TITLE:

IDEXX ColilertTest Method For The Simultaneous Detection Of Total Coliform and E. Coli In
Water
OBJECTIVE:
To detect and confirm of total coliform and E. Coli in water.
INTRODUCTION:
IDEXX Colilert reagent is used around the world for the detection of coliforms and E. coli in
water. Colilert is used for the simultaneous detection, specific identification, and confirmation
of total coliforms and E. Coli in water. Colilert based on the Defined Substrate
Technology(DST
TM
). DST utilizes indicator nutrients which cause target microbes contained in
the sample and incubated in the DST reagent system to produce a colour change (or another
signal, ie: fluorescence), both indicating and confirming their presence. The indicator-nutrient is
cleaved by the target microbe which metabolizes the nutrient and frees the indicator to express
a specific color. Total coliform and E. Coli are specifically and simultaneously detected and
identified the 1 CFU/100ml of sample, in 24 hours or less, by inoculating the reagent with the
water sample and incubating it.
APPARATUS:
IDEXX Colilert Reagent
E. Coli
Paddy Water Sample





PROCEDURE:
1. Contents of one pack to a 100ml water sample was added to a sterile vessel.

2. The vessel was cap and shake until dissolved.

3. The reagent mixture was poured into a Quanti tray and sealed in an INDEXX Quanti Tray
Sealer.

4. Next, the sealed tray was placed in a 35C incubator for 24 hours.

5. Then, the result was read according to the result interpretation table.

6. If no yellow color is observed, the test is negative.

7. If the sample has a yellow color equal to or greater than the comparator, the presence of
total coliforms is confirmed.

8. The number of positive wells was counted and MPN table provided was referred.










RESULTS:

Group

Type of Bacteria
Number of box
MPN of Bacteria Large Small

W1
Coliform 46 48 533.5
E. coli 30 9 58.8

W2
Coliform 47 47 665.3
E. coli 27 15 60.8

W3
Coliform 48 48 1011.2
E. coli 28 5 47.3

W4
Coliform 47 48 691.0
E. coli 42 39 227.7

W5
Coliform 47 48 691.0
E. coli 47 38 472.1

W6
Coliform 47 48 691.0
E. coli 47 38 472.1









QUESTIONS:
MICROBIOLOGICAL ANALYSIS DETERMINATION OF E. COLI AND TOTAL COLIFORM
1. What is the family name of coliform and E. Coli group?
Coliform bacteria and E. Coli are the type of bacteria that are part of the
enterobacteriaceae family.


2. What is the use of UV light? Why that bacteria must be observed under UV light?
The use of UV light is to detect the presence of bacteria. Because bacteria tend to
congregate on surfaces where high levels of flavin are present. Flavin is a material that
emits a fluorescent glow when exposed to UV light.


3. What to do if water sample has some background color? How to make the dilution?
If there is color, you should examine again with more dilution. Dilute the samples well
with PBS of 0.9% NaCl. PBS or 0.9% NaCl is recommendable for the dilution. (If you must
use water, apply 0.9 mL or 1.0 mL of sample solution and incubate at 35 or 30,
respectively.)

DISCUSSION:
The Colilert-18 system for enumeration of total coliforms and Escherichia coli is
approved by the U.S. Environmental Protection Agency for use in drinking water analysis and is
also used by various agencies and research studies for enumeration of indicator organisms in
fresh and saline waters. Coliform bacteria are widely used as indicators of fecal contamination
of both fresh and marine waters. Certain members of the coliform group live outside of the
gastrointestinal tract in the environment and may create a false indication of fecal
contamination. Index colilert test method is used for the simulteneous detection oftotal
coliform and E.Coli in water. From our experiment, MPN of bacteria of coliform in water sample
3 is very higher than other 5 water sample. It is 1011.2. we can see the present of coliform by
look at the water sample that turn into yellow flourescent. MPN of bacteria for water sample
1,2, is 533.5, 665.3 while water sample 4,5,6 is in the same value. It is 691.0.For MPN of
bacteria for E.coli is very higher in water sample 5 and 6. The value is 472.1. other MPN of
bacteria in water sample 1,2,3 and 4 are 58.8, 60.8, 47.3, and 227.7. from this experiment, we
can conclude that the water sample that has higher number of MPN of bacteria for E.coli is very
polluted.

REFERENCES:
1. Avocet Environmental Testing. Total Coliform Bacteria Testing. Retrieved April 15, 2013,
from http://www.avocetlab.com/drinking-water/what-mean/interpreting-results/

2. John Lindquist. (2009). Bacteriology 102. A General Overview of Coliforms. Retrieved April
15, 2013, from http://www.splammo.net/bact102/102coliforms.html

3. Ian L. Pepper, Charles P. Gerba. (2011). Experiment 17: Defined Substrate Technology for
Detection of Coliforms and Fecal Coliforms. Environmental Microbiology: A Laboratory
Manual (pp. 129-133). Academic Press. Retrieved April 15, 2013 from
http://books.google.com.my/books?id=0cFbAoJ_c7UC&pg=PA132&dq=coliform+e.+coli+col
ilert+test+experiment&hl=en&sa=X&ei=zK1rUZKmH4HIrQeji4GwAQ&ved=0CC0Q6AEwAA#v
=onepage&q=coliform%20e.%20coli%20colilert%20test%20experiment&f=false