INDUSTRIAL TRAINING

UNDERTAKEN AT
TUDAKA ANALYTICAL CONSULTANTS LIMITED
NO 1, ONORIJI STREET, EKPAN, DELTA STATE
BY
AKPOJOTOR VANESSA EMMANUELLA
MATRIC NO. FOS/17/18/250200
FACULTY OF SCIENCE
DEPARTMENT: SCIENCE LABORATORY TECHNOLOGY
SUB UNIT: BIOCHEMISTRY TECHNOLOGY
DELTA STATE UNIVERSITY ABRAKA DELTA STATE
MICROBIAL ANALYSIS

DETERMINATION OF TOTAL
COLIFORM IN WATER SAMPLES
 OUTLINE
INTRODUCTION
DETERMINATION OF TOTAL COLIFORM BACTERIA IN WATER
REAGENTS/APPARATUS
PROCEDURES OF ANALYSIS
STANDARD MPN CHART
RESULTS
CONCLUSION
 INTRODUCTION
WHAT IS COLIFORM?
Coliform bacteria is also known as "indicator organisms", which is used to detect the
potential presence of disease causing bacteria in water. Coliform is a name that
qualifies a group of bactaria.
WHAT IS TOTAL COLIFORM?
The term “total coliforms” refers to a large group of Gram-negative, rod-shaped
bacteria that share several characteristics. The group includes thermotolerant
coliforms and bacteria of faecal origin, as well as some bacteria that may be
isolated from environmental sources
They are provisionally identified by the production of acid and gas from the
fermentation of lactose
 DETERMINATION OF TOTAL COLIFORM BACTERIA IN WATER
METHOD: Multiple Fermentation Technique, using Most Probable Number (MPN)
Method
The technique has been used for the analysis of drinking-water for many years
with satisfactory results. It is the only procedure that can be used if water
samples are very turbid or if semi-solids such as sediments or sludges are to be
analysed. The procedure followed is fundamental to bacteriological analyses
and the test is used in many countries.
It is customary to report the results of the multiple fermentation tube test for
coliforms as a most probable number (MPN) index. This is an index of the
number of coliform bacteria that, more probably than any other number, would
give the results shown by the test. 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.
 REAGENTS/APPARATUS
1. Nutrient/Growth media (Mac Conkey broth)
2. Distilled Water
3. McCartney Bottles
4. Durham Tube
5. Syringe
6. Beakers
7. Autoclave
McCartney Bottle

Durham Tube
Syringe

Autoclave
Beaker
 PROCEDURE FOR THE ANALYSIS
1. A certain gram of Mac Conkey Broth (based on manufacturer's description) is weighed into
a preparation bottle and dissolved with 1000ml of Distilled water.
2. All apparatus, material and growth medium were sterilized using the autoclave at a 121C .
3. A Sets of Three Test tubes /mc cartney bottles and then 9ml of McConkey broth is then
added into each one of the bottles.
Also a control is prepared which is just broth in a bottle.
4. The sample were then introduced into the bottles (following serial dilution) except that of
the control.
1ml of the water sample is innoculated into first three set of mc cartney bottles/test-tubes.
1ml of the water sample is then diluted with 9ml of sterilized distilled water. Pour 0.1ml to
second set, and 0.01ml to the third set.
5. The inoculums is mixed with the growth medium (broth) in the bottles by agitating gently.
6. The Durham tube is placed inversely in each bottle and covered and properly Labeled.
7. The bottles are incubated at 37°C for 24 hours
8. Observe each tube for acid and gas production, which signify microorganism growth
through color change.
9. The number of positive tubes in each set is Collated and combined and arranged orderly.
10. The number combination was then read out from the standard MPN table to obtain the
estimated number of coliform cells present in 100ml of the original water sample (using
McCrady's Statistical table).
STANDARD MPN CHART
 RESULTS

MPN for coliform bacteria for 2/1/1 is 3.6, (from the above table)It means 9 coliform
bacteria per 100 ml of water sample are present.
 CONCLUSION
From the above Analysis The Result were observed through acid and gas production,
which signify microorganism growth through color change which were then
compared with control
SUMMARY OF MY FINDINGS
From the above result it was determined that there is the presence of coliform in the
water sample(3.6) which is slightly above the WHO limit which is stated to be 2.2
RECOMMENDATIONS
The Presence of Total coliform in Water means it is unhealthy and cannot be consumed.
The following recommendations should be cosidered
 Terminal Disinfection: The Water should undergo Treatment immediately before
consumption. Chlorine can be easily monitored and controlled as a drinking water
disinfectant
 Cleaning of Storage: The Water Storage should be cleaned regularly to avoid growth
of Micro organisms in them and excess turbidity
REFERENCE
Guidelines for Drinking-Water quality vol.3 WHO Geneva 1997

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