Tryptose Agar

EMD Cat. No. 1.10237.0500
500 g


For the cultivation and differentiation of various fastidious organisms, particularly Brucella, but also for isolating streptococci, pneumococci, meningococci, Listeria, and Pasteurellae.

Tryptose culture media are recommended by HAUSLER and KOONTZ (1970) in Diagnostic Procedures.






Mode of Action Preparation
Experimental Procedure and Evaluation Quality Control
Typical Composition (g/liter) Literature
Additives  


Mode of Action
 Isolation of Brucella strains from mixed cultures: addition of crystal violet inhibits the gram-positive bacterial flora (HAUSLER and KOONTZ 1970). Differentiation of Brucella species according to their sensitivity towards certain dyes: addition of thionine and basic fuchsin. Detection of Brucella melitensis with blood cultures: addition of sodium citrate as an anticoagulant, addition of agar-agar and cultivation in a 10 % CO2 atmosphere increases the yields of Brucella. Other possible applications: preparation of Brucella antigens (SILVERMAN and ELBERG 1950), isolation of Listeria monocytogenes from brain (GRAY et al. 1948), preparation of a Listeria Selective Agar by adding potassium tellurite (GRAY et al. 1950). Tryptose Agar also serves as a satisfactory base for preparing blood agar.
Typical Composition (g/liter)
 Tryptose 20.0; D(+)glucose 1.0; sodium chloride 5.0; thiaminium dichloride 0.005; aga-agar 13.0.
Preparation
 Suspend 39 g in 1 liter of purified water. Heat to boiling to dissolve completely. Autoclave (15 min at 121°C).

pH 7.3 ± 0.2 at 25°C.
 The prepared plates are clear and yellowish-brown.
Preparation of tryptose crystal violet agar: before autoclaving, add 1.4 ml of an aqueous 1 % crystal violet solution/liter, mix homogeneously.
Preparation of tryptose blood agar: sterile, liquified Tryptose Agar cooled to 45-50°C, add 5% sterile defibrinated blood and mix gently so as not to form any bubbles.
Experimental Procedure and Evaluation
 Tryptose crystal violet agar is particularly for the selective cultivation of Brucella. Spread the sample material thinly on the surface. A pre-enrichtment with Tryptose Broth should be carried out if only small numbers of Brucella are expected. Incubation should be carried out, in each case, for up to 5 days at 35°C in a 10% carbon dioxide atmosphere. This can be achieved using Anaerocult® C or Anaerocult® C mini.
 For the cultivation of other microorganisms, Tryptose Agar and Tryptose Broth are used. The incubation should be carried out, in each case, under optimum conditions.
Appearance of Colonies
 		Microorganisms
Pink, translucent, smooth-edged with a smooth suface, hemispherical, diameter 1-5 mm
 		Brucella
Pale pink, opaque, rough surface, large
 		streptococci


Further differentiation is possible, if Brucella Differential Agar is inoculated with pure Crucella colonies. Instead of employing culture media containing dyes, differentiation can also be performed with strips of paper (CRUICKSHANK 1948) or filter paper discs (PICKETT et al. 1953, SCHINDLER 1955) soaked in the dye solutions and placed on the surface of Tryptose Agar.

Additives


EMD Cat. No. Product Pack Size



Quality control of Tryptose Agar (spiral plating method)

Test strains
 Inoculum (cfu/ml)
 		Revovery rate %
Streptococcus pyogenes ATCC 12344
 		10³-105
 		70
Streptococcus pneumoniae ATCC 6301
 		10³-105
 		70
Pasteurella multocida ATCC 43137
 		10³-105
 		70
Listeria monocytogenes ATCC 19118
 		10³-105
 		70
Shigella flexneri ATCC 12022
 		10³-105
 		70
Literature

CRUICKSHANK, J.C.: A Simple Method for Testing Dye Sensitivity of Brucella Species. - J. Path. Bact., 60; 328-329 (1948).
 BORMAN a. WEST: Diagnostic Procedures and Reagents, 3rd ed.; 246 (1951).
 FAO/WHO: 4 Techn. Rep. Export Panel on Brucellosis (WHO Techn. Rep. Ser. 289; Genf (1964)).
 GRAY, M.L., STAFSEHT, H.J., THORP, F., a. RILEY, W.F.: A new technique for isolation of Listerella from bovine brain. - J. Bact., 55; 471-476 (1948).
 GRAY, M.L., STAFSEHT, H.J., a. THORP, F. jr.: The use of potassium tellurite, sodium azide and acetic acid in a selective medium for the isolation of Listeria monocytogenes. - J. Bact., 59; 443-444 (1950).
 HAUSLER, W.J., a. KOONTZ, F.P.: Brucellosis (in Diagnostic procedures for Bacterial, Mycotic and Parasitic Infections; 5th ed., APHA, New York (1970).
 JONES, L.M., a. WUNDT, W.: International Committee on Nomenclature of Bacteria, Subcommittee on the Taxonomy of Brucella. - Int. J. Syst. Bact., 21; 126-128 (1971).
 PICKET, M.J., NELSON, E.L., a. LIBERMAN, J.D.: Specification within ghe Genus Brucella. II. Evaluation of Differential Dye, Biochemical, and Serological Tests. - J. Bact., 66; 210-219 (1953).
 SCHINDLER, R.: Untersuchungen über die Differenzierung von Brucellatypen. - Zbl. Bakt., I. Orig., 164; 93-95 (1955).
 SILVERMAN, S.J., a. ELBERG, S.S.: The antigenic relationships of native antigens of species of Brucella. - J. Immunol., 65; 163-174 (1950).




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