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Brilliant Green HiCynth™ Agar Base, Modified

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SKU:
MCD016
Used for selective isolation of Salmonellae other than Salmonella Typhi from faeces and other materials.

Composition**

Ingredients Gms / Litre
HiCynth™ Peptone No.5* 3.000
HiCynth™ Peptone No.4* 10.000
Lactose 10.000
Sucrose 10.000
Sodium chloride 5.000
Phenol red 0.080
Brilliant green 0.0125
Agar 20.000
Final pH (at 25°C) 6.9±0.2

**Formula adjusted, standardized to suit performance parameters

*Chemically defined peptones.

Directions

Suspend 29.05 grams in 500 ml 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. For more selectivity, aseptically add rehydrated contents of 1 vial of Sulpha Supplement (FD068). Mix well and pour into sterile Petri plates.

Principle And Interpretation

Salmonella species cause many types of infections, from mild self-limiting gastroenteritis to life threatening typhoid fever. The most common form of Salmonella disease is self-limiting gastroenteritis with fever lasting less than 2 days and diarrhoea lasting less than 7 days.

Brilliant Green HiCynth™ Agar Base, Modified, as a primary plating medium for isolation of Salmonella species was first described by Kristensen et. al. (1) and further modified by Kauffmann (2). Brilliant Green Agar is also recommended by APHA (3,4) FDA (5) and described in EP, BP and IP (6,7,8).

This medium contains brilliant green, which inhibits growth of majority of Gram-negative and Gram-positive bacteria. Salmonella Typhi, Shigella species, Escherichia coli, Pseudomonas species, Staphylococcus aureus are mostly inhibited. Clinical specimens can be directly plated on this medium. 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.

The medium contains HiCynth™ Peptone No.4 and HiCynth™ Peptone No.5 as sources of carbon, nitrogen, long chain amino acids, vitamins and essential nutrients. The two sugars namely lactose and sucrose serve as energy sources. Fermentation of lactose and/or sucrose in the medium results in the formation of acidic pH which is detected by phenol red indicator. Sodium chloride maintains the osmotic equilibrium. Brilliant green helps to inhibit the contaminating microflora. The medium can further supplemented with sulphacetamide (1g/l) and sodium mandelate (0.25g/l) to inhibit contaminating microorganisms when the sample is suspected to contain large number of competing organisms along with Salmonella species.

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.

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

Reaction of 5.81% w/v aqueous solution at 25°C. pH: 6.9±0.2

pH

6.70-7.10

Cultural Response

Cultural response was carried out 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.

Organism Inoculum (CFU) Growth Lot value (CFU) Recovery Colour of Colony
Salmonella Typhimurium ATCC 14028 50-100 good-luxuriant 25-100 >=50% pinkish white
Salmonella Abony NCTC 6017 50-100 good-luxuriant 25-100 >=50% pinkish white
Salmonella Enteritidis ATCC 13076 50-100 luxuriant 25-100 >=50% pinkish white
Salmonella Typhi ATCC 6539 50-100 fair-good 15-40 30-40% reddish pink
Escherichia coli ATCC 25922 50-100 none-poor 0-10 0-10% yellowish green
Escherichia coli ATCC 8739 50-100 none-poor 0-10 0-10% yellowish green
Escherichia coli NCTC 9002 50-100 none-poor 0-10 0-10% yellowish green
Staphylococcus aureus ATCC 25923 >=10³ inhibited 0 0%
Staphylococcus aureus ATCC 6538 >=10³ inhibited 0 0%

Storage and Shelf Life

Store below 30°C in a tightly closed container and the prepared medium between 2 - 8°C. Use before expiry date on the label.

More Information
Product Name Brilliant Green HiCynth™ Agar Base, Modified
SKU MCD016
Product Type HiCynth™
Physical Form Powder
Origin Chemically defined (HiCynth™), Lactose
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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