Growth and Laboratory Maintenance of Campylobacter jejuni

Lindsay Davis1, Victor DiRita2

1 Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, 2 Unit for Laboratory Animal Medicine, University of Michigan, Ann Arbor, Michigan
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 8A.1
DOI:  10.1002/9780471729259.mc08a01s10
Online Posting Date:  August, 2008
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Abstract

Campylobacter jejuni is a fastidious organism, growing in microaerophilic conditions with a temperature range between 37° and 42°C. Multiple types of media can be used to cultivate it; however, Mueller Hinton broth and agar support the best C. jejuni growth. Optimum atmosphere for C. jejuni is 85% N2, 10% CO2, and 5% O2. Curr. Protoc. Microbiol. 10:8A.1.1‐8A.1.7. © 2008 by John Wiley & Sons, Inc.

Keywords: campylobacteriosis; microaerophilic growth; Mueller Hinton

     
 
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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Growth of C. jejuni from a Frozen Stock
  • Basic Protocol 2: Preservation of C. jejuni
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Growth of C. jejuni from a Frozen Stock

  Materials
  • C. jejuni frozen stock ( protocol 2)
  • 100 × 15–mm Mueller Hinton (MH) agar plates (BD Biosciences, cat. no. 22520; plates are poured in lab) containing 10 µg/ml trimethoprim (antibiotics added in lab)
  • Mueller Hinton (MH) broth (BD Biosciences, cat. no. 275730) containing 10 µg/ml trimethoprim
  • Equipment for maintaining Campylobacter‐specific microaerophilic atmosphere (see )
  • Additional reagents and equipment for streaking bacteria ( appendix 4A)

Basic Protocol 2: Preservation of C. jejuni

  Materials
  • C. jejuni organisms (cannot be purchased from ATCC or similar sources; must be obtained from individual labs)
  • 100 × 15–mm Mueller Hinton (MH) agar plates (BD Biosciences, cat. no. 22520; plates are poured in lab) containing 10 µg/ml trimethoprim (antibiotics added in lab)
  • Mueller Hinton (MH) broth (BD Biosciences, cat. no. 275730) containing 20% (v/v) glycerol
  • Sterile cotton swabs
  • 2‐ml cryotubes
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Figures

Videos

Literature Cited

Literature Cited
   Bacon, D. 2000. Involvement of a plasmid in virulence of Campylobacter jejuni 81‐176. Infect. Immun. 68:4384‐4390.
   Gaynor, E. 2004. The genome‐sequenced variant of Campylobacter jejuni NCTC11168 and the original clonal clinical isolate differ markedly in colonization, gene expression, and virulence‐associated phenotypes. J. Bacteriol. 186:503‐517.
   Hazeleger, W.C. 1998. Physiological activity of Campylobacter jejuni far below the minimal growth temperature. Appl. Environ. Microbiol. 64:3917‐3922.
   Joshua, G.W., Guthrie‐Irons, C., Karlyshev, A.V., and Wren, B.W. 2006. Biofilm formation in Campylobacter jejuni. Microbiology 152:387‐396.
   Leach, S. 1997. Changes with growth rate in the membrane lipid composition of and amino acid utilization by continuous cultures of Campylobacter jejuni. J. Appl. Microbiol. 82:631‐640.
   Ng, L.‐K. 1985. Comparison of basal media for culturing Campylobacter jejuni and Campylobacter coli. J. Clin. Microbiol. 21:226‐230.
   Young, K., Davis, L.M., and DiRita, V.J. 2007. Campylobacter jejuni: Molecular biology and pathogenesis. Nat. Rev. Microbiol. 5:665‐679.
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