Please refer disclaimer Overleaf.

M499Cooke Rose Bengal Agar Base

Ingredients Gms / LitreSoya Peptone 5.000Dextrose (Glucose) 10.000Potassium dihydrogen phosphate 1.000Magnesium sulphate 0.500Rose Bengal 0.035Agar 20.000Final pH ( at 25°C) 6.0±0.2**Formula adjusted, standardized to suit performance parameters

DirectionsSuspend 36.54 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. To increase the selectivity of the medium, add 35µg chlortetracycline per ml of the medium. Mix well and pour into sterile Petri plates.

Principle And InterpretationCooke Rose Bengal Agar is a selective medium formulated as per Cooke (2,3). A variety of inhibitory agents have been used

to inhibit bacteria in an attempt to isolate fungi from mixed flora. The Kingdom Fungi includes some of the most important organisms, both in terms of their ecological and economic roles. By breaking down dead organic material into simpler forms, they continue the cycle of elements through ecosystems. In addition, most vascular plants could not grow without a symbiotic association with fungi, or mycorrhizae, that inhabit their roots and supply essential nutrients. Other fungi provide numerous drugs (such as penicillin and other antibiotics), foods like mushrooms, truffles and morels, and the bubbles in bread, champagne, and beer (7). Waksman (11) described an acid medium consisting of peptone, dextrose, inorganic salts and agar for the isolation of fungi from soil. Cooke (2) used the Waksman medium without adjustment for isolation of fungi from sewage. It was discovered that soya pepton was particularly suitable for use in this medium and that the combination of chlortetracycline, or oxytetracycline, with rose bengal increased the selectivity of the medium.

Smith and Dawson (9) used rose bengal for the inhibition of bacteria in media which has almost neutral reaction concerned with retardation of the development of fungi. Martin (6) used 1: 30,000 Rose bengal and 30µg Streptomycin per ml and found that a wide variety of bacteria are inhibited at reactions between pH 5.5 to 6.5 without inhibiting fungi.

The medium should not be exposed to light as photo-degradation of rose bengal yields compounds that are toxic to fungi (1,8).

Microscopic examination coupled with biochemical testing using pure cultures is recommended for complete identification. Due to the selective properties of this medium and the type of specimen being cultured, some strains of fungi may be encountered that fail to grow or grow poorly on the complete medium; similarly, some strains of bacteria may be encountered that are not inhibited or partially inhibited.

Soya Peptone provides nitrogen, carbon and vitamins. Dextrose is an energy source. Rose bengal and chlortetracycline selectively inhibit bacterial growth and restrict the size and height of colonies of more rapidly growing moulds. Potassium

dihydrogen phosphate provides buffering capability. Magnesium sulfate is a source of divalent cations.

Composition**

Intended Use:Recommended for selective isolation and cultivation of fungi.

Type of specimen Food samples; Soil samples; Industrial fermentation samples.

Specimen Collection and Handling:

HiMedia Laboratories Technical Data

Quality ControlAppearanceLight yellow to light pink homogeneous free flowing powderGellingFirm, comparable with 2.0% Agar gelColour and Clarity of prepared mediumPink-red coloured, slightly opalescent forms in Petri platesReactionReaction of 3.65% w/v aqueous solution at 25°C. pH : 6.0±0.2pH5.80-6.20Cultural ResponseCultural characteristics observed after an incubation at 25-30°C for 1-4 days.

Organism Inoculum(CFU)

Growth(Plain)

Recovery(Plain)

Growth with chlortetracycline

Recovery with chlortetracycline

Candida albicans ATCC10231 (00054*)

50-100 luxuriant >=50% luxuriant >=50%

Saccharomyces cerevisiae ATCC 9763 (00058*)

50-100 luxuriant >=50% luxuriant >=50%

#Aspergillus brasiliensisATCC 16404 (00053*)

50-100 good good

Escherichia coli ATCC25922 (00013*)

50-100 luxuriant >=50% inhibited 0%

Enterococcus faecalis ATCC29212 (00087*)

>=104 inhibited 0% inhibited 0%

Key :*Corresponding WDCM numbers.#- Formerly known as Aspergillus niger

For soil samples follow appropriate techniques for handling specimens as per established guidelines (4,5). For food samples, follow appropriate techniques for sample collection and processing as per guidelines (8). For industrial fermentation samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(7) After use, contaminated materials must be sterilized by autoclaving before discarding.

Please refer disclaimer Overleaf.

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.This medium is general purpose medium and may not support the growth of fastidious organisms.

Performance and EvaluationPerformance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.

Storage and Shelf LifeStore 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. Use before expiry date on the label.Product performance is best if used within stated expiry period.

Disclaimer :

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

HiMedia Laboratories Technical Data

Reference

Revision :02 / 2019

HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate office : A-516,Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147 1919 Email: techhelp@himedialabs.com Website: www.himedialabs.com

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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).

Disposal

1. Banks, Board and Paton, 1985, Lett. Appl. Microbiol., 1:7.2. Cooke, 1954, Antibiot. Chemother., 4:657.3. Eaton A. D., Clesceri L. S., Rice E. W., and Greenberg A. E., (Eds.) , 2005, Standard Methods for the Examination of

Water and Wastewater, 21st Ed., APHA, Washington, D.C.4. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.5. 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. 1.6. Martin, 1950, Ibid., 69:215.7. Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of ClinicalMicrobiology, 8th Ed., American Society for Microbiology, Washington, D. C.8. 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.9. Smith and Dawson, 1944, Soil Sci., 58:467.10. Subba Rao N. S., 1977, Soil Microorganisms and Plant Growth, Oxford and IBH Publishing Co., New Delhi.11. Waksman, 1922, J. Bacteriol., 7:339.