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Thane(W)-400604, MH, India.
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Hugh Leifson Medium
Recommended for detecting aerobic and anaerobic breakdown of glucose. It is recommended by BIS committee under the specifications IS:5887 (Part V) -1976.
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Intended use
Recommended for detecting aerobic and anaerobic breakdown of glucose. It is recommended by BIS committee under the specifications IS:5887 (Part V)-1976, Reaffirmed 2005.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone | 2.000 |
| Sodium chloride | 5.000 |
| Dipotassium hydrogen phosphate | 0.300 |
| Glucose (Dextrose) | 10.000 |
| Bromothymol blue | 0.030 |
| Agar | 3.000 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 20.33 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Dispense into test tubes in duplicate for aerobic and anaerobic fermentation. Sterilize by autoclaving at 10 lbs pressure (115°C) for 20 minutes. Cool the tubed medium in an upright position.
Note: In an additional set of tubes 5 mm paraffin oil may be layered on surface of medium for the differentiation of oxidative and fermentative organisms.
Principle And Interpretation
Hugh Leifson Medium was formulated by Hugh and Leifson (1). They described the taxonomic significance of fermentative and oxidative metabolism of carbohydrates in gram-negative intestinal bacteria.
It is recommended by BIS (2) for the isolation and cultivation of Vibrio cholerae and other Vibrio species which cause food poisoning.
The medium contains a high concentration of carbohydrate and low concentration of peptone to avoid the possibility of an aerobic organism utilizing peptone and producing an alkaline condition which would neutralize slight acidity produced by an oxidative organism (3). Dipotassium phosphate promotes fermentation and acts as pH controlling buffer. Agar concentration enables the determination of motility and aids in distribution of acid throughout the tube produced at the surface of medium. Oxidative organisms produce acid in unsealed tube with little or no growth and no acid formation in sealed tube while fermentative organisms produce acid in both sealed and unsealed tubes.
Type of specimen
Food samples
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 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 guidleines should be followed while handling clincal specimens. Saftey guidelines may be referred in individual safety data sheets
Limitations
- Further biochemical and serological testing is required for complete identification.
- Certain strains may show poor growth due to nutritional variations.
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
Appearance
Light yellow to bluish green homogeneous free flowing powder
Gelling
Semisolid,comparable with 0.2% Agar gel.
Colour and Clarity of prepared medium
Greenish blue coloured, clear to slightly opalescent gel forms in tubes as butts
Reaction
Reaction of 2.03% w/v aqueous solution at 25°C. pH: 7.1±0.2
pH
6.90-7.30
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
| Organism | Aerobic fermentation | Anaerobic fermentatoin |
|---|---|---|
| Vibrio cholerae ATCC 15748 | acid (yellow) and gas production, positive reaction | acid (yellow) and gas production, positive reaction |
| Vibrio parahaemolyticus ATCC 17802 (00037*) | greenish blue, negative reaction | acid (yellow) and gas production, positive reaction |
Key- (*) coresponding WDCM number
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°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 (5,6).
| Product Name | Hugh Leifson Medium |
|---|---|
| SKU | M826S |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1.Hugh and Leifson, 1953, J. Bacteriol., 66:24.2.MacFaddin J.F., 1985, Media for Isolation-Cultivation-Identification- Maintenance of Medical Bacteria,Vol. I, Williams andWilkins, Baltimore.3.Bureau of Indian Standards, IS:5887 (Part V) 1976, reaffirmed 1986Co., St. Louis. |
| Customized Product Available | No |
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