TRYPTIC SOY BROTH (TSB)

Cat. no. K85 TSB, Filtered, 15x103mm Tube, 1ml 20 tubes/box
Cat. no. K88 TSB, 15x103mm Tube, 9ml 20 tubes/box
Cat. no. K89 TSB, 15x103mm Tube, 5ml 20 tubes/box
Cat. no. K285 TSB, Filtered, 15x103mm Tube, 2ml 20 tubes/box
Cat. no. R30 TSB, 13x100mm Tube, 5ml 20 tubes/box
Cat. no. R31 TSB, 13x100mm Tube, 2ml 20 tubes/box

INTENDED USE

Hardy Diagnostics Tryptic Soy Broth (TSB) is recommended for use as a general purpose medium for the isolation and cultivation of a wide variety of bacteria and fungi. It is the recommended medium for use in antibiotic susceptibility testing and disk diffusion testing.(5)

SUMMARY

Tryptic Soy Broth is widely used for the isolation of bacteria from clinical specimens, supporting the growth of the majority of pathogenic bacteria. It is also frequently used in blood culture work. Tubes of this medium may be used for preparing dilutions of organism for colony counts, preparation of standard inocula, and antibiotic sensitivity testing for both fastidious and non-fastidious microorganisms. Tryptic Soy Broth is also recommended for use in sterility testing for the detection of contamination with low incidence fungi and aerobic bacteria.(14)

Tryptic Soy Broth, also known as Soybean-Casein Digest, conforms to the formula given by the U.S. Pharmacopeia.(14) It was originally developed for testing the sensitivity of pneumococci and other microorganisms to sulfonamides without adding blood or serum to the medium.(1) Hamilton and Spink in the early 1950's, reported the use of Tryptic Soy Broth as suitable for the growth of aerobic and facultative microorganisms including Brucella species.(7,13) Later, Garrison and Hedgecock both demonstrated the use of this medium to promote the growth of pathogenic fungi.(6,8)

This medium contains digests of soybean meal and casein, which provide amino acids and other nitrogenous substances, making it a highly nutritious medium for a variety of organisms, including most pathogenic bacteria. Sodium chloride is added to maintain the osmotic equilibrium. Dextrose is incorporated as an energy source. The dipotassium phosphate is included in the formulation as a buffer to maintain the pH.

Product K85, TSB Filtered 1ml, and K285, TSB Filtered 2ml, are filtered using a 0.2 micron filter to remove non-viable organisms and is recommended for preparing suspensions of organisms for staining.

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digest of Casein 17.0gm
Sodium Chloride 5.0gm
Papaic Digest of Soybean Meal 3.0gm
Dextrose 2.5gm
Dipotassium Phosphate 2.5gm

Final pH 7.3 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. (2-8ºC. for Cat. no. K85) away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection.(2-4,9) Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.

Method of Use: Inoculate the medium as soon as possible after the specimen has been collected. Incubate with caps loosened, in the appropriate atmospheric environment and incubation temperature for 18-24 hours, or up to seven days. Refer to the current CLSI (formerly NCCLS) guidelines for sensitivity testing protocols, and other listed references for additional procedures using this media.(2-4,9,11,14)

INTERPRETATION OF RESULTS

Consult listed references for interpretation criteria and further biochemical testing of growth in Tryptic Soy
Broth.(2-4,9,11,14)

LIMITATIONS

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Tryptic Soy Broth: Cat. Nos. K85, K88, K285, R30, and R31
Staphylococcus aureus
ATCC ® 25923
A 1-3 days 35°C Aerobic Growth
Escherichia coli
ATCC ® 25922
A 1-3 days 35°C Aerobic Growth
Tryptic Soy Broth: Cat. No. K89
Staphylococcus aureus
ATCC ® 25923
A 1-3 days 35°C Aerobic Growth
Escherichia coli
ATCC ® 25922
A 1-3 days 35°C Aerobic Growth
Candida krusei
ATCC ® 14243
J 1-5 days 35°C Aerobic Growth
Bacillus subtilis
ATCC ® 6633
J 1-3 days 35°C Aerobic Growth

USER QUALITY CONTROL

Note: K85 is filtered using a 0.2 micron filter to remove non-viable organisms and is recommended for preparing suspensions of organisms for staining.

PHYSICAL APPEARANCE

Tryptic Soy Broth should appear clear, and light amber in color.

REFERENCES

1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Bauer, A.W., W.M.M. Kirby, et al. 1966. Am. J. Clin. Pathol.; 45:493-496.

5. Garrison, L.G. 1961. Studies of the respiratory activity of Histoplasma capsulatum. J. Infect. Disease; 108:120-124.

6. Hamilton, P.K. 1954. Am. J. of Clin. Pathol.; 24:580.

7. Hedgecock, L.W. 1971. Effect of vaccines prepared from Histoplasma capsulatum and other yeasts on experimental tuberculosis, J. Bact.; 82:115-123.

8. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

9. McCullough, N.B. 1949. Laboratory tests in the diagnosis of Brucellosis, Am. J. of Public Health; 39:866.

10. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams & Wilkins, Baltimore, MD.

11. Performance Standards for Antimicrobial Disk Susceptibility Tests, 6th ed., M2-A11, Vol. 33, No. 1. 2013. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.

12. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

13. Spink, W.W. 1952. Am. J. of Clin. Pathol.; 22:201.

14. The Official Compendia of Standards. USP-NF. United States Pharmacopeial Convention, Rockville, MD.

ATCC is a registered trademark of the American Type Culture Collection.

012617gr