Orientador: Fernado Antônio de Ávila / Banca: José Moacir Marin / Banca: Luiz Florencio Franco Margatho / Resumo: O objetivo desse estudo foi isolar cepas de estafilococos resistentes a meticilina (MRS) de humanos e cães e verificar os seus respectivos perfis de suscetibilidade a diversos antimicrobianos em um hospital veterinário de ensino no Brasil. Foram analisados espécimes de 50 pessoas e de 50 cães para isolamento de estafilococos que foram identificados através de características morfotintoral, reações bioquímicas e resistência a antimicrobianos. Foi verificado o perfil de suscetibilidade a oxacilina, penicilina, ciprofloxacina, gentamicina, clindamicina, eritromicina, sulfametoxazol+trimetoprim e vancomicina. Não foram isolados Staphylococcus aureus resistentes a meticilina (MRSA) e Staphylococcus intermedius resistentes a meticilina (MRSI). Foram isoladas duas cepas de estafilococos coagulase-negativas resistentes a meticilina (MRCoNS) de cães (4%) e 18 cepas de MRCoNS de seres humanos (36%). Todas as cepas de estafilococos isoladas foram sensíveis a vancomicina. O isolamento de MRCoNS merece atenção devido à hipótese que estes podem transferir o gene mecA para estafilococos coagulase-positivos. A ausência de cepas de MRSA resistentes a vancomicina é importante por ser este um antibiótico que pode ser usado como alternativa extrema. Programas de vigilância de MRS devem ser estimulados em unidades de saúde veterinárias. / Abstract: The aim of this study was to isolate methicillin-resistant staphylococci (MRS) strains from humans and dogs and verify their susceptibility profile to several antimicrobials in a veterinary teaching hospital in Brazil. Specimens from 50 people and 50 dogs were analyzed to the isolation of staphylococci. They were identified trough gram staining, biochemical reactions and resistance to antimicrobials. Susceptibility profile to oxacillin, penicillin, ciprofloxacin, gentamicin, clindamycin, erythromycin, trimethoprim + sulfamethoxazole and vancomycin was verified. It was not isolated any strain of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus intermedius (MRSI). Two strains of methicillin-resistant coagulasenegative staphylococci (MRCoNS) were isolated from dogs (4%) and 18 strains were isolated from humans (36%). MRCoNS isolation is important because there is the hypothesis that mecA transferring from MRCoNS to coagulase-positive staphylococci could occur. The absence of MRSA strains resistant to vancomycin is important because this antimicrobial could be used as an important alternative against MRSA infections. Surveillance programs aiming MRS should be stimulated in veterinary health units. / Mestre

Orientador: Maria de Lourdes Ribeiro de Souza da Cunha / Banca: Maria de Lourdes Ribeiro de Souza da Cunha / Banca: Carlos Alberto Magalhães Lopes / Banca: Elisa Yoko Hirooka / Resumo: Não disponível. / Abstract: The Staphylococcus spp. are often found in the hospital ambient, being associated with a large infection variety. Several factors of virulence are responsible for the Staphylococcus pathology, and the enter toxin originated from Staphylococcus and the TTST -1 (toxin of toxic shock syndrome) are accentuated among them. In this study, ninety Staphylococcus aureus patterns and ninety negative-coagulation of Staphylococcus patterns, isolated from different clinical materials, were examined by the PCR technique for the gene's research which encode the toxins originated from Staphylococcus A (sea), B (seb), C (sec-1 ), D (sed) and TTST -1 (tst). The patterns that appeared positives for the presence of one or more genes were tested by the production capacity of the respective toxins by the RT -PCR technique. In relation to ECN species, the S. epidermidis was isolated to a large frequency, corresponding to 71, 1% of the total of ECN examined patterns, as well as S. warneri (6, 7%), S. haemolyticus (5, 6%), S. hominis (5, 6%), S.lugdunensis (5, 6%)J S. simulans (3, 3%), S. saprophyticus (1, 1 % ) and S. Xylosus (1, 1 %). The results revealed a total of 108 toxicological patterns, corresponding to 59 (54, 6%) S. aureus patterns and 49 (45, 4%) ECN patterns with one or more toxicological genes. Among the ECN studied species, only the S. simulans type didn't present genes that encode toxins originated from Staphylococcus. TheS.aureus showed toxicological genes for ali the classes of examined toxins, while the ECN presented genes to the A (sea), B (seb) C (sec-1) and TSST -1 (tst) toxins. The sec-1 gene was the most found as well in S.aureus patterns as in ECN, while the seb gene was the most frequent in S. aureus. The detection of the toxin production by the RT -PCR technique revealed43 positive patterns, corresponding to 37 (86, 0% ) S.aureus patterns producers of EEA, EEB, EEC, EED, and/or TSST -1 and six (14, 0%) ECN patterns producers of EEA and EEC. / Mestre

Orientador: Antonio Nader Filho / Banca: Angela Cleusa de Fatima Banzatto de Carvalho / Banca: Luiz Francisco Zafalon / Resumo: O leite é um alimento altamente nutritivo e excelente substrato para a multiplicação de microrganismos. Os estafilococos estão entre os principais contaminantes do leite, seja em decorrência da mastite ou de falhas de higienização. A contaminação do leite pode favorecer a adesão bacteriana sobre superfícies com a formação de biofilmes, cujos fragmentos podem se desprender e contaminar o produto durante o processo de beneficiamento, o que representa um risco à saúde do consumidor. Tendo isto em foco, objetivou-se o presente estudo, em uma micro-usina do Estado de São Paulo, a fim de investigar a presença e formação de biofilme por Staphylococcus spp, antes e após o processo de higienização. Colheu-se o total de 60 amostras por meio de suabes, antes e após o processo de higienização, das superfícies do tanque de recepção, do tanque de estocagem de leite cru, da tubulação de saída do pasteurizador, do tanque de estocagem de leite pasteurizado, e da tubulação da máquina de envase. Foram colhidas, ainda, amostras de leite no tanque de recepção, no tanque de estocagem de leite cru, na tubulação de saída do pasteurizador e amostras de leite envasado, assim como das embalagens plásticas vedadas e vazias, utilizadas para o envase do leite pasteurizado. Dentre 41 estirpes de Staphylococcus spp isoladas, 16 (39,0%) mostraram-se positivas na prova da coagulase, enquanto que 25 (61,0%) foram negativas. Por meio de análise genotípica, com a técnica de Reação em Cadeia da Polimerase (PCR), pode-se observar que, dentre as 16 estirpes coagulase positivas, quatro (25%) apresentavam o gene icaA e 16 (100%) possuíam o gene icaD. Dentre as 25 estirpes coagulase negativas, 11 (44%) possuíam o gene icaA e 25 (100%) possuíam o gene icaD. Visualizou-se, por meio da microscopia eletrônica de varredura... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Milk is a highly nutritious food and excellent substrate for the multiplication of microorganisms. Staphylococci are one of the major contaminants of milk whether due to mastitis or hygiene failures in cleaning. Milk contamination may encourage bacterial adherence on surfaces with the formation of biofilms, whose fragments can detach and contaminate the product during beneficial processing, which represents a health risk to the consumers. With this in focus as the objective of this present study, to investigate the presence and formation of biofilm of Staphylococcus spp before and after the cleaning process in a micro-dairy plant in São Paulo State. A total of 60 swab samples were collected before and after the cleaning process from the reception tank surfaces, the raw milk storage tank storage, the pasteurizer outlet pipe, the pasteurized milk storage tank, and the filling machine. Further samples were collected of the milk in the receiving tank, the raw milk storage tank, from the pasteurizer outlet pipe, and packaged milk, as well as the empty sealed plastic packaging used for packaging the pasteurized milk. Of the 41 strains of isolated Staphylococcus spp, 16 (39.0%) indicated positive in the coagulase test, while 25 (61.0%) were negative. Through genetic analysis, using the Polymerase Chain Reaction (PCR) technique, it was observed that within the 16 strains of coagulase-positive, four (25%) presented the gene icaA and 16 (100%) had the gene icaD. Of the 25 strains of coagulase-negative, 11 (44%) had the gene icaA and 25 (100%) had the gene icaD. It was seen using a scanning electron microscope the start of bacteria adhesion and the formation of biofilm for all the isolated strains. From the obtained results it was possible to see evidence to the potential risk to the health of the consumer represented... (Complete abstract click electronic access below) / Mestre