Campylobacter coli (Doyle) Veron and Chatelain
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Using a single tube of #1423 or #1115 or #177 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a #260 slant, and/or plate.
5. Incubate tubes and plate at 37oC, under microaerophilic conditions, for 48 hours. Use an anaerobe jar with an active catalyst and a Campylobacter microaerophilic gas generator pack, or other acceptable method. Incubate slant with cap loose.
This is a slower growing organism that requires moist conditions for best growth. To observe growth, examine a wet mount of the broth under phase microscopy. The organism is a small curved motile rod. Colonies are circular, entire, gray, and a raised center. Once good growth is present, these organisms tend to lose viability, especially if exposed to air for lengthy periods. The cells do not Gram stain well using traditional procedures. To obtain the best results, use a basic fuchsin counterstain in place of the safranin.
Storage at liquid nitrogen temperatures, with 10% sterile glycerol as the cryoprotectant, is recommended for long-term preservation.
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Grajewski BA, et al. Development of a bacteriophage typing system for Campylobacter jejuni and Campylobacter coli. J. Clin. Microbiol. 22: 13-18, 1985. PubMed: 4019737