Dehydrated Culture Media

TRYPTONE SOYA AGAR (Casein soya bean digest agar) EP/USP/JP/BP

Code: CM0131

a general purpose medium for the growth of a wide variety of organisms

Formula gm/litre
Pancreatic digest of casein 15.0
Enzymatic* digest of soya bean 5.0
Sodium chloride 5.0
Agar 15.0

pH 7.3 0.2 @ 25C

*Contains papain

Directions
Add 40g to 1 litre of distilled water (purified as required). Bring to the boil to dissolve completely. Sterilise by
autoclaving at 121C for 15 minutes.

Description
A general purpose agar medium, containing two peptones, which will support the growth of a wide variety of
organisms. It is suitable for the cultivation both of aerobes and anaerobes, the latter being grown either in deep
cultures or by incubation under anaerobic conditions. The medium may also be used as a blood agar base - for
this purpose 7% of sterile blood should be added to the sterile molten medium which has been cooled to
approximately 45C. Tryptone Soya Agar can also be used for the preparation of `chocolate agar.

Since Tryptone Soya Agar contains no added carbohydrate it may be used, with added blood, in the
determination of haemolysis. Horse blood agar plates prepared with Tryptone Soya Agar are used for the
colicine typing of Shigella sonnei 1,2,3,4.

The Oxoid medium has also been used as a replacement for yeastrel-milk agar plates in the Lisboa test 5 and
for bacterial counts on eviscerated poultry6.

When supplemented with 0.7g lecithin and 5g Polysorbate (Tween 80) per litre of Tryptone Soya Agar, the
medium can be used as Microbial Content Test Agar for testing quaternary ammonium compounds 7.

Tryptone Soya Agar is recommended as a reference medium when testing selective media, to measure the
degree of inhibition8.

A medium for isolation of Bacteroides gracilis is prepared from Tryptone Soya Agar by adding formate,
fumarate and nitrate. The medium is made selective using nalidixic acid and teicoplanin 9.
Enhanced haemolysis agar (EHA) used to improve detection of Listeria monocytogenes when present amongst
other listeriae has been modified to optimise its performance by substituting Tryptone Soya Agar for Columbia
Agar in the original formulation10. Tryptone Soya Agar conforms to formulations detailed in various international
pharmacopoeia11,12,13,14.

Storage conditions and Shelf life

Store the dehydrated medium at 10-30C and use before the expiry date on the label.
Store the prepared plates of medium at 2-8C.

Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Light straw to straw coloured gel

Quality control - non-pharmaceutical testing

Positive controls: Expected results

Staphylococcus aureus ATCC 25923 *
Good growth; straw coloured colonies

Streptococcus pyogenes ATCC 19615 * Good growth; pale straw coloured colonies

Negative control:

Uninoculated medium No change

* This organism is available as a Culti-Loop

Quality control - pharmaceutical testing

Growth promotion and

suitability of the
Micro-organism Preparation of Strain
counting method in
presence of product

Staphylococcus
aureus ATCC 6538*
Pseudomonas <100 CFU in TSA
aeruginosa ATCC TSA (casein soya bean digest agar) at (casein soya bean
9027* 30-35C for 18-24 hours digest agar) at 30-35C
for 3 days
Bacillus subtilis ATCC
6633*

Candida Sabouraud Dextrose Agar (R454462) <100 CFU in TSA

albicans ATCC or Sabouraud (casein soya bean
10231* Dextrose Liquid Medium (CM0147) at digest agar) at 30-35C
20-25C for 2-3 days
for 5 days
Aspergillus Sabouraud Dextrose Agar (R454462)
brasiliensis ATCC or Potato Dextrose Agar(CM0139) at
16404* 20-25C for 5-7 days, or until good
sporulation is achieved

* This organism is available in Quanti-Cult format

Precautions
It should be noted that haemolytic reactions of streptococci on Tryptone Soya Agar can vary according to the
origin of the blood e.g. horse or sheep. Tryptone Soya Agar designed for sheep blood show significant
differences when used with horse blood and vice versa.

References
1. Abbott J. D. and Graham J. M. (1961) Mon. Bull. Min. Hlth Pub. Hlth Lab. Serv. 20. 51-58.
2. Barrow G. I. and Ellis C. (1962) Mon. Bull. Min. Hlth Pub. Hlth Lab. Serv. 21. 141-147.
3. Cooke G. T. and Daines C. F. (1964) Mon. Bull. Min. Hlth Publ. Hlth Lab. Serv. 23. 81-85.
4. Gillies R. R. (1964) J. Hyg. Camb. 62. 1-9.
5. Mitchell T. G. (1964) J. Appl. Bact. 27. 45-52.
6. Barnes Ella M. and Shrimpton D. H. (1958) J. Appl. Bact. 2. 313-329.
7. American Public Health Association (1978) Standard Methods for the Examination of Dairy Products. 14th
Edn. APHA Inc. Washington DC.
8. Anon. (1987) J. Food Microbiol. 5. 291-296.
9. Lee K., Baron E.J., Summanen P. and Finegold S. (1990) J. Clin. Microbiol. 28. 1747-1750.
10. Beumer R.R., te Giffel M.C. and Cox L.J. (1997) Lett. Appl. Microbiol. 24. 421-425.
11. British Pharmacopoeia Volume II (2000)
12. US Pharmacopoeia XXX, (2008)
13. European Pharmacopoeia. 6.1 Edition (2008)
14. Japanese Pharmacopoeia. 15th Edition. (2006)

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