Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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BLE Broth Base, Granulated (Buffered Listeria Enrichment Broth Base, Granulated)
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$163.90
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SKU:
GM1578
Recommended by FDA Committee for enrichment and isolation of Listeria monocytogenes.
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Intended Use
Recommended by FDA Committe for enrichment procedure for isolation of Listeria monocytogenes.
Composition**
| Ingredients | g/L |
|---|---|
| Tryptone | 17.000 |
| Soya peptone | 3.000 |
| Sodium chloride | 5.000 |
| Dipotassium hydrogen phosphate | 2.500 |
| Dextrose (Glucose) | 2.500 |
| Yeast extract | 6.000 |
| Potassium dihydrogen phosphate | 1.350 |
| Disodium hydrogen phosphate | 9.600 |
| Sodium pyruvate | 1.11 |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 24.03 grams in 500 ml purified/distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45 - 50°C. Aseptically add the rehydrated contents of 1 vial of ANC Selective Supplement (FD063I). Mix well and dispense into sterile tubes or flasks as desired.
Principle And Interpretation
Listeria monocytogenes is the only species of the Listeria genus that causes Listeriosis in human, however occasionally L.seeligeri, L. welshimeri and L. ivanovii have been related with human diseases. Microbiological and epidemiological evidence from both sporadic and epidemic cases of listeriosis has show that the principal route of transmission is via the consumption of foodstuffs contaminated with L. monocytogenes (1).
L. monocytogenes is a well-documented food borne pathogen because of its high morbidity on infection to animals and humans and also due to its psychrotrophic nature exhibiting high tolerance to heat, cold and desiccation.The organism has been isolated from commercial dairy and other food processing plants, and is ubiquitous in nature, being present in a wide range of unprocessed foods and in soil, sewage, silage and river water (2). Listeria species grow over a pH range of 4.4-9.6, and survive in food products with pH levels outside these parameters (3). Listeria species are microaerophilic, gram-positive, asporogenous, non-encapsulated, non-branching, regular, short, motile rods. Motility is most pronounced at 20°C. Food samples are often contaminated with organisms other than Listeria, which makes its isolation difficult (4). To recover low numbers of L. monocytogenes from food samples, initial enrichment is required. Listeria Enrichment Broth was modified by adding buffering strength thereby making it possible for the medium to be used successfully in conjunction with DNA probe and other methods that are more sensitive than conventional culture procedure. This medium is also recommended by APHA for the selective enrichment of L.monocytogenes.
Tryptone and soya peptone provide amino acids and other complex nitrogenous substances. Dextrose is the energy source. Sodium pyruvate aids in resuscitation of organisms. The phosphates provide buffering capacity. Sodium chloride maintains the osmotic equilibrium. Yeast extract provides vitamin B complex. The medium is rendered selective due to the inclusion of antimicrobial agents. Cycloheximide inhibits the growth of saprophytic fungi. Nalidixic acid inhibits the growth of gram-negative organisms, whereas acriflavin suppresses growth of gram-positive microorganisms.
Type of specimen
Food and Dairy samples
Specimen Collection and Handling
According to FDAs enrichment procedure (5) for isolation of L. monocytogenes from dairy products, the sample to be tested is inoculated in enrichment broth and incubated at 30°C for 4 hours without the selective supplement. After 4 hours the selective supplement is added and further kept for incubation for additional 44 hours at 30°C. After 24 hours and 48 hours the enriched culture is streaked on Oxford Listeria Medium Base (M1145) and LPM Agar (M1228) / Listeria Identification Agar Base, PALCAM (M1064) and incubated at 35°C for 24-48 hours. Presumptive Listeria colonies are selected and colonies are further purified on Tryptone Soya Yeast Extract Agar (M1214). Purified isolates are then subjected to a variety of biochemical tests to confirm the presence of L. monocytogenes. 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 biochemical tests must be carried out for complete identification.
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
Cream to yellow coloured granular medium
Colour and Clarity of prepared medium
Yellow coloured, clear to slightly opalescent solution with slight precipitate
Reaction
Reaction 4.8% w/v aqueous solution at 25°C. pH : 7.3±0.2
pH
7.10-7.50
Cultural Response
Cultural characteristics observed with added ANC Selective Supplement (FD0631 )after an incubation at 30°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited |
| Listeria innocua ATCC 33090 (00017*) | 50-100 | good to luxuriant |
| Listeria monocytogenes ATCC 19111 (00020*) | 50-100 | good to luxuriant |
| Listeria monocytogenes ATCC 19112 | 50-100 | good-luxuriant |
| Listeria monocytogenes ATCC 19118 | 50-100 | good-luxuriant |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | none-poor |
| Saccharomyces cerevisiae ATCC 9763 (00058*) | 50-100 | none-poor |
Key: *Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°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 (6,7).
| Product Name | BLE Broth Base, Granulated (Buffered Listeria Enrichment Broth Base, Granulated) |
|---|---|
| SKU | GM1578 |
| Product Type | Granulated |
| Physical Form | Granular |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Downes F. P. and Ito K., (Ed.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th Ed.,American Public Health Association, Washington, D.C. |
| Customized Product Available | No |
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