Successful control of foodborne pathogens requires placement of chemical and physical hurdles in the preharvest and postharvest food production sectors. Pathogens may also encounter indigenous antimicrobials in foods including certain botanical compounds that have historically been used for flavor enhancement as well as preservation. Chemical additives have traditionally included organic acids to control microbial contamination in foods and feeds. However, there is some concern that continuous application of certain chemical antimicrobials can lead to a buildup of microbial resistance. This creates problems if foodborne pathogens survive and develop resistance to a variety of environmental stressors encountered in pre- and postharvest animal production. To expand the diversity of potential antimicrobials that have practical application to food animal production requires exploring the interaction between the food matrix and foodborne pathogens. There is potential for isolating antimicrobial compounds that exhibit mechanisms unrelated to conventional antimicrobial compounds. However, understanding the potential for novel antimicrobial compounds in foods and feeds will require the physiological examination of foodborne pathogen response under experimental conditions comparable to the environment where the pathogen is most likely to occur. Research on foodborne Salmonella pathogenesis is extensive and should provide a model for detailed examination of the factors that influence antimicrobial effectiveness. Analysis of pathogen response to antimicrobials could yield clues for optimizing hurdle technologies to more effectively exploit vulnerabilities of Salmonella and other foodborne pathogens when administering antimicrobials during food and feed production.
Spent media is nutritionally exhausted media due to extensive previous growth of bacteria species and is known to influence the gene expression patterns of Salmonella Typhimurium in in vivo and in vitro environments. Comparative C T -based real time quantitative polymerase chain reaction (RT-PCR) requires a baseline reference gene whose expression is stable throughout experimental conditions. The normalization to a baseline gene is necessary to eliminate sample-to-sample variations of the RNA isolation and reverse transcription steps. The current study explores the suitability of rsmC to serve as a reference gene in RT-PCR gene expression quantification of S. Typhimurium during growth in spent media. In our study, although the growth range response of S. Typhimurium as measured either by optical density or colony forming units (CFU) over time in rich and spent media was significantly different, the expression of rsmC remained constant when evaluated by the corresponding C T values. rsmC could be successfully used as a reference gene in the comparative C T RT-PCR method to quantify S. Typhimurium gene expression throughout the growth range response to this nutritionally limited environment.
Salmonella is a foodborne pathogen causing severe gastroenteritis. Three types of Maillard reaction products (MRP) generated by heat sterilization of D-glucose and L-lysine, L-histidine, and L-arginine were studied at 2 different levels of supplementation (0.5% and 1.0%) for their influence on growth and virulence of Salmonella. Two methods, namely, real-time polymerase chain reaction (RT-PCR) and a beta-galactosidase gene fusion assay, were used to determine the expression of hilA, a regulatory gene for Salmonella pathogenicity. Neither the type of MRP nor their quantities up to 1.0% affected the growth rates of S. Typhimurium EE658 (P > 0.05). When determined by beta-galactosidase assay, lysine MRP in both levels of supplementation were not found to have any effect on the hilA expression compared to the control. The addition of histidine and arginine MRP to M9 media (0.5%) increased by 2-fold hilA induction and up to 6-fold at the higher level (1%) supplementation of these compounds. Although somewhat inconsistent, RT-PCR analyses of hilA expression confirmed the greater induction effect of arginine MRP on hilA compared to lysine MRP. In contrast to beta-galactosidase assay results, however, lysine MRP were found to increase hilA expression compared to the control in both supplementation levels in all trials. The potential of MRP serving as a bacterial virulence modulator may be a factor to be considered in food thermal processing when assessing Salmonella risk for causing foodborne disease.
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