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BACTERIOLOGICAL ANALYSIS OF DRINKING WATER

Conference Paper · March 2013

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1. Title of the paper
BACTERIOLOGICAL ANALYSIOS OF DRINKING WATER

2. Author(s) name(s)
CHANDRIMA ROY1, ANUBHA JOSHI2, VINITA KATIYAR3

3. Author(s) affiliations
1
Student, M.Sc. (DFSM-IGNOU)
2
Academic counselor-Food Microbiology (IGNOU)
3
Assistant Regional Director
IGNOU Ragional Center KARNAL

4. Short running title


Bacteriological analysis……………………….

5. Postal address, fax and phone numbers of the Corresponding Author


VINITA KATIYAR
Assistant Regional Director
IGNOU Ragional Center KARNAL
0184-2260075 Fax Number: 0184- 2255738

6. E-mail ID of the Corresponding Author


Email: vinita.katiyar@gmail.com

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BACTERIOLOGICAL ANALYSIOS OF DRINKING WATER

Abstract

The aim of this study is to analyze the microbiological quality of the available
drinking water collected directly from various sources, supply outlets, as well as
drinking water kept/stored in various containers or storage pots in order to check
the drinking water quality and evaluate the awareness of people for maintaining
cleanness and hygiene conditions for storage of drinking water.

Hence, the water samples were collected and analyzed for Most Probable
Number (MPN) as well as prevalence of various water borne pathogens,
indicators organisms to check for presence of fecal contaminants using various
media.

Out of 57 samples analyzed, 42 had contamination, with MPN ranged


from 7.4 to 1100 per 100 ml. Prevalence of E. coli, Pseudomonas was on higher
side as compare to the Enterobacters and Klebsiella. Vendors’ water, hand pump
supply water was positive for E. coli.

Present study indicate that water testing would ensure the supply and
availability of contamination-free drinking water and awareness among the people
towards sanitation and hygienic conditions for storage of drinking water is needed
to keep away the use of contaminated water.

Key words: Bacteria, drinking water quality (DW), fecal contaminants, MPN.

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Introduction

The human body has 55% to 78% water depending on body size. The
percentage of water observed in different body parts are as – muscular tissues
75%, brain contains 90% water, bones 22%, and blood 83%. Since, water is one
of the essential components required to our body, it is important to assess the
quality of water, which is being used for household activities as well as
consumption whether it is actually reliable and safe for health of the consumers.
Water potability refers to the quality of water that can be safe for consumption
and use with no risk of adverse health effects.
http://en.wikipedia.org/wiki/Drinking_water

Many studies have been carried out worldwide for checking the quality
and safety of drinking water. Kurup et al (2010) have carried out the microbial
and physiochemical analysis of water samples by taking biofilm samples from
residential areas in Georgetown, Guyana, discovering the most prevalent species
to be Lactobacillus and the least prevalent species to be Salmonella sp. Zvidzai et
al (2007) carried out a study on microbiological assessment of rural drinking
water in Zimbabwe.

Smeets et al (2008) have found that absence of indicator organisms in


drinking water does not guarantee microbial safety. So the water utilities are
implementing water safety plans (WSP) to safeguard drinking water quality.
Quantitative microbial risk assessment (QMRA) can provide objective
quantitative input for Water Safety Plans. Prasai et al (2007) have evaluated
quality of water from different sources (tap, stone spout, tube well and well) from
Kathmandu valley by utilizing Heterotrophic plate counts and Coliform counts
extensively as basis for regulating microbial quality of drinking water. Regulatory
parameters were found to be much above the WHO guideline values. This study
showed that most natural water sources are highly contaminated. The detection of
pathogenic enteric bacteria in different sources of drinking water depicted for
water borne epidemics situations there.

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Nagpal et al (2011) have examined the drinking water for the presence or
absence of Salmonella, Citrobacter, E. coli and Vibrio species in the Indira
Sagar/Omkeshwar project affected areas and rehabilitation/ resettlement colonies
of Sardar project in Madhya Pradesh. Tambekar et al (2008) revalidated the
testing methods for assessing microbial safety of drinking water in the villages of
Amrawati district of Maharashtra for using bacteriological analysis with the help
of Mutiple Tube fermentation technique to determine most probable number
(MPN), Membrane filter techniques, Eijekamn’s test for thermotolerant coliform
and Manja’s Rapid hydrogen sulphide test for detection of fecal contaminations in
drinking water.

Hence, it is essential to check the quality of the available drinking water


from various sources. In view of this present study was designed to analyze the
microbiological quality of the available drinking water from various sources like
food stalls, tea stalls, supply water, packaged water to facilitate the examination
of level of contamination and finally hence the risk associated with their
consumption.

Since, most of earlier undertaken studies in India have dealt with physical
and chemical parameters of drinking water quality. Hence the study has been
designed to examine the microbiological quality of available drinking water..

Material & method

The study was designed to analyze the microbiological quality of the


drinking water from various sources like food stalls, tea stalls, supply water,
packaged water etc in 3 different regions in order to check the level of
contamination and compare one another. Accurate and reliable laboratory
methods and analyses are a prerequisite to ensure potable drinking water
characteristics and to provide an abatement strategy to supply contamination free
water. Hence, in order to assess the quality of the drinking water collected
directly from various sources, supply outlets, as well as drinking water stored in
various containers or storage vessels in the three regions selected, and thereby

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survey the incidence of the different water-borne bacterial pathogens the
following experimental plan was undertaken.

Sampling sites

The DW samples were collected for testing from different areas of Delhi and
NCR,

Sampling strategy

DW water samples have been collected in sterile sampling bottles. A total


number of 57 samples were collected randomly from Delhi and NCR.

Sampling location

The drinking water samples were collected mainly from three areas namely-
NOIDA, West Delhi and Meerut.

Kind of samples collected for analysis

The samples were collected from various sites like restaurants, hospitals, schools,
various street vendors and also packaged mineral water, distilled water, domestic
water supply, office vending machines and the main water supply, from hospitals,
water vendors, food and tea stalls, fruit stalls, handpumps, tubewells, packaged
water, reverse osmosis treated water from all the 3 regions.

Sampling technique

Water samples were collected in sterile bottles as per the APHA (1992)
standard protocol.

Processing of drinking water samples

Collected drinking water samples were immediately stored in a chilled insulation


container preferably at a temperature between 1°C and 4°C in order to prevent the
overgrowth of bacteria which may result in false bacterial counts.

Analysis of drinking water

Bacteriological analysis of drinking water samples was done within 48 hours of


collection. Drinking water samples were analyzed for Most Probable Number

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(MPN) test using the method given by McCrady in 1915 as well as prevalence of
various water borne pathogens and indicators organisms examined using
specialized media and biochemical tests for identification of bacteria.

Results & discussion

A total of 57 drinking water samples were tested in 3 regions namely - West


Delhi, NOIDA and Meerut. Samples were collected from restaurant; fruit shop;
tea stall; hand pump; tube well; tube well (120 feet); outside hospital; school;
hospital kitchen; pouched; roadside vendors; supply (tap)

Plate 1 shows the results of MPN test after 24hrs incubation in lactose
broth (LB). Yellow color tubes indicate the positive result of presence of bacterial
contamination whereas no change in color of Lactose Broth medium shows no
bacterial growth after 24hrs incubation.

Plate 1

Of these, 42 samples were found to be contaminated with different bacterial


contamination. The MPN value was ranged between 7.4-1100 per 100ml. Area
wise comparative results are presented in graphical forms (Figure 1-3).

Local food shop

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The samples from west Delhi show TC of 24.66 as against 32.67 of NOIDA and
Meerut, which indicates that the samples in the latter regions are more
contaminated.

Fruit shop
Samples from NOIDA have shown a TC of 1100 as against 170 of Meerut and
131 of West Delhi. this indicates the samples from NOIDA have higher
contamination level.
Tea stall
Samples from Meerut show a TC of 25.67 and NOIDA having a tc of 28, as
against west Delhi’s sample having a TC of 32.67.The samples from West Delhi
hence show higher contamination level.
Tap water
Samples from Meerut show a TC of 780, as against West Delhi and NOIDA’s
sample TC of 1100. West Delhi and NOIDA’s samples have a higher
contamination level than that of Meerut samples.
Hand pump
Samples from West Delhi showed up a TC of 112, as against the samples of
NOIDA with a TC of 64.33 and that of Meerut samples with a TC of 90. This
reflects that the West Delhi samples have shown a greater number of microbes
compared to the other 2 areas.

Tube well water


Samples from West Delhi show a TC of 25.67, as against NOIDA sample’s TC of
10.8 and Meerut sample’s TC is 18.33. The TC scores show that West Delhi
samples are having higher levels of contamination compared to the other 2 areas.
Tube well water (120 feet) deep
Samples from NOIDA have a TC value of 1100 as against Meerut and West
Delhi’s TC value of 386.67. So, NOIDA tube samples have shown a higher levels
of contamination compared to the other 2 areas.
Tap water outside the hospital

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Samples from West Delhi show a TC value of 8.47, as against NOIDA and
Meerut sample’s TC of 8.6, hence West Delhi samples are less contaminated than
the other 2 areas.

School drinking water


Samples from West Delhi have a TC of 22.33,as against samples from NOIDA
with a TC 16.67 and Meerut with TC 16. So the West Delhi samples are having
higher levels of contamination compared to the other 2 areas.
Hospital kitchen
Samples from West Delhi have a TC of 31.33,as against that of NOIDA with a
TC of 16.67 and that of Meerut’s sample the TC is 28,showing West Delhi
samples are having higher levels of contamination compared to the other 2 areas.
Pouched (50)
Samples from NOIDA show a TC of 190 as against Meerut’s sample with a TC of
112 and West Delhi’s sample with a TC of 131. So NOIDA samples have
exhibited higher levels of contamination compared to the other 2 areas.
Fifty paise water vendor
Samples from NOIDA show up a TC of 170 as against West Delhi’s sample with
a TC of 112 and Meerut’s sample with a TC of 64.33 thus showing NOIDA
samples are having higher levels of contamination compared to the other 2 areas.
Figure 1. Most probable number of organisms (MPN) per 100 ml of water sample

West Delhi Area

1500
MPN/100ml

1000

500

0
1 2 3 4 5 6 7 8 9 10 11 12
Water sources

1-Restaurant; 2-Fruit shop; 3-Tea stall; 4-Hand pump; 5-Tube well; 6-Tube well (120 feet); 7-
Outside hospital (Tap); 8-School; 9-Hospital kitchen; 10-Pouch; 11-Road side vendor; 12-Supply
(Tap).

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Figure 2. Most probable number of organisms (MPN) per 100 ml of water sample

NOIDA Area
1500

MPN/100ml
1000

500

0
1 2 3 4 5 6 7 8 9 10 11 12
Water sources

1-Restaurant; 2-Fruit shop; 3-Tea stall; 4-Hand pump; 5-Tube well; 6-Tube well (120 feet); 7-
Outside hospital (Tap); 8-School; 9-Hospital kitchen; 10-Pouch; 11-Road side vendor; 12-Supply
(Tap).

Figure 3. Most probable number of organisms (MPN) per 100 ml of water


sample
Meerut Area
1500
MPN/100ml

1000

500

0
1 2 3 4 5 6 7 8 9 10 11 12
Water sources

1-Restaurant; 2-Fruit shop; 3-Tea stall; 4-Hand pump; 5-Tube well; 6-Tube well (120 feet); 7-
Outside hospital (Tap); 8-School; 9-Hospital kitchen; 10-Pouch; 11-Road side vendor; 12-Supply
(Tap).
Prevalence of bacterial isolates in different water samples
The prevalence of different isolates was checked in the samples obtained from
each of the three areas - Delhi, NOIDA and Meerut. Identification of various
kinds of bacteria have revealed that E. coli and Pseudomonas were predominant
in comparison to Enterobacter and Klebsiella.

Sohani et al (2012) carried out the microbiological analysis of surface


water in Indore (Madhya Pradesh, India) by using samples from variable sources

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of different regions wherein they observed a high prevalence of coliform group of
bacteria namely E. coli, Enterobacter, Klebsiella, Salmonella and Shigella on
EMB agar. Therefore our study corroborates their studies that E. coli,
Enterobacter, Klebsiella, Pseudomonas are the predominant water-borne
microbes.

Bacteriological study of drinking water during an epidemic of cholera in


Delhi was done by Baveja et al (1989). Sixty samples from various sources like
hand pumps, taps and tube wells and from different areas of Delhi during
epidemics of Cholera and Gastroentritis were collected. Only 27 (45%) of them
were found to be fit for human consumption and the rest showed presence of
coliform organisms with MPN values ranging from 10 to 1800 + per 100 ml.

As against the studies discussed above, present study analyses the sources
of drinking water from supply, point of use in three areas: West Delhi, NOIDA
and Meerut respectively. Such a comparative study has not been performed
earlier. Fifty seven samples were taken from various sources and point of use
from the three areas. It has helped in understanding the prevalence of pathogens
and Total coliforms present in different samples from the selected areas, using the
MPN counts. It also gave us an insight as to which micro-organisms are present in
the samples area wise.

In accordance with few of the earlier studies, the water from tube well
showed a higher percentage of Psuedomonas instead of E. coli in all the three
areas. Our studies also showed presence of Klebsiella in most of the samples. In
this study MPN values ranging from 7.4 to >1100 per 100 ml were obtained
indicating greater prevalence of fecal pollutants in deference to values of 10 to
1000 per 100 ml cited in earlier studies.

Based on the sample analysis, it can be deduced that out of the three
regions –

 NOIDA’s drinking water samples were found to be the most contaminated


followed by West Delhi and then Meerut’s drinking water samples, being
the least contaminated.

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 Higher MPN index indicates the presence of greater number of bacteria in
Drinking Water (DW).

 Presence of E. coli, in DW samples confirmed the presence of fecal


contaminants in DW. E. coli is considered to be more closely associated
with fecal contamination.
Concise study reveals that, water testing is a significant approach to ensure the
supply and availability of contamination free DW. According to Indian standard
IS 1622 (1981), no sample should contain E. coli as well as other coli form
organisms more than 10 per 100 ml. Simultaneously, awareness among the people
towards sanitation and hygienic conditions for storage of DW is needed to
encourage the use of contamination free water.
Acknowledgement
Study has been done as part of M.Sc Dissertation under self finance scheme.

References

1. American Public Health Association [APHA] (1992). Standard Methods for


the Examination of Water and Wastewater, 18th Edition, Washington, D.C.
Article URL: http://www.webmedcentral.com/article_view/2084
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Commun. Dis. 21 (1): 59-61.
3. Kurup R, Persaud R, Caesar J (2010). Microbiological and physiochemical
analysis of drinking water in Georgetown, Guyana. Nat & Sci 8(8): 261-264.
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6. Prasai T, Lekhak B, Joshi DR, Baral MP (2007). Microbiological Analysis of
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