Brachyspira alvinipulli Stanton et al.

1. Thaw vial at room temperature.  Under anaerobic conditions, aseptically withdraw entire contents of vial and transfer into a tube of #1827 broth.
2. Additional tubes of #1827 broth or slants can be inoculated with 0.5 ml each of the suspension, making a biphasic culture in the slants.  Inoculate two sheep blood agar plates.
3. Incubate the tubes under an anaerobic atmosphere at 37°C for 2-3 days.  One of the sheep blood agar plates is incubated under anaerobic conditions and the second plate is incubated aerobically to check for purity.
4. Growth can be observed as light turbidity in the broth and as a thin film on the slants and #260 plate. Viable growth can also be observed by looking for actively motile helical cells when examining a wet mount of a drop from the broth portion of the biphasic slant under phase contrast.  The aerobic plate should show no signs of growth.

ANAEROBIC CONDITIONS:

Anaerobic conditions for transfer may be obtained by either of the following:

• Use of an anaerobic gas chamber, or
• Placement of test tubes under a gassing cannula system hooked to anaerobic gas.

Anaerobic conditions for incubation may be obtained by any of the following:

• Loose screw caps on test tubes in anaerobic chamber,
• Loose screw caps on test tubes in an activated anaerobic gas pack jar, or   
• Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.  

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 If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.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.