Trichosel Broth, Modified

Intended Use

Trichosel Broth, Modified is used for the isolation and cultivation of Trichomonas species.

Summary and Explanation

Trichosel Broth, Modified is a modification of the Simplified Trypticase™ Serum (STS) Medium of Kupferberg et al. for the cultivation of Trichomonas spp.¹ The classical formula has been modified by the addition of beef extract and horse serum and an increased amount of yeast extract to improve performance. Chloramphenicol, a relatively stable antibiotic, replaces the penicillin and streptomycin recommended for addition to the STS base.

Principles of the Procedure

Trichosel Broth, Modified contains casein peptone, cysteine, beef extract and yeast extract as sources of amino acids, nitrogen, sulfur, carbon, vitamins and trace ingredients. Maltose is an energy source for the metabolism of microorganisms including Trichomonas spp. Chloramphenicol is a broad spectrum antibiotic which inhibits a wide range of gram-positive and gram-negative bacteria. Horse serum is added to provide growth factors required by Trichomonas.

User Quality Control

Identity Specifications
Trichosel Broth, Modified
Dehydrated Appearance: Fine, homogeneous, free of extraneous material.
Solution:                       23.1 g soluble in 950 mL purified water upon
                                    boiling. Solution is light to medium, yellow to tan,
                                   with blue-green to green ring at top, hazy.
Prepared Appearance:    Light to medium, yellow to tan, with blue-green
                                   to green ring at top, hazy.
Reaction of 23.1 g/950 mL
Solution at 25°C:           pH 6.0 ± 0.2
Cultural Response
Trichosel Broth, Modified
Prepare the medium per label directions. Inoculate with fresh cultures and incubate at 35 ± 2°C for 5 days

Formula

Trichosel Broth, Modified
Approximate Formula* Per Liter
Pancreatic Digest of Casein........................................ 12.0 g
Beef Extract............................................................... 2.0 g
Yeast Extract............................................................. 5.0 g
L-Cysteine HCl........................................................... 1.0 g
Maltose.................................................................... 2.0 g
Agar......................................................................... 1.0 g
Chloramphenicol........................................................ 0.1 g
Methylene Blue......................................................... 3.0 mg
*Adjusted and/or supplemented as required to meet performance criteria.

Directions for Preparation from Dehydrated Product

1. Suspend 23.1 g of the powder in 950 mL of purified water. Mix thoroughly.
2. Heat with frequent agitation and boil for 1 minute to completely dissolve the powder.
3. Autoclave at 118°C for 15 minutes. Immediately tighten caps. 4. Cool and aseptically add 5% horse serum.
5. Test samples of the finished product for performance using stable, typical control cultures.

Procedure

Inoculate specimens suspected of containing Trichomonas organisms into the broth medium using swabs containing the specimen or by alternative methods, as appropriate. Incubate tubes at 35 ± 2°C in an aerobic atmosphere. After 48 hours and 5 days of incubation, prepare a wet mount from the broth and examine microscopically under low power for the presence of flagellate protozoans.

Expected Results

If growth of trichomonads has occurred, organisms of typical morphology will be seen in the broth medium when an aliquot is examined microscopically using low power magnification.

Limitations of the Procedure

1. The test organism used for inoculating an assay medium must be cultured and maintained on media recommended for this purpose.
2. For successful results of these procedures, all conditions of the assay must be followed precisely. 3. Aseptic technique should be used throughout the assay procedure.
4. The use of altered or deficient media may cause mutants having different nutritional requirements that will not give a satisfactory response.

*Store at 2-8° C. 

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