scholarly journals Detection of Neospora caninum and Toxoplasma gondii in Semen of Naturally Infected Rams

2019 ◽  
Vol 47 (1) ◽  
Author(s):  
Marilia De Oliveira Koch ◽  
Rosangela Locatelli Dittrich ◽  
Romildo Romualdo Weiss ◽  
Tacia Bergstein-Galan ◽  
Daniel Moura Aguiar ◽  
...  

Background: Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle. Moreover, abortion cases attributed to N. caninum and T. gondii infection have been occasionally reported in sheep. Due to the relatively scarce information on the molecular detection of N. caninum in the semen of naturally infected rams, this study aimed to detect parasitic DNA in fresh semen samples and in frozen extended semen straws from male sheep from artificial inseminations centers in Southern Brazil.Materials, Methods & Results: Semen samples of 38 rams from artificial insemination centers were evaluated. Eleven rams were naturally infected (seropositive for anti-N. caninum and/or anti-T. gondii IgG) and were selected for fresh semen collection. We tested all the samples for the closely related protozoan T. gondii to detect a possible cross-reaction and co-infection, due to the close similarity with N. caninum. The indirect fluorescent antibody test was used to detect IgG antibodies in the 11 serum samples from rams. Fresh semen samples were collected from 11 rams on days 1, 50, 55, and 58 using an artificial vagina and ewe in estrus. Other 27 rams had their frozen extended semen straws analyzed. A total of 20 fresh semen samples and 27 frozen extended semen straws samples were used to detect the presence of N. caninum and T. gondii DNA by polymerase chain reaction (PCR). Nc-5 and B1 genes were used as target regions to detect N. caninum and T. gondii DNA, respectively. The presence of N. caninum DNA was confirmed in the third collection of a fresh semen sample of one seropositive ram. T. gondii DNA was detected in a fresh semen sample of one seropositive ram. The DNA sequences of 186 bp from N. caninum (GenBank accession: MH806393) and 492 bp from T. gondii (GenBank accession: MH793503) were obtained by sequencing, and analysis revealed 99% and 100% identity, respectively, compared with other sequences deposited at GenBank. N. caninum and T. gondii DNAs were not detected in any of the 27 frozen extended semen straws used for artificial insemination.Discussion: This study demonstrated the presence of N. caninum and T. gondii DNA in fresh semen samples of naturally infected rams. The non-detection of N. caninum and T. gondii DNA in frozen semen samples of rams could be due to the dilution that was used to prepare the semen straws (GGL diluent and 5% glycerol), since fresh semen samples were not diluted prior to the test. Moreover, in our study, the volume of frozen semen samples (0.25 mL) used for PCR was lower than the volume of sediment obtained from fresh semen (0.5 mL), and the fresh semen centrifugation to obtain the sediment may have grouped the tachyzoites, increasing the sensitivity of the technique employed. No high IgG serological titers were detected in the rams at the time they were eliminating the parasite through fresh semen. The final titer of anti-N. caninum and anti-T. gondii IgGs in serum was 1:100, suggesting chronic infection. It is suggested that a new parasite elimination pathway is occurring among rams used for reproduction, due to the presence of N. caninum and T. gondii DNA in fresh semen samples from seropositive animals. Although the detection of genomic DNA of N. caninum and T. gondii in semen does not necessarily imply the presence of infectious stages of the parasites and does not determine their viability, these results demonstrate the need for further studies. Our study also indicates the need to reinforce preventive measures for sheep in artificial insemination centers until the risks are evaluated, by performing serological examinations with anti-N. caninum and anti-T. gondii antibodies, for instance, to select the rams that will be used for breeding.

2016 ◽  
Vol 25 (1) ◽  
pp. 116-118 ◽  
Author(s):  
Juliana Aizawa Porto de Abreu ◽  
Felipe da Silva Krawczak ◽  
Fernanda Passos Nunes ◽  
Marcelo Bahia Labruna ◽  
Hilda Fátima de Jesus Pena

Abstract Capybaras (Hydrochoerus hydrochaeris) are the largest rodents found in South America. The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in 170 free-living capybaras in a residential park area in Itu Municipality, São Paulo State, Brazil. Serum samples were tested by indirect fluorescent antibody test (IFAT) for T. gondii (IFAT ≥ 1:16) and N. caninum (IFAT ≥ 1:50). Among the 170 samples analyzed, 10% (17/170) and 0% (0/170) were seropositive for T. gondii and N. caninum, respectively. This study confirms the widespread presence of T. gondii and reinforces the role of capybaras in the life cycle of this parasite. Capybaras may not be important as intermediate hosts of N. caninum in the studied environment.


2016 ◽  
Vol 36 (10) ◽  
pp. 947-950 ◽  
Author(s):  
Luiza P. Portella ◽  
◽  
Gustavo C. Cadore ◽  
Marcelo de Lima ◽  
Luís A. Sangioni ◽  
...  

ABSTRACT: The presence of antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii was evaluated in buffaloes (Bubalus bubalis) from Rio Grande do Sul state (RS), southern Brazil. Serum samples (n=220) were analyzed for antibodies by indirect fluorescent antibody test (IFAT). Antibody presence was considered when the titers were equal or higher than 100 for these protozoa. A total of 60.5% (133/220) buffalo serum samples were positive for at least one of the protozoa evaluated in this study. Antibodies for N. caninum, Sarcocystis spp. and T. gondii were found in 36.4% (80/220), 25.5% (56/220) and 16.8% (37/220) of the buffaloes respectively, indicating a higher frequency of N. caninum infection (p=0.0133). The IFAT is a suitable method to diagnose N. caninum, Sarcocystis spp. and T. gondii infection in buffaloes for detecting IgG antibodies. This study demonstrates the presence of these three protozoa in buffalo herds in RS, Brazil, which may be source of infection to other animals. The high frequency of animals positive for N. caninum is important and could be related to reproductive problems. Additionally, the presence of Sarcocystis spp. and T. gondii in buffaloes can be a possible public health issue.


Author(s):  
Marilia De Oliveira Koch ◽  
Luciane Maria Laskoski ◽  
Daniel Moura de Aguiar ◽  
Bianca Ressetti da Silva ◽  
Reinaldo Ramos Régio ◽  
...  

The occurrence and distribution of antibodies against Sarcocystis neurona, Neospora caninum and Toxoplasma gondii was investigated in horses, dogs and cats from Curitiba, Paraná state, Brazil. Serum samples were selected from 100 horses, 100 dogs and 100 cats from the routine of the Veterinary Clinical Pathology Laboratory in the Veterinary Hospital of the University of Paraná (UFPR). The 100 dog samples were divided into two groups: 35 samples from dogs with neurological sign (convulsion) and 65 samples from dogs without neurological signs. The animals were adults of different breeds, males and females. Samples were analyzed by indirect fluorescent antibody test (IFAT) for protozoa S. neurona, N. caninum andT. gondii at the following cut-off dilutions: horses: 1:50, 1:50 and 1:16; dogs: 1:50, 1:50 and 1:16; cats: 1:50, 1:50 and 1:50, respectively. The obtained results were 42% of horses, 7% of dogs and 5% of cats seropositive for S. neurona; 58% of horses, 68% of dogs and 42% of cats seropositive to N. caninum, and 36% of horses, 20% of dogs and 21% of cats seropositive for T. gondii. Among the dogs with neurological signs, 8.6%, 68.6% and 25.7% were seropositive for S. neurona, N. caninum and T. gondii, respectively. Among the dogs without neurological signs, 6.2% 67.7% and 16.9% were seropositive for S. neurona, N. caninum and T. gondii, respectively. No statistical difference was found between groups of seropositive dogs for the three protozoa with neurological signs and without neurological signs. Co-infection and high antibody titers were detected. The antibodies against Sarcocystis neurona, Neospora caninum and Toxoplasma gondii were found widely distributed among horses, dogs and cats in the region of Curitiba, state of Paraná, Brazil.   


2013 ◽  
Vol 22 (1) ◽  
pp. 58-63 ◽  
Author(s):  
Fernanda Evers ◽  
João Luis Garcia ◽  
Italmar Teodorico Navarro ◽  
Dauton Luiz Zulpo ◽  
Beatriz de Souza Lima Nino ◽  
...  

This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16), T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15%) of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed toT. gondii.


2016 ◽  
Vol 37 (3) ◽  
pp. 1397 ◽  
Author(s):  
Maiara Sanitá Tafner Ferreira ◽  
Fernanda Silveira Flores Vogel ◽  
Luis Antonio Sangioni ◽  
Alfredo Skrebsky Cezar ◽  
Fernanda Rezer de Menezes

Neospora caninum and Toxoplasma gondii are coccidian parasites (protozoa: Apicomplexa) that cause reproductive losses in ruminants. Although, in sheep, abortion is generally related to T. gondii, potential damage caused by N. caninum must be considered. The aims of this study were to evaluate the frequencies of antibodies against Neospora spp. and T. gondii, and to identify the risk factors related to neosporosis and toxoplasmosis in sheep from ten flocks located in Central-Western, Northwestern, Southwestern, and Southeastern mesoregions of the Rio Grande do Sul state, southern Brazil. Blood samples were collected from 300 sheep (30 per flock) and an epidemiological questionnaire was completed by the farmers. The presence of canids, felids, and rodents, the sources of water for the animals, and their reproductive history were investigated as potential epidemiological factors related to the studied infections. Serum samples were tested by the indirect fluorescent antibody test for the presence of antibodies against Neospora spp. (cutoff 1:50) and T. gondii (cutoff 1:64), with respective frequencies detected being 16.3% (49/300) and 41.3% (124/300). Seropositivity to both parasites was observed in 8% (24/300) of the animals. Neospora spp. and T. gondii were present in 90% (9/10) and 100% (10/10) of the flocks, respectively. The presence of cats in the pastures, pens, and barns was significantly associated (p=0.047) with high frequencies (53.3% to 70%) of antibodies against T. gondii in the flocks. However, no associations were detected (p>0.05) between each of the other risk factors and the frequency of antibodies against Neospora spp. or T. gondii. Results of this study showed a broad distribution of both protozoa in evaluated sheep flocks.


2014 ◽  
Vol 23 (4) ◽  
pp. 449-455 ◽  
Author(s):  
Keyla Carstens Marques de Sousa ◽  
Heitor Miraglia Herrera ◽  
Iara Helena Domingos ◽  
João Bosco Vilela Campos ◽  
Isabela Maria Campanelli dos Santos ◽  
...  

An investigation was made into the occurrence of antibodies to Toxoplasma gondii, Leishmania infantum and Neospora caninum in 151 domestic cats, based on the indirect fluorescent antibody test (IFAT). Serum samples were collected from 151 domestic cats (65 free-roaming and 86 domiciled cats; 55 males and 96 females) in Campo Grande, Mato Grosso do Sul, Brazil between January and April 2013. IgG antibodies to T. gondii, L. infantum and N. caninum were found, respectively, in 49 (32.5%), 34 (22.5%) and 10 (6.6%) sampled cats. A positive correlation was found between T. gondii and N. caninum, T. gondii and L. infantum, and N. caninum and L. infantum (p <0.05) infections. Also, a significant interaction was identified between gender and area of activity on the probability of T. gondii (p = 0.0324) infection. However, no significant interaction was observed between gender and area of activity on infections by either N. caninum or L. infantum. This study showed that cats from an area endemic for visceral leishmaniasis in Brazil are exposed to three different protozoans, two of which are causal agents of important zoonosis.


2011 ◽  
Vol 20 (3) ◽  
pp. 202-206 ◽  
Author(s):  
Nicolle Fridlund Plugge ◽  
Fabiano Montiani Ferreira ◽  
Rosária Regina Tesoni de Barros Richartz ◽  
Adriana de Siqueira ◽  
Rosangela Locatelli Dittrich

This study aimed to evaluate occurrences of antibodies against Neospora caninum and Toxoplasma gondii in dogs with neurological signs. Blood samples from 147 dogs were collected: 127 from owned dogs (attended at the Veterinary Teaching Hospital of the Federal University of Paraná (HV-UFPR) and at private veterinary clinics in the city of Curitiba), and 20 from stray dogs found in Curitiba's metropolitan region. The dogs presented one or more of the following neurological signs: seizures, paresis or paralysis, ataxia, behavioral abnormalities, sensory and somatic disorders and chorioretinitis. The samples were analyzed by means of the indirect fluorescent antibody test (IFAT), at a cutoff dilution of 1:50. Out of the 147 samples obtained, 17 (11.56%) were seropositive for N. caninum, 31 (21.08%) for T. gondii and four (2.72%) for both protozoa. Serum titration on the positive animals showed that 54.83% (17/31) and 41.18% (7/17) had titers > 1:200 against T. gondii and N. caninum, respectively. A significant difference in seropositivity for T. gondii (P = 0.021; OR = 2.87; CI = 1.1 > 2.8 > 7.4) was observed between owned dogs (18.11%) and stray dogs (40%). Inclusion of serological tests for neosporosis and toxoplasmosis is recommended in diagnosing neurological diseases in dogs.


2011 ◽  
Vol 20 (2) ◽  
pp. 111-114 ◽  
Author(s):  
Marcos Gomes Lopes ◽  
Ivete Lopes Mendonça ◽  
Kleber Portela Fortes ◽  
Marcos Amaku ◽  
Hilda de Fátima Jesus Pena ◽  
...  

This study aimed to evaluate the presence of antibodies against Neospora caninum, Toxoplasma gondii and Leishmania infantum in dogs attended at the Veterinary Hospital of the Federal University of Piauí, Northeastern Brazil, where there are no reports of the occurrence of N. caninum and T. gondii in dogs. Serum samples from 530 dogs of genders, different ages and breeds from the municipality of Teresina and nearby towns were analyzed using three indirect fluorescent antibody tests, each one targeting one of the three agents. The associations between the parasites and gender, breed and age of the dogs were assessed by the chi-square test (p > 0.05). The occurrence of antibodies to N. caninum, T. gondii and L. infantum was 3.2, 18.0 and 78.1%, respectively. Toxoplasma gondii was more frequently found in older dogs (p < 0.05) whereas L. infantum was more common in animals aged between 1 to 3 years (p < 0.05). In order to evaluate potential associations between the presence of anti-N. caninum and anti-T. gondii antibodies and Leishmania infection, 240 dogs were selected (120 positive and 120 negative for Leishmania spp.), based on serological and parasitological diagnoses. No association was found between Leishmania spp. and the coccidian parasites (p > 0.05). The results confirm the exposure of dogs to these parasites in the State of Piauí.


2010 ◽  
Vol 19 (4) ◽  
pp. 254-255 ◽  
Author(s):  
Lívia Saab Muraro ◽  
João Garcia Caramori Júnior ◽  
Maria Regina Reis Amendoeira ◽  
Joyce Alves Pereira ◽  
João Xavier de Oliveira Filho ◽  
...  

This report aimed to assess the seroprevalence of Toxoplasma gondii infection in 708 swine matrices in Nova Mutum and Diamantino in the state of Mato Grosso, Central-West Brazil. Serum samples were examined by indirect fluorescent antibody test (IFAT). It was found a seroprevalence of 12.8%, considering titers >64. Therefore, the data reinforce the need for appropriate management of swine raising to minimize the risk of infection of pigs with T. gondii.


2015 ◽  
Vol 24 (4) ◽  
pp. 459-463 ◽  
Author(s):  
Arthur Willian de Lima Brasil ◽  
Roberta Nunes Parentoni ◽  
Thais Ferreira Feitosa ◽  
Camila de Sousa Bezerra ◽  
Vinicius Longo Ribeiro Vilela ◽  
...  

Abstract The aims of this survey were to determine the frequency of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and to identify the risk factors associated with seropositivity among buffaloes in the state of Paraíba, Brazil. This survey included 136 buffaloes belonging to 14 herds. To detect anti-T. gondii and anti-N. caninum antibodies, the indirect fluorescent antibody test (IFAT) was used. Among the 136 samples analyzed, 17 (12.5%) were positive for anti-T. gondii antibodies with titers ranging from 64 to 1,024, and 26 (19.1%) for anti-N. caninum with titers from 200 to 1,600. Animals seropositive for both T. gondii and N. caninum were found in 10 of the 14 herds (71.4%). Semi-intensive management systems (odds ratio = 2.99) and presence of pigs (odds ratio = 4.33) were identified as risk factors for T. gondii and N. caninum, respectively. It can be suggested that T. gondii and N. caninum are widespread in buffaloes in Paraíba, and that additional surveys are needed in order to ascertain the importance of these agents for this species and for pigs, and the influence of the farming type on occurrences of seropositive animals.


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