Thioglycollate Broth (THIO) is an enriched liquid medium for the growth of microaerophilic and anaerobic bacteria, including fastidious organisms, isolated from clinical specimens. Thioglycollate Broth is an excellent growth medium for most sporeforming and non-sporeforming anaerobes.  It is especially useful for the cultivation of non-sporeforming anaerobes. Thioglycollate Broth basal medium is supplemented with hemin and vitamin K1, both of which are heat stable and are autoclaved in the medium with other ingredients.  A small amount of agar is added to slow the diffusion of oxygen within the media.  A marble chip (calcium carbonate) is added to the medium to provide additional pH buffering capacity and a constant supply of carbon.  This medium is prepared, dispensed and packaged under oxygen-free conditions to prevent the formation of oxidized products prior to use.

  Anaerobe Systems PRAS Thioglycollate
                   Broth w/ Hemin and
             Vitamin K w/o indicator (THIO)

 

Formula

Pancreatic Digest of Casein, 15.0 g

Yeast Extract, 5.0 g

Animal Tissue, 3.0 g

Sodium Chloride, 2.5 g

Dextrose, 6.0 g

Sodium Thioglycolate, 0.5 g

Agar, 0.7 g

L-Cystine, 0.25 g

Sodium Sulfite, 0.1 g

Hemin (0.1% soln), 5.0 ml

Vitamin K₁ (1% soln), 0.1 ml

Marble Chip, 1 piece each

Distilled Water, 1000.0 ml

 

Final pH 7.3 +/- 0.3 at 25 degrees C.

Final volume 7.0 mL +/- 0.7 for AS-801.

Final volume 5.0 mL +/- 0.5 for AS-805.

 

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, evaporation, container damage, or contamination.  The expiration date applies to the product in its original outer packaging or stored anaerobically as directed.  The shelf-life of each tube is five months after outer packaging is opened if stored aerobically and as otherwise 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.  Consult appropriate references for detailed instructions concerning collection and transport of anaerobes.

Methods for Use:  Thioglycollate Broth should be inoculated directly with clinical material.  Inoculated tubes should be immediately placed in an anaerobic atmosphere and incubate 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

Thioglycollate Broth will not provide complete information for identification of bacterial isolates.  Additional test procedures and media are required for complete identification.  Consult reference materials for additional information (1-5).  Do not use tubes from opened packages that have been stored under aerobic conditions for more than five months.

 

Quality Control

If used as described (1), this medium should support good growth of essentially all commonly isolated anaerobes from clinical materials, including known strains of Clostridium perfringens, Bacteroides fragilis, Prevotella melaninogenica, and Fusobacterium necrophorum.

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

Organism Tested	ATCC #	Results	Time (Hours)
Bacteroides fragilis	25285	Growth	24 h
Prevotella melaninogenica	25845	Growth	24 – 48 h
Fusobacterium necrophorum	28286	Growth	24 – 48 h
Fusobacterium nucleatum	25586	Growth	24 – 48 h
Clostridium perfringens **	13124	Growth	24 h
Peptostreptococcus anaerobius **	27337	Growth	24 h
Staphylococcus aureus **	25923	Growth	24 h
Bacteroides vulgatus **	8482	Growth	24 h
Clostridium novyi	7659	Growth	48 h

                     ** Organisms specified by NCCLS (5) for quality control testing of anaerobic broth.

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
C. perfringens	24 hrs
P. anaerobius	24 hrs
B. vulgatus	24 hrs

 

Physical Appearance: Thioglycollate Broth should appear as a clear, golden-yellow liquid within the 16mm x 100mm glass tube with a submerged marble chip.

  

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.        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.