Microbiological water quality monitoring of four ponds from Lagoa do Sino Farm located in São Paulo State, Brazil

This study aimed to evaluate the microbiological quality of the water of four ponds used for irrigation on the Lagoa do Sino Farm, as well as to perform the genotypic characterization of virulence factors in Escherichia coli isolates. Sampling was conducted for 11 months, between 2015 and 2016. Samples were analyzed for the presence of thermotolerant coliforms, E. coli and heterotrophs. DNA was extracted from E. coli isolates, followed by genotypic characterization by polymerase chain reaction. Agricultural activities and pesticides used in the sampling period were documented in order to assess possible relationships between agricultural activities and microbiological water quality. The absence of suitable riparian vegetation around all the ponds was observed, benefiting the entry of organic matter and contaminants in the water body. A high index of thermotolerant coliforms in some months indicated the possibility of the transmission of pathogenic microorganisms in these ponds. The values found in some months were above the regulatory limits for water potability and water intended for irrigation. The agrochemicals used in the period seem to influence the results obtained. All 17 E. coli isolates showed at least one of the virulence genes estA, stx1, stx2, and aatA, indicating enterotoxigenic, enterohaemorrhagic or enteroaggregative nature. The presence of E. coli in the waters may be associated with the presence of animals. The water samples analyzed are not suitable for irrigation of vegetables that are consumed raw and/or low lying fruits ingested without skin removal. It is essential to broaden the control of the use of chemicals, as well as the preservation of riparian vegetation to improve the quality of water used in the farm's agricultural activities.

Microbiological water quality monitoring of four ponds from Lagoa do Sino Farm located in São Paulo State, Brazil

This study aimed to evaluate the microbiological quality of the water of four ponds used for irrigation on the Lagoa do Sino Farm, as well as to perform the genotypic characterization of virulence factors in Escherichia coli isolates. Sampling was conducted for 11 months, between 2015 and 2016. Samples were analyzed for the presence of thermotolerant coliforms, E. coli and heterotrophs. DNA was extracted from E. coli isolates, followed by genotypic characterization by polymerase chain reaction. Agricultural activities and pesticides used in the sampling period were documented in order to assess possible relationships between agricultural activities and microbiological water quality. The absence of suitable riparian vegetation around all the ponds was observed, benefiting the entry of organic matter and contaminants in the water body. A high index of thermotolerant coliforms in some months indicated the possibility of the transmission of pathogenic microorganisms in these ponds. The values found in some months were above the regulatory limits for water potability and water intended for irrigation. The agrochemicals used in the period seem to influence the results obtained. All 17 E. coli isolates showed at least one of the virulence genes estA, stx1, stx2, and aatA, indicating enterotoxigenic, enterohaemorrhagic or enteroaggregative nature. The presence of E. coli in the waters may be associated with the presence of animals. The water samples analyzed are not suitable for irrigation of vegetables that are consumed raw and/or low lying fruits ingested without skin removal. It is essential to broaden the control of the use of chemicals, as well as the preservation of riparian vegetation to improve the quality of water used in the farm's agricultural activities.(AU)

Subinhibitory concentrations of silver nanoparticles and silver nitrate on the adaptative and cross-resistance to antibiotics on bovine mastitis pathogens / Concentrações subinibitórias de nanopartículas de prata e nitrato de prata na resistência adaptativa e cruzada com antibióticos em patógenos de mastite bovina

Biocides and/or antibiotics used in subinhibitory concentrations can promote the development of adaptive resistance or even cross-resistance in microorganisms. However, studies on these responses following silver treatments are scarce in the literature. Silver-based compounds, including silver nanoparticles (Ag-NPs), can be an alternative in the prevention and treatment of bovine mastitis. Thus, this research evaluated the effect of subinhibitory dosages of Ag-NPs and Ag+ ions from silver nitrate (AgNO3) on Staphylococcus aureus and Escherichia coli isolated from milk of cows with mastitis. Ag-NPs were synthesized by chemical reduction using AgNO3 and sodium citrate and the minimum inhibitory concentration (MIC) of Ag-NPs and Ag+ ions on the mastitis pathogens were determined. Isolates were exposed to subinhibitory concentrations of Ag-NPs or AgNO3 for 10 consecutive days to verify the development of adaptive resistance evaluated by changes in the MIC values. The development of cross-resistance with antibiotics was also studied, being verified by comparing the sensitivity profile of treated cells with non-treated cells. AgNO3 was more effective against all isolates. There was no change in the MIC values or in the antibiotic sensitivity profile for both bacteria following consecutive exposure to subinhibitory dosages of Ag-NPs or AgNO3, indicating that silver was not able to select adaptive resistance or cross resistance to the tested antibiotics. The potential of silver presented by these results is favorable to the continuity of studies aiming to elaborate silver-based therapies for the treatment of bovine mastitis.

Biocidas e/ou antibióticos em concentrações sub-inibitórias podem promover o desenvolvimento de resistência adaptativa ou mesmo resistência cruzada nos micro-organismos. Entretanto, estudos destas respostas após o tratamento com a prata são escassos na literatura. Compostos a base de prata, incluindo as nanopartículas de prata (Ag-NPs), podem ser uma alternativa na prevenção e/ou tratamento de mastite bovina. Assim, este trabalho objetivou determinar o efeito de doses sub-inibitórias de Ag-NPs e dos íons Ag+ provenientes do nitrato de prata (AgNO3) sobre isolados de Staphylococcus aureus e de Escherichia coli, provenientes de leite de vacas com mastite. As Ag-NPs foram sintetizadas por redução química utilizando AgNO3 e citrato de sódio e a Concentração Mínima Inibitória (CMI) das Ag-NPs e íons Ag+ nos patógenos da mastite foi determinada. Os isolados foram expostos a concentrações sub-inibitórias de Ag-NPs ou de AgNO3 por 10 dias consecutivos para verificar o desenvolvimento de resistência adaptativa à prata pela mudança no valor da CMI, e de resistência cruzada com antibióticos pela mudança no perfil de sensibilidade em relação ao controle. AgNO3 apresentou-se mais efetivo contra todos os isolados. Não foi verificada alteração no valor da CMI nem do perfil de sensibilidade aos antibióticos, indicando que não houve seleção de resistência adaptativa à prata e de resistência cruzada aos antibióticos pelos micro-organismos estudados. O uso potencial da prata apresentado nos resultados é favorável à continuidade dos estudos objetivando a elaboração de terapias à base de prata para o tratamento da mastite bovina.

Subinhibitory concentrations of silver nanoparticles and silver nitrate on the adaptative and cross-resistance to antibiotics on bovine mastitis pathogens / Concentrações subinibitórias de nanopartículas de prata e nitrato de prata na resistência adaptativa e cruzada com antibióticos em patógenos de mastite bovina

ABSTRACT: Biocides and/or antibiotics used in subinhibitory concentrations can promote the development of adaptive resistance or even cross-resistance in microorganisms. However, studies on these responses following silver treatments are scarce in the literature. Silver-based compounds, including silver nanoparticles (Ag-NPs), can be an alternative in the prevention and treatment of bovine mastitis. Thus, this research evaluated the effect of subinhibitory dosages of Ag-NPs and Ag+ ions from silver nitrate (AgNO3) on Staphylococcus aureus and Escherichia coli isolated from milk of cows with mastitis. Ag-NPs were synthesized by chemical reduction using AgNO3 and sodium citrate and the minimum inhibitory concentration (MIC) of Ag-NPs and Ag+ ions on the mastitis pathogens were determined. Isolates were exposed to subinhibitory concentrations of Ag-NPs or AgNO3 for 10 consecutive days to verify the development of adaptive resistance evaluated by changes in the MIC values. The development of cross-resistance with antibiotics was also studied, being verified by comparing the sensitivity profile of treated cells with non-treated cells. AgNO3 was more effective against all isolates. There was no change in the MIC values or in the antibiotic sensitivity profile for both bacteria following consecutive exposure to subinhibitory dosages of Ag-NPs or AgNO3, indicating that silver was not able to select adaptive resistance or cross resistance to the tested antibiotics. The potential of silver presented by these results is favorable to the continuity of studies aiming to elaborate silver-based therapies for the treatment of bovine mastitis.

RESUMO: Biocidas e/ou antibióticos em concentrações sub-inibitórias podem promover o desenvolvimento de resistência adaptativa ou mesmo resistência cruzada nos micro-organismos. Entretanto, estudos destas respostas após o tratamento com a prata são escassos na literatura. Compostos a base de prata, incluindo as nanopartículas de prata (Ag-NPs), podem ser uma alternativa na prevenção e/ou tratamento de mastite bovina. Assim, este trabalho objetivou determinar o efeito de doses sub-inibitórias de Ag-NPs e dos íons Ag+ provenientes do nitrato de prata (AgNO3) sobre isolados de Staphylococcus aureus e de Escherichia coli, provenientes de leite de vacas com mastite. As Ag-NPs foram sintetizadas por redução química utilizando AgNO3 e citrato de sódio e a Concentração Mínima Inibitória (CMI) das Ag-NPs e íons Ag+ nos patógenos da mastite foi determinada. Os isolados foram expostos a concentrações sub-inibitórias de Ag-NPs ou de AgNO3 por 10 dias consecutivos para verificar o desenvolvimento de resistência adaptativa à prata pela mudança no valor da CMI, e de resistência cruzada com antibióticos pela mudança no perfil de sensibilidade em relação ao controle. AgNO3 apresentou-se mais efetivo contra todos os isolados. Não foi verificada alteração no valor da CMI nem do perfil de sensibilidade aos antibióticos, indicando que não houve seleção de resistência adaptativa à prata e de resistência cruzada aos antibióticos pelos micro-organismos estudados. O uso potencial da prata apresentado nos resultados é favorável à continuidade dos estudos objetivando a elaboração de terapias à base de prata para o tratamento da mastite bovina.

Subinhibitory concentrations of silver nanoparticles and silver nitrate on the adaptative and cross-resistance to antibiotics on bovine mastitis pathogens / Concentrações subinibitórias de nanopartículas de prata e nitrato de prata na resistência adaptativa e cruzada com antibióticos em patógenos de mastite bovina

Biocides and/or antibiotics used in subinhibitory concentrations can promote the development of adaptive resistance or even cross-resistance in microorganisms. However, studies on these responses following silver treatments are scarce in the literature. Silver-based compounds, including silver nanoparticles (Ag-NPs), can be an alternative in the prevention and treatment of bovine mastitis. Thus, this research evaluated the effect of subinhibitory dosages of Ag-NPs and Ag+ ions from silver nitrate (AgNO3) on Staphylococcus aureus and Escherichia coli isolated from milk of cows with mastitis. Ag-NPs were synthesized by chemical reduction using AgNO3 and sodium citrate and the minimum inhibitory concentration (MIC) of Ag-NPs and Ag+ ions on the mastitis pathogens were determined. Isolates were exposed to subinhibitory concentrations of Ag-NPs or AgNO3 for 10 consecutive days to verify the development of adaptive resistance evaluated by changes in the MIC values. The development of cross-resistance with antibiotics was also studied, being verified by comparing the sensitivity profile of treated cells with non-treated cells. AgNO3 was more effective against all isolates. There was no change in the MIC values or in the antibiotic sensitivity profile for both bacteria following consecutive exposure to subinhibitory dosages of Ag-NPs or AgNO3, indicating that silver was not able to select adaptive resistance or cross resistance to the tested antibiotics. The potential of silver presented by these results is favorable to the continuity of studies aiming to elaborate silver-based therapies for the treatment of bovine mastitis.(AU)

Biocidas e/ou antibióticos em concentrações sub-inibitórias podem promover o desenvolvimento de resistência adaptativa ou mesmo resistência cruzada nos micro-organismos. Entretanto, estudos destas respostas após o tratamento com a prata são escassos na literatura. Compostos a base de prata, incluindo as nanopartículas de prata (Ag-NPs), podem ser uma alternativa na prevenção e/ou tratamento de mastite bovina. Assim, este trabalho objetivou determinar o efeito de doses sub-inibitórias de Ag-NPs e dos íons Ag+ provenientes do nitrato de prata (AgNO3) sobre isolados de Staphylococcus aureus e de Escherichia coli, provenientes de leite de vacas com mastite. As Ag-NPs foram sintetizadas por redução química utilizando AgNO3 e citrato de sódio e a Concentração Mínima Inibitória (CMI) das Ag-NPs e íons Ag+ nos patógenos da mastite foi determinada. Os isolados foram expostos a concentrações sub-inibitórias de Ag-NPs ou de AgNO3 por 10 dias consecutivos para verificar o desenvolvimento de resistência adaptativa à prata pela mudança no valor da CMI, e de resistência cruzada com antibióticos pela mudança no perfil de sensibilidade em relação ao controle. AgNO3 apresentou-se mais efetivo contra todos os isolados. Não foi verificada alteração no valor da CMI nem do perfil de sensibilidade aos antibióticos, indicando que não houve seleção de resistência adaptativa à prata e de resistência cruzada aos antibióticos pelos micro-organismos estudados. O uso potencial da prata apresentado nos resultados é favorável à continuidade dos estudos objetivando a elaboração de terapias à base de prata para o tratamento da mastite bovina.(AU)