Formula

Protein Mixture, 10.0 g

Agar, 14.0 g

Yeast Extract, 5.0 g

Sodium Bicarbonate, 5.0 g

Sodium Formate, 0.5 g

Sodium Fumarate, 1.0 g

Sodium Succinate, 0.5 g

Potassium Phosphate, dibasic, 2.0 g

Sodium Chloride, 5.0 g

Magnesium Sulfate, 0.1 g

Dextrose, 1.0 g

Volatile Fatty Acid Mix, 3.0 ml

Vitamin K₁ (1% soln), 1.0 ml

Sodium Pyruvate, 0.8 g

Thiamine Pyrophosphate, 0.025 g

L-Cysteine, 0.5 g

Serum, 50.0 ml

Distilled Water, 940.0 ml

 

Final pH 7.3 +/- 0.2 at 25 degrees C.

Final weight 16.0 g +/- 1.6.

 

Precautions

For IN VITRO DIAGNOSTIC USE only.  Approved biohazard precautions and aseptic techniques should be observed when using this product.  This product is for use only by properly-trained and qualified personnel.  Sterilize all biohazard waste prior to disposal.

 

Storage and Shelf Life

Storage:  Upon receipt, store at room temperature (13°C - 27°C) in original container until use.  Avoid overheating or freezing.  Do not use medium if there are signs of deterioration (shrinking, cracking or discoloration due to oxidation of media) or contamination.  The expiration date applies to the product in its original packaging and stored as directed.  Do not use product past the expiration date shown on the container.

Shelf Life:  90 days from date of manufacture.

 

Procedure

Specimen Collection:  Specimens for anaerobic culture should be protected from air (oxygen) during collection, transport and processing.  Consult appropriate references for detailed instructions concerning collection and transport of anaerobes.

Methods for Use:  MTGE should be inoculated directly with clinical material or a broth that has been previously inoculated from clinical material.  Inoculated plates should be streaked to obtain isolated colonies, immediately placed in an anaerobic atmosphere and incubated at 35-37^oC for 18-48 hours.  Additional periods of incubation may be necessary to recover some anaerobes.  Detailed instructions for processing anaerobic cultures can be found in the appropriate references.

 

Materials Required But Not Provided

Standard microbiological supplies and equipment such as loops, saline blanks, slides, staining supplies, microscope, incinerator or autoclave, incubators, anaerobic chamber or anaerobic jar, disinfectant, other culture media and serological and biochemical reagents.

 

Limitations

MTGE will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  In some cases, MTGE may be overgrown with swarming Proteus spp. or Clostridium spp.  It is recommended that selective media such as Anaerobic Brucella Laked Blood Agar with Kanamycin and Vancomycin (LKV) and/or Anaerobic Brucella Blood Agar with Phenylethyl Alcohol (PEA) also be inoculated from clinical specimens to prevent such overgrowth and thus provide isolated colonies.  Consult reference materials for additional information.

 

Quality Control

MTGE should support good growth of many fastidious and non-fastidious anaerobes isolated from clinical specimens.

The following organisms are routinely used for quality control testing at Anaerobe Systems. 

Organism Tested	ATCC #	Results	Time (Hours)
Bacteroides forsythus	1592	Growth	24 – 72 h
Bacteroides ureolyticus	33387	Growth	24 h
Bacteroides fragilis	25285	Growth	24 h
Bacteroides vulgatus	8482	Growth	24 h
Prevotella melaninogenica	25845	Growth	24  h
Fusobacterium necrophorum	25286	Growth	24 h
Fusobacterium nucleatum	25586	Growth	24 h
Clostridium perfringens	13124	Growth	24 h
Peptostreptococcus anaerobius	27337	Growth	24 h
Clostridium novyi, Type A	7659	Growth	24  h
Propionibacterium acnes	6919	Growth	24 h
Staphylococcus aureus	25923	Growth	24 h
Porphyromonas gingivalis	33277	Growth	24 - 48 h
Prevotella intermedia	25611	Growth	24 - 48 h

 

User Quality Control: The final determination to the extent and quantity of user laboratory quality control must be determined by the end user.

If sterility testing is to be performed on this product, the appropriate percentage of the original shipment amount should be incubated anaerobically and aerobically for 48 – 96 hours.

If the nutritive capacity of this medium is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth.

 

Organism	Expected Growth
B. fragilis	24 hrs
P. melaninogenica	24 hrs
F. necrophorum	24 hrs
C. perfringens	24 hrs
P. anaerobius	24 hrs
S. aureus	24 hrs

 

Physical Appearance:  Anaerobic Enrichment Agar should appear opaque to translucent yellow in color.

 

References

1.        Dowell, V. R., Jr. and T. M. Hawkins.  1974.  Laboratory Methods in Anaerobic Bacteriology.  CDC Laboratory Manual.  USDHEW C. D. C. Atlanta, GA 30333.

 

2.        Dowell, V. R., Jr. and G. L. Lombard.  1977.  Presumptive Identification of Anaerobic Non-sporeforming Gram-negative Bacilli.  USDHEW, CDC.  Atlanta, GA 30333.

 

3.        Holdeman, L. V., F. P. Cato and W. E. C. Moore.  1977.  Anaerobe Laboratory Manual.  Virginia Polytechnic Institute and State University.  Blacksburg, VA 24061.

 

4.        Jousimies-Somer, H. R., Summanen, P., Citron, D. M., Baron, E. J., Wexler, H. M. and S. M. Finegold.  2002.  Wadsworth – KTL Anaerobic Bacteriology Manual.  Star Publishing Co., Belmont, CA 94002.

 

5.        Engelkirk, P. G., Duben-Engelkirk, J. and Dowell, V. R.  1992.  Principles and Practices of Clinical Anaerobic Bacteriology.  Star Publishing Co., Belmont, CA 94002.

 

6.        Isenberg, H. D.  1992.  Clinical Microbiology Procedures Handbook.  American Society for Microbiology Publishing, Washington, D.C.

 

7.        NCCLS.  Quality Control for Commercially Prepared Microbiological Culture Media; Approved Standard- Third Edition.  (2004).  NCCLS document M22-A3.  NCCLS, 940 West Valley Road, Suite 1400, Wayne, PA 19087-1898.