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M-FC Agar Base, Granulated

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GM1122
For detection and enumeration of faecal coliforms using membrane filter technique at higher temperature (44.5°C).

Composition**

Ingredients Gms / Litre
Tryptose 10.000
Proteose peptone 5.000
Yeast extract 3.000
Lactose 12.500
Bile salts mixture 1.500
Sodium chloride 5.000
Aniline blue 0.100
Agar 15.000
Final pH (at 25°C) 7.4±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 52.1 grams in 1000 ml distilled water containing 10 ml 1% Rosolic Acid (FD058). Heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

M-FC Agar Base, designed by Geldreich et al (2) is used for the detection and enumeration of faecal coliforms using the membrane filter technique. This medium is based on the property of faecal coliforms to grow at 44-45°C (1). M-FC Agar Base is recommended by APHA (3) and by various other standards for detection of faecal coliforms (4-6). APHA recommends the membrane filtration procedure and delayed incubation for faecal coliforms.

Proteose peptone, tryptose and yeast extract provide necessary nutrients for the growth of faecal coliforms. Lactose is the carbon source as well as fermentable carbohydrate in the medium. Bile salts inhibit the growth of contaminating gram-positive microorganisms. Aniline blue is a triphenyl methane dye which suppresses the growth of many gram-positive microorganisms. Aniline blue along with rosolic acid forms the indicator system of the medium.

Membrane filters, through which water sample is passed are aseptically placed onto M-FC Agar base plates. If total coliforms are to be estimated, incubation is carried out at 35-37°C whereas if faecal coliform count is to be estimated, incubation is done at 44-45°C. Coliforms will form blue colonies whereas non-coliforms will form gray coloured colonies on M-FC Agar Base.

Quality Control

Appearance
Light yellow to greyish yellow coloured granular medium, may have slight green or blue tinge

Gelling
Firm, comparable with 1.5% Agar gel

Colour and Clarity of prepared medium
After Addition of 1% Rosolic Acid : Red coloured slightly opalescent gel forms in Petri plates

Reaction
Reaction of 5.21% w/v aqueous solution at 25°C. pH: 7.4±0.2

pH
7.20-7.60

Cultural Response
Cultural characteristics observed with added 1% Rosolic Acid (FD058) after an incubation at different temperatures for 22-24 hours.

Organism Inoculum (CFU) Growth at 35-37°C Growth at 45.5°C Colour of colony (on membrane filter)
Enterococcus faecalis ATCC >=10³ 29212 inhibited inhibited
Escherichia coli ATCC 25922 50-100 luxuriant luxuriant light blue
Salmonella Typhimurium ATCC 14028 50-100 luxuriant inhibited pinkish
Shigella flexneri ATCC 12022 50-100 luxuriant inhibited pinkish

Storage and Shelf Life

Store below 30°C in tightly closed container and use freshly prepared medium. Use before expiry date on the label.

Reference

  1. Collee J. G., Fraser A. G., Marmion B. P., Simmons A., (Eds.) Mackie and McCartney, Practical Medical Microbiology, 1996, 14th Edition, Churchill Livingstone.
  2. Geldreich E. E., Clark H. F., Huff E. E. and Bert M., 1965, J. Am. Water Works Assoc., 57:208.
  3. Rice E.W., Baird R.B., Eaton A. D., and Clesceri L. S. (Eds.), 2012, Standard Methods for the Examination of Water and Wastewater, 22nd Ed., APHA, Washington, D.C.
  4. Official Methods of Analysis of AOAC International, 2000, 17th Ed., AOAC International, Gaithersburg, Md.
  5. U.S. Environmental Protection Agency, 1992, EPA-814B-92-2002, Office of Ground Water and Technical Support Division, USEPA, Cincinnati, Ohio.
  6. Bordner R. H., Winter J. A. and Scarpino P. V. (Eds.), 1978, EPA-600/8-78-017, Environmental Monitoring and Support Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Cincinnati, Ohio.
More Information
Product Name M-FC Agar Base, Granulated
SKU GM1122
Product Type Granulated
Physical Form Granular
Origin Animal
Packaging type HDPE
References 1. Collee J. G., Fraser A. G., Marmion B. P., Simmons A., (Eds.) Mackie and McCartney, Practical Medical Microbiology,1996, 14th Edition, Churchill Livingstone.2.Geldreich E. E., Clark H. F., Huff E. E. and Bert M., 1965, J. Am. Water Works Assoc., 57:208.3.Eaton A. D., Clesceri L. S. and Greenberg A. W., (Eds.), 2005, Standard Methods for the Examination of Water andWastewater, 21st Ed., APHA, Washington, D.C.4.Official Methods of Analysis of AOAC International, 2000, 17th Ed., AOAC International, Gaithersburg, Md.5.U.S. Environmental Protection Agency, 1992, EPA-814B-92-2002, Office of Ground Water and Technical Support Division,USEPA, Cincinnati, Ohio.6.Bordner R. H., Winter J. A. and Scarpino P. V. (Eds.), 1978, EPA-600/8-78-017, Environmental Monitoring and SupportLaboratory, Office of Research and Development, U.S. Environmental Protection Agency, Cincinnati, Ohio.
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