Isolation and Whole-Genome Sequencing of Environmental Campylobacter

Brittni R. Kelley, J. Christopher Ellis, Doug Hyatt, Dan Jacobson, Jeremiah Johnson

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

As a leading cause of bacterial-derived gastroenteritis worldwide, Campylobacter has a significant impact on human health. In the developed world, most campylobacteriosis cases are attributed to the consumption of undercooked, contaminated poultry; however, it has been shown that Campylobacter can be transmitted to humans through contaminated water and other types of food, including beef and milk. As such, high-resolution microbial source-tracking is essential for health department officials to determine the source(s) of Campylobacter outbreaks. For these reasons, this protocol provides the techniques needed for isolation of Campylobacter from agricultural and environmental sources, as well as human clinical specimens. Additionally, we describe a simple method for preparing high-quality genomic DNA that can be used for whole-genome sequencing and downstream bioinformatics analyses of Campylobacter genotypes.

Original languageEnglish
Article numbere64
JournalCurrent Protocols in Microbiology
Volume51
Issue number1
DOIs
StatePublished - Nov 2018

Funding

This research was supported by a Joint Directed Research and Development (JDRD) program grant R013318090 in collaboration between the University of Tennessee and Oak Ridge National Laboratory. This research used resources of the Oak Ridge Leadership Computing Facility at the Oak Ridge National Laboratory, which is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC05-00OR22725. This research was also supported by the Plant-Microbe Interfaces Scientific Focus Area in the Genomic Science Program, the Office of Biological and Environmental Research (BER) in the U.S. Department of Energy Office of Science, and by the Department of Energy, Laboratory Directed Research and Development funding (ProjectID 8321), at the Oak Ridge National Laboratory. This work was also supported by the Center for Bioenergy Innovation, a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the US DOE under contract DE-AC05-00OR22725. The authors would also like to acknowledge the Tennessee Department of Health for providing human isolates and the University of Tennessee College of Veterinary Medicine for assistance with obtaining animal isolates.

Keywords

  • campylobacter
  • environmental isolation
  • whole-genome sequencing

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