Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.(AU)

Enterococcus Spp. Survival Through the Use of Standard Protocol for Clostridium Sp. Isolation

The genera Clostridium and Enterococcus are very different from each other, both morphologically and physiologically. Due to the high resistance by the sporulation capacity of Clostridium species, the thermal shock is a characteristic tool used for the isolation and identification of these microorganisms, this way, it would eliminate any other bacteria that did not present spores. The objective of this work is to show that Enterococcus sp. resist the temperature treatment and grow in culture media used for the isolation of Clostridium sp. For this, the present study initially attempted to identify reducing sulfite clostridia in poultry products, through the use of specific culture media and heat shock treatment. However, the PCR did not detect the presence of Clostridium sp. Then, sequencing of the 16S rDNA region was performed, which showed that the reducing sulfite colonies that were being isolated were, actually, Enterococcus spp. With this, some tests were carried out using different temperature and time combinations in the thermal shock, as well as the use of five different selective and differential culture media, in an attempt to eliminate any contaminants, but all without success, because these bacteria resisted to all modification. Therefore, the standard protocol for the isolation of bacteria of the genus Clostridium does not eliminate Enterococcus, which can lead to failures in the quantification and qualification of sulfite reducing microorganisms, a fact that can significantly affect food safety and animal health.

Microbiological profile of donor corneas stored for tectonic transplantation purposes in rabbits / Microbiologia de amostras de bancos de córneas destinadas a transplantes, em coelhos

This study aimed to evaluate the microbiota of donor rabbit corneas stored for tectonic transplantation purposes. Swabs from both corneas of 20 rabbits were carefully collected and submitted to microorganism isolation and identification. After this first swab collection, rabbits were euthanized for reasons other than this project and the eyes were enucleated. The corneas were collected and stored to compose the cornea tissue bank. Corneas were stored in a 0.3% tobramycin solution at -20ºC. After 30 days, the corneas were thawed at room temperature and removed from the antibiotic. New swabs were obtained from the corneas and submitted to microorganism isolation and identification. Gram positive organisms were predominant in the rabbit corneal flora before storage and the Staphylococcus sp. was the most common microorganism isolated from those samples. No growth was observed on the samples collected after storage. The methods used for collection and storage of the corneas were efficient to constitute a sterile donor corneal tissue bank.

Analisaram-se córneas armazenadas para transplantes tectônicos usando-se suabes coletados de 20 coelhos, visando ao isolamento e à identificação de microrganismos. Após a coleta das amostras, os coelhos foram submetidos à eutanásia, por razões alheias ao estudo, e enucleados. As córneas foram coletadas e armazenadas a fim de se constituir o banco de córneas. O armazenamento deu-se em solução de tobramicina 0,3% a -20ºC, por 30 dias. Após esse período, as córneas foram descongeladas à temperatura ambiente e removidas da solução de antibiótico. Novos suabes foram coletados e submetidos ao isolamento e à identificação dos microrganismos. A flora corneal mostrou-se predominantemente composta por bactérias Gram positivas, sendo o Staphylococcus sp. o mais identificado. Não se verificou crescimento de colônias bacterianas ou fúngicas nas amostras após o armazenamento. Considerando-se a maneira como a pesquisa foi concebida e as injunções do meio em que ela foi realizada, há como admitir, pela ausência de crescimento microbiano nas amostras armazenadas, que a técnica de armazenamento empregada é segura para a estocagem de córneas destinadas a transplantes.

Presença de Clostridium perfringens em frangos de corte provenientes de aviários da região de Ribeirão Preto SP / Presence of clostridium perfringens in broiler from chicken farms in Ribeirão Preto-SP

Visando estudar a presença de Clostridium perfringens em frangos de cortes provenientes de aviários da região de Ribeirão Preto-SP, amostras de conteúdo cecal de aves foram pesquisadas quanto a presença desse microrganismo. As amostras foram semeadas em meios seletivos para clostrídios e incubadas anaerobica­mente. As culturas foram identificadas e caracterizadas pelo método de Gram e séries bioquímicas. Posteriormente realizou-se teste de inoculação em camundongos para confirmar patogenicidade. De um total de 560 amostras coletadas, 374(66,78%) foram positivas para o gênero Clostridium, sendo 94 (16,78 %) Clostridium perfringens. Verificou–se que 19 (3,4%) amostras foram positivas para outras espécies de clostrídios patogênicos como Clostridium chauvoei (3 - 0,54 %), Clostridium sordelli (9 - 1,61%), Clostridium bifermentans (3 - 0,54%), Clostridium septicum (3 - 0,54%) e Clostridium tetani (1 - 0,18%).(AU)

The aim of the present study was to detect Clostridium perfringens in broiler chickens from intensive poultry farms in Ribeirão Preto-SP. We collected 516 samples of caecal content that were cultivated onto selective medium for Clostridium and incubated under anaerobic conditions. Gram smears were carried out from the positive samples and identified using biochemical tests. Mice inoculation test was performed in order to confirm pathogenicity. The results showed that 66.78% of the samples were positive for Clostridia, of which 16.78% were C. perfringens. It was confirmed that 19 (3.4%) of the samples were positive to other Clostridia pathogenic species such as Clostridium chauvoei (3 - 0.54%), Clostridium sordelli (9 - 1.61%), Clostridium bifermentans (3 - 0.54%), Clostridium septicum (3 - 0.54%) and Clostridium tetani (1 - 0.18%).(AU)

Microbiological profile of donor corneas stored for tectonic transplantation purposes in rabbits / Microbiologia de amostras de bancos de córneas destinadas a transplantes, em coelhos

This study aimed to evaluate the microbiota of donor rabbit corneas stored for tectonic transplantation purposes. Swabs from both corneas of 20 rabbits were carefully collected and submitted to microorganism isolation and identification. After this first swab collection, rabbits were euthanized for reasons other than this project and the eyes were enucleated. The corneas were collected and stored to compose the cornea tissue bank. Corneas were stored in a 0.3% tobramycin solution at -20ºC. After 30 days, the corneas were thawed at room temperature and removed from the antibiotic. New swabs were obtained from the corneas and submitted to microorganism isolation and identification. Gram positive organisms were predominant in the rabbit corneal flora before storage and the Staphylococcus sp. was the most common microorganism isolated from those samples. No growth was observed on the samples collected after storage. The methods used for collection and storage of the corneas were efficient to constitute a sterile donor corneal tissue bank.(AU)

Analisaram-se córneas armazenadas para transplantes tectônicos usando-se suabes coletados de 20 coelhos, visando ao isolamento e à identificação de microrganismos. Após a coleta das amostras, os coelhos foram submetidos à eutanásia, por razões alheias ao estudo, e enucleados. As córneas foram coletadas e armazenadas a fim de se constituir o banco de córneas. O armazenamento deu-se em solução de tobramicina 0,3% a -20ºC, por 30 dias. Após esse período, as córneas foram descongeladas à temperatura ambiente e removidas da solução de antibiótico. Novos suabes foram coletados e submetidos ao isolamento e à identificação dos microrganismos. A flora corneal mostrou-se predominantemente composta por bactérias Gram positivas, sendo o Staphylococcus sp. o mais identificado. Não se verificou crescimento de colônias bacterianas ou fúngicas nas amostras após o armazenamento. Considerando-se a maneira como a pesquisa foi concebida e as injunções do meio em que ela foi realizada, há como admitir, pela ausência de crescimento microbiano nas amostras armazenadas, que a técnica de armazenamento empregada é segura para a estocagem de córneas destinadas a transplantes.(AU)

Presença de Clostridium perfringens em frangos de corte provenientes de aviários da região de Ribeirão Preto – SP / Presence of clostridium perfringens in broiler from chicken farms in Ribeirão Preto-SP

Visando estudar a presença de Clostridium perfringens em frangos de cortes provenientes de aviários da região de Ribeirão Preto-SP, amostras de conteúdo cecal de aves foram pesquisadas quanto a presença desse microrganismo. As amostras foram semeadas em meios seletivos para clostrídios e incubadas anaerobica­mente. As culturas foram identificadas e caracterizadas pelo método de Gram e séries bioquímicas. Posteriormente realizou-se teste de inoculação em camundongos para confirmar patogenicidade. De um total de 560 amostras coletadas, 374(66,78%) foram positivas para o gênero Clostridium, sendo 94 (16,78 %) Clostridium perfringens. Verificou–se que 19 (3,4%) amostras foram positivas para outras espécies de clostrídios patogênicos como Clostridium chauvoei (3 - 0,54 %), Clostridium sordelli (9 - 1,61%), Clostridium bifermentans (3 - 0,54%), Clostridium septicum (3 - 0,54%) e Clostridium tetani (1 - 0,18%).

The aim of the present study was to detect Clostridium perfringens in broiler chickens from intensive poultry farms in Ribeirão Preto-SP. We collected 516 samples of caecal content that were cultivated onto selective medium for Clostridium and incubated under anaerobic conditions. Gram smears were carried out from the positive samples and identified using biochemical tests. Mice inoculation test was performed in order to confirm pathogenicity. The results showed that 66.78% of the samples were positive for Clostridia, of which 16.78% were C. perfringens. It was confirmed that 19 (3.4%) of the samples were positive to other Clostridia pathogenic species such as Clostridium chauvoei (3 - 0.54%), Clostridium sordelli (9 - 1.61%), Clostridium bifermentans (3 - 0.54%), Clostridium septicum (3 - 0.54%) and Clostridium tetani (1 - 0.18%).