Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Gelatin Liquefaction Test#CC293D
Intended Use
Recommended for detection of Clostridium species from food samples.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Lactose | 10.000 |
| Disodium hydrogen phosphate | 5.000 |
| Gelatin | 120.000 |
| Phenol red | 0.050 |
| Final pH (at 25°C) | 7.5±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 135.0 grams in 1000 ml warm purified / distilled water. Heat if necessary to dissolve the medium completely and dispense 10 ml amounts in 15x150 mm screw capped tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Just before use, heat to boiling to remove dissolved oxygen and cool rapidly to incubation temperature.
Principle And Interpretation
Members of the genus Clostridium are distributed widely in nature and are found in soil as well as in fresh water and marine water sediments throughout the world (5). Clostridial species are one of the major causes of food poisoning / gastro-intestinal illnesses. They are gram-positive, spore-forming rods that occur naturally in soil (1). Among the family are: Clostridium botulinum, which produces one of the most potent toxins in existence; Clostridium tetani, causative agent of tetanus; and Clostridium perfringens, commonly found in wound infections and diarrhoea cases. The use of toxins to damage the host is a method deployed by many bacterial pathogens including Clostridium. Lactose Gelatin Medium is prepared as per APHA (6) for detecting Clostridium species from food samples. The medium contains lactose which is fermented by the Clostridium species, mainly by Clostridium perfringens yielding acid and gas. Phenol red is the pH indicator which turns yellow at acidic pH. Gelatin is usually liquefied by Clostridium perfringens within 24-48 hours (2). Disodium phosphate buffers the medium.
Type of specimen
Food samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (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
- Further isolation on selective media is required.
- For complete identification, biochemical and serological tests must be carried out.
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 pink coloured homogeneous free flowing slightly coarse powder
Colour and Clarity of prepared medium
Red coloured clear to slightly opalescent gel forms in tubes.
Reaction
Reaction of 13.5% w/v aqueous solution at 25°C. pH: 7.5±0.2
pH
7.30-7.70
Cultural Response
Cultural characteristics observed under an anaerobic condition after an incubation at 35-37°C for 48-72 hours.
| Organism | Lactose Fermentation | Gelatin liquefaction |
|---|---|---|
| Clostridium perfringens ATCC 12924 | Positive reaction, yellow colour | Positive |
| Clostridium sporogenes ATCC 11437 | Negative reaction, no colour change or red | Positive reaction |
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°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 (3,4).
| Product Name | Lactose Gelatin Medium |
|---|---|
| SKU | M628 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C. 2.Czeczulin J. R., Hanna P. C., Mcclane B. A., 1993, Cloning, nucleotide sequencing, and expression of the Clostridiumperfringens enterotoxin gene in Escherichia coli. Infect. Immun. 61: 3429-3439. 3.Speck M.(Ed.), 1984, Compendium of Methods for the Microbiological Examination of Foods, 2nd ed., APHA, Washington,D.C. 2.Hauschild A.H.W. and Hilscheimer R., 1974, Appl. Microbiol., 27:78. |
| Customized Product Available | No |
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