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Anti-Neospora caninum antibodies in beef cattle from the northern region of Parana state, Brazil/Anticorpos anti-Neospora caninum em bovinos de corte da regiao norte do estado do Parana, Brasil.


Neosporosis, caused by Neospora caninum, is one of the major reasons for reproductive failure in cattle and is responsible for fetal mummification, still births, and miscarriages (MARQUES et al., 2011; DUBEY & SCHARES, 2011; IBRAHIM et al., 2012). It has been estimated that the disease is responsible for worldwide economic losses of around 1 billion US dollars per year, and a loss of more than 100 million US dollars in the Brazilian beef industry (REICHEL et al., 2013).

Neospora caninum is prevalent in all continents (DUBEY & SCHARES, 2011), including Brazil, where different studies have been conducted to detect antibodies in beef cattle, with the prevalence ranging from 9.1 % to 97.2 % (HASEGAWA et al., 2004; VIANNA et al., 2008; ANDREOTTI et al., 2010; AMARAL et al., 2012; SILVA et al., 2017). In the state of Parana, different regions have been studied; the north central and southwest have a prevalence of antibodies against the parasite in beef cattle of up to 30.3 % and 13.2 %, respectively (MARQUES et al., 2011; MOURA et al., 2012; NASCIMENTO et al., 2014); however, different diagnostic techniques and cut-offs were used in these studies, which could explain the difference in prevalence rates. Moreover, seroprevalence of neosporosis may vary according to the type of cattle (beef versus dairy), mainly due to the differences between the production systems (DUBEY & SCHARES, 2011).

As there are few studies on neosporosis in beef cattle from the northern region of Parana state, the aim of the present study was to evaluate the occurrence of anti-N. caninum antibodies in beef cattle from slaughterhouses in the northern region of Parana state.


Location and sampling

The sampling size was determined as 384, using the software EpiInfo, version 6, using a prevalence of 50 % and a 5 % confidence interval. Between April 2015 and November 2016, blood samples from 401 animals were collected, 221 samples from an abattoir in Rolandia and 180 samples from a slaughterhouse in Borrazopolis, both located at northern region of the state of Parana, Brazil. The samples from beef cattle used in this study were from different municipalities in the northern region of Parana State, including Borrazopolis, Cambe, Conselheiro Mairink, Florai, Florestopolis, Guaraci, Ibipora, Jaguapita, Kalore, Lobato, Londrina, Nova Londrina, Rolandia, Sabaudia, Sapopema, and Tamarana. Blood samples were collected in the slaughter line and serum samples stored at -20 [degrees]C until serological analysis.


Serum samples were analyzed using an indirect fluorescent antibody test to detect anti-N. caninum antibodies according to the methodology described by CONRAD et al (1993). Tachyzoites of the Nc-1 strain previously cultivated in Vero cells in our lab were used as the antigen. Serum samples obtained from experimentally infected cattle with N. caninum and from healthy cattle were used as positive and negative controls, respectively, and included in all slides. The conjugate used was anti-bovine IgG FITC (whole molecule, Sigma-Aldrich[R], St. Louis, Missouri, USA) and serum samples that showed fluorescence through all parasite contours and titers [greater than or equal to] 100 were considered positive. Positive samples were titrated until the maximum dilution titer was reached.

Statistical analysis

Association between the variables was analyzed using a chi-square test ([x.sup.2]) with a significance level of 5 % using the Epi Info software, version 6.04b (DEAN et al., 1994). P-values [less than or equal to] 0.05 were considered significant.


Out of 401 analyzed samples, 37 (9.3%) were positive according to IFAT, with titers ranging from 100 to 400 (Table 1). Most of these samples were from the north central region of the state, with 8.8 % of animals positive for N. caninum (32/362, including the municipalities of Cambe, Florai, Florestopolis, Guaraci, Ibipora, Jaguapita, Lobato, Londrina, Rolandia, Sabaudia, Tamarana, and Kalore); the Pioneer north, with 14% of animals positive (4/29, including the municipalities of Conselheiro Mairink, and Sapopema) and northwest of Parana State, with 10% of animals positive (1/10, including the municipality of Nova Londrina).

Previous studies of N. caninum seroprevalence performed in Brazil have shown a prevalence ranging from 9.1% to 97.2% (CERQUEIRA-CEZAR et al., 2017). Although, RAGOZO et al. (2003) in Minas Gerais State and AGUIAR et al. (2006) in Rondonia State observed a similar of prevalence using IFAT of 9.5% and 11.1%, respectively, these studies used a cut-off of 25, which is different to our study. Other studies, also using IFAT and beef cattle, have described higher prevalences, ranging from 29.6% to 62.5%; however, care should be taken when comparing the studies, since the animals were from different regions, and might have had different management, and the cut-off used in each study might have varied (MELO et al., 2006; ANDREOTTI et al., 2010; SILVA et al., 2017). Epidemiological studies have indicated considerable differences in the prevalence of neosporosis between different countries, regions, beef and dairy cattle, and types of production. However, it is difficult to compare these results, mainly due to the differences in serological techniques and cut-offs used (DUBEY & SCHARES, 2011).

Analyzing the results according to sex revealed that 9.7% (28/289) of females and 8.0% (9/112) of males were positive; however, there was no statistical difference (Table 1), which corroborates many seroprevalence studies previously performed (CERQUEIRA-CEZAR et al., 2017).

The age of animals used in this study ranged from one and a half to eight years old, with most of them less than three years old (Table 1). The age of animals did not statistically influence the occurrence of N. caninum antibodies (p>0.05); although, the occurrence differed numerically (Table 1). These results do not allow us to identify whether horizontal or vertical transmission is the main route of infection and maintenance of the parasite in beef cattle herds in the region.

In a previous study performed in Para State, it was not possible to observe a relationship between age or seropositivity and neosporosis (SILVA et al., 2015). In another study performed in Peru, the authors did not find a correlation between age or the origin of the animals and seropositivity (p>0.05). Risk factors associated with N. caninum infection in cattle were the presence of more than three dogs in the herd, the disposal of animal waste in the environment, and the herd being composed of more than one hundred animals (PORTOCARRERO et al., 2015). MARQUES et al. (2011) analyzed 159 samples obtained from the northern region of Parana State and observed that the prevalence of N. caninum in dairy cattle did not proportionally increase with the age of the infected animals, corroborating the results of the present study.

Some of the municipalities had a prevalence below the average (9.2 %), including the cities Florestopolis, Ibipora, Londrina, Sapopema, and Tamarana, where no sero-occurrence of the parasite was observed (Table 1). However, no statistical significance was observed. This could be due to the lower number of collected samples. In contrast, in the municipality Sabaudia and Conselheiro Mairink, a high percentage of positive animals was observed. Geographical differences in the distribution of animals infected by N. caninum has already been reported in other studies and may be related to the relative abundance of the definitive host (canids) and the type of production (NASIR et al., 2012).

In the present study we did not evaluate reproductive status of the animals. Previous studies have shown that N. caninum infection in beef cattle causes economic losses. In Brazil, ANDREOTTI et al (2010) observed that N. caninum seroprevalence in animals with disorders during pregnancy was higher than in cows with normal pregnancies, and pregnancy rate for seropositive heifers was lower than for uninfected animals.


Our results showed a sero-occurrence of N. caninum in beef cattle slaughtered in the northern region of the state of Parana, showing that epidemiological studies performed in slaughterhouses could be used as a model for regional serum occurrence of N. caninum in beef cattle.

Received 10.23.18 Approved 03.22.19 Returned by the author 04.10.19


We would like to thank Fundacao Araucaria, Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) and Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) for financial support. J.L. Garcia is recipient of CNPq fellowship.


The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.


The present study was approved by Animal Ethical Committee of the Universidade Norte do Parana (n[degrees] 007/14).


All authors contributed equally for the conception and writing of the manuscript. All authors critically revised the manuscript and approved of the final version.


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Joice Loures Guerra (1) Werner Okano (1) Alexey Leon Gomel Bogado (1) Beatriz de Souza Lima Nino (2) Felippe Danyel Cardoso Martins (2) Sergio Tosi Cardim (2) Luiz Daniel de Barros (2) * (iD) Joao Luis Garcia (2)

(1) Universidade Norte do Parana (UNOPAR), Arapongas, PR, Brasil.

(2) Departamento de Medicina Veterinaria Preventiva, Universidade Estadual de Londrina (UEL), 86057-970, Londrina, PR, Brasil. E-mail: * Corresponding author.
Table 1--Results of indirect immunofluorescence antibody test
(IFAT) for Neospora caninum in beef cattle from the north
region of the state of Parana, Brazil.

Variables                    Animals      Positive

                           N      %     N       %

Municipalities     A      95    23.69   6     6,32
                   B      10    2.49    1     10.00
                   D      26    6.48    3     11.54
                   E       4    1.00    0       0
                   F      51    12.72   7     13.73
                   G       3    0.75    0       0
                   H      37    9.23    6     16.22
                   I      85    21.20   4     4.71
                   F       7    1.75    1     14.29
                   G      14    3.49    0       0
                   I      18    4.49    2     11.11
                   J      11    2.74    2     18.18
                   L       1    0.25    0       0
                   C      22    5.49    4     18.18
                   K       7    1.75    0       0
                   H      10    2.49    1     10.00
Sex               Male    112   27.93   9     8.04
                 Female   289   72.07   28    9.69
                   <3     307   76.56   30    9.77
Age (years)      3 a 5    73    18.20   6     8.22
                   >5     21    5.24    1     4.76
Total              --     401    100    37    9.23

Variables                 Negative      P-value

                           N      %

Municipalities     A      89    93.68   0.6020
                   B       9    90.00
                   D      23    88.46
                   E       4     100
                   F      44    86.27
                   G       3     100
                   H      31    83.78
                   I      81    95.29
                   F       6    85.71
                   G      14     100
                   I      16    88.89
                   J       9    81.82
                   L       1     100
                   C      18    81.82
                   K       7     100
                   H       9    90.00
Sex               Male    103   91.96   0.7483
                 Female   261   90.31
                   <3     277   90.23
Age (years)      3 a 5    67    91.78   0.7056
                   >5     20    95.24
Total              --     364   90.77   --

A: Borrazopolis, B: Cambe, C: Mairink, D: Florai, E: Florestopolis,
F: Guaraci, G: Ibipora, H: Jaguapita, I: Kalore, F: Lobato, G:
Londrina, H: Nova Londrina, I: Rolandia, J: Sabaudia, K: Sapopema,
L: Tamarana
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Title Annotation:PARASITOLOGY
Author:Guerra, Joice Loures; Okano, Werner; Bogado, Alexey Leon Gomel; de Souza Lima Nino, Beatriz; Martins
Publication:Ciencia Rural
Date:May 1, 2019
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