Prevalência de Clostridios sulfito redutores e Clostridium perfringens na mucosa intestinal de frangos de corte
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Data
2014-06-27
Autores
Beraldo Massoli, Mariana Casteleti [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O gênero Clostridium é amplamente distribuído na natureza, está presente no solo e conteúdo intestinal dos animais, produzem toxinas que são capazes de provocar doenças e intoxicações. O Clostridium perfringens está envolvido com prejuízos na avicultura, pois a doença (Enterite Necrotica) na maioria das vezes é subclínica, e o produtor só observa na hora do abate que a ave não ganhou o peso esperado. O controle dessa e de outras enfermidades e agentes patogênicos na avicultura de corte tem grande importância uma vez que o Brasil está entre os principais países exportadores de carne. O objetivo deste trabalho primeiramente foi pesquisar Clostridium perfringens em frangos de corte sadios, provenientes de dois grandes frigoríficos com Inspeção federal, um no Estado de São Paulo e outro no Estado de Minas Gerais. Foram obtidos 300 raspados intestinais, os quais foram semeados em Agar SPS e incubados em anaerobiose. A identificação dos tipos A, B, C, D e E de C. perfringens foi realizada mediante o emprego da PCR multiplex por amplificação dos genes codificadores das toxinas alfa, beta, épsilon e iota do mesmo. A partir das amostras positivas na PCR (37), procedeu-se o isolamento do agente. Depois de muitas tentativas, verificou-se por meio do sequenciamento da região 16S rDNA, que as colônias sulfito redutoras que estavam sendo isoladas eram Enterococcus spp, microrganismo coexistente na microbiota intestinal de frango de corte, que compete com o C. perfringens pelo crescimento das colônias no mesmo meio de cultura, ainda que este seja seletivo e diferencial para clostrídios sulfito redutores. Dessa forma, notou-se que o crescimento de colônias de enterococos interferia muito na identificação e enumeração do C. perfringens e ainda apresentavam colônias muito semelhantes às de C. perfringens nos oito meios de cultura testados. Visto isto, foram feitos alguns testes para avaliar o comportamento dos ...
Clostridium genus is widely distributed in nature and is present in soil and intestinal contents of animals, produce toxins which are capable of causing diseases and intoxication. Clostridium perfringens is involved with losses in the poultry industry, because the disease is most often subclinical, and producer only observed at the time of slaughter the bird has not gained the expected weight. The control of this and other diseases and pathogens in poultry production is very important since Brazil is among the major meat exporting countries. The aim of this study was to evaluate the presence of Clostridium perfringens in poultry coming from two large slaughter-house, one from São Paulo state and another from Minas Gerais. Were obtained 300 intestinal scraped and cecal contents, which were sown onto SPS agar and incubated anaerobically. The identification of C. perfringens types A, B, C, D and E was performed by the use of multiplex PCR. After identification of the presence of C. perfringens in 37 of 300 samples by amplification of the cpa gene, alpha toxin encoder, the isolation of pure colonies was proceeded. However, through the sequencing of 16S rDNA region, it was also found the presence of Enterococcus spp. that lives in the intestinal microbiota of poultry and disputes in growth on the same culture medium, even been selective and differential for clostridia. Thus, it was observed that colony morphology of enterococci is very similar to C. perfringens in this medium what make isolation of pure colonies difficult. Having on mind the difficulty of C. perfringens isolation under these circumstances, the PCR is an rapid and secure alternative for detection of C. perfringens and its different types, being effective for diagnostics
Clostridium genus is widely distributed in nature and is present in soil and intestinal contents of animals, produce toxins which are capable of causing diseases and intoxication. Clostridium perfringens is involved with losses in the poultry industry, because the disease is most often subclinical, and producer only observed at the time of slaughter the bird has not gained the expected weight. The control of this and other diseases and pathogens in poultry production is very important since Brazil is among the major meat exporting countries. The aim of this study was to evaluate the presence of Clostridium perfringens in poultry coming from two large slaughter-house, one from São Paulo state and another from Minas Gerais. Were obtained 300 intestinal scraped and cecal contents, which were sown onto SPS agar and incubated anaerobically. The identification of C. perfringens types A, B, C, D and E was performed by the use of multiplex PCR. After identification of the presence of C. perfringens in 37 of 300 samples by amplification of the cpa gene, alpha toxin encoder, the isolation of pure colonies was proceeded. However, through the sequencing of 16S rDNA region, it was also found the presence of Enterococcus spp. that lives in the intestinal microbiota of poultry and disputes in growth on the same culture medium, even been selective and differential for clostridia. Thus, it was observed that colony morphology of enterococci is very similar to C. perfringens in this medium what make isolation of pure colonies difficult. Having on mind the difficulty of C. perfringens isolation under these circumstances, the PCR is an rapid and secure alternative for detection of C. perfringens and its different types, being effective for diagnostics
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Palavras-chave
Ave - Criação, Frango de corte, Microbiota, Reação em cadeia de polimerase, Clostridium perfringens, Enterococcus, Frigorificos, Cold storage
Como citar
BERALDO MASSOLI, Mariana Casteleti. Prevalência de Clostridios sulfito redutores e Clostridium perfringens na mucosa intestinal de frangos de corte. 2014. xii, 46 p. Tese (doutorado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Ciências Agrárias e Veterinárias, 2014.