THIOGLYCOLLATE BROTH WITHOUT INDICATOR WITH HEMIN & VITAMIN K

Technical Data #2028a / 2013.11.01

Thioglycollate Broth without indicator with Hemin and Vitamin K is a liquid medium used for cultivation of fastidious microorganisms. Mixed peptones and Hemin-Vitamin Supplement provide the essential nutrients. Thioglycollate and agar maintain a low oxygen of the medium. Vitamin K enhances Bacteroides recovery.

 


 

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

5,5

Sodium Thioglycollate

0,5

Agar

0,75

Enrichments

 

Hemin

0,005

Vitamin K

0,001

pH 7,0 +/- 0,2 25 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 2-8 C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25 C.



SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 29,5 grams in 1000 ml of purified water. Heat gently with frequent agitation and boil for one minute. Add 2x5 ml of Vitamin K-Hemin (#8751). Mix gently and dispense into culture tubes. Sterilize at 121 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 35 C for 24-72 hours. Reincubate tubes for up to 4 weeks if highly fastidious microrganisms are suspected.
  3. Examine tubes daily for visual signs of growth.


QUALITY CONTROL

Expected results after 72 hrs at 35C

Organisms

ATCC

Growth

Bacteroides fragilis

25285

+

Bacteroides levii

29147

+

Bacteroides vulgatus

8482

+

Clostridium novyi

7659

+

Clostridium perfringens

13124

+

Clostridium sporogenes

11437

+

Staphylococcus aureus

25923

+


LIMITATIONS OF METHOD
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 grams QB-39-4706
     
Vitamin K-Hemin Supplement, 5ml 10/pkg 8751
     
Prepared media    
Tubes 16x125 mm, 5 ml 10/pkg 2028
Tubes 16x125 mm, 10 ml 10/pkg 2420
Tubes 20x125 mm, 15 ml 10/pkg 2393

 


 

 

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