Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for selective isolation and cultivation of Listeria monocytogenes, from food stuffs and other samples.
Composition
| Ingredients | g / L |
|---|---|
| HiVeg™ hydrolysate | 5.000 |
| HiVeg™ peptone | 5.000 |
| HiVeg™ extract | 3.000 |
| Sodium chloride | 5.000 |
| Glycine anhydride | 10.000 |
| Lithium chloride | 0.500 |
| Phenyl ethanol | 2.500 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 46.00 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. Before gelling, aseptically add sterile rehydrated contents of 1 vial of McBride Selective Supplement (FD070). Mix well and pour into sterile Petri plates.
Principle And Interpretation
The disease listeriosis is a frequent cause of abortions in cattle and sheep. In human, symptoms are manifested as septicemia, encephalitis and circulatory monocytosis (1). In women, Listeria may be harboured in the genital tract and may be transmitted transplacentally from infected amniotic fluid and vaginal discharge to infants (2). Listeria was first definitively described by Murray et al (3) in connection with an epizootic disease among laboratory-raised guinea pigs and rabbits. Since Listeria species are generally associated with animals, outbreaks of listeriosis are usually associated with contaminated dairy products. Listeria multiplies over a wide range of temperatures, from 3°C to 45°C, and over a pH range of 5.0 to 9.6. It also survives in food products with pH levels outside these parameters (4). Because of these properties, Listeria survives the various food processing techniques (5).
Modified McBride Listeria Agar Base differ from McBride Listeria Agar Base in the nutrient source available to Listeria species. Modified McBride Listeria HiVeg™ Agar Base is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. HiVeg™ hydrolysate, HiVeg™ peptone and HiVeg™ extract in the medium supply nitrogen, carbon, sulphur and trace nutrients required for the growth of Listeria. Phenyl ethyl alcohol is bacteriostatic for gram-negative bacteria as it selectively inhibits DNA synthesis (6). Sodium chloride maintains the osmotic balance of the medium. Glycine inhibits certain gram-negative and gram-positive bacteria including Escherichia coli and Enterococcus faecalis, the common accompanying contaminants. Lithium chloride also has antibacterial activity. Further selectivity is achieved by the addition of McBride Selective Supplement (FD070).
Type of specimen
Food and Dairy samples
Specimen Collection and Handling
The detection of L.monocytogenes is greatly improved by pre-enrichment in liquid media either by one step or two steps. In one step method (7), infected material is inoculated directly in Listeria Selective HiVeg™ Broth Base (MV889), while in two steps method (5), infected material is inoculated in Listeria Enrichment HiVeg™ Medium Base (UVM) (MV890A) and incubated at refrigeration temperature of 4°C for few weeks (cold enrichment), as the organism has the ability to grow in low temperature. It is then inoculated in Fraser Secondary Enrichment HiVeg™ Broth Base (MV1083), followed by plating onto selective agar such as Modified McBride Listeria Agar. The presumptive Listeria colonies are selected under 45° transillumination. Listeria colonies are dense white to iridescent white appearing as crushed glass. Small colonies tend to be blue, while non-Listeria show yellowish orange colonies that are further purified. 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 confirmation.
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 homogeneous free flowing powder
Gelling: Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium: Light amber clear to slightly opalescent gel forms in Petri plates
Reaction: Reaction of 4.6% w/v aqueous solution at 25°C. pH: 7.3±0.2
pH: 7.10-7.50
Cultural Response
Cultural characteristics observed under anaerobic condition with added McBride Listeria Supplement (FD070) after an incubation at 35-37°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | none - poor | 0-10% |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none - poor | 0-10% |
| Listeria monocytogenes ATCC 19112 | 50-100 | good-luxuriant | >=50% |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | none - poor | 0-10% |
* 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 (9,10).
| Product Name | Modified McBride Listeria HiVeg™ Agar Base |
|---|---|
| SKU | MV891 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1.Hyslop N., St. G. and Osborne A. D., 1959, Vet. Rec. 71 : 10822.Gray M. L., and McWade D. H., 1964, J. Bacteriol., 68:6343.Murray E. G. D., Webb R. A. and Swann M. B. R., 1926, J. Pathol. Bacteriol., 29:407.4.Conner D. E., Brackett R. E., and Beuchat L. R., 1986, Appl. Environ. Microbiol., 52: 595.Doyle M. P., Meske L. M. and Marth E. H., 1985, J. Food Prot., 48:7406.Dowell, Hill and Altemeier, 1964, J. Bacteriol., 88:18 |
| Customized Product Available | No |
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