Tryptic Soy Blood Agar (TSBA) is intended for the isolation and cultivation of many fastidious and non-fastidious microorganisms found in clinical specimens.  This medium is also suitable for use in the CAMP test for the presumptive identification of group B. Streptococci. TSBA is an enriched non-selective medium, supplemented with sheep blood for the hemolytic reactions.  It is supplemented with hemin and vitamin K1 to improve the recovery of fastidious anaerobes. This medium is prepared, stored and dispensed under oxygen-free conditions to prevent the formation of oxidized products prior to use.

     Anaerobe Systems PRAS Tryptic Soy Blood
                        Agar (TSBA) AS-542

Formula

Pancreatic Digest of Casein, 15.0 g

Soy Peptone, 5.0 g

Sodium Chloride, 5.0 g

Hemin (0.1% soln), 5.0 ml

Vitamin K₁ (1% soln), 1.0 ml

Agar, 15.0 g

Sheep Blood, 45.5 ml

Distilled Water, 1000.0 ml

 

Final pH 7.1 +/- 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:  TSBA 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 / autoclave, incubators, anaerobic chamber or anaerobic jars, disinfectant, other culture media and serological and biochemical reagents.

 

Limitations

TSBA will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  This medium may not support the growth of all clinically significant anaerobes.  Consult reference materials for additional information.

 

Quality Control

TSBA should support good growth of fastidious microorganisms found in clinical infections.  In addition, this media should support typical double zone of hemolysis around colonies of Clostridium perfringens.

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

Organism Tested	ATCC #	Results	Time (Hours)	Special Reaction
Bacteroides fragilis **	25285	Growth	24 h
Prevotella melaninogenica **	25845	Growth	48 h
Fusobacterium nucleatum **	25586	Growth	24 h
Clostridium perfringens **	13124	Growth	24 h	Double Zone of hemolysis
Peptostreptococcus anaerobius **	27337	Growth	24 h
Staphylococcus aureus   or    Enterococcus faecalis	25923     29212	Growth	24 h
Escherichia coli	25922	Growth	24 h
Proteus mirabilis	12453	Growth	24 h
Propionibacterium acnes   or  Clostridium difficile	6919      9686	Growth	24 – 48 h 24 h

** Organisms specified by NCCLS for quality control of Anaerobic Blood Agars

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	Growth in Hours	Expected Results
B. fragilis	24 hrs	-
C. perfringens	24 hrs	Double zone of b -hemolysis
P. anaerobius	24 hrs	-
E. faecalis	24 hrs	-

 

Physical Appearance: TSBA should appear opaque burgundy red 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.        Holdeman, L. V., F. P. Cato and W. E. C. Moore.  1977.  Anaerobe Laboratory Manual.  Virginia Polytechnic Institute and State University.  Blacksburg, VA 24061.

 

3.        Somer- Jousimies, 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.

 

4.        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.

 

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.