Please refer disclaimer Overleaf.

Tryptone Salt Agar, w/1% NaCl M1877

Composition**Ingredients Gms / LitreTryptone 10.000Sodium Chloride 10.000Agar 20.000**Formula adjusted, standardized to suit performance parameters

DirectionsSuspend 40.0 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. For slants, dispense into tubes Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Solidify tubes as slants or mix well and pour into sterile Petri plates.

Principle And InterpretationMembers of the genus Vibrio are defined as Gram-negative, asporogenous rods that are straight or have a single, rigid curve. They are motile; most have a single polar flagellum, when grown in liquid medium. Different methods used for the confirmation of Vibrio species include physical, biochemical and serological assays (5). Tryptone salt agar w/ 1% NaCl is used for the growth of Vibrio sp. in accordance with FDA BAM (1) for differentiation of El Tor and Classical biotypes.

Blend the food sample to be analysed with Alkaline peptone water (APW) in appropriate ratio and incubate as per the recommendation by FDA BAM. Pure cultures can be isolated from APW by plating a loopful of the inoculum into TCBS

agar. For biochemical and serological identification of Vibrio, colonies from crowded plates must be streaked to Tryptone salt agar, w/1% NaCl for purity. Incubate overnight at 35 ±2° C and proceed with identification using a single isolated colony for differentiation of Classic and El Tor biotypes. Further biochemical tests can also be done using colonies from this medium.

Intended Use:Recommended for differentiation of El Tor and Classical biotypes of Vibrio in accordance with FDA BAM, 1998.

Type of specimen Food samples - Raw, undercooked, contaminated, or recontaminated seafood.

Specimen Collection and Handling

For food samples, follow appropriate techniques for sample collection and processing as per guidelines (4). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and PrecautionsRead the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations :

1. Further biochemical and serological tests must be carried out for complete identification.

expiry period Performance and EvaluationPerformance of the medium is expected when used as per the direction on the label within thewhen stored at recommended temperature.

HiMedia Laboratories Technical Data

Disclaimer :

User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained inthis and other related HiMedia™ publications. The information contained in this publication is based on our research and developmentwork and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes tospecifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use butfor laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should notbe considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.

5.Vera. 1944. J. Bact., 47.

1. FDA, U.S. 1998. Bacteriological Analytical Manual. 8 ed. Gaithersburg, Md. : AOAC International.

Revision :02 / 2019

Quality ControlAppearanceCream to yellow homogeneous free flowing powderGellingFirm, comparable with 2.0% Agar gelColour and Clarity of prepared mediumLight yellow coloured clear to slightly opalescent gel forms in Petri platesCultural ResponseCultural characteristics observed after an incubation at 35-37°C for 18-48 hours.

Organism Inoculum(CFU)

Growth Recovery

Vibrio cholerae ATCC15748

50-100

Vibrio parahaemolyticus ATCC 17802 (00037*)

50-100

good-luxuriant >=50%

good-luxuriant >=50%

Reference

Key : (*) Corresponding WDCM numbers.

Storage and Shelf LifeStore between 10-30°C in a tightly closed container and the prepared medium at 20-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition Seal the container tightly after use. Product performance is best if used within stated expiry period.

Disposal

User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (2,3).

4. Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.

2. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.3. 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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