Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the detection of Escherichia coli in water and food samples by using a combination of chromogenic and fluorogenic substrate.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg™™ hydrolysate | 20.000 |
| Synthetic detergent | 1.500 |
| Tryptophan | 1.000 |
| Lactose | 5.000 |
| Sodium chloride | 5.000 |
| Dipotassium hydrogen phosphate | 4.000 |
| Potassium dihydrogen phosphate | 1.500 |
| Flurogenic substrate | 0.070 |
| Chromogenic substrate | 0.100 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.0±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 53.17 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. Mix well and pour in sterile Petri plates.
Principle And Interpretation
HiCrome™ ECD HiVeg™™ Agar w/ MUG is prepared by completely replacing animal based peptones with vegetable peptones thereby making the medium BSE/TSE risk free. HiCrome™ ECD HiVeg™™ Agar w/ MUG is a slight modification of HiCrome™ ECD Agar w/ MUG and is recommended for rapid detection of Escherichia coli by using a combination of chromogenic and fluorogenic substrates. The presence of Escherichia coli is indicated by blue coloured colony formation due to cleavage of chromogenic substrate. Fluorogenic substrate permits rapid detection of Escherichia coli when medium is observed for fluorescence using UV light (1,2). Fluorogenic substrate also detects anaerogenic strains, which may not be detected in conventional procedure (1). It is hydrolysed by enzyme beta-D-glucuronidase, possessed by Escherichia coli to yield a fluorescent end product. The reaction is indicated by a blue fluorescence under UV light.
HiVeg™™ hydrolysate provides carbon, nitrogen substances, long chain amino acids, vitamins and other essential nutrients. Lactose is the fermentable carbohydrate. Sodium chloride maintains osmotic equilibrium. The medium has a strong buffering system to control the pH in the presence of fermentative action. Synthetic detergent inhibits gram-positive bacteria especially Bacillus species and faecal Streptococci.
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 (3).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (4).
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
- B-glucuronidase especially present in 97% of E.coli strains, however few E.coli may be negative.
- Slight colour variation may be observed depending upon the utilization of the substrate by the organism.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
AppearanceCream to yellow homogeneous free flowing powder
GellingFirm, comparable with 1.5% Agar gel
Colour and Clarity of prepared mediumLight amber coloured clear to slightly opalescent gel forms in Petri plates
ReactionReaction of 5.32% w/v aqueous solution at 25°C. pH: 7.0±0.2
pH6.80-7.20
Cultural Response
Cultural characteristics observed after an incubation at 43 - 45°C for 18 - 24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony | Fluorescence (under uv) | Indole |
|---|---|---|---|---|---|---|
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | 0% | |||
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good | >=50% | bluish-green | Positive | Positive, red ring at the interface of the medium |
| Klebsiella pneumoniae ATCC 13883 (00097*) | 50-100 | good | >=50% | colourless | Negative | Negative, no colour development / cloudy ring |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good | >=50% | colourless | Negative | Negative, no colour development / cloudy ring |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% |
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 (5,6).
| Product Name | HiCrome™ ECD HiVeg™ Agar w/ MUG |
|---|---|
| SKU | MV1488 |
| Product Type | HiVeg™, HiCrome™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1.Feng PCS and Hartman PAS, (1982), Appl. Environ. Microbiol. 43:132.2.Robinson (1984), Appl. Environ. Microbiol., 48:285. |
| Customized Product Available | No |
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