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Tryptic Soy Agar (Soybean-Casein Digest Agar)

Name: Tryptic Soy Agar (Soybean-Casein Digest Agar)
SKU: TSAP
Availability: InStock

Tryptic Soy Agar (Soybean-Casein Digest Agar)

$15.00 – $25.00

5 plates/pack, 10 plates/pack
Number of sizes: 2
Type: Agar Gel

Prepared Media – Plates

No sub-products available for this product.

SKU: TSAP Category: Ready To Use - Plates

Description

Intended Use and Principle of Procedure

Tryptic (Trypticase) Soy Agar (TSA) is used for the isolation and cultivation of non fastidious and fastidious microorganisms. It is not the medium of choice for anaerobes. The 150 × 15 mm-style plates of Trypticase Soy Agar are convenient for use with Taxo factor strips in the isolation and differentiation of Haemophilus species. Sterile Pack and Isolator Pack plates are useful for monitoring surfaces and air in clean rooms, Isolator Systems, and other environmentally-controlled areas when sterility of the medium is of importance.

Hycheck hygiene contact slides are used for assessing the microbiological contamination of surfaces and fluids.

The combination of casein and soy peptones in TSA renders the medium highly nutritious by supplying organic nitrogen, particularly amino acids and longer-chained peptides. The sodium chloride maintains osmotic equilibrium. Agar is the solidifying agent.

Haemophilus species may be differentiated by their requirements for X and V factors. Paper strips impregnated with these factors are placed on the surface of the medium after inoculation with the test organism. Following incubation, a zone of growth around the strip indicates a requirement for the factor(s).

Summary and Explanation

The nutritional composition of TSA has made it a popular medium for many years. It is the medium specified as Soybean-Casein Digest Agar Medium in the USP for the total aerobic microbial count portion of the microbial limit testing procedures. The medium is used for a multitude of purposes, including maintenance of stock cultures, plate counting, isolation of microorganisms from a variety of specimen types, and as a base for media containing blood. It is included in the Bacteriological Analytical Manual of Methods for the Examination of Water, Wastewater, and Foods and is used for testing bacterial contaminants in cosmetics.

Formulae

Pancreatic Digest of Casein	15.0g
Enzymatic Digest of Soybean Meal	5.0g
Sodium Chloride	5.0g
Agar	15.0g

Procedure

Use standard procedures to obtain isolated colonies from specimens. Since many pathogens require carbon dioxide on primary isolation, plates may be incubated in an atmosphere containing approximately 3-10% CO₂. Incubate plates at 35 ± 2°C for 18-24 hours.

Trypticase Soy Agar (150 mm plates) for Haemophilus The initial specimens should be inoculated onto Chocolate II Agar or another suitable medium and incubated for 18-24 hours in an aerobic atmosphere supplemented with carbon dioxide. Choose one or two well-isolated colonies that resemble Haemophilus species and perform a Gram stain to confirm that the isolate is a gram-negative rod or coccobacillus.

Suspend 1-2 colonies in 5 mL sterile, purified water or Trypticase Soy Broth and vortex to mix. Dip a swab in the suspension and inoculate the entire surface of the plate with the swab. With sterile forceps, place a Taxo X factor strip, a V factor strip, and an XV strip on the plate, at least 20 mm apart.

Incubate plates at 35 ± 2°C for 24 hours in an aerobic atmosphere supplemented with carbon dioxide.

Directions for Preparation from Dehydrated Product

1. Suspend 40 g of the powder in 1 L of purified water. Mix thoroughly.

2. Heat with frequent agitation and boil for 1 minute to completely dissolve the powder.

3. Autoclave at 121°C for 15 minutes. DO NOT OVERHEAT.

4. For the preparation of blood plates, add 5-10% sterile, defibrinated blood to the sterile agar, which has been cooled to 45-50°C.

5. Test samples of the finished product for performance using stable, typical control cultures.

Identification Specification and Cultural Response

Dehydrated Appearance:	Light beige, free-flowing, homogeneous.
Prepared Appearance: Solution:	Plain – Light amber, slightly opalescent. With 5% sheep blood – Bright red, opaque. 4.0% solution, soluble in purified water upon boiling. Solution is light amber, slightly opalescent.
pH:	7.3 ± 0.2
Cultural Response:	Prepare the medium per label directions, without (plain) and with 5% sheep blood (SB). Inoculate and incubate at 35  2°C for 18-48 hours under 5-10% CO₂.
Reaction	4.0%
Solution	25°C

Microorganism	ATCC	CFU	Recovery Plain/SB	Hemolysis
Escherichia coli	25922	102-103	Good	Beta
Neisseria meningitidis	13090	102-103	Good	None
Staphylococcus aureus	25923	102-103	Good	Beta
Streptococcus pneumoniae	6305	102-103	Good	Alpha
Streptococcus pyogenes	19615	102-103	Good	Beta

Expected Results

After incubation, it is desirable to have isolated colonies of organisms from the original sample. Subculture colonies of interest so that positive identification can be made by means of biochemical and/or serological testing.

Consult appropriate texts for the growth patterns produced by the various strains of Haemophilus.

References

1. United States Pharmacopeial Convention, Inc. 2001. The United States pharmacopeia 25/The national formulary 20 – 2002. United States Pharmacopeial Convention, Inc., Rockville, Md.

2. MacFaddin. 1985. Media for isolation-cultivation-identification-maintenance of medical bacteria, vol. 1. Williams & Wilkins, Baltimore, Md.

3. Forbes, Sahm and Weissfeld. 1998. Bailey & Scott’s diagnostic microbiology, 10th ed. Mosby Inc., St. Louis, Mo.

4. Nash and Krenz. 1991. In Balows, Hausler, Herrmann, Isenberg and Shadomy (ed.), Manual of clinical microbiology, 5th ed. American Society for Microbiology, Washington, D.C.

5. Clesceri, Greenberg and Eaton (ed.). 1998. Standard methods for the examination of water and wastewater, 20th ed. American Public Health Association, Washington, D.C.

6. Downes and Ito (ed.). 2001. Compendium of methods for the microbiological examination of foods, 4th ed. American Public Health Association, Washington, D.C.

7. U.S. Food and Drug Administration. 1995. Bacteriological analytical manual, 8th ed. AOAC International, Gaithersburg, Md.

8. Curry, Joyce, and McEwen. 1993. CTFA microbiology guidelines. The Cosmetic, Toiletry and Fragrance Association, Inc., Washington, D.C.

9. Association for the Advancement of Medical Instrumentation. 1984. Process control guidelines for gamma radiation sterilization of medical devices. AAMI, Arlington, Va.

10. Murray, Baron, Pfaller, Tenover, and Yolken (ed.). 1999. Manual of clinical microbiology, 7th ed. American Society for Microbiology, Washington, D.C.

11. Holt, Krieg, Sneath, Staley, and Williams (ed.). 1994. Bergey’s Manual of determinative bacteriology, 9th ed. Williams & Wilkins, Baltimore, Md.

Campos. 1999. In Murray, Baron, Pfaller, Tenover and Yolken (ed.), Manual of clinical microbiol- ogy, 7th ed. American Society for Microbiology, Washington, D.C.

Additional information

Size	5 plates/pack, 10 plates/pack

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Tryptic Soy Agar (Soybean-Casein Digest Agar)

Tryptic Soy Agar (Soybean-Casein Digest Agar)

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