ENRICHED THIOGLYCOLLATE BROTH W/O INDICATOR W/ MARBLE CHIPS

Technical Data #2501a / 98.06.26

Enriched Thioglycollate Broth without indicator and with Marble chips is a liquid medium used for cultivation of aerobes, microaerobes and obligate anaerobes. This general utility medium supports a minimal inoculum with early visibility of growth. Absence of indicator prevents possible toxicity of indicator. Various enrichments and special peptone mixture enhance fastidious anaerobes growth. Cystine and sodium thioglycollate reduce the Eh of the medium; agar decreases oxygen tension, marble chips neutralize acid production during growth and prevent rapid death of acid-sensitive bacteria .

FORMULA
in grams per liter purified filtered water

Casein Peptone	17,0
Soy Peptone	3,0
Sodium Chloride	2,5
L-Cystine	0,25
Dextrose	6,0
Sodium Thioglycollate	0,5
Sodium Sulfite	0,1
Agar	0,7
Enrichments
Hemin	0,005
Vitamin K1	0,0001

pH 7,0 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-25º C protected from direct light. Medium must not be used if there are signs of contamination or if the expiry date has passed. Store dehydrate powder in a dry place in tightly-sealed containers at 2-25º C. Do not use dehydrated media if it is caked and if expiry date has passed.

DIRECTIONS
Suspend 30 grams in 1000 ml of purified water. Heat gently with frequent agitation and boil for one minute. Add 0,005 g of Hemin and 0,0001 g of Vitamin K1. Mix gently and dispense into culture tubes containing a marble chip. Sterilize at 121º C for 15 minutes.

PROCEDURE
Boil medium for 15 minutes and cool to room temperature before use.

1. Inoculate specimens as soon as possible. Tighten the caps.
2. Incubate at 35º C for 24-72 hours.
3. Examine tubes daily for visual signs of growth.

QUALITY CONTROL
Expected results after 48 hrs at 35ºC

Organisms	ATCC	Growth
Bacteroides fragilis	25285	+
Bacteroides levii	29147	+
Bacteroides vulgatus	8482	+
Clostridium novyi	7659	+
Clostridium perfringens	13124	+
Staphylococcus aureus	25923	+

LIMITATIONS OF METHOD
Do not heat tubes more than once to drive off absorbed oxygen. This is only a part of the identification. Other tests may be required.

REFERENCES

1. Dowell, V.R. Jr., G.L. Lombard, F.S. Thompson, A.Y. Armfield 1977 Media for isolation, characterization and identification of obligately anaerobic bacteria. Center for Disese Control (CDC) U.S.DHEW, Atlanta, Ga
2. Lennette, E.H., A. Balows, W.S. Hausler Jr., H.J. Shadomy 1985 Manual of Clinical Microbiology 4th ed. American Society for Microbiology, Washington, D.C.
3. Mc Faddin, J.F. 1985 Media for isolation-cultivation-identification-maintenance of medical bacteria. Williams and Wilkins, Baltimore
4. Sutter, V.L., D.M. Citron, M.A.C. Edelstein, S.M. Finegold 1986 Wadsworth Anaerobic Bacteriology Manual. 4th ed. Star Publishing Co. Belmont. Calif.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-4706
Prepared media
Tubes 20x125 mm, 20 ml	10/pkg	2501

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