Isolamento, caracterização molecular e detecção por PCR em tempo real (qPCR) de Salmonella spp. em amostras fecais de potros com e sem diarreia.
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Data
2021-11-29
Autores
Braga, Pollyana Rennó Campos
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O presente estudo investigou a ocorrência de Salmonella spp. em fezes de 200 potros com até 1 ano de idade, através das técnicas de cultivo bacteriológico e PCR em tempo real (qPCR) pelos métodos de corante fluorescente (SYBR® Green) e sonda específica (Taqman®) para o gene invA com e sem etapa de pré-enriquecimento seletivo em tetrationato. Sorotipificação, testes de sensibilidade antimicrobiana e análise genética para a presença de genes produtores de β-lactamases foram realizados. Houve a identificação dos sorovares: S. Infantis, S. Minessota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. I.4,12:d:-, S. Braenderup, S. Give, S. Newport, S. I.6,8:-:-, S.IIIb 61:c:z35, S.IO9:-:1,5. O cultivo bacteriológico revelou 15% (n=30) de animais positivos (21 animais com diarreia e 9 de animais sem diarreia). Multirresistência foi constatada em 43,33% (n=13) dos isolados. A análise genética revelou 10 isolados (33,3%) produtores de β-lactamases. Houve superioridade dos resultados positivos pela qPCR em relação ao cultivo bacteriológico. O método de qPCR SYBR® com tetrationato apresentou 22,5% (n=45) de animais positivos, qPCR Taqman® com tetrationato identificou 22% de animais positivos (n=44), qPCR SYBR® sem tetrationato obteve 24% (n=48) de animais positivos e a qPCR Taqman® sem tetrationato evidenciou 20% (n=40) de animais positivos. Os resultados confirmaram a utilidade do método qPCR sem a etapa de pré-enriquecimento em tetrationato, como um teste rápido para detecção de Salmonella spp. em animais portadores ou em animais com sinais clínicos de diarreia, associado ao cultivo bacteriológico para a realização do teste de sensibilidade aos antimicrobianos e sorotipificação.
This study has investigated the ocurrence of Salmonella spp. in fecal samples from 200 foals up to one year of age. Feces have been tested for Salmonella spp. by microbiologic culture and TaqMan® and SYBR® Green real time polymerase chain reaction (qPCR) methods for invA gene with and without pre-enrichement in tetrathionate. Salmonella spp. strains have been serotyped, antimicrobial susceptibility profiles have been analyzed and the presence of β-lactamase genes have been evaluated. Salmonella spp. was isolated in 30 (15%) horses (21 horses with diarrhea and nine horses without diarrhea. Salmonella spp. serotypes have been identified: S. Infantis, S. Minessota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. enterica subsp enterica 4,12:d:-, S. Braenderup, S. Give, S. Newport, S. enterica subsp. enterica 6,8:-:-, S.IIIb 61:c:z35, S.IO9:-:1,5. Multiresistance has been observed in 43,33% (n=13) of bacterial isolates. Genetic analyses have revealed 10 bacterial isolates (33,3%) β-lactamases producers. There were high superiority of positive qPCR results compared to bacterial culture results. The qPCR SYBR® Green with tethrationate has showed 22,5% (n=45) of positive animals, qPCR Taqman® with tethrationate has exhibited 22% (n=44) of positive animals, qPCR SYBR® Green without tetrathionate has identified 24% (n=48) of positive animals and the qPCR Taqman® without tetrathionate has identified 20% (n=40) of positive animals. The results have confirmed the utility of qPCR method as a rapid test for detection of Salmonella spp. in animal carriers or in animals with clinical signs of diarrhea, associated with bacterial culture for antimicrobial susceptibility testing and serotyping data.
This study has investigated the ocurrence of Salmonella spp. in fecal samples from 200 foals up to one year of age. Feces have been tested for Salmonella spp. by microbiologic culture and TaqMan® and SYBR® Green real time polymerase chain reaction (qPCR) methods for invA gene with and without pre-enrichement in tetrathionate. Salmonella spp. strains have been serotyped, antimicrobial susceptibility profiles have been analyzed and the presence of β-lactamase genes have been evaluated. Salmonella spp. was isolated in 30 (15%) horses (21 horses with diarrhea and nine horses without diarrhea. Salmonella spp. serotypes have been identified: S. Infantis, S. Minessota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. enterica subsp enterica 4,12:d:-, S. Braenderup, S. Give, S. Newport, S. enterica subsp. enterica 6,8:-:-, S.IIIb 61:c:z35, S.IO9:-:1,5. Multiresistance has been observed in 43,33% (n=13) of bacterial isolates. Genetic analyses have revealed 10 bacterial isolates (33,3%) β-lactamases producers. There were high superiority of positive qPCR results compared to bacterial culture results. The qPCR SYBR® Green with tethrationate has showed 22,5% (n=45) of positive animals, qPCR Taqman® with tethrationate has exhibited 22% (n=44) of positive animals, qPCR SYBR® Green without tetrathionate has identified 24% (n=48) of positive animals and the qPCR Taqman® without tetrathionate has identified 20% (n=40) of positive animals. The results have confirmed the utility of qPCR method as a rapid test for detection of Salmonella spp. in animal carriers or in animals with clinical signs of diarrhea, associated with bacterial culture for antimicrobial susceptibility testing and serotyping data.
Descrição
Palavras-chave
Fezes, Sorovares, Equinos, Salmonella spp., invA, Multirresistência, Horses, Personality, Cognition