Trypticase Soy Broth

Intended Use

Tryptic (Trypticase) Soy Broth (Soybean-Casein Digest Medium) is a general purpose medium used in qualitative procedures for the cultivation of fastidious and nonfastidious microorganisms from a variety of clinical and nonclinical specimens.
Tryptic (Trypticase) Soy Broth meets United States Pharmacopeia (USP), European Pharmacopoeia (EP) and Japanese Pharmacopoeia (JP)1-3 performance specifications, where applicable.

Summary and Explanation

Tryptic (Trypticase) Soy Broth (TSB) is a nutritious medium that will support the growth of a wide variety of microorganisms, including common aerobic, facultative and anaerobic bacteria and fungi.^4-7 This formulation is included in the USP as a medium for use in performing microbial enumeration tests and tests for specified microorganisms when testing nonsterile pharmaceutical products.¹
TSB was chosen by the USDA Animal and Plant Health Inspection Service for detecting viable bacteria in live vaccines.⁸ TSB is recommended for testing bacterial contaminants in cosmetics^9,10 and complies with established standards in the food industry.^10-16
Because of its capacity for growth promotion, TSB is also recommended for use as the inoculum broth for disc diffusion and agar dilution antimicrobial susceptibility testing as standardized by the Clinical and Laboratory Standards Institute (CLSI).^17,18

Principles of the Procedure

Enzymatic digests of casein and soybean provide amino acids and other complex nitrogenous substances. Dextrose is an energy source. Sodium chloride maintains the osmotic equilibrium. Dibasic potassium phosphate acts as a buffer to control pH.
Tryptic Soy Broth and Trypticase Soy Broth are provided as prepared media in a variety of bottle styles. In addition, Tryptic Soy Broth is provided as a Sterile Pack Bottle; i.e., the bottle has been terminally sterilized inside of autoclavable double-bags. All varieties of bottled TSB conform with requirements for Ready- To-Use Media as described in the USP.

User Quality Control

NOTE: Differences in the Identity Specifications and Cultural Response testing for media offered as both Difco™/Bacto™ and BBL™ brands may reflect differences
in the development and testing of media for industrial and clinical applications, per the referenced publications.

Identity Specifications
Bacto™ Tryptic Soy Broth
Dehydrated Appearance: Light beige, free-flowing, homogeneous.
Solution:                        3.0% solution, soluble in purified water upon
                                     warming. Solution is light amber, clear.
Prepared Appearance:     Light amber, clear.
Reaction of 3.0%
Solution at 25°C:            pH 7.3 ± 0.2

BBL™ Trypticase™ Soy Broth
Dehydrated Appearance: Fine, homogeneous, free of extraneous material.
Solution:                        3.0% solution, soluble in purified water upon warming. Solution is light,

                                     tan to yellow, clear to slightly hazy.
Prepared Appearance:     Light, tan to yellow, clear to slightly hazy.
Reaction of 3.0%
Solution at 25°C:            pH 7.3 ± 0.2

Cultural Response

Bacto™ Tryptic Soy Broth
Prepare the medium per label directions. Inoculate and incubate at 30-35°C for 18-72 hours (up to 5 days for A. brasiliensis and C. albicans). Prepare duplicate cultures of A. brasiliensis, B. subtilis and C. albicans and incubate at 20-25°C for up to 3 days (up to 5 days for A. brasiliensis and C. albicans).

BBL™ Trypticase™ Soy Broth
Prepare the medium per label directions. Inoculate tubes and incubate at 30-35°C for up to 3 days (up to 5 days for A. brasiliensis and C. albicans). Prepare duplicate cultures of A. brasiliensis, B. subtilis and C. albicans and incubate at 20-25°C for up to 3 days (up to 5 days for A. brasiliensis and C. albicans).

Formula

Bacto™ Tryptic Soy Broth (Soybean-Casein Digest Medium)
Approximate Formula* Per Liter
Pancreatic Digest of Casein......................................... 17.0 g
Papaic Digest of Soybean............................................. 3.0 g
Dextrose..................................................................... 2.5 g
Sodium Chloride.......................................................... 5.0 g
Dipotassium Phosphate................................................ 2.5 g
BBL™ Trypticase™ Soy Broth (Soybean-Casein Digest Broth)
Approximate Formula* Per Liter
Pancreatic Digest of Casein........................................ 17.0 g
Papaic Digest of Soybean............................................ 3.0 g
Sodium Chloride......................................................... 5.0 g
Dipotassium Phosphate............................................... 2.5 g
Dextrose.................................................................... 2.5 g
*Adjusted and/or supplemented as required to meet performance criteria.

Directions for Preparation from Dehydrated Product

1. Suspend the powder in 1 L of purified water:
Tryptic Soy Broth – 30 g;
Trypticase Soy Broth – 30 g; Mix thoroughly.
2. Warm gently until solution is complete.
3. Autoclave at 121°C for 15 minutes.
4. Test samples of the finished product for performance using stable, typical control cultures.

Sample Collection and Handling

For clinical specimens, refer to laboratory procedures for details on specimen collection and handling.^5,7,17-19
For food, dairy or cosmetic samples, follow appropriate standard methods for details on sample collection and preparation according to sample type and geographic location.^9-16
For pharmaceutical samples, refer to the USP for details on sample collection and preparation for testing of nonsterile products.¹

Procedure

For clinical specimens, refer to appropriate standard references for details on testing protocol to obtain isolated colonies from specimens using Tryptic/Trypticase Soy Broth.^17-19
For food, dairy or cosmetic samples, refer to appropriate standard references for details on test methods using Tryptic/ Trypticase Soy Broth.^9-16
For pharmaceutical samples, refer to USP General Chapters and for details on the examination of nonsterile products and performing microbial enumeration tests and tests for specific organisms using Tryptic/Trypticase Soy Broth.¹
Swab specimens may be inserted into the medium after inoculation of appropriate plated media. For liquid specimens, use a sterile inoculating loop to transfer a loopful of the specimen to the broth medium. Specimens known or suspected to contain obligate anaerobes should be inoculated near the bottom of the tube.
Incubate the tubes and bottles with loosened caps at 35 ± 2°C aerobically with or without supplementation with carbon dioxide. Tubed and bottled media intended for the cultivation of anaerobes should be incubated under anaerobic conditions. An efficient and easy way to obtain suitable anaerobic conditions is through the use of BD GasPak™ EZ anaerobic systems or equivalent alternative system. Examine for growth after 18-24 hours and 42-48 hours of incubation.

Expected Results

Growth in broth media is indicated by the presence of turbidity compared to an uninoculated control. Broth cultures should be held for at least a week before discarding as negative.

*Store at 2-8° C.
†QC testing performed according to USP/EP/JP performance specifications.

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