Helicobacter bilis Fox et al.

1. Open vial according to enclosed instructions. Rehydrate contents of vial with 0.5 mL of #18 broth. 
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.
4. Within three to five days of incubation, good growth should be obtained in the broth pool at the bottom of the slant. Secondary subcultures to fresh biphasic slants require shorter incubation times, 24 to 48 hours. Additional incubation may be required for colonies to appear on the plate. Further subcultures can be made using broth pool as the inoculum source.

Individual colonies will not be observed.  Growth appears as a film on the surface of fresh, moist plates, extending out from original inoculation site to fill entire plate, usually within 48 to 72 hours.

The cells do not Gram stain well using traditional procedures.  For best results, use a basic fuchsin counterstain in place of the 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.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.