This study genetically Toxoplasma gondii isolates obtained from pigs intended for human consumption in northeastern Brazil; multilocus PCR-RFLP and sequencing techniques were utilized. Bioassays were conducted using the brain and tongue of 20 pig heads purchased at butcher shops in the city of Ilheus, Bahia, Brazil. Overall, 11 T. gondii isolates designated TgPgBr06-16 were identified. Application of multilocus PCR-RFLP with seven molecular markers (SAG1, SAG2, SAG3, BTUB, C22-8, PK1 and Apico) identified six different genotypes. Isolates TgPgBr 06, 08, 11, 12, 14 and 15 were indistinguishable by this technique, forming a single genotype; the remaining isolates were characterized as distinct genotypes. However, when five genetic markers (SAG1, SAG2, SAG3, BTUB and c22-8) were employed in multilocus PCR-sequencing, all eleven strains of T. gondii were shown to be different. All isolates differed from Type I, II and III clonal genotypes using both genotyping techniques. These results demonstrate that the multilocus PCR-RFLP assay underestimated the true diversity of the T. gondii population in this study. Thus, DNA sequencing is the preferred technique to infer the genetic diversity and population structure of T. gondii strains from Brazil. Moreover, it is necessary to develop new molecular markers to group and characterize atypical T. gondii isolates from South America.

ABSTRACT. The objective of this study was to verify whether Toxoplasma gondii is excreted in the milk of naturally infected sheep. In order to accomplish this, 275 lactating ewes were used; these were bred extensively in 17 estates distributed across nine cities. Polymerase chain reaction amplification was used to detect T. gondii DNA in milk samples, and the indirect immunofluorescence test was employed for the detection of anti-T. gondii IgG antibodies in the sera, with a cut-off value of 1:64. It was possible to verify the presence of the parasite DNA in 6.5% (18/275) of the studied animals. Anti-T. gondii antibodies were present in 41.5% of the animals studied (114/275). There was no correlation between parasite excretion in milk and the presence of IgG in 38.9% of the studied animals (7/18). The high seropositivity and the presence of parasite DNA in the milk led to the conclusion that T. gondii infection is present in the sheep population in southern and southwestern Bahia, and that there is a risk of the human population becoming infected due to the consumption of raw, in natura milk.

This study aimed to compare the sensitivity of five diagnostic methods commonly used for the detection of Toxoplasma gondii in tissues of naturally infected pigs. We purchased 20 heads of pigs in butcher shops in the city of Ilhéus, Bahia. Brain and tongue fragments were taken from each animal for the performance of PCR against T. gondii. The rest of these two tissues were processed and inoculated into three mice. These rodents were observed for 42 days and euthanized. We prepared slides with brain and lungs of each mouse for the visualization of T. gondii. From the tissues of mice, we carried out polymerase chain reaction (PCR), histopathology, and immunohistochemistry in an attempt to identify the parasite. The PCR direct from the tissue of pigs showed 10% (2/20) of positive samples, all from the brain. PCR in tissue from mice found that 55% (11/20) of pigs were positive: 55% (11/20) and 45% (9/20) for brains and tongues, respectively. Mice were inoculated with material obtained from the samples and examined by various methods for resulting Toxoplasma infection (bioassay). Cyst detection in bioassay mice identified 25% (5/20) and immunohistochemistry 30% (6/20) of the samples pigs as positive for T. gondii. Histopathology of mice tissue could not detect parasite; only suggestive pathological changes such as inflammation with foci of necrosis were seen. The results indicated PCR of mice tissue as the most sensitive among those tested.

In this study, we aimed to determine the prevalence of Toxoplasma gondii antibodies and identify risk factors associated with this infection in sheep from the southern region of Bahia state. Between February and December 2010, 795 sheep from 31 farms located in nine municipalities were tested. We found seroprevalence of 30.2% (240/795), with titers of 64 (38.3%), 256 (34.2%), 1,024 (18.3%), and 4,096 (9.2%) by Indirect Fluorescent Antibody Test (IFAT). Seropositive sheep were detected in all farms sampled. Univariate statistical analysis detected association between T. gondii seropositivity and the variables age, use of fresh food mainly, water source, stocking rate, production system, presence and number of cats on the farm, and transit of cats (p < 0.05). In the logistic regression model, transit of cats (p = 0.001), production system (p = 0.007), and age (p = 0.027) were identified as risk factors associated with T. gondii infection.Keywords: Epidemiology, ovine, toxoplasmosis, zoonosis, risk factors. ResumoObjetivou-se com este estudo determinar a prevalência de anticorpos anti-Toxoplasma gondii e identificar os fatores de risco associados à infecção em ovinos no sudeste do Estado da Bahia. De fevereiro a dezembro de 2010, 795 ovinos de 31 propriedades localizadas em nove municípios foram analisados. A soroprevalência foi de 30,2% (240/795), com títulos de 64 (38,3%), 256 (34,2%), 1.024 (18,3%) e 4.096 (9,2%) pela Reação de Imunoflorescência Indireta (RIFI). Ovinos positivos foram detectados em todas as fazendas estudadas. Na análise estatística univariada detectou-se associação entre a soropositividade e idade, uso de alimentação fresca, fonte de água, sistema de produção, presença e número de gatos na fazenda e o transito de gatos (p < 0,05). No modelo de regressão logística, transito de gatos (p = 0,001), sistema de produção (p = 0,007) e idade (p = 0,027) foram identificados como fatores de risco associados à infecção por T. gondii.

ResumoRealizou-se um levantamento sorológico de Brucella ovis em ovinos do Estado de Sergipe, com o objetivo de determinar a positividade e a distribuição da infecção em propriedades rurais e analisar os possíveis fatores associados à infecção. Foram analisadas 54 propriedades criadoras de ovinos, das quais foram colhidas 932 amostras de soro sanguíneo de animais com idade superior a seis meses, nas três regiões do Estado. Todos os soros foram examinados por Imunodifusão em Gel de Agar (IDGA). De acordo com o teste realizado, 46,30% (25/54) das propriedades apresentaram evidência sorológica de infecção por B. ovis, com uma positividade de 4,40% (41/932) dos animais. Como fatores associados à infecção por B. ovis, observaram-se a presença de tratador de ovinos (OR=2,31) e propriedades com área superior a 50 ha (OR=1,98) e como fator de proteção, a utilização de cabanha (OR=0,37). Assim, verificou-se a presença de anticorpos contra Brucella ovis nos ovinos do Estado e salienta-se a importância de estudos complementares para determinação de medidas sanitárias específicas para prevenir os rebanhos desta enfermidade. Palavras-chave:Brucelose; doença; epididimite ovina. AbstractSeroepidemiological study of Brucella ovis in sheep from Sergipe State was carried out to determine the seropositivity and infection distribution in rural properties and possible factors associated with infection. A total of 54 sheep properties were studied and 932 blood serum samples from animals older than six months were collected in the three regions of Sergipe State. All sera samples were examined by agar gel immunodiffusion (AGID). According to the serological tests, 46.30% (25/54) of the properties had serologic evidence of infection by B. ovis, with 4.40% of positive animals (41/932). Presence of sheep handler (OR=2.31) and properties with an area bigger

The aim of this study was to determine the prevalence and factors associated with the occurrence of antibodies against Neospora caninum in sheep from Sergipe, northeastern Brazil. A total of 932 sheep serum samples from 54 properties in 19 municipalities from the State of Sergipe, Brazil were collected and assayed using an indirect fluorescent antibody test (IFAT) to assess antibodies against N. caninum. A cutoff point of 1:50 was adopted and results showed that 12.45% (116/932) of sheep were serum-reactive. Based on an unconditional logistic regression, the presence of dogs on the property was associated with protection (OR= 0.323), whereas the use of exchanged or borrowed breeding males was associated with infection (OR= 22.287). These results indicate that the occurrence of antibodies against N. caninum is endemic in the State municipalities.