THIOGLYCOLLATE MEDIA

Cat. no. K21 Thioglycollate with Indicator, 16x125mm Tube, 10ml 20 tubes/box

Cat. no. K22 Thioglycollate with H & K, 16x100mm Tube, 5ml 20 tubes/box

Cat. no. K24 Thioglycollate with H & K, 16x125mm Tube, 10ml 20 tubes/box

Cat. no. K23 Thioglycollate with Supplements, 16x125mm Tube, 10ml 20 tubes/box

Cat. no. K28 Thioglycollate with Supplements, 20x125mm Tube, 15ml 20 tubes/box

Cat. no. K29 Thioglycollate without Indicator, 16x125mm Tube, 10ml 20 tubes/box

INTENDED USE
Hardy Diagnostics Thioglycollate Media is recommended for the cultivation of aerobic, microaerophilic, and anaerobic
microorganisms.

SUMMARY
The addition of a small amount of agar in Thioglycollate Medium aids in the initiation and growth of small inocula and
anaerobes by impeding the diffusion of oxygen into the medium. It also retards the dispersion of CO2 and the reducing
substance from the microenvironment surrounding the inoculum. Sodium Thioglycollate is a reducing agent which
maintains a low oxygen tension by removing molecular oxygen from the environment. Peroxides, which may be lethal
to many anaerobic organisms, are not formed under this condition. Cystine and casein supply carbon and nitrogenous
compounds, dextrose is added as another energy source, and sodium chloride maintains osmotic equilibrium.

Certain additives are incorporated into the Thioglycollate Medium as desired. Resazurin is an oxidation-reduction
indicator that turns pink when increased oxidation has occurred. Yeast extract or papaic digest of soybean meal are
added as growth enhancers. Hemin is incorporated to supply X-factor for the stimulated growth of many fastidious
organisms, and vitamin K because it is a growth requirement for some gram-positive spore-formers and Bacteroides
species. The calcium carbonate chip is added to act as a buffer for the medium and prevents a buildup of toxic acid.

FORMULA
Ingredients per liter of deionized water:*

Thioglycollate with Indicator:

Pancreatic Digest of Casein 15.0gm

Dextrose 5.5gm

Yeast Extract 5.0gm

Sodium Chloride 2.5gm

 Sodium Thioglycollate 0.5gm

L-Cystine 0.5gm

Resazurin 1.0mg

Agar 0.75gm

Final pH 7.1 +/- 0.3 at 25ºC.

Thioglycollate without Indicator, Thioglycollate with H & K,

Thioglycollate with Supplements:

Pancreatic Digest of Casein 15.0gm

Dextrose 5.0gm

Yeast Extract 5.0gm

Sodium Chloride 2.5gm

Sodium Thioglycollate 0.5gm

L-Cystine 0.25gm

Agar 0.75gm

In addition;

Thioglycollate with H & K contains:

Hemin 5.0mg/L

Vitamin K 1.0mg/L

Thioglycollate with Supplements contains:

Hemin 5.0mg/L

Vitamin K 1.0mg/L

Calcium Carbonate Chip 1/tube

Final pH 7.2 +/- 0.3 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Media should not be used if there are any signs of
deterioration, discoloration, contamination, or if the expiration date has passed. Product is light and temperature
sensitive; protect from light, excessive heat, moisture, and freezing.

The expiration date on the product label applies to the product in its intact packaging when stored as directed. The
product may be used and tested up to the expiration date on the product label and incubated for the recommended
incubation times as stated below.

Refer to the document "Storage" for more information.

PRECAUTIONS
This product may contain components of animal origin. Certified knowledge of the origin and/or sanitary state of the
animals does not guarantee the absence of transmissible pathogenic agents. Therefore, it is recommended that these
products be treated as potentially infectious, and handle observing the usual Universal Precautions for blood. Do not
ingest, inhale, or allow to come into contact with skin.

This product is for in vitro diagnostic use only. It is to be used only by adequately trained and qualified laboratory
personnel. Observe approved biohazard precautions and aseptic techniques. All laboratory specimens should be
considered infectious and handled according to "standard precautions." Refer to the document "Guidelines for Isolation
Precautions" from the Centers for Disease Control and Prevention.

For additional information regarding specific precautions for the prevention of the transmission of all infectious agents
from laboratory instruments and materials, and for recommendations for the management of exposure to infectious
disease, refer to CLSI document M29: Protection of Laboratory Workers from Occupationally Acquired Infections.

Sterilize all biohazard waste before disposal.

Refer to the document "Precautions When Using Media" for more information.

PROCEDURE
Specimen Collection: Consult listed references for information on specimen collection.(1-3,6) Infectious material should
be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a
delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until
inoculation.

In some tubes, the media may become oxidized within the tube during shipment. Resazurin, an oxidation-reduction
indicator causes the entire medium to turn pink when oxidation has occurred. These tubes may be restored to their
proper condition by bringing the media up to 100ºC. in a boiling waterbath. Loosen screw caps slightly before heating,
and tighten during cooling to room temperature. The boiling serves to reduce media intended for the culture of
anaerobic organisms.

Method of Use: Consult the listed references for the appropriate cultivation techniques using this medium.(1-3,6) It is
recommended that liquid media for anaerobic incubation should be reduced prior to inoculation by placing tubes (with
loosened caps) under anaerobic conditions for 18-24 hours. Alternatively, the boiling method described above may be
used. Thioglycollate Medium should be incubated at 35-37ºC., checking daily, as needed. Growth or turbidity should
be confirmed by gram stain and subculture onto an appropriate growth medium.

Note: Thioglycollate with Indicator Media contains a resazurin indicator which will cause the upper layer of this broth
to be pink, due to the exposure to oxygen. Containers that have been agitated recently (as during shipping) will turn
pink throughout. This can be reversed by allowing the container to stand still for a few hours or by putting the
containers in a boiling waterbath for 10 minutes with loosened caps. The caps are then tightened firmly before the
media cools. Also note that it is common for a whitish precipitate to form in this medium due to the agar content. This
does not affect the performance of this medium.

INTERPRETATION OF RESULTS
Consult listed references for the interpretation of growth and other identification tests to identify growth of organism in
this medium.(1-3,6)

LIMITATIONS
It is recommended that biochemical, immunological, molecular, or mass spectrometry testing be performed on colonies
from pure culture for complete identification of bacteria and/or fungi.
In test samples, the proper surface to volume ratio of the Thioglycollate Medium must be maintained to avoid oxidation
of the medium, making it unsuitable for microaerophilic and anaerobic growth.

A slight turbidity or haziness may be present due to the small amount of agar in the medium. When the media has been
boiled it appears clear.

Do not boil media more than once, as frequent boiling may lead to toxic products forming in the medium.(7) If it is
suspected that the medium has more than 30% oxidation after boiling, it should be discarded.

Refer to the document "Limitations of Procedures and Warranty" for more information.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks,
incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL
Hardy Diagnostics tests each lot of commercially manufactured media using appropriate quality control
microorganisms and quality specifications as outlined on the Certificate of Analysis (CofA) and the CLSI document
M22-A3 Quality Assurance for Commercially Prepared Microbiological Culture Media. The following
microorganisms are routinely used for testing at Hardy Diagnostics:

Incubation
Inoculation
Test Organisms Results
Method*
Time Temperature Atmosphere

Thioglycollate with Indicator, Thioglycollate without Indicator:

Clostridium novyi A
A 24-48hr 35°C Aerobic** Growth
ATCC® 7659***

Staphylococcus aureus
A 24hr 35°C Aerobic** Growth
ATCC® 25923

Candida albicans
A 24hr 35°C Aerobic** Growth
ATCC® 10231

Bacillus subtilis
A 24hr 35°C Aerobic** Growth
ATCC® 6633

Thioglycollate with H & K, Thioglycollate with Supplements:

Clostridium perfringens
A 48hr 35°C Aerobic** Growth
ATCC® 13124***

Bacteroides levii
A 48hr 35°C Aerobic** Growth
ATCC® 29147

Bacteroides vulgatus
A 48hr 35°C Aerobic** Growth
ATCC® 8482

* Refer to the document "Inoculation Procedures for Media QC" for more information.

** Tubes are incubated in an aerobic incubator with the caps screwed down tightly to create an atmosphere of low
oxygen tension within the tube.

*** Recommended QC strains for User Quality Control according to the CLSI document M22 when applicable.

USER QUALITY CONTROL

End users of commercially prepared culture media should perform QC testing in accordance with applicable
government regulatory agencies, and in compliance with accreditation requirements. Hardy Diagnostics recommends
end users check for signs of contamination and deterioration and, if dictated by laboratory quality control procedures or
regulation, perform quality control testing to demonstrate growth or a positive reaction and to demonstrate inhibition or
a negative reaction, if applicable. Hardy Diagnostics quality control testing is documented on the certificate of analysis
(CofA) available from Hardy Diagnostics Certificate of Analysis website. Also refer to the document "Finished Product
Quality Control Procedures," and the CLSI document M22-A3 Quality Assurance for Commercially Prepared
Microbiological Culture Media for more information on the appropriate QC procedures. See the references below.

REFERENCES
1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed.,
A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Brewer, J.H. 1940. J. Amer. Med. Assoc.; 115:598.

5. Federal Security Agency, Food and Drug Administration, Compilation of Regulations for Test and Methods of
Assay and Certification of Antibiotic Drugs.

6. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology,
Washington, D.C.

7. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams &
Wilkins, Baltimore, MD.

8. National Formulary, 9th ed. p.768, 1950.

9. National Institutes of Health Circular: Culture Media for the Sterility Test, 2nd rev. Feb. 5, 1946.

10. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory
Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

IFU-10792[A]

1430 West McCoy Lane, Santa Maria, CA 93455, USA

Phone: (805) 346-2766 ext. 5658
Fax: (805) 346-2760
Website: HardyDiagnostics.com
Email: TechnicalServices@HardyDiagnostics.com
Ordering Information

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