., Rocha, C.M.B.M., Oliveira, T.M.F.S., Rosado, I.R., Morais, L.G., Santos,
327
R.R.D., 2009. Fatores associados à soropositividade para Babesia, Toxoplasma, Neospora e
328
Leishmania em cães atendidos em nove clínicas veterinárias do município de Lavras, MG. Rev.
329
Brazil. Parasitol. Vet. 18, 49-53.
AC C
326
330 331
Hemphill, J., Gottstein, B., 2000. A European perspective on Neospora caninum. Int. J. Parasitol.
332
30, 877-924.
333 14
ACCEPTED MANUSCRIPT 334
Kuruca, L., Spasojević-Kosić, L., Simin, S., Savović, M., Lauš, S., Lalošević, V., 2013. Neospora
335
caninum antibodies in dairy cows and domestic dogs from Vojvodina, Serbia. Parasite 20, 40.
336
Lindsay, D.S., Dubey, J.P., Duncan, R.B., 1999. Confirmation that the dog is a definitive host for
338
Neospora caninum. Vet. Parasitol. 82, 327-333.
RI PT
337
339
Martins, J., Kwok, O.C.H., Dubey, J.P., 2011. Seroprevalence of Neospora caninum in free-range
341
chickens (Gallus domesticus) from the Americas. Vet. Parasitol. 182, 349-351.
SC
340
342
McAllister, M.M., Dubey, J.P., Lindsay, D.S., Jolley, W.R., Wills, R.A., McGuire, A.M., 1998.
344
Dogs are definitive hosts of Neospora caninum. International J Parasitol. 28, 1473-1478.
M AN U
343
345
Packham, A.E., Sverlow, K.W., Conrad, P.A., Loomis, E.F., Rowe, J.D., Anderson, M.L., Marsh,
347
A.E., Cray, C., Barr, B.C., 1998. A Modified Agglutination Test for Neospora caninum:
348
Development, Optimization, and Comparison to the Indirect Fluorescent-Antibody Test and
349
Enzyme-Linked Immunosorbent Assay. Clin. Diagn. Lab. Immunol. 5, 467-473.
EP
350
TE D
346
Paradies, P., Capelli, G., Testini, G., Cantacessi, C., Trees, A.J., Otranto, D., 2007. Risk factors for
352
canine neosporosis in farm and kennel dogs in southern Italy. Vet. Parasitol. 145, 240-244.
353
AC C
351
354
Poglayen, G., Giannetto, S., Foti, S., Foti, M., Finocchiaro, B., Genchi, C., 2000. Serological survey
355
of Neospora caninum in dogs from Sicily. Parassitol. 42, 186.
356 357
Rasmussen, K., Jensen, A.L., 1996. Some epidemiologic features of canine neosporosis in
358
Denmark. Vet. Parasitol. 62, 345-349.
359 15
ACCEPTED MANUSCRIPT 360
Romanelli, P.R., Freire, R.L., Vidotto, O., Marana, E.R.M., Ogawa, L., De Paula, V.S.O., Garcia,
361
J.L., Navarro, I.T., 2007. Prevalence of Neospora caninum and Toxoplasma gondii in sheep and
362
dogs from Guarapuava farms, Paraná State. Brazil. Res. Vet. Sci. 82, 202-207.
363
Václavek, P., Sedlák, K., Hůrková, L., Vodrázka, P., Sebesta, R., Koudela, B., 2007. Serological
365
survey of Neospora caninum in dogs in the Czech Republic and a long-term study of dynamics of
366
antibodies. Vet. Parasitol. 143, 35-41.
RI PT
364
SC
367
Wanha, K., Edelhofer, R., Gabler-Eduardo, C., Prosl, H., 2005. Prevalence of antibodies against
369
Neospora caninum and Toxoplasma gondii in dogs and foxes in Austria. Vet. Parasitol. 128, 189-
370
193.
371
Wouda,
Dijkstra,
T.,
Kramer,
A.M.,
373
Seroepidemiological evidence for a relationship between Neospora caninum infections in dogs and
374
cattle. Inter. J. Parasitol. 29, 1677-1682.
AC C
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W.,
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van
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C.,
Brinkhof,
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1999.
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Table 1.
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Prevalence of anti-Neospora caninum antibodies in farm dogs according to the variables age, sex, breed, reproductive
378
status, feeding habits, pregnancy, reproductive disorders. Variables “Age and Feeding habits” were recorded for each
379
animal according to the owners knowledge.
380 381 % positivity 44.8% 382 42.1%
13 14
11 12
45.8% 46.2%
Pure breed Mixed breed
Reproductive status
Intact Neutered
Feeding habits (fetuses/placenta)
Yes No
Pregnancy
Yes No
Reproductive disorders
Yes No
10 13
76.9% 35.1%
21 6
16 7
43.2% 53.8%
24 3
18 4
42.9% 57.1%
4 23
8 15
66.7% 39.5%
0 27
2 21
100% 43.8%
3 24
M AN U
Breed
Male Female
SC
Sex
3 years
RI PT
16 11
IFAT + 13 8
IFAT -
Variables Age
383 384 385 386 387 388 389
EP
393
Pure breed: Malamute, Newfoundland, Boxer, German Shepherd, Bull Terrier.
AC C
392
TE D
390
17
391
ACCEPTED MANUSCRIPT 394 Table 2. 395 Prevalence of anti-Neospora caninum antibodies in dogs breeding according to the variables age, sex, breed, reproductive 396 status, pregnancy, reproductive disorders. Variables “Reproductive status, Pregnancy and Reproductive disorders” have 397 been evaluated only for female dogs.
IFAT -
400 401 402 403
Sex
Male Female
14 26
3 6
Breed
Malamute Newfoundland Boxer German Shepherd Hunting dogs Bull Terrier
13 10 4 7 2 5
Reproductive status
Intact Neutered
Pregnancy
None Normal Patological
Reproductive disorders
Yes No
17.6% 18.8%
4 1 0 1 3 0
23.5% 9.1% 0% 12.5% 60.0% 0%
39 1
9 0
18.8% 0%
15 7 3
3 1 0
16.7% 12.5% 0%
1 25
0 6
0% 19.4%
M AN U
TE D
% positivity 16.7% 19.2%
SC
20 21
IFAT + 4 5
AC C
399
3 years
EP
Variables Age
RI PT
398
404 405 406 407 408 18
ACCEPTED MANUSCRIPT 409
Table 3.
410
Results of the multivariate analysis of potential risk factors.
411
Provenance Breed by provenance Age Sex Constant
B
S.E.
Wald
df
Sig.
Exp(B)
0.365 0.999 0.012 -0.068 -0.151
0.264 0.377 0.251 0.255 0.379
1.918 7.002 0.002 0.072 0.158
1 1 1 1 1
0.166 0.008 0.960 0.788 0.691
1.441 2.715 1.013 0.934 0.860
412
AC C
EP
TE D
M AN U
SC
413
19
95.0% C.I.for EXP(B) Lower Upper 0.859 1.296 0.620 0.567
RI PT
Independent variables
2.415 5.690 1.655 1.538
ACCEPTED MANUSCRIPT Highlights • The seropositivity for Neospora caninum in breeding and farm dogs was determine. • Seropositivity was performed by indirect immunofluorescent antibody test (IFAT). • Dogs living in the cattle farms showed a higher seropositivity for N. caninum.
AC C
EP
TE D
M AN U
SC
RI PT
• The potential risk of horizontal transmission of N. caninum between dogs and cattles was confirmed.
