THIOGLYCOLLATE BROTH with INDICATOR and VITAMIN K-HEMIN

Technical Data #2978a / 2013.07.20

Enriched fluid Thioglycollate Broth with indicator is a liquid medium used for cultivation of aerobes, microaerobes and anaerobes microorganisms. Vitamin K-Hemin and special peptone mixture enhance fastidious anaerobes growth. Cystine and sodium thioglycollate reduce the Eh of the medium; agar decreases oxygen tension and resazurin is the redox indicator.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

15,0

Yeast Extract

5,0

Sodium Chloride

2,5

L-Cystine

0,5

Dextrose

5,5

Sodium Thioglycollate

0,5

Resazurin

0,001

Agar

0,75

Supplements

 

Vitamin K

0,0005

Hemin

0,005

pH 7,1 +/- 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 29,8 grams in 1000 ml of purified water. Heat gently with frequent agitation and boil for one minute. Add 2x5 ml of Vitamin K-Hemin Supplement (#8751). Mix gently and dispense into culture tubes. 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 or for up to 4 weeks depending on the microorganism to be recovered.
  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

+

Bacillus subtilis

6633

+

Clostridium sporogenes

11437

+


LIMITATIONS OF METHOD
Boil medium if more than 1/3 of the volume is oxidized. 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.
  5. The United States Pharmacopeia, 21st rev 1985 United States Pharmacopeial Convention, Rockville, Md


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-1806
     
Vitamin K-Hemin Supplement, 5 ml 10/pkg 8751
     
Prepared media    
Tubes 16x125 mm, 10 ml 10/pkg 2473
Tubes 20x125 mm, 20 ml 10/pkg 2978

 


 

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