Phenotypic and genotypic characterization of Shigella spp. with reference to its virulence genes and antibiogram analysis from river Narmada.

Water samples of the river Narmada from origin to end were analyzed for the presence of shigellae. Analysis of 40 water samples by biotyping, serotyping, and molecular typing were done. Out of all 40 isolates, 23 were identified as Shigella flexneri, 10 as Shigella sonnei, and seven as Shigella dysenteriae. Serotyping was found to be a better identification method than biotyping since biotyping was not found to be very sharp. In the present investigation, amplified ribosomal DNA restriction analysis (ARDRA) with a probe complementary to 16S rRNA was performed. Repeated ARDRA analysis establishes the similarities between the isolates and thus suggested ARDRA as authentic and precise detection protocol. The isolates were also analyzed for the presence of virulence genes including ipaBCD, ipaH, and stx1 and all the 40 isolates of Shigella showed positive result for ipaH gene while the plasmid encoded invasion-associated genes ipaBCD was present only in S. flexneri and stx1 gene was present only in S. dysenteriae. This study demonstrated the existence of Shigella in the river Narmada and dispersion of different virulence genes among these isolates, which appear to constitute an environmental reservoir of Shigella-specific virulence genes.