Salmonella Typhimurium in Iran: Contribution of molecular and IS200 PCR methods in variants detection.

Salmonella Typhimurium, a zoonotic pathogen, is regarded as a major health and economic concern worldwide. Recently, monophasic variants of this serovar have been significantly associated with human gastroenteritis outbreaks globally, making its accurate identification essential for epidemiological and control purposes. We have identified and analyzed 150 S. Typhimurium from 884 Salmonella genus isolated from humans, domestic animals, poultry, food items and abattoirs origins. The Salmonella isolates were obtained from Iranian National Veterinary Reference Laboratories of 9 provinces during 2007-2016, and from five hospitals in Tehran in 2015. The isolates were evaluated biochemically, serologically, and by PCR amplification of invA, mdh, STM4492, fliC, fljA, fljB, hin genes, IS200 and DT104. invA and mdh genes were used to confirm the S. Typhimurium serotype, fliC and fljB genes for determination of monophasic variants and amplification of IS200 to discriminate the monophasic variants from the closely related serotypes. We identified 78.6% (118/150) as classical S. Typhimurium (fliC, fljB and IS200 positive), 12.6% (19/150) were IS200 negative from all isolates. DT104 is another marker for S.Typhimurium serovar typing. Contrary to EFSA guidelines 20.6% (19/29) of human isolates that lacked IS200 insertion sequence, were confirmed as S.Typhimurium. Compared to the North American/European isolates the low prevalence of fljB negative 6% (9/150) and the high abundance of fliC negative 23.3% (35/150) isolates also were indicative of a different regional atypical population. Studies have shown that the prevalence of monophasic (fljB-) S. Typhimurium worldwide is promoted by the Swine industry. Thus, one reason for this high number of different atypical strains could be inhibition of swine breeding system (house hold and industry) in Iran. These results demonstrate a need for a modified identifying protocol to overcome the regional differences.

Immune responses to oral and IM administration of M2e-Hsp70 construct.

The eukaryotic expression plasmid of M2e-Hsp70 is a candidate M2e-based DNA vaccine. In order to evaluate the immunization potential of this construct, Specific Pathogen Free chickens were immunized either intramuscularly or orally. Mutant Salmonella typhimurium was used as carrier for oral delivery of the M2e-Hsp70 construct. M2e-specific humoral and cellular responses were tested by ELISA and Lymphocyte Proliferation Assay, respectively. Our results indicate that both humoral and cellular immune responses are conferred against M2e-Hsp70 plasmid in either of the intramuscular or oral routes of administration; however, these responses are significantly higher in intramuscular injection in contrast to oral administration. When it comes to mass vaccination of commercial chicken flocks oral administration is preferred due to the ease of application as well as its capability of eliciting mucosal, humoral and cellular immune responses; so measurements should be taken to improve the immunization potency of our orally delivered DNA vaccine.