Protocol Optimization of qPCR for Analysis of the Effects of Antibiotics on Multidrug-Resistant Salmonella Populations in Swine

Abstract

Antibiotic resistance is a prominent food safety concern. Antibiotics given to food animals may promote the development of antibiotic resistance within the present bacterial populations, which then poses a threat to both workers in the food animal operation and consumers. A previous study was performed which sought to analyze the effects of ceftiofur and chlortetracycline on pan-susceptible and multi-drug resistant (MDR) resistant strains of Salmonella within a test group of swine. The purpose of this study is to optimize protocols for quantitative PCR analysis of the presence of antibiotic resistance genes, specifically qnrB and blaSHV, found within fecal samples extracted over the course of the previous trial. The gene invA was also analyzed to determine the presence of pan-susceptible bacteria within the samples as well, allowing for comparison as to how the pan-susceptible strains fared against the antibiotic treatments compared to the MDR ones. The protocols created for qnrB and blaSHV were successfully optimized and readied for use in sample analysis, but further work still remains to be done on the protocol created for invA gene analysis.