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
Ammonium Formate, 1.8
g
Fumaric Acid, free acid, 1.8 g
Distilled Water, 1000.0
ml
Final pH 7.2 +/- 0.3 at 25 degrees C.
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-FF
FA/GLC should be inoculated with
pure cultures of an organism. Inoculated
tubes should be immediately placed in an anaerobic atmosphere and incubate at
35 – 37oC for 24 – 48 hours.
Detailed instructions for processing anaerobic cultures can be found in
the MIDI instruction manual.
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-FF 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
This medium, when cultured with Bacteroides ureolyticus should show characteristic cellular fatty
acid products when analyzed using the MIDI MIS system. For interpretation of chromatographic results
consult the MIDI manual.
The following organisms are routinely used for
quality assurance testing, for growth, at Anaerobe Systems. Formate/fumarate utilization is determined on
the basis of growth stimulation when compared to Peptone Yeast Extract Broth with
Glucose (PYG).
|
Organism Tested
|
ATCC #
|
Results
|
Time (Hours)
|
Growth Stimulation
|
|
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
|
|
|
Bacteroides ureolyticus *
|
33481
|
Growth
|
24 – 48 h
|
+
|
|
Peptoniphilus asaccharolyticus
|
29743
|
Growth
|
24 – 48 h
|
|
* Organisms recommended by
MIDI MIS Manual for quality control.
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/stimulation capacity of this medium is to be tested for
performance, it is recommended that the following ATCC organisms be evaluated
for growth/stimulation.
|
Organism
|
Growth in
|
Growth
|
|
|
Hours
|
Stimulation
|
|
B. fragilis
|
24 hrs
|
|
|
C. perfringens
|
24 hrs
|
|
|
F. necrophorum
|
24 - 48 hrs
|
|
|
B. ureolyticus
|
24 - 48 hrs
|
+
|
Physical Appearance: PYG-FF 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.
