RESUMO

The life cycle of synanthropic flies and their behavior, allows them to serve as mechanical vectors of several pathogens. Given that flies can carry multidrug-resistant (MDR) bacteria, this study aimed to investigate the spread of genes of antimicrobial resistance in Escherichia coli isolated from flies collected in two dairy farms in Brazil. Besides antimicrobial resistance determinants, the presence of virulence genes related to bovine colibacillosis was also assessed. Of 94 flies collected, Musca domestica was the most frequently found in the two farms. We isolated 198 E. coli strains (farm A=135 and farm B=63), and >30% were MDR E. coli. We found an association between blaTEM and phenotypical resistance to ampicillin, or chloramphenicol, or tetracycline; and blaCTX-M and resistance to cefoperazone. A high frequency (86%) of phylogenetic group B1 among MDR strains and the lack of association between multidrug resistance and virulence factors suggest that antimicrobial resistance possibly is associated with the commensal bacteria. Clonal relatedness of MDR E. coli performed by Pulsed-Field Gel Electrophoresis showed wide genomic diversity. Different flies can carry clones, but with distinct antimicrobial resistance pattern. Sanger sequencing showed that the same class 1 integron arrangement is displayed by apparently unrelated strains, carried by different flies. Our conjugation results indicate class 1 integron transfer associated with tetracycline resistance. We report for the first time, in Brazil, that MDR E. coli is carried by flies in the milking environment. Therefore, flies can act as carriers for MDR strains and contribute to dissemination routes of antimicrobial resistance.

Assuntos

RESUMO

A produção e o consumo de leite e derivados vêm aumentando anualmente no Brasil. A emergência do consumo de leite cru está relacionada à busca por produtos que mantenham preservadas suas propriedades nutricionais. No entanto, este hábito aumenta os riscos do consumo de leite e derivados contaminados por micro-organismos patogênicos. As bactérias do gênero Enterobacter são amplamente distribuídas na natureza e fazem parte da microbiota do trato gastrintestinal humano e animal. Enterobacter cloacae, Enterobacter aerogenes, Pantoea agglomerans e Cronobacter sakazakii são considerados patógenos oportunistas. Nesse sentido, o presente estudo investigou a ocorrência e a diferenciação bioquímica de espécies de Enterobacter isolados de leite bovino cru, a caracterização do perfil de sensibilidade aos antimicrobianos e a pesquisa de genes de resistência. Do total de 104 cepas de Enterobacter spp. foram identificadas E. cloacae (n=39), Pantoea agglomerans (n=23), Cronobacter sakazakii (n=18) e E. aerogenes (n=9). Destas, 26% apresentaram resistência a três ou mais antimicrobianos. O gene blaTEM foi encontrado em 27% dos isolados. Este achado alerta quanto ao risco de infecções por micro-organismos ambientais resistentes, associados ao consumo de leite bovino cru.

The production and consumption of milk and dairy products is growing fast in Brazil. The increase consumption of raw milk is related to the interest for products that keep conserved their nutritional properties. However, this habit increases the risks of consumption of milk and dairy products contaminated with pathogenic microorganisms. Enterobacter spp. is widely distributed in nature and is part of the commensal microbiota of the intestinal tracts of humans and animal. Enterobacter cloacae, Enterobacter aerogenes, Pantoea agglomerans and Cronobacter sakazakii are recognized as opportunistic pathogens. The current study investigated the occurrence and the differentiation of Enterobacter species isolated from raw cow milk, the characterization of sensitivity profile to antimicrobials and presence of resistant genes. From a total of 104 strains of Enterobacter spp. were identified E. cloacae (n=39), Pantoea agglomerans (n=23), Cronobacter sakazakii (n=18) and E. aerogenes (n=9). Among these, 26% were resistant to three or more antimicrobials. The blaTEM gene was found in 27% of the isolates. Here, we highlighted the risk of infection by potential environmental microorganisms, resistant to some antimicrobials, associated with consumption of raw cow milk.

La producción y el consumo de leche y productos lácteos está aumentando anualmente en Brasil. El surgimiento del consumo de leche cruda se relaciona con la búsqueda de productos que conservan sus propiedades nutricionales. Esta conducta aumenta riesgos del consumo de leche y productos lácteos contaminados con microorganismos patógenos. Las bacterias del género Enterobacter son ampliamente distribuidas en la naturaleza y forma parte de los microorganismos comensales del aparato gastrointestinal humano y animal. Enterobacter cloacae, Enterobacter aerogenes, Pantoca agglomerans y Cronobacter sakazakii son citadas como patógenos oportunistas. De esta manera, este estudio investigó la ocurrencia y diferenciación bioquímica de Enterobacter spp. aislados de leche cruda de ganado bobino, la caracterización del perfil de sensibilidad frente a los antimicrobianos y la detección de los genes de resistencia. Del total de 104 cepas de Enterobacter spp. se identificaron E. cloacae (n=39), Pantoea agglomerans (n=23), Cronobacter sakazakii (n=18), E. aerogenes (n=9). De estos, 26% mostraron resistencia a tres o más antimicrobianos. En 27% de las cepas el gen blaTEM fue detectado. Por lo tanto, este hallazgo advierte sobre el riesgo de infecciones por microorganismos resistentes ambientales, asociados con el consumo de leche cruda de vaca.

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RESUMO

Milk that is adequate for consumption must be of hygienic quality, nutritional value, and should maintain its organoleptic properties. Isolation of fecal and/or total coliforms from bovine milk is considered an indicator of hygiene and good management practices, and can be used as a quality indicator. This study aimed to isolate, identify, and assess the resistance profile of coliforms isolated from collective bulk tanks and individual milk tanks. A total of 89 milk samples were collected from collective bulk tanks and, from these, 21 Klebsiella spp., one E. coli, and 29 Enterobacter spp. were isolated, whereas 102 milk samples from individual tanks showed isolation of one Klebsiella spp. and seven Enterobacter spp. In collective bulk tanks, at least 47% of Klebsiella spp. and Enterobacter spp. were resistant to cephalexin and 30% to ampicillin. From these, at least 24% showed multidrug resistance. Among the microorganisms isolated from the individual tanks, 85% or more were resistant to ampicillin. The ESBL phenotype and the blaTEM gene were detected in strains of Klebsiella spp. isolated from both tanks. It was concluded that contamination of milk with resistant total coliforms, and especially the storage of raw milk from several small producers in the collective bulk tank increase the risk of contamination.

O leite adequado ao consumo deve possuir qualidade higiênica, valor nutritivo e a manutenção das propriedades organolépticas. O isolamento de coliformes totais e/ou fecais de leite bovino é considerado um indicador de sanidade e de boas práticas de manejo, podendo ser usado como indicador de qualidade. Este estudo teve como objetivo isolar, identificar e observar o perfil de resistência de coliformes presentes em tanques de refrigeração coletivos e individuais de acondicionamento de leite. Foram colhidas 89 amostras de leite de tanques coletivos e isolados 21 Klebsiella spp., uma Escherichia coli (E. coli) e 29 Enterobacter spp., e 102 amostras de leite de tanques individuais das quais foram isolados uma Klebsiella spp. e sete Enterobacter spp. Dos tanques coletivos, aproximadamente 47% de Klebsiella spp. e Enterobacter spp. foram resistentes para cefalexina e 30% para ampicilina. Destes, no mínimo 24% apresentaram multirresistência. Entre os isolados de tanques individuais, 85% ou mais apresentaram resistência para ampicilina. O fenótipo ESBL e o gene blaTEM foram detectados em linhagens de Klebsiella spp. isoladas de ambos os tanques. Constatou-se contaminação do leite com coliformes totais resistentes e, principalmente, que o armazenamento do leite cru de vários pequenos produtores em tanques coletivos potencializa o risco de contaminação.

Assuntos

RESUMO

Milk that is adequate for consumption must be of hygienic quality, nutritional value, and should maintain its organoleptic properties. Isolation of fecal and/or total coliforms from bovine milk is considered an indicator of hygiene and good management practices, and can be used as a quality indicator. This study aimed to isolate, identify, and assess the resistance profile of coliforms isolated from collective bulk tanks and individual milk tanks. A total of 89 milk samples were collected from collective bulk tanks and, from these, 21 Klebsiella spp., one E. coli, and 29 Enterobacter spp. were isolated, whereas 102 milk samples from individual tanks showed isolation of one Klebsiella spp. and seven Enterobacter spp. In collective bulk tanks, at least 47% of Klebsiella spp. and Enterobacter spp. were resistant to cephalexin and 30% to ampicillin. From these, at least 24% showed multidrug resistance. Among the microorganisms isolated from the individual tanks, 85% or more were resistant to ampicillin. The ESBL phenotype and the blaTEM gene were detected in strains of Klebsiella spp. isolated from both tanks. It was concluded that contamination of milk with resistant total coliforms, and especially the storage of raw milk from several small producers in the collective bulk tank increase the risk of contamination.(AU)

O leite adequado ao consumo deve possuir qualidade higiênica, valor nutritivo e a manutenção das propriedades organolépticas. O isolamento de coliformes totais e/ou fecais de leite bovino é considerado um indicador de sanidade e de boas práticas de manejo, podendo ser usado como indicador de qualidade. Este estudo teve como objetivo isolar, identificar e observar o perfil de resistência de coliformes presentes em tanques de refrigeração coletivos e individuais de acondicionamento de leite. Foram colhidas 89 amostras de leite de tanques coletivos e isolados 21 Klebsiella spp., uma Escherichia coli (E. coli) e 29 Enterobacter spp., e 102 amostras de leite de tanques individuais das quais foram isolados uma Klebsiella spp. e sete Enterobacter spp. Dos tanques coletivos, aproximadamente 47% de Klebsiella spp. e Enterobacter spp. foram resistentes para cefalexina e 30% para ampicilina. Destes, no mínimo 24% apresentaram multirresistência. Entre os isolados de tanques individuais, 85% ou mais apresentaram resistência para ampicilina. O fenótipo ESBL e o gene blaTEM foram detectados em linhagens de Klebsiella spp. isoladas de ambos os tanques. Constatou-se contaminação do leite com coliformes totais resistentes e, principalmente, que o armazenamento do leite cru de vários pequenos produtores em tanques coletivos potencializa o risco de contaminação.(AU)

Assuntos