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United States Department of Agriculture

Agricultural Research Service

Research Project: Pre-Harvest Interventions For Application During Poultry Production To Reduce Food-Borne Bacterial Pathogens

Location: Poultry Microbiological Safety Research

Title: Characterization and Reactivity of Broiler Chicken Sera to Selected Recombinant Campylobacter jejuni Chemotactic Proteins

Authors
item Yeh, Hung-Yueh
item Hiett, Kelli
item Line, John
item Seal, Bruce

Submitted to: Archives Of Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: February 24, 2014
Publication Date: May 1, 2014
Citation: Yeh, H., Hiett, K.L., Line, J.E., Seal, B.S. 2014. Characterization and Reactivity of Broiler Chicken Sera to Selected Recombinant Campylobacter jejuni Chemotactic Proteins. Archives Of Microbiology. 196:375-383.

Interpretive Summary: Campylobacter jejuni is an important food-borne and water-borne pathogen, which causes human acute bacterial gastroenteritis worldwide. Poultry products are the major source of this bacterium for human infection. Vaccination of chickens is one of powerful means to reduce this bacterium in broiler chicken gut and minimize contamination in poultry. In order to survive in environments, C. jejuni develop a sense system with flagella to guide themselves toward or away from various locations. There are many proteins involving in this complicated process, and thus, these proteins can be targets for vaccine development. Molecular technique was used to produce a large amount of these proteins. A total of 15 proteins were successfully produced. These recombinant proteins were confirmed by nucleotide sequencing, SDS-PAGE analysis and immunoblot analysis of six-His and hemagglutinin (HA) tags. Twelve recombinant proteins were further tested whether they were antigenic using sera from broilers older than four weeks. The immunoblot results show that each chicken serum reacted to various numbers of recombinant proteins, but all sera reacted to the Cjj0473 (annotated as methyl-accepting chemotaxis protein), suggesting that anti-Campylobacter antibodies may be prevalent in the poultry population. These antibody screening results provide a rationale for further evaluation of the Cjj0473 protein as a potential vaccine for broilers to improve food safety. This study is related to the goal of ARS National Program 108-Food Safety, Component 1D-Intervention and Control Strategies.

Technical Abstract: Campylobacter jejuni, a Gram-negative rod bacterium, is the leading causative agent of human acute bacterial gastroenteritis worldwide. Consumption and handling of raw or undercooked poultry are regarded as a major source for human infection. Because bacterial chemotaxis guides microorganisms to colonization and invasion in the host cells, proteins involved in chemotactic process can be novel targets for vaccine development. In this communication, we report amplification, cloning and expression of the C. jejuni chemotactic proteins in an Escherichia coli expression system. A total of 15 chemotactic protein genes were successfully expressed. These recombinant proteins were confirmed by nucleotide sequencing, SDS-PAGE analysis and immunoblot analysis of six-His and hemagglutinin (HA) tags. Twelve recombinant chemotactic proteins were further tested whether they were antigenic using sera from broiler chickens older than four weeks. The immunoblot results show that each chicken serum reacted to various numbers of recombinant proteins, but all sera reacted to the Cjj0473 (annotated as methyl-accepting chemotaxis protein), suggesting that anti-Campylobacter antibodies may be prevalent in the poultry population. These antibody screening results provide a rationale for further evaluation of the Cjj0473 protein as a potential vaccine for broilers to improve food safety.

Last Modified: 11/24/2014
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