Search

Introduction: The objective of this study was to evaluate the involvement of the ompR gene in the acid adaptation and thermal resistance of S. Enteritidis SE86, responsible agent of more than 95 % of investigated food-borne diseases, throughout the last decade in Southern Brazil. In this study, we constructed a mutant strain of S. Enteritidis SE86 (ΔompR) that was attenuated by a knockout technique. The OmpR protein expression was determined in a tagged (3XFLAG) strain of S. Enteritidis SE86. Methodology: The mutant strains were cultivated separately in nutrient broth and nutrient broth supplemented with 1% glucose (NBG) to induce acid adapted cells. The organisms were exposed to different temperature such as 37 °C, 52 °C, and 60 °C. The survival of the SE86 wild type (WT) and attenuated strain was determined by bacterial count, and the tagged protein (ompR::3XFLAG cat::FLAG) was detected by SDS-PAGE and immunoblotting with anti-FLAG antibodies Results: Results showed that when exposed at 52°C, the acid-adapted SE86 WT cells were completely inactivated after 300 minutes; however, non-adapted cells (WT and ΔompR) and acid-adapted ΔompR demonstrated higher thermal sensitivity, since they were completely inactivated in 240 minutes. At 60°C, the acid-adapted SE86 ΔompR also demonstrated higher sensitivity that SE86 WT, being totally inactivated after 15 minutes, while the WT cells were inactivated in 20 minutes. Conclusion: The acid adapted cells showed increased expression of OmpR when exposed to 52°C and 60°C, this confirmed the requirement of acid adaptation for S. Enteritidis SE86 to resist elevated temperatures. [ABSTRACT FROM AUTHOR]

In Rio Grande do Sul, southern Brazil, Salmonella Enteritidis is one of the principal microorganisms responsible for foodborne disease. The present study was conducted to compare the sodium hypochlorite resistance of Salmonella Enteritidis SE86 with that of other strains of Salmonella Enteritidis isolated from different regions of the world and to investigate the involvement of the rpoS and dps genes in resistance to this disinfectant. We tested five Salmonella Enteritidis wild-type (WT) strains isolated from different countries, two mutant strains of Salmonella Enteritidis SE86, and two tagged (3XFLAG) strains of Salmonella Enteritidis SE86 for their resistance to sodium hypochlorite (200 ppm). The survival of the WT and attenuated strains was determined based on bacterial counts, and tagged proteins (Dps and RpoS) were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with anti-FLAG antibodies. None of the WT strains of Salmonella Enteritidis were totally inactivated after 20 min. The SE86 strain lacking dps was more sensitive to sodium hypochlorite than was the WT SE86 strain, with a 2-log reduction in counts after 1 min. The RpoS and Dps proteins were actively expressed under the conditions tested, indicating that in Salmonella Enteritidis SE86 these genes, which are expressed when in contact with sodium hypochlorite, are related to oxidative stress. [ABSTRACT FROM AUTHOR]