Detecção de Campylobacter jejuni em produtos de frango utilizando separação imunomagnética / Detection of Campylobacter jejuni in chicken products using immunomagnetic separation

Campylobacter jejuni é o principal causador de gastroenterite bacteriana aguda, e a carne de frango tem se mostrado uma importante fonte de transmissão. Este microrganismo é de difícil isolamento e os métodos convencionais muitas vezes não são eficientes, podendo levar a resultados errôneos. O objetivo deste trabalho foi desenvolver e testar a técnica de separação imunomagnética (IMS) na detecção de C. jejuni em produtos de frango. Micropartículas magnéticas ligadas a anticorpos policlonais anti-C. jejuni foram utilizadas para concentrar C. jejuni antes da semeadura em ágar. O protocolo foi comparado com o método convencional. C. jejuni foi recuperado do alimento experimentalmente contaminado por ambos os métodos, entretanto, quando foi usada a IMS, a presença de microrganismos contaminantes nos meios de cultura foi menor. C. jejuni foi isolado de 7% das amostras de alimento naturalmente contaminadas, usando IMS, e de 3% pelo método convencional. C. coli foi isolado de uma amostra pelo método convencional, mas não foi detectado pelo protocolo com IMS. A técnica de IMS pode ser usada para isolamento de C. jejuni de alimentos, oferecendo a vantagem de detectar em amostras o microrganismo cujo isolamento não é obtido por meio do método convencional.(AU)

Campylobacter jejuni is the main cause of acute bacterial gastroenteritis and chicken meat has shown to be an important source of infection. This microorganism is difficult to isolate and the conventional methods are often inefficient and may lead to erroneous results. This study aimed at developing and testing the technique of immunomagnetic separation (IMS) in the detection of C. jejuni in chicken products. Microparticles magnetically connected anti-C. jejuni polyclonal antibodies were used to concentrate C. jejuni before agar seeding. The protocol was compared with the conventional method. C. jejuni was recovered from experimentally contaminated food for both methods, however, when the IMS was used, the presence of contaminating microorganisms in the means of culture was smaller. C. jejuni was isolated from 7% of samples of food naturally contaminated, using IMS, and 3% by conventional method. C. coli was isolated from a sample by conventional method, but it was not detected by protocol with IMS. The IMS technique can be used for isolation of C. jejuni in food, offering the advantage of detecting the microorganism in samples from which the isolation is not obtained with the use of the conventional method.(AU)

Detecção de Campylobacter jejuni em produtos de frango utilizando separação imunomagnética / Detection of Campylobacter jejuni in chicken products using immunomagnetic separation

Campylobacter jejuni é o principal causador de gastroenterite bacteriana aguda, e a carne de frango tem se mostrado uma importante fonte de transmissão. Este microrganismo é de difícil isolamento e os métodos convencionais muitas vezes não são eficientes, podendo levar a resultados errôneos. O objetivo deste trabalho foi desenvolver e testar a técnica de separação imunomagnética (IMS) na detecção de C. jejuni em produtos de frango. Micropartículas magnéticas ligadas a anticorpos policlonais anti-C. jejuni foram utilizadas para concentrar C. jejuni antes da semeadura em ágar. O protocolo foi comparado com o método convencional. C. jejuni foi recuperado do alimento experimentalmente contaminado por ambos os métodos, entretanto, quando foi usada a IMS, a presença de microrganismos contaminantes nos meios de cultura foi menor. C. jejuni foi isolado de 7% das amostras de alimento naturalmente contaminadas, usando IMS, e de 3% pelo método convencional. C. coli foi isolado de uma amostra pelo método convencional, mas não foi detectado pelo protocolo com IMS. A técnica de IMS pode ser usada para isolamento de C. jejuni de alimentos, oferecendo a vantagem de detectar em amostras o microrganismo cujo isolamento não é obtido por meio do método convencional.(AU)

Campylobacter jejuni is the main cause of acute bacterial gastroenteritis and chicken meat has shown to be an important source of infection. This microorganism is difficult to isolate and the conventional methods are often inefficient and may lead to erroneous results. This study aimed at developing and testing the technique of immunomagnetic separation (IMS) in the detection of C. jejuni in chicken products. Microparticles magnetically connected anti-C. jejuni polyclonal antibodies were used to concentrate C. jejuni before agar seeding. The protocol was compared with the conventional method. C. jejuni was recovered from experimentally contaminated food for both methods, however, when the IMS was used, the presence of contaminating microorganisms in the means of culture was smaller. C. jejuni was isolated from 7% of samples of food naturally contaminated, using IMS, and 3% by conventional method. C. coli was isolated from a sample by conventional method, but it was not detected by protocol with IMS. The IMS technique can be used for isolation of C. jejuni in food, offering the advantage of detecting the microorganism in samples from which the isolation is not obtained with the use of the conventional method.(AU)

DNA profiles of Salmonella spp. isolated from chicken products and from broiler and human feces

Salmonella species have been isolated from various kinds of food and are accountable for outbreaks of foodborne diseases in humans. This study aimed at identifying the similarities between the DNA profiles of Salmonella isolated from chicken feces, chicken products, and human feces in southern Brazil. Six hundred samples were collected (200 from chicken products, 200 from broiler chicken feces, and 200 from human feces) and tested for the presence of Salmonella. Isolates proven to be Salmonella compatible by biochemical and serological tests were tested by the Polymerase Chain Reaction. Their DNA profiles were then analyzed by PFGE and rep-PCR. Salmonella was isolated from 16 out of 600 analyzed samples, with Schwarzengrund serotype presenting the highest incidence, followed by Mbandaka in chicken meat and fecal samples, and Panama in human fecal samples. Some strains isolated from chicken fecal and product samples were indistinguishable by the molecular methods used in the study, suggesting that that the contamination of the broilers on the farm can be transmitted the processed products.(AU)

DNA profiles of Salmonella spp. isolated from chicken products and from broiler and human feces

Salmonella species have been isolated from various kinds of food and are accountable for outbreaks of foodborne diseases in humans. This study aimed at identifying the similarities between the DNA profiles of Salmonella isolated from chicken feces, chicken products, and human feces in southern Brazil. Six hundred samples were collected (200 from chicken products, 200 from broiler chicken feces, and 200 from human feces) and tested for the presence of Salmonella. Isolates proven to be Salmonella compatible by biochemical and serological tests were tested by the Polymerase Chain Reaction. Their DNA profiles were then analyzed by PFGE and rep-PCR. Salmonella was isolated from 16 out of 600 analyzed samples, with Schwarzengrund serotype presenting the highest incidence, followed by Mbandaka in chicken meat and fecal samples, and Panama in human fecal samples. Some strains isolated from chicken fecal and product samples were indistinguishable by the molecular methods used in the study, suggesting that that the contamination of the broilers on the farm can be transmitted the processed products.

Ocorrência de Campylobacter em carne de frango, fezes de frango e humanas e pesquisa dos genes cdt / Occurrence of Campylobacter in poultry, meat chicken and human feces, and cdt GENES research

Foram coletadas 100 amostras de conteúdo fecal de aves de corte, 100 de produtos de frango (coxa, sobrecoxa, asa, dorso, carne moída e fígado) e 100 de fezes de humanos, e analisadas para pesquisa de Campylobacter. Realizou-se a determinação da espécie e da presença dos genes cdt, responsáveis pela codificação da toxina citoletal distensiva (CDT), através da técnica da PCR. A bactéria foi isolada de 61% das amostras de fezes de frango, 20% de produtos de frango e 3% de fezes de humanos. A maioria dos isolados foi determinada como C. jejuni . Destes, 93,5% apresentaram os genes para a toxina CDT. Apesar de a ocorrência de Campylobacter em fezes de humanos ter sido baixa, a prevalência em frangos de corte e produtos de frango foi elevada, fato que, aliado à presença dos genes cdt na maioria dos isolados, representa risco potencial para os consumidores. Esses resultados são indicativos da necessidade de medidas preventivas no sistema de produção e de boas práticas de fabricação na indústria, de forma a minimizar a contaminação dos produtos e diminuir o risco para os consumidores.

A hundred chicken fecal samples, a hundred samples of retail poultry products and a hundred samples of human feces were collected and tested for the presence of Campylobacter. The species identification and the analysis for the presence of cdt genes, responsible for encoding the cytolethal distending toxin, were performed by PCR. Campylobacter was found in 61% of the chicken fecal samples, in 20% of the poultry products and in 3% of the human feces. Most isolates were identified as C. jejuni. In 93.5% of these isolates, the cdt genes have been detected. Despite the occurrence of Campylobacter in feces of humans has been low, the prevalence in broilers and poultry products was high, which, combined with the presence of cdt genes in most isolates, represents a potential risk to consumers. These results suggest there is a need for preventive measures in the production system and good manufacturing practices in the industry so as to minimize contamination of products and reduce the risk to consumers.

Ocorrência de Campylobacter em carne de frango, fezes de frango e humanas e pesquisa dos genes cdt / Occurrence of Campylobacter in poultry, meat chicken and human feces, and cdt GENES research

Foram coletadas 100 amostras de conteúdo fecal de aves de corte, 100 de produtos de frango (coxa, sobrecoxa, asa, dorso, carne moída e fígado) e 100 de fezes de humanos, e analisadas para pesquisa de Campylobacter. Realizou-se a determinação da espécie e da presença dos genes cdt, responsáveis pela codificação da toxina citoletal distensiva (CDT), através da técnica da PCR. A bactéria foi isolada de 61% das amostras de fezes de frango, 20% de produtos de frango e 3% de fezes de humanos. A maioria dos isolados foi determinada como C. jejuni . Destes, 93,5% apresentaram os genes para a toxina CDT. Apesar de a ocorrência de Campylobacter em fezes de humanos ter sido baixa, a prevalência em frangos de corte e produtos de frango foi elevada, fato que, aliado à presença dos genes cdt na maioria dos isolados, representa risco potencial para os consumidores. Esses resultados são indicativos da necessidade de medidas preventivas no sistema de produção e de boas práticas de fabricação na indústria, de forma a minimizar a contaminação dos produtos e diminuir o risco para os consumidores.(AU)

A hundred chicken fecal samples, a hundred samples of retail poultry products and a hundred samples of human feces were collected and tested for the presence of Campylobacter. The species identification and the analysis for the presence of cdt genes, responsible for encoding the cytolethal distending toxin, were performed by PCR. Campylobacter was found in 61% of the chicken fecal samples, in 20% of the poultry products and in 3% of the human feces. Most isolates were identified as C. jejuni. In 93.5% of these isolates, the cdt genes have been detected. Despite the occurrence of Campylobacter in feces of humans has been low, the prevalence in broilers and poultry products was high, which, combined with the presence of cdt genes in most isolates, represents a potential risk to consumers. These results suggest there is a need for preventive measures in the production system and good manufacturing practices in the industry so as to minimize contamination of products and reduce the risk to consumers.(AU)

Mastite subclínica por Staphylococcus coagulase negativa em ovinos de corte / Subclinical mastitis caused by Staphylococcus coagulase negative in meet-producing sheep

Microorganisms that cause mastitis were isolated in a herd of sheep and their sensitivity to antimicrobials was tested. Twenty-one sheep had their mammary glands monitored during lactation. Eighty-eight milk sample from CMT-positive sheep were analyzed in order to isolate the etiologic agent of mastitis. Coagulase negative Staphylococcus were isolated from 29.5% of samples. Ampicillin and penicillin were the antimicrobials with higher number of resistant strains. The importance of coagulase negative staphylococci has been demonstrated, especially S. lentus and S. sciuri as a cause of ovine mastitis.

Mastite subclínica por Staphylococcus coagulase negativa em ovinos de corte

Microorganisms that cause mastitis were isolated in a herd of sheep and their sensitivity to antimicrobials was tested. Twenty-one sheep had their mammary glands monitored during lactation. Eighty-eight milk sample from CMT-positive sheep were analyzed in order to isolate the etiologic agent of mastitis. Coagulase negative Staphylococcus were isolated from 29.5% of samples. Ampicillin and penicillin were the antimicrobials with higher number of resistant strains. The importance of coagulase negative staphylococci has been demonstrated, especially S. lentus and S. sciuri as a cause of ovine mastitis.

Qualidade higiênico-sanitária de carne bovina moída e de embutidos frescais comercializados no sul do rio grande do sul, brasil

ABSTRACT Meat is an excellent medium for the growth of microorganisms and it can be responsible for the transmission of pathogenic bacteria to human. The aim of this work was to analyze the sanitary-hygienic quality of ground beef and fresh sausages commercialized in southern Brazil and to evaluate the resistance profile of Salmonella isolates to antimicrobial agents. The methods used in the thermotolerant coliforms counting and in the Salmonella isolation were according to Instrução Normativa nº 62/2003 from Brazils Ministério da Agricultura, Pecuária e Abastecimento. The Salmonella isolates were tested in regard to susceptibility to ampicilin (10 mcg), norfloxacin (10 mcg), tetracycline (30 mcg), nalidixic acid (30 mcg) and chloramphenicol (30 mcg) antimicrobials. From 67 samples analized, 12 were not within the limits established by the current legislation to thermotolerant coliforms and Salmonella. Salmonella Typhimurium was isolated from 1 (4.2%) sample of ground beef. Salmonella serotypes Typhimurium and Infantis were isolated from 2 (9.5%) samples of pork meat sausages, and serotypes Typhimurium, Infantis, Derby from 3 (13.6%) samples of poultry sausages. The 6 (100%) isolates were sensitive to norfloxacina and ampicilina. Three (50%) isolates were sensitive to tetracycline, 3 (50%) to chloramphenicol and 2 (33.3%) to nalidixic acid. The results highlight the importance of meat products from swine, cattle and chicken as sources of Salmonella contamination.

RESUMO A carne constitui excelente meio para a multiplicação de microrganismos, podendo ser responsável pela transmissão de doenças para o homem através de bactérias patogênicas. O trabalho teve por objetivo analisar a qualidade higiênico-sanitária da carne bovina moída e de embutidos frescais comercializados no sul do Brasil e avaliar o perfil de resistência dos isolados de Salmonella a agentes antimicrobianos. Os métodos utilizados na contagem de coliformes termotolerantes e no isolamento de Salmonella foram realizados de acordo com a Instrução Normativa nº 62/2003 do Ministério da Agricultura, Pecuária e Abastecimento. Os isolados de Salmonella foram testados quanto à sensibilidade aos antimicrobianos ampicilina (10 mcg), norfloxacina (10 mcg), tetraciclina (30 mcg), ácido nalidíxico (30 mcg) e cloranfenicol (30 mcg). De 67 amostras analisadas, doze não atenderam aos limites estabelecidos pela legislação vigente para coliformes termotolerantes e Salmonella. Salmonella Typhimurium foi isolada de uma (4,2%) amostra de carne moída. Salmonella sorotipos Typhimurium e Infantis foram isoladas de duas (9,5%) amostras de lingüiça suína e sorotipos Typhimurium, Infantis e Derby de três (13,6%) amostras de lingüiça de frango. Os seis (100%) isolados foram sensíveis à norfloxacina e à ampicilina. Três (50%) isolados foram sensíveis à tetraciclina, três (50%) ao cloranfenicol e dois (33,3%) ao ácido nalidíxico. Os resultados obtidos ressaltam a importância dos produtos cárneos de origem suína, bovina e avícola como fonte de contaminação por Salmonella.