Swine cysticercosis and associated risk factors in non-technified pig breeding in semi-arid region of Sergipe state, Brazil.

The aim of this study was to determine the frequency of Taenia solium anti-metacestode antibodies in slaughtered pigs in a semi-arid region of the "Alto Sertão" of Sergipe state, Brazil, and verify the risk factors associated with swine cysticercosis. For this, 230 samples of swine blood from two slaughterhouses were collected and analyzed by indirect ELISA. The pigs came from five non-technical properties in the semi-arid region of the Alto Sertão of Sergipe state. Searches for cysts in the skeletal muscles of the pigs were performed during slaughter. In addition, an epidemiological questionnaire was applied to the pigs' original properties to determine risk factors. Besides that, the official health services database was evaluated for confirmed cases of neurocysticercosis and taeniasis in humans in the last 5 years, living in the studied region. Seropositivity in pigs was 12.6%, with no significant difference between males and females. No cysts were found in the carcasses of the slaughtered pigs. A positive association was found for properties that discharge domestic sewage into the environment, in river or streams, increasing the risk of positivity by 5.72 times. When analyzing the database of official agencies, there were no records of cases of neurocysticercosis or taeniasis in the resident population in the last 5 years. However, there were frequent cases of idiopathic epilepsy. The results demonstrate that study area is endemic for swine cysticercosis and serves as a warning of the possibility of the occurrence of the taeniasis-cysticercosis complex.

Validation of real-time PCR technique for detection of Mycobacterium bovis and Brucella abortus in bovine raw milk.

Brucellosis and tuberculosis are diseases of great economic impact in cattle herds and are controlled by governmental programs in many countries. The validation of a diagnostic technique is fundamental for its application in official control programs of these diseases. The aim of the present study was to validate a polymerase chain reaction in real time (qPCR) for detection of Mycobacterium bovis and Brucella abortus in samples of artificially contaminated raw milk. The technique was evaluated using tests of analytical sensitivity and specificity, repeatability, internal reproducibility, and robustness. Initially, five DNA extraction methodologies were tested, and the DNeasy Mericon Food Kit-Qiagen and the Maxwell® 16 Tissue DNA Purification Kit-Promega presented the best analytical specificity of all the commercial kits tested and were used exclusively in subsequent tests. The lowest limits of detection obtained in the qPCR were 2.3 pg for M. bovis DNA and 20.7 fg for B. abortus DNA. The repeatability and reproducibility associated with the robustness indicate that the evaluated methods are applicable as rapid tools for the official in vivo diagnosis of bovine tuberculosis and brucellosis in raw milk from dairy herds in Brazil.

Inhibition of Escherichia coli invasion into bovine mammary epithelial cells previously infected by Mycobacterium avium subsp. paratuberculosis.

Background: The coinfection process of Escherichia coli, an etiological agent of clinical mastitis and Mycobacterium avium subsp. paratuberculosis (MAP), a non-mastitic etiological agent in the bovine mammary gland is not fully known.Objective: Verify the ability of MAP to interfere with the invasion and translocation of E. coli in bovine mammary epithelial cell line (MAC-T).Methods: For the invasion assay, MAC-T cells were challenged with MAP K10 for 2 h and then challenged with E. coli for 10, 30 and 120 min. For the translocation assay, the trans well plates were used and the challenge sequence was repeated as previously described. The amount of E. coli in the assays was determined by counting colony forming units (CFU) in Luria-Bertani medium. Quantitative real-time PCR was used to quantify MAP in MAC-T cells. To verify the viability of the MAC-T cells, the MTT assay was performed. MAP culture supernatant was also evaluated at different percentages for E. coli growth.Results: Previous MAP infection in MAC-T cells inhibited E. coli invasion in 10, 30 and 120 min. No significant interference of MAP in the translocation of E. coli from the apical-basal direction was verified. Quantity of MAP DNA inside the MAC-T cells was statistically similar. Neither reduction in MAC-T cells viability was detected during the experiment nor MAP-released factor in the supernatant inhibited E. coli invasion.Conclusion: These findings suggest that MAP-positive cows could be more resistant to E. coli infection, but when infected, could rapidly translocate E. coli to the subepithelial region.