Zoonoses ocupacionais: inquérito soro-epidemiológico em estudantes de Medicina Veterinária e análise de risco para leptospirose, brucelose e toxoplasmose

Pós-graduação em Doenças Tropicais - FMB

Aspectos epidemiológicos, clínicos e avaliação dos métodos diagnósticos nas fases de evolução da brucelose em ovinos inoculados experimentalmente com Brucella ovis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Epidemiologia molecular aplicada ao estudo de estirpes de Staphylococcus aureus na produção de queijo tipo Minas Frescal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detecção molecular de Paracoccidioides brasiliensis em amostras de tecido e sangue periférico de pacientes portadores de Pacoccidioidomicose: correlação com perfis histopatológico, sorológicos e produção de TNF-α

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da acurácia do teste imunoenzimático e de sua contribuição no seguimento de pacientes com paracoccidioidomicose em tratamento e no diagnóstico de doença reativada

Pós-graduação em Doenças Tropicais - FMB

Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina

Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE.

Construção de um indicador qualitativo (status de saúde) para avaliação da qualidade de saúde animal de propriedades rurais

Aiming at developing a tool for assessing the magnitude of farm animal health, an indicator named herd “animal health status” was built. To illustrate the construction of the indicator, serum samples were taken from 923 goats on 17 goat farms in the state of Sao Paulo, Brazil, where three diseases were evaluated along with their impact percentages: caprine arthritis encephalitis, toxoplasmosis and neosporose – all of an infectious and transmissible nature and the cause of considerable economic losses. The mathematical rationale underlying the building of the “animal health status” indicator ranks properties on a 0 - 100% scale in terms of disease positivity in the herd, with the lowest indicator value indicating the highest cumulative disease frequency (0% = all animals tested positive for the three diseases, 100% = all animals tested negative for all diseases). Anti-T. gondii, anti-N. caninum and anti-CAEV antibodies were tested using indirect immunofluorescence reaction (IIFR ≥ 16), Neospora agglutination test (NAT ≥ 25) and agarose gel immunodiffusion (AGID positive or negative), respectively. The animal health status of the farms ranged from 32.38% to 96.40% according to disease positivity. The animal health status value will have a direct reflection on the production chain both on and off the farm, providing not only great market advantages, due to how the expression of animal health will add value to livestock farms, as well as on the consumer who can check the quality of animal health at the beginning of the production chain.

Assessment of the risk of raw milk consumption related to staphylococcal food poisoning

This study aimed to identify the risks of staphylococcal food poisoning due to the consumption of raw milk. Fifty-one farms in Londrina (PR) and 50 in Pelotas (RS) were analyzed, to determine the population of coagulase-positive staphylococci (UFC/ mL), as well as to verify the ability of producing Staphylococcal Enterotoxin A (SEA) by immunodifusion (OSP), the presence of the gene for the production of SEA (PCR) in the cultures, and the research of enterotoxin (SEA to SEE) in milk samples using ELISA commercial kit. Considering the 101 farms analyzed, 19 (18.8%) presented coagulase-positive staphylococci count above 105 UFC/mL. For the evaluation of the enterotoxigenic ability (SEA) by the OSP technique, six cultures coagulase-positive (5.5%) were positive to the test and identified as S. aureus. From the coagualse-negative sample, one (5.5%) was OSP positive. For the evaluation of the presence of the gene for EEA synthesis, 51 cultures of staphylococci were tested. From this total, 14 (27.45%) presented the gene, and from that, only 5 (9.81%) cultures were capable of expressing it in the technique of the OSP. The morphologic characteristic of the evaluated cultures that had enterotoxigenic capacity, from the 14 (33,3%) cultures that presented the gene for EEA production, 05 (11.9%) were characterized as typical cultures of S.aureus in Baird Parker agar. All the 12 milk samples studied for the presence of EEA to EEE in milk were negative. Thus, it can be concluded that there is extensive contamination of raw milk for staphylococci coagulase, however, most of the isolated strains were not enterotoxigenic or did not express such a characteristic. Only 9.81% of the tested colonies expressed the gene and effectively produced SEA. None of the samples had sufficient counts to produce detectable amounts of SEA. The milk samples did not present risk to cause staphylococcal food poisoning if consumed in natura until the collection moment.