Helicobacter pullorum Stanley et al.
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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.
single tube of #18 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.
2. To obtain a biphasic culture, add 0.4 ml of the suspension
to a #260 slant. Add remaining 0.1 ml of the suspension
to a #260 plate and streak for isolation.
3. Incubate at 37°C under microaerophilic conditions using
an anaerobe jar with an active catalyst and a
microaerophilic gas generator pack, or other acceptable
method to obtain microaerophilic conditions. Incubate
slant with cap loose.
using broth pool as the inoculum source.
Growth on agar takes longer than in the biphasic culture. Colonies appear as a haze to pinpoint in size. More mature colonies will appear slightly larger with some spreading. Once good growth is present, these organisms tend to lose viability, especially if exposed to air for lengthy periods. Viability also decreases with repeated subculturing. The cells do not Gram stain well using traditional procedures. To obtain the best results, use a basic fuchsin counterstain in place of safranin.
Once good growth is obtained, transfer or freeze the culture. Adding an equal amount of 20% sterile glycerol to pooled broth from several biphasic slants, followed by freezing in liquid nitrogen or "ultra-low temperature" freezer is recommended.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.
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Stanley J, et al. Helicobacter pullorum sp. nov.-genotype and phenotype of a new species isolated from poultry and from human patients with gastroenteritis. Microbiology 140: 3441-3449, 1994. PubMed: 7533595
feces of human with gastroenteritis, Berne