Prevotella melaninogenica (Oliver and Wherry) Shah and Collins emend. Wu et al.

1. Open vial according to enclosed instructions.                            
2. Under anaerobic conditions, withdraw 0.5 mL of #2863 from a single test tube (5 to 6 mL) and rehydrate the entire vial contents.   
3. Aseptically transfer this aliquot back into the broth tube. A brucella blood plate and a secondary broth maybe inoculated.  An aerobic blood plate may also be streaked to check for purity.
4. Incubate tubes and one plate under anaerobic conditions at 37°C.  Incubate one blood plate aerobically at 37°C.
5. Within 24 to 48 hours, growth is evident by turbidity in the broth and colonies on an agar plate.  No growth should occur on the blood agar plate incubated aerobically.

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 connected 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.   

If grown on Anaerobic Brain Heart Infusion agar with 5% Laked (hemolyzed) Blood and supplemented with Vitamin K and Hemin, the black pigment production will be stronger and seen earlier. 

Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions.  Resazurin in the media is a color indicator for anaerobic conditions.  Observance of a pink color in medium before use, or during incubation, shows anaerobic conditions have not been met and oxidation has occurred.  Medium should be discarded.

Purified genomic DNA of this strain is available as ATCC^® 25845D-5™.

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.