Contribution of the gyrA and waaG mutants to fluoroquinolones resistance, biofilm development, and persister cells formation in Salmonella enterica serovar Typhi

Abstract

Fluoroquinolone resistance in Salmonella has been reported worldwide and poses a serious public health threat in developing countries. Multiple factors contribute to fluoroquinolone resistance, including mutations in DNA gyrase and the acquisition of antimicrobial resistance genes. Salmonella enterica serovar Typhi (S. Typhi) causes typhoid fever in humans, which is highly prevalent in counties with poor sanitation and hygiene standards. Here, we reported S. Typhi clinical isolates that showed varying degrees of susceptibility to fluoroquinolones and were characterized by Analytical Profile Index 20E test kit and 16S rRNA sequencing. S. Typhi strain S27 was resistant to fluoroquinolones and had multiple mutations in the gyrA gene. The gyrA lies in the quinolone resistance determining region of S. Typhi and has mutations at codon 83 (Ser83Phe), codon 87 (Asp87Gly), codon 308 (Lys308Glu), and codon 328 (Val328Ile). S. Typhi strain S6 has no gyrA mutations and is sensitive to fluoroquinolones but forms a strong biofilm relative to S. Typhi S27. Transcriptional analysis of biofilm associated genes revealed that the waaG gene was significantly downregulated. The ΔwaaG mutant showed a significant decrease in persister cells and a strong biofilm formation relative to wild type and gyrA mutant. The gyrA tetra mutant persister assay revealed a significant increase in persister cells compared to wild type and ΔwaaG. Collectively, this is the first report of S. Typhi's two key genes and their roles in antibiotic tolerance, biofilm formation, and fluoroquinolone resistance that can help in understanding the mechanism of persister formation and eradication.