17171 Mineral modified Glutamate Broth (Base)
For the enumeration of coliform bacteria in water and waste water samples.
Composition (double strength):
Ingredients Grams/Litre
Part A (Cat. No.28226): Lactose 20.0
Dipotassium phosphate 1.8
Sodium formate 0.5
L-Cystine 0.04
L-Aspartic 0.048
L-Arginine 0.04
Thiamine 0.002
Nicotinic acid 0.002
Pantothenic 0.002
Magnesium sulfate 0.2
Ferric ammonium citrate 0.02
Calcium chloride 0.02
Bromocresol Purple 0.02
Part B (Cat. No. 28227): Sodium glutamate 12.7
Store prepared media below 8°C, protected from direct light. Store dehydrated powder, in a dry place,
in tightly-sealed containers at 2-25°C.
Directions :
Suspend 22.7 g of Part A and 12.7 g of Part B in 1 litre distilled water containing 5 g of ammonium
chloride (Cat. No. 09702). Heat if necessary to dissolve the medium completely. Dispense in tubes
containing inverted Durham‘s tubes. Sterilize by autoclaving at 115°C for 10 minutes. For single
strength medium use 11.35 g of Part A and 6.35 g of Part B in 1 litre distilled water containing 2.5 g of
ammonium chloride (Cat. No. 09702).
Principle and Interpretation:
Folpmers first described a chemically defined medium based on glutamic acid for enumerating
coliforms in water (1). However in the early days following its discovery, it was seen that the medium
containing glucose gave many false positive results in 48 hours (2). It was then modified by Gray who
incorporated formate and lactose in the medium which gave improved performance (3). The major
feature of this medium is its superiority in initiating growth of Escherichia coli after exposure to
chlorine. It is better than Lauryl Tryptose Lactose Broth for the detection of small numbers of
Escherichia coli.
This medium contains sodium glutamate and formate as base nutrients. Lactose is the fermentable
carbohydrate and the amino acids, vitamins and magnesium ions allows an increased rate of
fermentation. Bromocresol purple is the pH indicator. Phosphate control the pH during the
fermentation of Lactose. The addition of ammonium chloride results in increased gas production.
Presumptive positive tubes must be subcultured into Lauryl Tryptose Mannitol Broth and Brilliant Green
Bile Broth (Cat. No. 16025) and incubated at 44°C to detect indole formation at this temperature
before identifying Escherichia coli
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Cultural characteristics after 18-24 hours at 37°C.
Organisms (ATCC) Growth Acid production Gas production
Escherichia coli (25922) +++ + +
Enterobacter aerogenes (13048) +++ + -
Salmonella typhi (6539) +++ - -
Shigella flexneri (12022) +++ - -
Staphylococcus aureus (25923) - - -
Enterococcus faecalis (29212) - - -
References:
1. T. Folpmers, Ant. V. Leeuwenhoek, J. Microbiol. Serol., 14, 58 (1948)
2. Public Health Laboratory Service, Water Committee, J. Hyg. Camb., 56, 377(1958)
3. R.D. Gray, J. Hyg. Camb., 57, 249 (1959)
Precautions and Disclaimer
This product is for R&D use only, not for drug, household, or other uses. Please consult the Material
Safety Data Sheet for information regarding hazards and safe handling practices.
The vibrant M, Millipore, and Sigma-Aldrich are trademarks of Merck KGaA, Darmstadt, Germany or
its affiliates. Detailed information on trademarks is available via publicly accessible resources.
© 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.
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