The isolation of multidrug-resistant Klebsiella pneumoniae in hospitals is a major public health threat, increasing patient hospitalization costs, morbidity and mortality. Therefore, this work investigated the resistance mechanisms that produced different carbapenems susceptibility profiles in two isogenic strains of K. pneumoniae isolated from the same patient in a public hospital in Recife, Pernambuco. The genes that encode the main porins in K. pneumoniae, ompK35 and ompK36, and several beta-lactamase genes were analyzed. The expression of these genes was evaluated by quantitative real time PCR (polymerase chain reaction) with reverse transcriptase (RT-qPCR). SDS-PAGE (sodium dodecyl sulphate–polyacrylamide gel electrophoresis) was performed to analyze the outer membrane proteins. The analysis of the ompK36 genetic environment disclosed an IS903 insertion sequence disrupting this gene in the ertapenem resistant isolate (KPN133). The blaKPC-2 gene showed down-regulated expression in both isolates. Our findings show that changes in porins, especially OmpK36, are more determinant to carbapenems susceptibility profile of bacterial isolates than variations in blaKPC gene expression.
O objetivo deste trabalho foi avaliar a produção de biofilme, o perfil de suscetibilidade e a detecção de genes de resistência presentes em isolados de Salmonella spp de carcaças frescas de frango comercializadas em uma metrópole brasileira. De um total de 61 amostras de carcaças frescas de aves, 21 foram positivas para a presença de Salmonella spp. Em relação ao teste de suscetibilidade aos antimicrobianos, (13/21) os isolados testados foram resistentes a pelo menos um antibiótico, correspondendo a 61,9%, e 38% (21/08) foram resistentes a múltiplas drogas. Pelo menos dois genes de resistência foram identificados em todos os isolados, especialmente os genes relacionados às β-lactamases e à resistência às quinolonas. Observou-se também que alguns isolados de Salmonella spp apresentaram padrões genéticos idênticos. E todos os 21 isolados foram capazes de formar biofilme. A identificação do biofilme de Salmonella spp. formando e transportando diferentes genes β-lactamase e determinantes da resistência às quinolonas demonstra a capacidade dessas bactérias de acumular vários mecanismos de virulência e resistência aos antimicrobianos. Portanto, a disseminação de diferentes grupos clonais de Salmonella spp. MDR em carcaças de carne de aves de capoeira expressas neste atestam a necessidade de controles efetivos para conter esse microrganismo, que além de ser um risco para a saúde pública, também é responsável por perdas econômicas consideráveis.
El objetivo de este estudio fue evaluar la producción de biofilm, el perfil de susceptibilidad y la detección de genes de resistencia presentes en aislados de Salmonella spp de canales frescas de pollo vendidas en una metrópoli brasileña. De un total de 61 muestras de canales frescas de aves de corral, 21 fueron positivas para la presencia de Salmonella spp. En cuanto a la prueba de sensibilidad antimicrobiana, (13/21) los aislamientos probados fueron resistentes a al menos un antibiótico, correspondiente al 61,9%, y el 38% (08/21) fueron resistentes a múltiples fármacos. Se identificaron al menos dos genes de resistencia en todos los aislamientos, especialmente los genes relacionados con la resistencia a β-lactamasas y quinolonas. También se observó que algunos aislados de Salmonella spp mostraron patrones genéticos idénticos. Y los 21 aislados fueron capaces de formar biopelícula. La identificación del biofilm de Salmonella spp. formando y portando diferentes genes de β-lactamasa y determinantes de resistencia a quinolonas demuestra la capacidad de estas bacterias para acumular diversos mecanismos de virulencia y resistencia a antimicrobianos. Por lo tanto, la propagación de diferentes grupos clonales de Salmonella spp. MDR en canales de carne de aves de corral expresada en este atestiguan la necesidad de controles efectivos para contener este microorganismo, que además de ser un riesgo para la salud pública, también es responsable de considerables pérdidas económicas.
This work investigated the resistance mechanisms that produce different susceptibility profiles in Klebsiella pneumoniae clinical isolates. The genes that encode the main porins in K. pneumoniae, ompK35 and ompK36, and several beta-lactamase genes were analyzed. The expression of these genes was evaluated by quantitative real time PCR (polymerase chain reaction) with reverse transcriptase (RT-qPCR). SDS-PAGE (sodium dodecyl sulphate–polyacrylamide gel electrophoresis) was performed to analyze the outer membrane proteins. The analysis of the ompK36 genetic environment found the presence of the IS903 insertion sequence interrupting this gene in the ertapenem resistant isolate (KPN133). The blaKPC-2 gene showed down-regulated expression in both isolates.
The aim of this study was to evaluate the biofilm production, the susceptibility profile and the detection of resistance genes present in Salmonella spp isolates from fresh chicken carcasses sold in a Brazilian metropolis. From a total of 61 samples of fresh poultry carcasses, 21 were positive for the presence of Salmonella spp. Regarding the antimicrobial susceptibility test, (13/21) isolates tested were resistant to at least one antibiotic, corresponding to 61.9%, and 38% (08/21) were Resistant to Multiple Drugs. At least two resistance genes were identified in all isolates, especially the genes related to β-lactamases and Quinolones resistance. It was also observed that some Salmonella spp isolates showed identical genetic patterns. And all 21 isolates were able to form biofilm. The identification of Salmonella spp. biofilm forming and carrying different β-lactamase genes and determinants of resistance to quinolones demonstrates the capacity of these bacteria to accumulate various mechanisms of virulence and resistance to antimicrobials. Therefore, the spread of different clonal groups of Salmonella spp. MDR in poultry meat carcasses expressed in this attest to the need for effective controls to contain this microorganism, which besides being a risk to public health, is also responsible for considerable economic losses.
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