Formula

Peptone, 5.0 g

Pepticase, 5.0 g

Yeast Extract, 10.0 g

L-Cysteine, 0.5 g

Hemin (0.1% Soln), 5.0 ml

Vitamin K₁ (1% Soln), 0.1 ml

Resazurin, 0.001 g

Calcium Chloride, anhydrous, 0.01 g

Magnesium Sulfate, anhydrous, 0.008 g

Potassium Phosphate, monobasic, 0.04 g

Potassium Phosphate, dibasic, 0.04 g

Sodium Chloride, 2.0 g

Sodium Bicarbonate, 0.4 g

Glucose, 10.0 g

Distilled Water, 1000.0 ml

 

Final pH 7.3 +/- 0.3 at 25^oC.

Final volume 10.0 ml +/- 1.0.

 

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 packaging until use.  Avoid overheating or freezing.  Do not use medium if there are signs of deterioration (discoloration or evaporation), oxygen exposure, 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:  1 year from date of manufacture.

Procedure

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

Methods for Use:  PYG FA/GLC should be inoculated with pure cultures of an organism.  Inoculated tubes should be immediately placed in an anaerobic atmosphere and incubated at 35 – 37^oC for 24 – 48 hours.  Detailed instructions for processing anaerobic cultures can be found in the MIDI instruction manual (8).

 

Materials Required But Not Provided

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

 

Limitations

PYG FA/GLC will not provide complete information for the identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  Consult reference materials for additional information.

 

Quality Control

Un-inoculated PY-based carbohydrate broth should show only trace amounts, if any, of volatile and nonvolatile fatty acids when analyzed with gas liquid chromatography as described (1, 4).  This medium, when cultured with appropriate control strains such as Bacteroides fragilis and Fusobacterium necrophorum should show characteristic cellular fatty acid products when analyzed using the MIDI MIS system.  For interpretation of chromatographic results consult the MIDI manual (8).

The following organisms are routinely used for quality assurance performance testing, for growth and the fermentation of glucose at Anaerobe Systems.  Glucose utilization is determined on the basis of pH with the production of acid.  A strong acid will result in a pH of 5.5 or below, weak acid results in a pH of 5.5 – 6.0 and no acid production results in a pH of 6.0 or above. 

Organism Tested	ATCC #	Results	Time (Hours)	Utilization of Glucose
Bacteroides fragilis *	25285	Growth	24 h	+
Prevotella melaninogenica	25845	Growth	24 – 48 h
Bacteroides vulgatus	8482	Growth	24 h
Fusobacterium nucleatum	25586	Growth	24 – 48 h
Fusobacterium necrophorum	25286	Growth	24 – 48 h	-
Clostridium perfringens	13124	Growth	24 h
Peptostreptococcus anaerobius	27337	Growth	24 h
Peptoniphilus asaccharolyticus	29743	Growth	24 – 48 h	-
Propionibacterium acnes	6919	Growth	24 – 48 h	+
Staphylococcus aureus	25923	Growth	24 h
Clostridium novyi	7659	Growth	48 h

                   * This organism is tested for characteristic fatty acids results using the MIDI MIS system.

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 media is to be tested for performance, it is recommended that the following ATCC organisms be evaluated for growth.  Glucose utilization is then determined by pH analysis.

 

Organism	Expected Growth	Special Reaction
B. fragilis *	24 hrs	+
F. necrophorum	48 hrs	-
P. asaccharolyticus	48 hrs	-
P. acnes	48 hrs	+

                          * This organism is tested for characteristic fatty acid results using the MIDI MIS system.

 

Physical Appearance:  PYG FA/GLC should appear as a clear, golden-yellow liquid within the 16mm x 100mm glass tube.

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.        Dowell, V. R., Jr., G. L. Lombard, F. S. Thompson and A. Y. Armfield.  1977.  Media for the Isolation, Characterization and Identification of Obligately Anaerobic Bacteria.  USDHEW, CDC, Atlanta, GA 30333.

 

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

 

5.        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.

 

6.        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.

 

7.        Isenberg, H. D.  1992.  Clinical Microbiology Procedures Handbook.  American Society for Microbiology Publishing, Washington DC.

 

8.        Sherlock Microbial Identification System (MIS). Operating Manual, Version 6.  MIDI, Inc.  Newark, DE 19713.