TCBS Agar (Vibrio Selective Agar)

TCBS Agar

Thiosulfate Citrate Bile Sucrose Agar

Cat. No. 1.10263.0500
(500 g)

Thiosulfate Citrate Bile Sucrose Agar proposed by NAKANISHI (1962), modified by KOBAYASHI et al. (1963) is used for the isolation and selective cultivation of Vibrio cholerae and other enteropahtogenic vibrios (V. parahaemolyticus, NAG vibrios).

This culture medium complies with the recommendations of the World Health Organization WHO (1965, 1967) and the APHA (1992).

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

Mode of Action
The high concentrations of thiosulfate and citrate and the strong alkalinity of this medium largely inhibit the growth of Enterobacteriaceae. Ox bile and cholate suppress primarily enterococci. Any coliform bacteria, which may grow, cannot metabolize sucrose. Only a few sucrose-positive Proteus strains can grow to form yellow, vibrid-like colonies. The mixed indicator thymol blue-bromothymol blue changes its color to yellow, when acid is formed, even in this strongly alkaline medium.

Typical Composition (g/liter)
Peptone from casein 5.0; peptone from meat 5.0; yeast extract 5.0; sodium citrate 10.0; sodium thiosulfate 10.0; ox bile 5.0; sodium cholate 3.0; sucrose 20.0; sodium chloride 10.0; iron(III) citrate 1.0; thymol blue 0.04; bromothymol blue 0.04; agar-agar 14.0.

Preparation
Suspend 88 g/liter and pour plates.
Do not autoclave.
pH: 8.6 ± 0.2 at 25°C.
The plates are clear and green-blue.

Experimental Procedure and Evaluation
Inoculate by spreading the sample or material from an enrichment culture, Alkaline Peptone water, on the surface of the plates.
Incubation: 18-24 hours at 35°C aerobically.

According to BURKHARDT (1969), it is advised to use, in addition to a liquid enrichment medium, two different solid culture media - a highly selective (e.g. TCBS Agar) and a less selective culture medium (e.g. Nutrient Agar: EMD Cat. No. 1.05450.).

Appearance of Colonies	Microorganisms
Flat, 2-3 mm in diameter, yellow	Vibrio cholerae, Vibrio cholerae type El Tor
Small, blue-green centre	Vibrio parahaemolyticus
Large, yellow	Vibrio alginolyticus
Blue	Pseudomonas, Aeromonas and others
Very small, translucent	Enterobacteriaceae and others

Further tests are necessary for complete identification (MUCKERJEE 1961, FINKELSTEIN and MUCKERJEE 1963, ROY et al. 1965, BOCKEMÜHL 1974 etc.).

Quality control

Test strains	Growth	color change to yellow
Vibrio alginolyticus	good / very good	+
Vibrio cholerae Inaba NIH 35	good / very good	+
Vibrio cholerae El Tor Inaba CH 38	good / very good	+
Vibrio cholerae Ogawa NIH 41	good / very good	+
Vibrio cholerae El Tor Ogawa CH 60	good / very good	+
Vibrio parahaemolyticus ATCC 17802	good / very good	-
Escherichia coli ATCC 25922	none / poor	-
Enterobacter cloacae ATCC 13047	none / poor	-
Proteus mirabilis ATCC 14273	none / poor	-
Pseudomonas aeruginosa ATCC 27853	none / poor	-

Additives

EMD Cat. No.	Product	Pack Size
1.01800.0500	Alkaline Peptone water	500 g

TCBS Agar - Vibrio chol. eltor ogava

Literature

American Public Health Association: Compendium of methods for the microbiological examination of foods. – 3 ^rd edition (1992).
BOCKEMÜHL, J.: Einfache Laboratoriumsdiagnostik der El Tor-Cholera. - Ärztl. Lab., 20; 32-41 (1974).
BURKHARDT, F.: Die bakteriologische Diagnose der Vibrio El Tor-Infektion. - Zbl. Bakt. I. Orig., 212; 177-189 (1969).
FINKELSTEIN, R.A., a. MUCKERJEE, S.: Haemagglutination a rapid method for differentiating V. cholerae and El Tor vibrios. - Proc. Soc. Exp. Biol. 112; 335-359 (1963).
KAMPELMACHER, E.H., MOSSEL, D.A.A., VAN NOORLE-JANSEN, a. VINCENTIE, H.: A survey on the occurrence of Vibrio parahaemolyticus on fish and shellfish, marketed in the Netherlands. - J. Hyg. Camp., 68; 189-196 (1970).
KOBAYASHI, T., ENOMOTO, S., SAKAZAKI, R., a. KUWAHARA, S.: A new selective isolation medium for pathogenic vibrios: TCBS-Agar. - Jap. J. Bact., 18; 391-397 (1963).
MUCKERJEE, S.: Diagnostic use of bacteriophage. - J. Hyg., 59; 109-115 (1961).
NAKANISHI, Y.: An isolation agar medium for cholerae and enteropathogenic halophilic vibrios. - Modern Media, 9; 246 (1963).
ROY, C., MRIDHA, K., a. MUCKERJEE, S.: Action of polymyxin on cholera vibrios. Techniques of determinatoin of polymyxin sensitivity. - Proc. Soc. Exp. Biol., 119; 893-896 (1965).
WHO Expert Committee on Cholera (2^nd Rep. Techn. Rep. Series No. 352, 1967). WHO: Cholera Information (1965).