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Brilliant Green Agar Medium

Name: Brilliant Green Agar Medium
Brand: HiMedia Laboratories
SKU: MU016
Availability: InStock

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MU016

For selective isolation of Salmonellae other than Salmonella typhi from faeces, food, dairy products etc. in accordance with US Pharmacopoeia.

Description

Intended Use:

Recommended for selective isolation of Salmonellae other than Salmonella typhi from faeces, food, dairy products etc. in accordance with USP.

Composition**

Ingredients	g/L
Peptone	5.000
Tryptone	5.000
Yeast extract	3.000
Lactose	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

Directions

Suspend 58.09 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. AVOID OVERHEATING. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

Brilliant Green Agar medium is recommended 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 (1). Kauffmann further modified it for isolation of Salmonella from stool samples (2). Brilliant green agar is also recommended by APHA (3,4) FDA (5) and is in accordance with United States Pharmacopoeia (6). 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. Brilliant Green Agar is used in the microbial limits test and with novobiocin for testing food and pharmaceutical products. Combination of peptone, tryptone and yeast extract makes the medium highly nutritious and supplies amino acids and long chains of peptides. Sodium chloride maintains the osmotic equilibrium. Lactose 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, Staphylococcus aureus are mostly inhibited.

However, 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, 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

Foodstuffs & dairy samples; Pharmaceutical samples.

Specimen Collection and Handling

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (3,4,7,8). 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

Though this medium is selective for Salmonella other species of Enterobacteriaceae may grow.
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.
Further confirmation has to be carried out on presumptive Salmonella isolates.

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

pH
6.70-7.10

Growth Promotion Test
Growth Promotion was observed in accordance with USP.

Cultural Response

Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.

Organism	Inoculum (CFU)	Growth	Observed Lot Recovery value (CFU)	Recovery	Colour of Colony	Incubation temperature
Growth Promotion Test
Salmonella Typhimurium ATCC 14028 (00031*)	50-100	good-luxuriant	25-100	>=50%	pinkish white	24-48 hrs
Salmonella Abony NCTC 6017 (00029*)	50-100	good-luxuriant	25-100	>=50%	pinkish white	24-48 hrs
Additional Microbiological testing
Salmonella Enteritidis ATCC 13076 (00030*)	50-100	Good-luxuriant	25 -100	>=50%	pinkish white	24-48 hrs
Salmonella Typhi ATCC 6539	50-100	poor-good	15-40	30-40%	reddish pink	24-48 hrs
Escherichia coli ATCC 25922 (00013*)	50-100	none-poor	0-10	0-10%	yellowish green	24 -48 hrs
Escherichia coli ATCC 8739 (00012*)	50-100	none-poor	0-10	0-10%	yellowish green	24-48 hrs
Staphylococcus aureus subsp. aureus ATCC 25923(00034*)	>=10³	inhibited	0	0%		24-48 hrs
Staphylococcus aureus subsp. aureus ATCC 6538 (00032*)	>=10³	inhibited	0	0%		24-48 hrs

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 (9,10).

Product Information

More Information
Product Name	Brilliant Green Agar Medium
SKU	MU016
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,9.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C. 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.
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