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Directions
Dissolve 37g in 1 litre of distilled water. Dispense into final containers and sterilize by autoclaving at
121°C for 15 minutes.
Description
The original formulation described by Perry and Hajna1 with Tryptose as the nitrogen source was
subsequently substituted for Tryptone . The medium has been shown to be a reliable medium producing
dependable results in 24 to 48 hours2.
EC Broth is a selective medium for the differentiation of faecal coliforms and the confirmatory test for
Escherichia coli from food and environmental samples3,4. It is also suitable for the enumeration of
presumptive E. coli in milk and milk products using the most probable number technique2.
Escherichia coli is a common Gram-negative micro-organism which may be present in food and water
samples. The presence of E. coli cells may be indicative of faecal contamination, although their ingestion
does not necessarily have adverse effects on health. Nevertheless, certain strains are responsible for
diarrhoeal disease and can also lead to more serious forms of illness.
The presence of E. coli in water is also regarded as a specific indicator of faecal pollution, but lack of
rapid identification methods means that the non-specific group - faecal coliforms - is also used as the
indicator of such pollution. In this regard, it is important to differentiate between coliforms of faecal origin
and those from other sources.
The identification criterion for E. coli and faecal coliforms is the fermentation of lactose to acid and gas
within 24-48 hours at 44-45.5°C, is dependent upon which method and/or material is used:
Technique
The formulation includes a buffered lactose broth and bile salts No.3. The presence of bile salts inhibits
the growth of spore formers and enterococci, but allows the growth of E.coli and coliforms. Gas
production from lactose fermentation is indicated by using inverted Durham tubes. Methods are
summarised below but refer to the appropriate standard for the detailed technique.
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For differentiation of faecal coliforms inoculate EC Broth (CM0853) fermentation tubes containing
inverted Durham tubes, and incubate in a water bath at 44.5°C for 24 ± 2°C hours. Gas production and
growth within 24 hours or less is considered a positive faecal coliform reaction. For full details please
refer to the APHA Standard Method3.
Appearance
Dehydrated medium: White, free-flowing powder
Prepared medium: Straw coloured solution
Quality control
References
1. Hajna, A. A. and Perry, C. A. (1943) Amer. J. Pub. Hlth. 33, 550-556.
2. Perry, C. A. and Hajna, A. A. (1944) Amer. J. Pub. Hlth. 34, 735-738.
3.International Organisation for Standardisation: Milk and Milk Products - Enumeration of presumptive
Escherichia coli. Part 1. Most probable number technique ISO 11866-1 : 1997.
4. American Public Health Association (1998) Standard Methods for the Examination of Water and
Wastewater. 20th Edn. APHA Inc. Washington DC.
5."Association of Official Analytical Chemists" F.D.A. Bacteriological Analytical Manual 8th Edition (1995)
AOAC, Arlington Va.
6. International Organization for Standardization. ISO 11133:2014 Microbiology of food, animal feed and
water - Preparation, production, storage and performance testing of culture media
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