Submitted By: Deepali Talwar M.Tech (Environmental Engg.)

Submitted By: Deepali Talwar M.Tech(Environmental Engg.
Water, of adequate quantity and quality, is essential for healthy life.
The approach that has been adopted is to analyse for indicator organisms that inhabit the gut in large numbers and are excreted in human faeces.
1.
2.
Total coliforms The term total coliforms refers to a large group of Gram-negative, rod-shaped bacteria. Thermotolerant (faecal) coliforms These are organisms which grow at 44 or 44.5 C and ferment lactose to produce acid and gas.
Two techniques are commonly used to detect the presence of coliforms in water:
Most probable number or Multiple fermentation tube technique. Membrane filter technique.
Multiple fermentation tube technique Slower: requires 48 hours for a positive. More labour-intensive. Requires more culture medium. Requires more glassware. More sensitive. Result obtained indirectly by statistical approximation. (low precision) Not readily adaptable for use in field. Applicable to all types of water.
Membrane filter technique More rapid: quantitative results in or presumptive positive about 18 hours. Less labour-intensive. Requires less culture medium. Requires less glassware. Less sensitive. Results obtained directly by colony count. (high precision) Readily adapted for use in the field. Not applicable to turbid waters.
Consumables readily available in most countries.

May give better recovery of stressed or damaged organisms in some.
Cost of consumables is high in many countries.
The MPN test is a method to estimate the concentration of viable microorganisms in drinking water. It is not a count of the actual number of indicator bacteria present in the sample.
The presence of coliforms is indicated by turbidity in the culture medium, by a pH change and/or by the presence of gas. The MPN index is determined by comparing the pattern of positive results with statistical tables. The tabulated value is reported as MPN per 100 ml of sample.
Medium (Isolation media)
Uses
Incubation temperature
48 hours at 35 0.5 C or 37 0.5 C for total coliforms and 24 hours at 44 0.25 C or 44.5 0.25 C for thermotolerant coliforms. 48 hours at 35 0.5 C or 37 0.5 C for total coliforms and 24 hours at 44 0.25 C or 44.5 0.25 C for thermotolerant coliforms 48 hours at 35 0.5 C or 37 0.5 C for total coliforms and 24 hours at 44 0.25 C or 44.5 0.25 C for thermotolerant coliforms 44.5 0.25 C for thermotolerant coliforms
Lactose broth (Prepare single Total or strength by diluting double thermotolerant strength medium with Coliforms. distilled water.Each tube should contain an inverted fermentation (Durham tube) MacConkey broth Total or thermotolerant coliforms
Lauryl tryptose (lactose) broth
Total or thermotolerant coliforms
Confirmatory media
Brilliant green lactose bile Broth 44.5 0.25 C for thermotolerant coliforms Total or thermotolerant coliforms (gas
Isolation media
Reactions of Total coliforms at 35 or 37 C
Reactions of Thermotolerant coliforms at 44 or 44.5 C
Lactose broth
Gas visible in the Same as total inverted coliforms at 35 or fermentation (Durham) 37C. tube plus turbidity of the medium.
Gas visible in the Same as total inverted coliforms at 35 or fermentation (Durham) 37C. tube plus turbidity of the medium
MacConkey broth
Confirmatory Media
Reactions of Total coliforms at 35 or 37 C
Reactions of Thermotolerant coliforms at 44 or 44.5 C
Brilliant green lactose bile broth
Gas visible in the Same as total inverted coliforms at 35 or 37 fermentation (Durham) C tube plus turbidity of the medium Gas visible in the Same as total inverted coliforms at 35 or 37 fermentation (Durham) C tube plus turbidity of the medium
EC medium
Combination of positives
0-0-0 0-0-1 0-1-0 0-2-0
MPN index per 100 ml

<2 2 2 4
95 % confidence Limits-Upper
1.0 1.0 1.0
95 % confidence limits-Lower
10 10 13
1-0-0
1-0-1 1-1-1
2
4 6
1.0
1.0 2.0
11
15 18
2-0-0
2-0-1
4
7
1.0
2.0
17
20
Waterborne pathogenic diseases (typhoid fever, viral and bacterial gastroenteritis and hepatitis A)
Decrease in quality of drinking water.
The basic concept for the MPN method is to dilute the sample to such a degree that inocula in the tubes will sometimes contain viable organisms.
This method is commonly used in the pharmaceutical industry for water testing. The possible applications of MPN are environmental monitoring, media growth promotion studies and aspects of the validation of rapid microbiological methods.

Submitted By: Deepali Talwar M.Tech (Environmental Engg.)

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Submitted By: Deepali Talwar M.Tech (Environmental Engg.)

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Submitted By: Deepali Talwar M.Tech (Environmental Engg.)

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Submitted By: Deepali Talwar M.Tech(Environmental Engg.

Water, of adequate quantity and quality, is essential for healthy life.

Consumables readily available in most countries.

Cost of consumables is high in many countries.

Medium (Isolation media)

Lauryl tryptose (lactose) broth

Total or thermotolerant coliforms

Reactions of Total coliforms at 35 or 37 C

Reactions of Thermotolerant coliforms at 44 or 44.5 C

Reactions of Total coliforms at 35 or 37 C

Reactions of Thermotolerant coliforms at 44 or 44.5 C

Brilliant green lactose bile broth

MPN index per 100 ml

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