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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Poultry Microbiological Safety and Processing Research Unit » Research » Publications at this Location » Publication #368983
Research Project: Novel Pre-harvest Interventions and Alternatives to Antibiotics to Reduce Foodborne Pathogens

Location: Poultry Microbiological Safety and Processing Research Unit

Title: Detection of Campylobacter jejuni diversity by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) from poultry sources

Author
item Yeh, Hung-Yueh
item AWAD, AMAL - Mansoura University
item Rothrock, Michael

Submitted to: Veterinary Medicine and Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/18/2021
Publication Date: 9/12/2021
Citation: Yeh, H., Awad, A., Rothrock Jr, M.J. 2021. Detection of Campylobacter jejuni diversity by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) from poultry sources. Veterinary Medicine and Science. https://doi.org/10.1002/vms3.622.
DOI: https://doi.org/10.1002/vms3.622

Interpretive Summary: Campylobacter jejuni is the leading bacterial pathogen that causes foodborne illness worldwide. Because of fastidious growth and sophisticated biochemical requirements of Campylobacter jejuni, several genotyping methods have been investigated to classify this bacterium during the outbreaks. One of such method is to use clustered regularly interspaced short palindromic repeats (CRISPR). The goal of this study was to apply CRISPR to explore the genetic diversity of Campylobacter jejuni isolates from the U.S. organic poultry farms. Seventy-seven Campylobacter jejuni isolates were used in this study. Genomic DNA were isolated using a commercial kit. The quality and quantity of DNA were determined by agarose gel electrophoresis and a spectrophotometer, respectively. The PCR amplification of CRISPR type 1 was previously described. The amplicons were sequenced by the Sanger dideoxy sequencing method. The direct repeats (DR) and spacers of the CRISPR sequences were identified using the CRISPRFinder. The CRISPR sequences were detected in all 77 isolates. One type of DR was identified in these 77 isolates. The lengths of the CRISPERs ranged from 100 to 560 nucleotides, while the number of spacers ranged from one to eight. Further analysis of spacer sequences, a total of 266 sequences were found and 67 distinctive sequences were identified in 77 Campylobacter jejuni isolates. However, by comparison with known spacer sequences, we observed that 18 from 67 sequences were known previously. In conclusions, our results provide a rationale for further evaluation of the CRISPR genotyping on Campylobacter jejuni for tracking outbreaks and evolution of this bacterium.

Technical Abstract: Background: Campylobacter jejuni is the leading bacterial pathogen that causes foodborne illness worldwide. Because of fastidious growth and sophisticated biochemical requirements of Campylobacter jejuni, several genotyping methods have been investigated to classify this bacterium during the outbreaks. One of such method is to use clustered regularly interspaced short palindromic repeats (CRISPR). Objectives: The goal of this study was to apply CRISPR to explore the genetic diversity of Campylobacter jejuni isolates from the U.S. organic poultry farms. Methods: Seventy-seven Campylobacter jejuni isolates were used in this study. Genomic DNA were isolated using a commercial kit. The quality and quantity of DNA were determined by agarose gel electrophoresis and a spectrophotometer, respectively. The PCR amplification of CRISPR type 1 was previously described. The amplicons were sequenced by the Sanger dideoxy sequencing method. The direct repeats (DR) and spacers of the CRISPR sequences were identified using the CRISPRFinder. Results: The CRISPR sequences were detected in all 77 isolates. One type of DR was identified in these 77 isolates. The lengths of the CRISPERs ranged from 100 to 560 nucleotides, while the number of spacers ranged from one to eight. Further analysis of spacer sequences, a total of 266 sequences were found and 67 distinctive sequences were identified in 77 Campylobacter jejuni isolates. However, by comparison with known spacer sequences, we observed that 18 from 67 sequences were known previously. Conclusions: Our results provide a rationale for further evaluation of the CRISPR genotyping on Campylobacter jejuni for tracking outbreaks and evolution of this bacterium.