HiCrome™ Coliform HiVeg™ Agar w/ SLS

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SKU:
MV1300
A selective agar for the simultaneous detection of total coliforms and Escherichia coli in water and food samples.


Intended use

Recommended for the simultaneous detection of Escherichia coli and total coliforms in water and food samples.

Composition**

Ingredients g / L
HiVeg™ special peptone 3.000
Sodium chloride 5.000
Dipotassium hydrogen phosphate 3.000
Potassium dihydrogen phosphate 1.700
Sodium pyruvate 1.000
L-Tryptophan 1.000
Sodium lauryl sulphate 0.100
Chromogenic mixture 0.200
Agar 12.000
Final pH (at 25°C) 6.8±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 27 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Add 5mg/l novobiocin before autoclaving the medium, when a high number of gram positive accompanying bacteria are expected. Mix well and pour into sterile Petri plates.

Principle And Interpretation

HiCrome™ Coliform Agar w/ SLS is a selective medium recommended for the simultaneous detection of Escherichia coli and total coliforms in water and food samples (1). HiCrome™ Coliform HiVeg™ Agar w/ SLS is prepared by completely replacing animal based peptones with vegetable peptones to avoid BSE/TSE risks associated with vegetable peptones.

HiVeg™ special peptone and sodium pyruvate provide essential growth nutrients to the organisms. The phosphates buffer the medium well. The medium composition helps even the sublethally injured coliforms to grow rapidly. Sodium lauryl sulphate inhibits gram-positive organisms. The chromogenic mixture contains two chromogenic substrates. The enzyme ß-galactosidase produced by coliforms cleaves one chromogen, resulting in the salmon red colouration of coliform colonies. The enzyme ß-glucuronidase produced by E. coli, cleaves X-glucuronide. E.coli forms dark blue to violet coloured colonies due to cleavage of both the chromogens (2,3,4). The addition of L-Tryptophan improves the indole reaction, thereby increasing detection reliability in combination with the two chromogens. To confirm E.coli, add a drop of Kovac's reagent (R008) on the dark-blue to violet colony. Formation of cherry-red colour indicates positive reaction.

Type of specimen

Food samples; Water samples

Specimen Collection and Handling:

For food samples, follow appropriate techniques for sample collection and processing as per guidelines (5).

For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (6).

After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

Read 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. B-glucuronidase is present in 97% of E.coli strains, however few E.coli may be negative.
  2. Certain species of Shigella and Salmonella are B-glucuronidase positive

Performance and Evaluation

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

Quality Control

Appearance
Light yellow to beige homogeneous free flowing powder

Gelling
Firm, comparable with 1.2% Agar gel.

Colour and Clarity of prepared medium
Light yellow, clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 2.7% w/v aqueous solution at 25°C. pH: 6.8±0.2

pH
6.60-7.00

Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24 hours (48 hours if necessary).

Organism Inoculum (CFU) Growth Recovery Colour of Colony Indole production
Citrobacter freundii ATCC 8090 50-100 good-luxuriant >=50% salmon to red negative reaction
Escherichia coli ATCC 8739 (00012)* 50-100 good-luxuriant >=50% dark blue/violet positive, confirmation of red colour around the colony by addition of Kovacs reagent (R008)
Enterobacter cloacae ATCC 23355 (00082*) 50-100 good-luxuriant >=50% salmon to red negative reaction
Enterococcus faecalis ATCC 29212 (00087*) >=104 inhibited 0%
Klebsiella pneumoniae ATCC 13883 (00097*) 50-100 good-luxuriant >=50% light pink negative reaction
Salmonella Enteritidis ATCC 13076 (00030*) 50-100 good 40-50% colourless negative reaction
Shigella flexneri ATCC 12022 (00126*) 50-100 good 40-50% colourless negative reaction

Key : *Corresponding WDCM numbers.

Storage and Shelf Life

Store between 15-25°C in a tightly closed container and the prepared medium at 2-8°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 (7,8).

More Information
Product Name HiCrome™ Coliform HiVeg™ Agar w/ SLS
SKU MV1300
Product Type HiVeg™, HiCrome™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water andWastewater, 23rd ed., APHA, Washington, D.C.2.Frampton E. W., Restaino L. and Blaszko N., 1988, J. Food Prot., 51:402.3.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.4.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. 1.5.Kilian M. and Bülow P., 1976, Acta. Pathol. Microbiol. Scand., Sect. B, 84:245.6.LeMinor L. and Hamida F., 1962, Ann. Inst. Pasteur (Paris), 102:2677.Manafi M. and Kneifel W., 1989, Zentralbl. Hyg., 189:225.8.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2001, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.
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