TINSDALE AGAR

Technical Data #1072a / 2013.04.19

The QUE-BACT Tinsdale Agar Base is used with QUE-BACT Tinsdale Supplement in isolating and differentiating Corynebacterium diphtheriae.

 


 

FORMULA
in grams per liter purified filtered water

Proteose Peptone No. 3

20.0

Sodium Chloride

5.0

Agar

20.0

Supplement

 

L-Cystine

0.24

Sodium Thiosulfate

0.425

Potassium Tellurite

0.3

Bovine Serum

100 ml

pH 7,4 +/- 0,2 25 C

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


PRECAUTIONS
This product is for laboratory use only.


STORAGE
Store prepared media at 2-8 C protected from direct light. Medium must not be used if there are signs of contamination or if the expiry date has passed. Store dehydrated powder in a dry place in tightly-sealed containers at 2-25 C. Do not use dehydrated media if it is caked and if expiry date has passed.


DIRECTIONS
Suspend 45 grams of powder in 1 liter of purified filtered water. Mix thoroughly. Heat to boiling to dissolve completely. Mix carefully. Autoclave for 15 minutes at 121 C (15 psi). Cool down to 50-55 C and aseptically add 150 ml of Tinsdale Supplement. Mix well. Dispense into sterile Petri dishes.


PROCEDURE
This prepared medium is light to medium amber, slightly opalescent to opalescent without precipitate.

  1. Bring to room temperature before use.
  2. Inoculate specimens in a manner to obtain discrete colonies and stab the medium several times with an inoculating needle.
  3. Incubate at 35 C for 18-48 hours.
  4. Examine for growth. Colonies of Corynebacterium diphtheriae are brown with halos.


QUALITY CONTROL

Expected results at 35C for 18-48 hours.

Organisms

ATCC

Growth

Appearance

Corynebacterium diphtheriae
type gravis

8028

+

Brown w/halos

Corynebacterium diphtheriae
type mitis

8024

+

Brown w/halos

Klebsiella pneumoniae

13883

-

-

Streptococcus pyogenes

19615

+

Brown w/o halos


LIMITATIONS OF METHOD

  1. C. ulcerans, C. pseudotuberculosis and (rarely) Staphylococcus Spp occasionally may produce colonies with characteristic halo similar to C. diphtheriae and required biochemical identification.
  2. Some strains may be encountered that fail to grow or grow poorly on this medium. Consequently, it is not suitable as a primary plating medium.
  3. Several organisms may exhibit slight browning on this medium in 18 hours. So, early reading must be avoided.
  4. Incubation at 5-10% CO2 retards the development of halos on this medium.
  5. This is only a part of the identification. Other tests may be necessary due to similar reactions on this medium.


REFERENCES

  1. Tinsdale, G.F.W. 1947. A new medium for the isolation and identification of C.diphtheriae based on the production of hydrogen sulphide. J. Pathol. Bacteriol. 59:461-466.
  2. Billings, E. 1956. An investigation of Tinsdale Tellurite medium: its usefulness and mechanisms of halo-formation. M.S. thesis. University of Michigan, Ann Arbor, MI.
  3. Moore, M.S. and E.I.Parsons. 1958. A study of a modified Tinsdale's medium for the primary isolation of C. diphtheriae. J. Infect. Dis. 102:88.
  4. Bailey, R.W. and E.G. Scott. 1966 Diagnostic microbiology, 2nd ed., p.213. The C.V. Mosby Company, St-Louis, MO.
  5. Isenberg, H.D. (ed) 1992. Clinical microbiology procedures handbook. American Society for Microbiology, Washington, D.C.
  6. Baron.E.J., L.R. Peterson and S.M. Finegold. 1994. Baily & Scott's diagnostic microbiology, 9th ed. Mosby-Year Book, Inc. St-Louis, MO.
  7. Claridge, J.E. and C.A. Spiegel. 1995. Corynebacterium and miscellaneous irregular gram-positive rod, Erysipelothrix, and Gardnerella, p.357-377. In P.R. Murray, E.J. Baron, M.a. Pfaller, F.C. Tenover, and R.H. Yolken (ed). Manual of clinical microbiology, 6th ed. American Society for Microbiology, Washington, D.C.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-4811
     
Tinsdale Supplement Bottle of 150 ml 8770
     
Prepared media    
Plates, 100x15 mm 10/pkg 1072

 


 





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