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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR BIOLOGY AND GENOMICS OF FOODBORNE PATHOGENS Title: Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulse-Field Gel Electrophoreis, MLST and rep-PCR Reveals Different Discriminatory Capabilities

Authors
item Wilson, Melissa - UNIV. OF ILLINOIS
item Lane, Allison - WASHINGTON STATE UNIV.
item Law, Bibiana - UNIV. OF ARIZONA
item Miller, William
item Joens, Lynn - UNIV. OF ARIZONA
item Konkel, Michael - WASHINGTON STATE UNIV.
item White, Bryan - UNIV. OF ILLINOIS

Submitted to: Microbial Ecology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: July 22, 2009
Publication Date: August 21, 2009
Citation: Wilson, M.K., Lane, A.B., Law, B.F., Miller, W.G., Joens, L.A., Konkel, M.E., White, B.A. 2009. Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulse-Field Gel Electrophoreis, MLST and rep-PCR Reveals Different Discriminatory Capabilities. Microbial Ecology. DOI 10, 1007/s00248-009-9571-3

Interpretive Summary: Campylobacter jejuni is one of the leading bacterial causes of food-borne illness in the United States. Typing methods are often used in food safety in source tracking sporadic illness and outbreaks. Moreover, the identification of genetically related strains may facilitate the development of intervention strategies for control and prevention of food-borne diseases. This study compared three different typing methods. Sixty C. jejuni isolates from chickens reared in three different production environments were analyzed. Specifically, the C. jejuni recovered from chickens reared in conventional, organic, and free-range poultry flocks were analyzed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and repetitive sequence polymerase chain reaction (rep-PCR). Comparison of these genotypic methods revealed that rep-PCR had the greatest ability to distinguish closely related strains. Regardless of the method used to genotype the C. jejuni isolates, chickens reared in the conventional, organic, and free-range environments were all found to exhibit a high level of genotypic diversity.

Technical Abstract: Campylobacter jejuni is one of the leading bacterial causes of food-borne illness in the United States. Molecular genotyping methods are often used in food safety for identifying sources of infection and pathways of transmission. Moreover, the identification of genetically related strains (i.e., clades) may facilitate the development of intervention strategies for control and prevention of food-borne diseases. The goal of this study was to determine if the environment used to raise chickens resulted in a difference in the genotypes of resident Campylobacter jejuni as determined by three different genotyping methods. To gain insight into the genetic diversity, and pan, core and variable genome distributions of isolates within a defined setting, we analyzed a total of sixty C. jejuni isolates from chickens reared in three different production environments. Specifically, the C. jejuni recovered from chickens reared in conventional, organic, and free-range poultry flocks were analyzed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and repetitive sequence polymerase chain reaction (rep-PCR). Comparison of the three genotypic methods revealed that rep-PCR had the greatest ability to distinguish closely related strains. Genetic fingerprints were subjected to computer-assisted pattern analysis. ERIC (Enterobacterial Repetitive Intergenic Consensus)- and BOX-PCR yielded the highest number of PCR products and greatest reproducibility. Regardless of the method used to genotype the C. jejuni isolates, chickens reared in the conventional, organic, and free-range environments were all found to exhibit a high level of genotypic diversity.

Last Modified: 11/23/2014
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