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Agar Medium L (Brilliant Green, Phenol Red, Lactose Monohydrate, Sucrose Agar)
As low as
$48.40
Availability: Out of stock
Availability: In stock
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SKU:
ME016
Used for selective isolation of Salmonellae other than Salmonella Typhi from faeces, food, dairy products etc in accordance with European Pharmacopoeia.
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Intended use
Brilliant Green, Phenol Red, Lactose Monohydrate, Sucrose Agar is used for selective isolation of Salmonellae other than Salmonella Typhi from foods, dairy products etc in accordance with European Pharmacopoeia.
Composition
| Ingredients | g/L |
|---|---|
| HMC Peptone ~ | 10.000 |
| Yeast extract | 3.000 |
| Lactose monohydrate | 10.000 |
| Sucrose | 10.000 |
| Sodium chloride | 5.000 |
| Phenol red | 0.080 |
| Brilliant green | 0.0125 |
| Agar | 20.000 |
| pH after sterilization(at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
~ - Equivalent to Peptones (meat and casein)
Directions
Suspend 57.59 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15lbs pressure (121°C) for 15 minutes or as per validated cycle. Cool to 45-50°C. Mix well before pouring into sterile Petri plates.
Principle And Interpretation
The composition of medium is as per European Pharmacopoeia and is cited as Agar medium L (1). It can also be used for food samples (2) and clinical samples (3,4). Brilliant Green, Phenol Red, Lactose Monohydrate, Sucrose Agar is used as a primary plating medium for isolation of Salmonella species was first described by Kristensen et al as medium for differentiation of paratyphoid B from other Gram negative enteric bacteria (5) It was further modified by Kauffmann for isolation of Salmonella from stool samples (6). Brilliant green agar is also recommended by APHA (7,8) FDA (2). This medium is employed in testing clinical specimens. Heavy inocula and heavily contaminated samples can be analyzed due to the outstanding selectivity of this medium. HMC Peptone and yeast extract supplies essential amino acids and long chains of peptides for enhanced growth. Sodium chloride maintains the osmotic equilibrium. Lactose monohydrate and sucrose are the fermentable carbohydrate sources. Phenol red serves as an acid base indicator giving yellow colour to lactose and or sucrose fermenting bacteria. This medium also contains brilliant green, which inhibits growth of majority of Gram-negative and Gram-positive bacteria. Salmonella Typhi, Shigella species, Escherichia coli, Proteus species, Pseudomonas species, and Staphylococcus aureus are mostly inhibited. Being highly selective, it is recommended that this medium should be used along with a less inhibitory medium to increase the chances of recovery. Often cultures enriched in Selenite or Tetrathionate Broth are plated on Brilliant Green Agar along with Bismuth Sulphite Agar, SS Agar, and MacConkey Agar. Non-lactose fermenting bacteria develop white to pinkish red colonies within 18-24 hours of incubation. Salmonella Typhi and Shigella species may not grow on this medium, moreover Proteus, Pseudomonas and Citrobacter species may mimic enteric pathogens by producing small red colonies.
Type of specimen
Food and dairy samples; Pharmaceutical samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (2,7,8). For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (1).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
- Salmonella Typhi and Shigella species may not grow on this medium, moreover Proteus, Pseudomonas and Citrobacter species may mimic enteric pathogens by producing small red colonies.
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 light pink homogeneous free flowing powder
Gelling: Firm, comparable with 2.0% agar gel.
Colour and Clarity of prepared medium: Greenish brown clear to slightly opalescent gel forms in Petri plates
Reaction: After sterilization, reaction of 5.8% w/v aqueous solution. pH: 6.9±0.2
pH: 6.70-7.10
Growth Promotion Test
Growth Promotion was observed in accordance with EP, after an incubation at 30-35°C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
Cultural Response
Cultural characteristics observed after incubation at 30-35°C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Growth Promoting | ||||
| Salmonella Typhimurium ATCC 14028 | 50-100 | good-luxuriant | >=50% | pinkish white |
| Salmonella Abony NCTC 6017 | 50-100 | good-luxuriant | >=50% | pinkish white |
| Additional Microbiological testing | ||||
| Salmonella Enteritidis ATCC 13076 | 50-100 | luxuriant | >=50% | pinkish white |
| Salmonella Typhi ATCC 6539 | 50-100 | fair-good | 30-40% | reddish pink |
| Escherichia coli ATCC 25922 | 50-100 | none-poor | 0-10% | yellowish green |
| Escherichia coli ATCC 8739 | 50-100 | none-poor | 0-10% | yellowish green |
| Staphylococcus aureus ATCC 25923 | >=104 | inhibited | 0% | |
| Staphylococcus aureus ATCC 6538 | >=104 | inhibited | 0% | |
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 (3,4).
| Product Name | Agar Medium L (Brilliant Green, Phenol Red, Lactose Monohydrate, Sucrose Agar) |
|---|---|
| SKU | ME016 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Kristensen M., Lester V, and Jurgens A., 1925, Brit.J.Exp.Pathol.,6:291. 2.Kauffman F., 1935, Seit F. Hyg. 177:26. 3.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C. 4.Standard Methods for the Microbiological Examination of Dairy Products, 1995, 19th Ed, APHA, Washington, D.C. 5.Bacteriological Analytical Manual, 5th Ed, 1978, AOAC, Washington D.C. 6.The European Pharmacopoeia, 2008, Council or Europe, Strasbourg. 7.The British Pharmacopoeia, 2008 vol. II, London. 8.Indian Pharmacopoeia, 2010, Ministry of Health and Family Welfare, Govt., of India, 10.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. 11. Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater,23rd ed., APHA, Washington, D.C. 12..Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.1 3.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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