TRYPTIC SOY AGAR (TSA)

Cat. no. G62 Tryptic Soy Agar (TSA) (without plate label, orientation tabs, and logo), 15x100ml Plate, 23ml 10 plates/bag
Cat. no. G151 Tryptic Soy Agar (TSA), 15x60mm Plate, 12ml 10 plates/bag
Cat. no. J108 TSA / TSA, 15x100mm Biplate, 10ml/10ml 10 plates/bag
Cat. no. L60 TSA, 16x100mm Tube, 5.5ml Slant 20 tubes/box
Cat. no. Q90 TSA w/NaCl 5%, 16x100mm Tube, 5ml 20 tubes/box
Cat. no. Q91 TSA w/NaCl 10%, 16x100mm Tube, 5ml 20 tubes/box
Cat. no. Q92 TSA w/NaCl 20%, 16x100mm Tube, 5ml 20 tubes/box

INTENDED USE

Hardy Diagnostics Tryptic Soy Agar is recommended for use as a general growth medium for the isolation and cultivation of microorganisms. This medium is also recommended for use in the cultivation, storage, maintenance and transportation of pure cultures of microorganisms.

SUMMARY

Tryptic Soy Agar contains digests of soybean meal and casein, making it suitable for the growth of a wide variety of fastidious and nonfastidious microorganisms.(2-6, 8) The combination of soy and casein peptones supply organic nitrogen in the form of amino acids and polypeptides, making the medium highly nutritious. Sodium chloride is added to maintain the osmotic equilibrium. Products containing additional salt (Cat. nos. Q90, Q91, and Q92) are recommended for use in determining the halotolerance levels of microorganisms. Agar is the solidifying agent.

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digest of Casein 15.0gm
Peptic Digest of Soybean Meal 5.0gm
Sodium Chloride 5.0gm
Agar  15.0gm

Final pH 7.3 +/- 0.2 at 25°C

In addition,

    TSA with NaCl 5% contains 50.0gm of NaCl.

    TSA with NaCl 10% contains 100.0gm of NaCl.

    TSA with NaCl 20% contains 200.0gm of NaCl.

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

STORAGE AND SHELF LIFE

Storage: Upon receipt store plates at 2-8°C away from direct light. Products in tubes and bottles (Cat. nos. Q90, Q91, Q92, and L60) should be stored at 2-30°C 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

For Cat. nos. G62, J108, and L60.

For Cat. nos. G151, H19, Q90, Q91, and Q92.

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection.(2-6, 8) 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 medium and refrigerated until inoculation.

Method of Use: Plates should be warmed to room temperature and the agar surface should be dry prior to inoculating. Inoculate and streak the specimen as soon as possible after collection. If the specimen to be cultured is on a swab, roll the swab over a small area of the agar surface. Streak for isolation with a sterile loop. Incubate plates aerobically at 35-37°C for 18-24 hours (some organisms may take longer than 24 hours for visible growth to appear). Examine for colonial morphology.

Spread Plate Method:

1. Prepare decimal dilutions in sterile diluent to obtain 30-300 CFU per plate.

2. Aseptically inoculate agar surface with 0.1ml of well mixed diluted sample.

3. Using a sterile spreader device, distribute the inoculum evenly over the agar surface.

4. Incubate plates aerobically for 1 to 5 days at 35°C.

INTERPRETATION OF RESULTS

Consult listed references for the identification of colony morphology and further biochemical tests required for identification.(1-5)

Following incubation, examine the plates for growth. Count the number of colonies and express in number of colony forming units (CFU) per gram or milliliter of sample. (Take into account the dilution factor.) If duplicate plates were set up, express the average for the two plates in terms of the number of microorganisms per gram or milliliter of sample. Consult listed references for additional information on interpretation and enumeration of microbial growth on this medium.(2-6,8)

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
Staphylococcus aureus
ATCC® 25923
A 24hr 35°C Aerobic Growth
Escherichia coli
ATCC® 25922
A 24hr 35°C Aerobic Growth
Candida albicans
ATCC® 10231
A 24hr 35°C Aerobic Growth
Bacillus subtilis
ATCC® 6633
A 24hr 35°C Aerobic Growth
Aspergillus brasiliensis
ATCC® 16404
G 1-5 days 20-25°C Aerobic Growth
For Cat. no. Q90:
Staphylococcus aureus
ATCC® 25923
A 24hr 35°C Aerobic Growth
Escherichia coli
ATCC® 25922
B 24hr 35°C Aerobic Inhibited
For Cat. no. Q91:
Staphylococcus aureus
ATCC® 25923
A 24-48 hrs 35°C Aerobic Growth
Escherichia coli
ATCC® 25922
B 24-48 hrs 35°C Aerobic Inhibited
For Cat. no. Q92:
Staphylococcus aureus
ATCC® 25923
A 24hr 35°C Aerobic Inhibited
Escherichia coli
ATCC® 25922
B 24hr 35°C Aerobic Inhibited

USER QUALITY CONTROL

PHYSICAL APPEARANCE

Tryptic Soy Agar should appear translucent, and light amber in color.

S. aureus growing on TSA

Staphylococcus aureus (ATCC® 25923) colonies growing on TSA (Cat. no. G62). Incubated aerobically for 24 hours at 35°C.

E. coli growing on TSA

Escherichia coli (ATCC® 25922) colonies growing on TSA (Cat. no. G62). Incubate aerobically for 24 hours at 35°C.

C. albicans growing on TSA

Candida albicans (ATCC® 10231) colonies growing on TSA (Cat. no. G62). Incubated aerobically for 48 hours at 35°C.

B. subtilis growing on TSA

Bacillus subtilis (ATCC® 6633) colonies growing on TSA (Cat. no. G62). Incubated aerobically for 24 hours at 35°C.



TSA

Uninoculated plate of TSA (Cat. no. G62).

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, J.H., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

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

4. American Public Health Association. Standard Methods for the Examination of Water and Wastewater, APHA, Washington, D.C.

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

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

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

8. APHA Technical Committee on Microbiological Methods for Foods. Compendium of Methods for the Microbiological Examination of Foods. APHA, Washington, D.C.

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

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