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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Poultry Microbiological Safety & Processing Research » Research » Publications at this Location » Publication #352790

Research Project: Novel Pre-harvest Interventions and Alternatives to Antibiotics to Reduce Foodborne Pathogens

Location: Poultry Microbiological Safety & Processing Research

Title: Immunological reactions to Salmonella FlgK and FliD flagellar proteins by broiler sera

Author
item Yeh, Hung-yueh
item Acosta, Amimee - University Of Puerto Rico
item Serrano, Katherine - University Of Puerto Rico

Submitted to: American Society for Microbiology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 2/22/2018
Publication Date: 6/9/2018
Citation: Yeh, H., Acosta, A.S., Serrano, K.V. 2018. Immunological reactions to Salmonella FlgK and FliD flagellar proteins by broiler sera [abstract]. American Society for Microbiology Meeting.

Interpretive Summary: none

Technical Abstract: Background: Salmonella is the leading foodborne pathogen that causes human acute bacterial gastroenteritis worldwide. Chickens are considered as one of major reservoirs of this bacterium. Because the bacterial flagellum is involved in motility, adhesion, quorum sensing and other virulence activities, the flagellum may be the targets for immune responses by chickens. The flagellum is composed of more than 35 proteins. Two flagellar proteins - FliD and FlgK- were selected for the study due to their exposure to the environment and involvement in colonization in the intestinal mucosa. This communication describes expression and characterization of these two proteins, and their antigenicity of the FliD and FlgK proteins in chickens. Methods: The fliD and flgK genes were amplified by PCR, and the proteins were over-expressed in an Escherichia coli Expression System. The recombinant proteins were purified by a nickel-chelating affinity chromatography, and confirmed by SDS-PAGE analysis, the His tag detection and MALDI-TOF analysis. The recombinant proteins was tested for their antigenicity with ELISA using chicken sera from several geographical locations. Results: The recombinant FliD and FlgK proteins were purified by a His-tag affinity chromatography and had a respective, relative mobility of relevant sizes and positions in SDS-PAGE. Sera from the FlgK and FliD immunized broilers reacted strongly to FlgK and FliD, respectively, indicating that these proteins are immunogenic. Further, we used FliD and FlgK as probe to survey prevalence of anti-Salmonella antibodies in broilers. The ELISA showed 66% of broiler sera reacted to FlgK, while about 38% to FliD. The results implicating that these anti-FlgK antibody may be prevalent in the poultry population. Conclusion: These results provide a rationale for further evaluation of these proteins as vaccine candidates for broiler chickens so that food safety for poultry can be improved. These proteins may also hold important insights for Salmonella commensalism in chickens and pathogenesis in humans.