8/10/2019 Water Analysis Final

1/1

Microbiological Analysis of Drinking Water

Test for Coliforms and E.coli

Group 3 3-A Biochemistry

Cua, G.A; Diego, J.L; Divina, T.R; Dulatre, E.A

Abstract

Water is one of the most important substances on earth. Drinking water must be free of disease-causing organisms called pathogens.Pathogens can be viruses, protozoa or bacteria. For this experiment three samples of drinking water from UST were taken from thefollowing locations: the drinking fountain beside the female washroom on the first floor of the Quadri Pavilion , the faculty room, and the

fountain in front of lab 19 located at the Main Building of the Faculty of Pharmacy. Moreover, the sources were then put through theprocess of serial dilution , yielding the dilution factors of 10-1,10-2and 10-3. Lastly, each dilution had three test tubes . Therefore, therewere three samples, each having three different dilutions , and three trials per dilution. In conclusion, each of the water samplesgathered were proven to be unsafe for consumption due to it having an MPN number greater than 1 in the test for E. coli, water shouldbe free of Total Coliforms to be considered totally safe to drink. Lastly, the main objectives of this experiment were the following: todetermine the potential presence of coliforms and E. coli in water , to skillfully perform multitube technique in technique in testing totalcoliforms.

IntroductionWater is one of the most important substances on earth. All plants andanimals must have water to survive. If there was no water there would be no

life on earth.[1] Moreover , an average adult body contains 42 liters of water

and with just a small loss of 2.7 liters he or she can suffer from dehydration,

displaying symptoms of irritability, fatigue, nervousness, dizziness, weakness,

headaches and consequently reach a state of pathology.[2]

Also, it serves as alubricant for digestion, regulation of body temperature, removes harmful

toxins from the body, and transports nutrients to the body.[3]

Drinking water must be free of disease-causing organisms called pathogens.

Pathogens can be viruses, protozoa or bacteria. Waterborne pathogens cause

diseases such as hepatitis, giardiasis, and dysentery. To actually test water for

specific harmful viruses, protozoa and bacteria is very time consuming and

expensive. In addition, not all water laboratories are equipped and approved

to do the testing required. Therefore, testing water for specific organisms is

limited to investigating specific waterborne disease outbreaks. Coliform

bacteria are used as water quality indicators for two main reasons: Coliforms

may be associated with the sources of pathogens contaminating water,

analysis of drinking water for coliforms is relatively simple, economical and

efficient.[5]

Escherichia coli which is in the large intestine of virtually all people has been

used as the indicator of human fecal contamination of water and food. Tests

for the presence of this organism (and

closely related types generally known as coliforms . [7] Coliforms are a broad

class of bacteria found in our environment, including the feces of man and

other warm-blooded animals. The presence of coliform bacteria in drinking

water may indicate a possible presence of harmful, disease-causing

organisms.[5]

The MPN method is a well-established and fully documented method that

utilizes the multitube technique in order to estimate the number of

viable microorganisms in product in which the microorganisms are randomly

distributed. Its principal uses are in the food and water industries where it is

used with liquid, powdered and semi-solid products or raw materials. The

method is particularly appropriate for product having bio burden of low meannumber. [7]

Lastly, the main objectives of this experiment were the

following: to determine the potential presence of coliforms and E. coli in

water , to skillfully perform multitube technique in technique in testing total

coliforms.

Accordingly, a 3-tube MPN table was used in order to determine the MPN

value of each sample for the tests. For the test for E. coli only the growth has

to be positive to be counted as a part of the MPN. Therefore , this yielded a

result of >24 for the Quadri Pavilion drinking fountain ,11 for the faculty

room and 2.4 for the lab 19 drinking fountain . (See source [4] for the 3-

tube MPN table)

The results showed that P. aeruginosa, P. mirabilis may exist in the QuadiPavilion drinking fountain, P. aeruginosa may exist in the faculty room

water , and P. aeruginosa , E. faecalis, E. aerogenes , B. subtilis may exist in

the lab 19 drinking fountain water.

Table 1.2 Results obtained from the Confirmation Test with MPN

For the Test for Coliforms, the Brilliant Green Bile broth was used in order

detect or confirm the presence of members of the coli-aerogenes group; the

brilliant green content suppresses anaerobic lactose fermenters. Also, a

Durham tube is provided to detect the production of gas.

Accordingly, a 3-tube MPN table was used in order to determine the MPN

value of each sample for the tests. For the test for coliforms both growth

and gas production have to be positive to be counted as a part of the MPN.

Therefore, this yielded a result of 0.094 for the Quadri Pavilion drinking

fountain, less than 0.03 for both the faculty room and the lab 19 drinking

fountain.

The results showed that E. faecalis possibly thrives in all the sample

sources.

Table 1.5 Results obtained from the Completed Test for E. coli

For the Completed Test for E. coli , Eosin methylene blue agar was used in

order to confirm the presence of E. coli in the water. It is used in water

quality tests to distinguish coliforms and fecal coliforms that signal possible

pathogenic microorganism contamination in water samples, contains

sucrose and lactose which serves as fermentable carbohydrate source to

encourage the growth of fecal coliforms and provide a means of

differentiating them.

There are several different types of growths that may be observed in the

agar. Bacteria that ferments lactose show up as colored colonies, those

which do not ferment lactose show up colorless colonies, Escherichia

coli shows up as a green metallic sheen with a dark center, Aerobacter

aerogenescolonies show up as brown centered colonies , and nonlactose-

fermenting gram-negative bacteria show up as pink colorations.

ConclusionIn conclusion, each of the water samples gathered were proven to be unsafe forconsumption due to it having an MPN number greater than 1 in the test for E.

coli, water should be free of Total Coliforms to be considered totally safe to

drink

References1. (2012, November ) . Water Its Importance and Source

http://www.health.gov.au/internet/publications/publishing.nsf/Content/ohp-

enhealth-manual-atsi-cnt-l~ohp-enhealth-manual-atsi-cnt-l-ch6~ohp-enhealth-

manual-atsi-cnt-l-ch6.1

2. (2010) . Water . Retrieved form http://www.laleva.cc/environment/water.html

3. (2011) . The Importance of Water and Your Health. Retrieved from

http://www.freedrinkingwater.com/water-education/water-health-page2.htm

4. (2012). Coliform Bacteria in Drinking Water Supplies. Retrieved from

https://www.health.ny.gov/environmental/water/drinking/coliform_bacteria.htm

5. (2012 ). Coliform Bacteria and Drinking Water . Retrieved from

http://www.bfhd.wa.gov/info/coliform.php

6. (2013). Most Probable Number (MPN) Method for Bioburden Testing ISO 117371.

Retrieved from http://www.novatx.com/most-probable-number-mpn/

7. (2014). Bacterial Test for Water . Retrieved from

http://www2.fiu.edu/~psharp01/Exp9WaterBact.pdf

Methodology

prepare necessarymedia for PresumptiveTest , Test for E. coli,

Test for Coliforms andstore

Gather water samples using

a sterile container andinoculate serially dilutedsamples to pre-preparedmedia and wait for 24-48

hours

Determineresults orgrowth

and recordresults

Results and Discussion

For this experiment three samples of drinking water from UST were takenfrom the following locations: the drinking fountain beside the female

washroom on the first floor of the Quadri Pavilion , the faculty room, and

the fountain in front of lab 19 located at the Main Building of the Faculty

of Pharmacy. Moreover, the sources were then put through the process of

serial dilution , yielding the dilution factors of 10-1,10-2and 10-3. Lastly,

each dilution had three test tubes . Therefore, there were three samples,

each having three different dilutions , and three trials per dilution.

Table 1.1 Results obtained from the Presumption Test with MPN

For the Presumption Test, due to the unavailability of the Mac Conkey

broth the EC broth was utilized for both this test and the Test for E. coli.

The EC broth is is used in the detection of fecal coliform bacteria as an

indication of fecal pollution of food and water from sources other than

potable water. A Durham tube is provided to detect the production of gas.

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