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Cystine Tryptone Agar, HiVeg™

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MV159
Recommended for maintenance, subculturing, detection of motility and fermentation studies with the addition of various carbohydrates.

Intended Use

Cystine Tryptone Agar, HiVeg™ is recommended for maintenance, subculturing, detection of motility etc. With added carbohydrates, it can be also used for fermentation reactions of fastidious organisms.

Composition

Ingredients Grams/Litre
HiVeg hydrolysate 20.0
L-Cystine 0.5
Sodium chloride 5.0
Sodium sulphite 0.5
Phenol red 0.017
Agar 2.5
Final pH (at 25°C) 7.3 ± 0.2

Formula adjusted, standardized to suit performance parameters.

Product Profile

Type Code Hydrolysate
Vegetable based MV159 HiVeg hydrolysate
Animal based M159 Casein enzymic hydrolysate

Technical Specifications

Reconstitution 28.5 g/l
Quantity on preparation (500g) 17.54 L
(100g) 3.5 L
pH (25°C) 7.3 ± 0.2
Supplement Carbohydrate
Sterilization 121°C / 15 minutes.
Storage Dry Medium - Below 30°C, Use freshly prepared medium

Directions

Suspend 28.5 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Dispense in tubes in 8 - 10 ml amounts. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 50°C and add appropriate carbohydrate. Mix well and allow the tubed medium to cool in an upright position.

Principle and Interpretation

This medium is prepared by completely replacing Casein enzymic hydrolysate by HiVeg hydrolysate that is free from BSE/TSE risks. Cystine Tryptone Agar, HiVeg is the modification of Cystine Tryptone Agar which can be used as a maintenance medium for many fastidious organisms like Brucella, Corynebacteria, Pasteurella, Pneumococci and Streptococci without added enrichments (1, 2, 3). Anaerobic organisms like Actinomyces bovis, Bacteroides funduliformis and Leptotrichia (4) grow well on this medium in presence of Carbon dioxide (CO₂). The medium contains cystine & peptone which supply the nutrients necessary to support the growth of fastidious microorganisms.

Motility can be detected in this medium by stabbing the cultures. Motility is indicated by diffuse growth throughout the medium after incubation. Non-motile organisms show growth only in the inoculated area, whereas surrounding area remains clear. This medium is recommended as a basal medium for studying fermentation reactions of fastidious organisms because it is free from fermentable carbohydrates. Carbohydrate fermentation is detected by the colour change of the medium from red to yellow due to the pH indicator dye, phenol red incorpoarted in the medium. Cystine Tryptone Agar, HiVeg like the conventional medium requires heavy inoculum and still many times gives delayed results. Inadequate growth in many carbohydrates is due to different strains having various nutritional requirements. Addition of more than 0.5% cabohydrates may necessiate pH adjustment. Some times sodium chloride in the medium has inhibitory effect on Neisseria gonorrhoeae. Medium should be freshly prepared and cooled just before inoculation.

Quality Control

Appearance of powder Pink coloured, homogeneous, free flowing powder.

Gelling Semisolid, comparable with 0.25% Agar gel.

Colour and Clarity Red coloured clear to slightly opalescent gel forms in tubes as butts.

Reaction Reaction of 2.85% w/v aqueous solution is pH 7.3 ± 0.2 at 25°C.

Cultural Response

Cultural characteristics observed after an incubation at 35-37°C for 4-18 hours or longer if necessary.

Organisms (ATCC) Inoculum (CFU) Growth Motility Acid*
Escherichia coli (25922) 102-103 good-luxuriant + +
Neisseria gonorrhoeae (19424) 102-103 good - +
Neisseria meningitidis (13090) 102-103 good - +
Streptococcus pneumoniae (6303) 102-103 good - +

Key:

  • + = positive reaction, yellow colouration for acid/ diffused growth for motility
  • - = negative, no colour change / non motile
  • * = in presence of dextrose

References

  1. Peterson and Hartsell, 1955, J. Inf. Dis., 96:75.
  2. Myers and Koshy, 1962, Am. J. Pub. Health, 96:75.
  3. Alford, Wiese and Gunter, 1955, J. Bact., 69:518.
  4. Kroeger and Sibal, 1961, J. Bact., 50:581.
More Information
Product Name Cystine Tryptone Agar, HiVeg™
SKU MV159
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Vera H. D., 1948, J. Bacteriol. 55:531.
2.Peterson and Hartsell, 1955, J. Inf. Dis., 96:75.
3.Myers and Koshy, 1962, Am. J. Public Health, 96:75.
4.Alford, Wiese and Gunter, 1955, J. Bacteriol. 69:518.
5.Kroeger and Sibal, 1961, J. Bacteriol. 50:581.
6.Vera and Petran, 1954, Bull. Nat. Assoc. Clin. Labs., 5:907.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification -Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore.
8.Faur, Weisburd and Wilson, 1975, J. Clin. Microbiol. 1:294.
9.Applebaum and Lawrence, 1979, J. Clin. Microbiol. 9:598.
10.Kellogg, 1974, Manual of Clinical Microbiology, 2nd Ed. American Society for Microbiology, Washington D.C.
11. Yu and Washington, 1985, Laboratory Procedures in Clinical Microbiology, 2nd Ed., Springer - Verlag, New York, N.Y.1
2.Morse and Knapp, 1987, 7th Ed., American Public Health Association,Washington D.C.1
3.Baron E. J., Peterson and Finegold S. M., Bailey & Scotts Diagnostic Microbiology, 9th Ed., 1994, Mosby-Year Book,Inc., St. Louis, Mo.1
4.Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition. 15.Jorgensen,J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1.
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