Molecular characterization of Salmonella spp. isolates from river and dam water, irrigated vegetables, livestock, and poultry manures in Jordan

Background and Aim: Salmonellosis is an important food-borne and zoonotic disease with high morbidity and mortality rates. The objectives of this study were to isolate, serotype, and genetically characterize Salmonella spp. from Zarqa river and King Talal dam waters, vegetables irrigated by such waters, and manure of poultry and livestock farms located in the Zarqa river basin in Jordan. In addition, certain virulence factors and antimicrobial resistance patterns of isolated Salmonella strains were determined. Materials and Methods: A total of 250 samples were cultured using routine microbiological methods. Suspected Salmonella spp. were identified based on colony morphology and confirmed using biochemical and molecular methods. Virulence genes including invA, stn, and pCT plasmid were detected using multiplex PCR. Phylogenetic analysis was performed using pulsed-field gel electrophoresis (PFGE). Results: In total, 32/250 (12.8%) Salmonella spp. isolates were recovered from different sources. Of these, the most common serotype was Salmonella subspecies 1 (23 isolates), followed by Salmonella enterica serovar Typhimurium (4 isolates), Salmonella enterica serovar Typhi (3 isolates), and finally Salmonella enterica serovar Enteritidis (2 isolates). The PFGE indicated that Salmonella enterica serovar Typhimurium isolated from poultry manure and from parsley were closely related (84.6%). Salmonella enterica serovar Enteritidis isolated from the dam water was closely related to Salmonella enterica serovar Enteritidis isolated from spearmint (73.8%). Salmonella enterica serovar Typhi isolated from the river and dam water were 100% related to Salmonella enterica serovar Typhi isolated from lettuce. In the antimicrobial sensitivity test, 14 out of 32 (43.8%) isolated Salmonella strains were resistant to two or more of the major antimicrobial agent groups. However, the majority of isolates were sensitive to ceftriaxone, ciprofloxacin, cefuroxime, and gentamicin (97%, 93.8%, and 87.5%, 84.4%, respectively). All isolates were resistant to erythromycin and amoxicillin. Conclusion: Results of this study indicate a serious potential threat to public health associated with consuming leafy green vegetables grown on the banks of Zarqa river and its dam because of widespread Salmonella spp. contamination. Appropriate monitoring of irrigation water must be applied to reduce the possibility of cross-contamination