Laboratory Maintenance of Fusobacteria

Yiping W. Han1

1 Case Western Reserve University, Cleveland, Ohio
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 13A.1
DOI:  10.1002/9780471729259.mc13a01s00
Online Posting Date:  January, 2006
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Abstract

This unit describes routine laboratory handling of fusobacteria. Different media that can be used to grow or enrich Fusobacterium nucleatum and other species of this genus are described. The growth and stock conditions as well as the susceptibility of F. nucleatum to oxygen in a pure culture are also discussed.

Keywords: fusobacteria; anaerobe; periodontal disease; preterm birth; adherence; coaggregation; dental plaque; oxygen sensitivity

     
 
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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Growth of Pure Fusobacterial Cultures
  • Basic Protocol 2: Enrichment of Fusobacteria from a Mixed Population
  • Basic Protocol 3: Storage of Fusobacteria
  • Reagents and Solutions
  • Commentary
  • Literature Cited
     
 
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Materials

Basic Protocol 1: Growth of Pure Fusobacterial Cultures

  Materials
  • Columbia blood agar plates (see recipe)
  • Columbia broth (see recipe)
  • Fusobacterium cultures (clinical isolates or ATCC strains)
  • Anaerobic incubator: 37°C anaerobic chamber, desiccant, palladium catalyst pellets, nitrogen gas tank, gas mix (90% N 2, 5% CO 2, and 5% H 2) tank—e.g., Coy or Forma or
  • Anaerobic jar system (e.g., 2.5‐liter Pack‐Rectangular Jar, AnaeroPack System; Mitsubishi Gas Chemical Co.; http://www.mgc‐a.com/Pages/anaeropac.html)
  • Sachets for anaerobic jar system (Mitsubishi Gas Chemical Co.)
  • Anaerobic indicator strips (Becton Dickinson)

Basic Protocol 2: Enrichment of Fusobacteria from a Mixed Population

  Materials
  • Reduced transport fluid (RTF; see recipe)
  • FEA or CVE plates (see reciperecipes)
  • Dental plaque (provided by a dental care professional)
  • Anaerobic incubator: 37°C anaerobic chamber, desiccant, palladium catalyst pellets, nitrogen gas tank, gas mix (90% N 2, 5% CO 2, and 5% H 2) tank—e.g., Coy or Forma or
  • Anaerobic jar system (e.g., 2.5‐liter Pack‐Rectangular Jar, AnaeroPack System; Mitsubishi Gas Chemical Co.; http://www.mgc‐a.com/Pages/anaeropac.html)
  • Sachets for anaerobic jar system (Mitsubishi Gas Chemical Co.)
  • Anaerobic indicator strips (Becton Dickinson)

Basic Protocol 3: Storage of Fusobacteria

  Materials
  • Fusobacterium cultures
  • Columbia broth (see recipe)
  • Glycerol (autoclaved)
  • Anaerobic chamber or anaerobic jar (see protocol 1)
  • 1.5‐ml microcentrifuge tubes, sterile
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Figures

Videos

Literature Cited

Literature Cited
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