Dichloran Rose Bengal Chloramphenicol Agar (DRBC Agar) M1881

Intended Use:
Recommended for selective isolation of fungi-yeasts and moulds of significance in food spoilage. The composition and
performance criteria are in accordance with ISO 21527-1:2008.

Composition**
Ingredients Gms / Litre
Peptone 5.000
Dextrose (Glucose) 10.000
Potassium dihydrogen phosphate 1.000
Magnesium sulphate 0.500
Rose Bengal 0.025
Chloramphenicol 0.100
Dichloran 0.002
Agar 15.000
Final pH ( at 25°C) 5.6±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 31.6 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. Mix well and pour into sterile Petri plates

Principle And Interpretation

Dichloran Rose Bengal Chloramphenicol Agar (DRBC Agar) is formulated by as described by King et.al (4) and
is recommended for selective isolation of yeasts and moulds especially in food samples. It is recommended by ISO (5)
This medium is a modification of Rose Bengal Chloramphenicol Agar which additionally contains dichloran.

Peptone provides nitrogeneous compounds, carbon, long chain amino acids, vitamins and other essential growth
nutrients. Dextrose (Glucose) is a carbohydrate source. Phosphate buffers the medium. Magnesium sulfate
provides divalent cations and sulfate. Dichloran is an antifungal agent, added to the medium to reduce colony
diameters of spreading fungi. Rose Bengal exhibits an improved inhibitory activity at pH 5.6 and hence the final
pH of the medium is maintained at 5.6 for the inhibition of spreading fungi (4) The presence of rose bengal in the
medium suppresses the growth of bacteria and restricts the size and colonies of the more rapidly growing
moulds. Chloramphenicol is included to inhibit the growth of bacteria present in environmental and food
samples. Inhibition of growth of bacteria and restriction of spreading of more-rapidly growing moulds aids in
the isolation of slow-growing fungi by preventing their overgrowth by more-rapidly growing species.
Additionally Rose Bengal is taken by yeast and moulds colonies, which allows these colonies to be easily recognized
and enumerated.
This medium should not be exposed to direct light as rose bengal undergoes photo-degradation leading to formation of
toxic chemicals for fungi (6,7).

Type of specimen
Food sample : Eggs, Meat, Dairy products (except milk powder), Fruits, Vegetables,Fresh pastes, etc.

Specimen Collection and Handling

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines
(1,5,6,8,9).
After use, contaminated materials must be sterilized by autoclaving before discarding.

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

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
1. Due to nutritional variations some strains may show poor growth.

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 pink homogeneous free flowing powder
Gelling
Firm,comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Pink coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 3.16% w/v aqueous solution at 25°C. pH : 5.6±0.2
pH
5.40-5.80

Cultural Response
Cultural characteristics observed after an incubation at 25-30°C for upto 6 days.

Organism Inoculum Growth Recovery

(CFU)
Bacillus subtilis subsp. >=104 inhibited 0%
spizizenii ATCC 6633
(00003)*
Candida albicans ATCC 50-100 good-luxuriant >=50%
10231 (00054)*
Escherichia coli ATCC >=104 inhibited 0%
25922 (00013)*
Escherichia coli ATCC >=104 inhibited 0%
8739 (00012)*
Mucor racemosus ATCC good-luxuriant
42647 (00181)*
Saccharomyces cerevisiae 50-100 good-luxuriant >=50%
ATCC 9763 (00058)*
Aspergillus brasiliensis ATCC good-luxuriant
16404 (00053)*
Key : (*) - Corresponding WDCM numbers

Storage and Shelf Life

Store the dehydrated powder and the prepared medium between 15-25°C in a tightly closed container. 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.

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

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 (2,3).

Reference
1. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,
Washington D.C.
2. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
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.
4. King D.A. Jr., Hocking A.D. and Pitt J.I., 1979, J. Appl. Environ. Microbiol., 37:959.
5. Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of yeasts and moulds -- Part 1:
Colony count technique in products with water activity greater than 0,95, ISO 21527-1:2008
6. Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination of
Foods, American Public Health Association, Washington, D.C.
7. Sharp A.N. and Jackson A.K., 1972, J. Appl. Bact., 24:175.
8. U.S. Food and Drug Administration, 1995, Bacteriological Analytical Manual, 8th Ed., AOAC International,
Gaithersburg, Md.
9. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,
APHA Inc., Washington, D.C.

Revision : 04/ 2019

Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in
this and other related HiMedia™ publications. The information contained in this publication is based on our research and development
work and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to
specifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use but
for laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should not
be considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.

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