ABSTRACT
The multidrug efflux system in bacteria can reduce antibiotic concentration inside the cell, leading to failure in the treatment of bacterial diseases. This study evaluated the influence of two efflux pump inhibitors (EPIs), phenylalanine arginyl ß-naphthylamide (PAßN) and 1-(1-naphthylmethyl)-piperazine (NMP), on the gene expression of three multidrug efflux systems, AcrAB, AcrEF and EmrAB in Escherichia coli bovine mastitis isolates resistant to ampicillin and sulfamethoxazole/trimethoprim simultaneously. Each isolate had at least three multidrug efflux system genes. The acrA and acrB had the lowest expression levels in all treatments, while the emrA or emrB showed the highest expression levels in the presence of ampicillin, sulfamethoxazole/trimethoprim, PAßN and NMP. EPIs also contributed to the decrease in arcF expression when used in combination with ampicillin treatment. Since PAßN showed stronger effects than NMP, it may serve as an alternative to assist in the antimicrobial therapy of mastitis.
Subject(s)
Dipeptides/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Gene Expression Regulation, Bacterial/drug effects , Mastitis, Bovine/microbiology , Piperazines/pharmacology , Ampicillin/pharmacology , Animals , Anti-Infective Agents/pharmacology , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/drug effects , Bacterial Proteins/genetics , Cattle , Colony Count, Microbial , Escherichia coli/isolation & purification , Female , Mastitis, Bovine/drug therapy , Membrane Transport Proteins/drug effects , Membrane Transport Proteins/genetics , Microbial Sensitivity Tests/methods , Microbial Sensitivity Tests/veterinary , Polymerase Chain Reaction , Trimethoprim, Sulfamethoxazole Drug Combination/pharmacologyABSTRACT
Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic granulomatous enteritis that affects all ruminants worldwide. Some researchers have indicated a possible role of MAP in Crohn's disease. Despite extensive research and large and important advances in the past few decades, the etiology of Crohn's disease remains indefinite. The most probable transmission route of MAP from animals to humans is milk and dairy products. Mycobacterium avium ssp. paratuberculosis has already been detected in milk samples worldwide, and some studies have reported that MAP is resistant to pasteurization. In Brazil, MAP has been reported in raw milk samples; however, Brazilian retail pasteurized milk has not yet been tested for viable MAP. The aim of this study was to investigate MAP in pasteurized milk in the region of Viçosa (Minas Gerais, Brazil). Thirty-seven samples were collected and processed for culture of MAP. One colony similar to MAP was observed and confirmed by IS900-nested PCR and sequencing. Analysis revealed 97 to 99% identity with the MAP K-10 strain. This study is the first report of the presence of MAP in retail pasteurized whole milk in Brazil.
