Dr.

Hari Singh Gour University

SAGAR (M.P.) 470003

Project Report

“ASSESSMENT OF WATER QUALITY OF

DIFFERENT WATER SOURCES WITH
SPECIAL REFERENCE TO FAECAL
CONTAMINATION”

Submitted to NAME OF
STUDENT
Prof. P.C. Jain Ashutosh Srivastava
B.Sc.III sem.
Roll no. 516115
 ACKNOWLEDGEMENT

I want to express my deepest gratitude to

my guide who has guided me faithfully through the

entire project.

I extend my heartfelt thanks and sincere

regards to Head of the department Prof. P.C. JAIN

for giving their valuable views for better

presentation of this project.

I am immensely grateful to my family

members and my friends for their support

throughout my endeavor.

Ashutosh Srivastsava
B.Sc. V sem.
Microbiology
INTRODUCTION
 Water Contamination
It is a well-known fact that clean water is absolutely essential for healthy
living. Adequate supply of fresh and clean drinking water is a basic need
for all human beings on the earth, yet it has been observed that millions
of people worldwide are deprived of clean water. Freshwater resources
all over the world are threatened not only by over exploitation and poor
management but also by ecological degradation. The main source of
freshwater pollution can be attributed to discharge of untreated waste,
dumping of industrial effluent, and run-off from agricultural fields.
Industrial growth, urbanization and the increasing use of synthetic
organic substances have serious and adverse impacts on freshwater
bodies. It is a generally accepted fact that the developed countries
suffer from problems of chemical discharge into the water sources
mainly groundwater, while developing countries face problems of
agricultural run-off in water sources. Polluted water like chemicals in
drinking water causes problem to health and leads to water borne
diseases, which can be prevented by adopting preventive measures.
Many areas of groundwater and surface water are now contaminated
with heavy metals, persistent organic pollutants (POPs) like dioxins,
pesticides etc and nutrients that have an adverse affect on health.
Water-borne diseases and water-caused health problems are mostly due
to inadequate and incompetent management of water resources. Safe
water for all can only be assured when access, sustainability, and equity
can be guaranteed.

Water-borne diseases constitute one of the major public health hazards

in developing countries. In 1995 at the global level, contaminated water
and food caused more than 3 million deaths, of which more than 80%
were among children under age five. Besides the conventional
pathogens, which are transmitted by water, several emerging water-
borne pathogens have become increasingly important during the last
decade or so.
These include Vibrio cholerae O139, Crypto-poridium parvum, shiga
toxin producing E. coli especially enterohaemorrhagic E. coli (EHEC),
Yersinia enterocolitica, Campylobacter jejuni, Calciviruses and
Microsporidia. In India, more than 70% of the epidemic emergencies are
either water-borne or are water related. Although a substantial amount
of work has been carried out on common water-borne pathogens in
India, unfortunately only a little information is available on the emerging
water-borne pathogens.

Centre for Science and Environment (CSE) in January 2003 reported

results of a study on analyses of pesticide residues in bottled water
samples and found the presence of lindane, DDT and its metabolites,
malathion and chlorpyrifos in majority of samples (70-94%). Endosulfan
was detected in few samples (8.8%). These findings had led to adoption
of European Union norms of 0.1 and 0.5ppb for individual and total
pesticide residues in packaged drinking water in India. During August
2003, CSE released another report on pesticide residues in soft drinks,
which indicates the presence of lindane, DDT & its metabolites,
chlorpyrifos and malathion in 81 to 100% of samples. The Government
of India constituted a Joint Parliamentary Committee, which agreed with
the concerns on pesticide residues in soft drinks and suggested adoption
of packaged water norms for water to be used for soft drinks as well.
These have since been formulated by Bureau of Indian Standards (BIS)
and notified under Prevention of Food Adulteration Act of India (PFA).
Recently after a gap of three years, CSE reported that the pesticide
residue status in soft drinks has still not changed. The government has
now constituted a National Level Expert Group (NLEG) to resolve the
issue to prescribe pesticide residues standards for carbonated
beverages, which the expert group is presently deliberating.
 Recommended Quality of Drinking Water – CPHEEO*
Guidelines

(A) Physico-Chemical Characteristics

S. No. Characteristics Acceptable Cause of Rejection

1 Turbidity (NTU) 1 10
2 Colour (Units on Pt-Co scale) 5 25
3 Taste and Odour Unobjectionable Objectionable
4 pH 7.0 to 8.5 <6.5 or >9.2
5 Total Dissolved Solids, mg/L 500 2000
6 Total Hardness (as CaCO3) mg/L 200 600
7 Chlorides (as Cl) mg/L 200 1000
8 Sulphates (as SO4), mg/L 200 400
9 Fluorides (as F), mg/L 1.0 1.5
10 Nitrates (as NO3), mg/L 45 45
11 Calcium (as Ca), mg/L 75 200
12 Magnesium (as mg), mg/L < 30 150
If there are 250 mg/L of sulphates, Mg content can be increased to a maximum of 125 mg/L with the
reduction of sulphates at the rate of 1 unit per every 2.5 units of sulphates
13 Iron (as Fe) mg/L 0.1 1.0
14 Manganese (as Mn), mg/L 0.05 0.5
15 Copper (as Cu) mg/L 0.05 1.5
16 Aluminium (as Al), mg/L 0.03 0.2
17 Alkalinity, mg/L 200 600
18 Residual Chlorine mg/L 0.2 >1.0
19 Zinc (as Zn), mg/L 5 15
20 Phenolic Compounds (as Phenol, mg/L 0.001 0.002
21 Anionic detergents (as MBAS) mg/L 0.2 1.0
22 Mineral Oil, mg/L 0.01 0.03

S. No. Characteristics Acceptable Cause of

Rejection
TOXIC MATERIALS
23 Arsenic (as As) mg/L 0.01 0.05
24 Cadmium (as Cd) mg/L 0.01 0.01
25 Chromium (as hexavalent Cr) mg/L 0.05 0.05
26 Cyanides (as CN) mg/L 0.05 0.05
27 Lead (as Pb), mg/L 0.05 0.05
28 Selenium (as Se) mg/L 0.01 0.01
29 Mercury (total as Hg) mg/L 0.001 0.001
30 Polynuclear aromatic hydrocarbons (PAH), μg/L 0.2 0.2
31 Pesticides (total) mg/L Absent Refer to WHO
guidelines for
drinking water
quality vol. 1-1993

(B) Bacteriological Quality of Drinking Water a

Organism Guidelines Value

All Water intended for drinking
E.Coli or thermotolerant coliform b.c Must not be detectable in any 100 mL
sample

Treated Water entering the distribution system

E.coli or thermotolerant coliform bacteria Must not be detectable in any 100 mL
sample

Total coliform bacteria Must not be detectable in any 100 mL

sample

Treated water in the distribution system

E.coli or thermotolerant coliform bacteriab Must not be detectable in any 100 mL
sample

Total coliform bacteria Must not be detectable in any 100 mL

sample. In case of large supplies,
where sufficient samples are
examined, must not be present in
95% of samples taken throughout any
12 month period

Notes

- The figures indicated under the column ‘Acceptable’ are the limits upto
which water is generally acceptable to the consumers

- Figures in excess of those mentioned under “Acceptable” render the

water not acceptable, but still may be tolerated in the absence of
an alternative and better source but upto the limits indicated
under column “Cause for Rejection” above which the sources will
have to be rejected.
+ It is possible that some mine and spring waters may exceed these radio activity limits and
in such cases it is necessary to analyze the individual radio-nuclides in order to assess the
acceptability or otherwise for public consumption.
* cpheeo :- Central Public Health & Environmental Engineering
Organization

Public & Private Water

Supply System:
-
Public water supply system is one that serves piped water to several
homes round the year. Water that does not come from a public water
supply, and which serves one or only a few homes, is called private
supply . How Does Water Get To Your Taps? In a typical community
water supply system, water is transported under pressure through a
distribution network of buried pipes. Smaller pipes, called house service
lines, are attached to the main water lines to bring water from the
distribution network to your house. In many community water supply
systems, water pressure is provided by pumping water up into storage
tanks that store water at higher elevations than the houses they serve.
The force of gravity then "pushes" the water into your home when you
open your tap. Houses on a private supply usually get their water from
a private well. A pump brings the water out of the ground and into a
small tank within the home, where the water is stored under pressure.
How Do Public Water Suppliers Treat My Water To Make It Safe?
Water suppliers use a variety of treatment processes to remove
contaminants from drinking water. These individual processes may be
arranged in a "treatment train" to remove undesirable contaminants
from the water. The most commonly used processes include filtration,
flocculation and sedimentation, and disinfection. Some treatment trains
also include ion exchange and adsorption. A typical water treatment
plant would have only the combination of processes needed to treat the
contaminants in the source water used by the facility. If you want to
know what types of treatment are used for your water supply, contact
your local water supplier or public works department. How Can You Help
To Protect Your Drinking Water Supply?Drinking water protection is a
shared responsibility, involving water suppliers, local and state
governments, business, and individuals. We all have an important role
to play, and as private citizens we have many opportunities.
Environmental protection activities such as watershed protection
projects are taking place across the Nation. Clean and healthy
watersheds are vital to safe drinking water. Other opportunities to be
involved in drinking water protection are discussed in the rest of this
section. Sole source aquifer protection programs also protect ground
water supplies, but usually over a much larger area than that covered
by wellhead protection programs. They focus on government-funded
projects that may affect the aquifer. Source water protection should be
a critical part of all community water programs. In the past, water
suppliers used most of their resources to treat water from rivers, lakes,
and underground sources of drinking water before supplying it to our
homes as drinking water. Now, we understand that if we place greater
emphasis on protecting our sources of drinking water, the need for
treatment can be reduced

THE GENERAL COMPONENT OF A

SOURCE WATER PROTECTION
PROGRAM INCLUDES:
 Delineation: Identifying the area of land that water passes
through to reach the drinking water intake.

 Contaminant source inventory: Mapping the locations of

potential sources of drinking water contamination.

 Source water protection area management: Using

regulatory controls, such as zoning or health ordinances, or non
regulatory controls, such as technical assistance to businesses and
public education, to keep contaminants out of drinking water supplies.

 Contingency planning: Plan special actions in case a sudden

event (for instance, a flood or spill) occurs that threaten the drinking
water supply.

How Can You Get Involved To Protect Water Supplies?

Many communities are in the process of implementing source water
protection programs. Your local water supplier can tell you whether your
community has a source water protection program. Source water
protection works by involving all members of the community. Citizens
can voice their support for controlling how land is used near drinking
water intakes. Citizens can also educate their neighbors about the
danger that household chemicals pose to drinking water supplies. Many
communities sponsor household hazardous waste disposal days to
promote proper handling of waste paints and thinners, pesticides, used
oil, and other hazardous materials. Your state or local environmental
agency should have information about such programs in your
community.
Successful Source Water Protection Elkhart, Indiana began developing a
source water protection program in the late 1980s, after being forced to
close one of its drinking water well fields when dangerous chemical
solvents were found in the drinking water supply. Officials in Elkhart
realized that they needed an efficient way to inventory and map all of
the potential sources of contamination that could reach their new
drinking water supply. They discovered that retired senior volunteers
are an excellent resource for conducting the inventories. By working
with the Service Corps of Retired Executives and the American
Association of Retired Persons, Elkhart recruited 20 senior volunteers,
from a list of over 400 potential volunteers, to conduct the inventories.
Working with city staff, the senior volunteers visited homes and
businesses throughout Elkhart and mapped 280 potential contaminant
sources for management. Management controls include zoning/land use
control and technical assistance. So far, Elkhart has been successful in
keeping its drinking water sources clean.

COLIFORM BACTERIA
Coliform is the name of a test adopted in 1914 by the Public Health
Service for the Enterobacteriaceae family. It is the commonly-used
bacterial indicator of sanitary quality of foods and water. They are
defined as rod-shaped Gram-negative non-spore forming organisms.
Some enteron forms can ferment lactose with the production of acid and
gas when incubated at 35-37°C. Coliforms are abundant in the feces of
warm-blooded animals, but can also be found in the aquatic
environment, in soil and on vegetation. In most instances, coliforms
themselves are the cause of many nosocomial illnesses, they are easy to
culture and their presence is used to indicate that other pathogenic
organisms of fecal origin may be present. Fecal pathogens include
bacteria, viruses,or protozoa and many multicellular parasites.

Typical genera include:

• Citrobacter,
• Enterobacter
• Escherichia
• Hafnia
• Klebsiella
• Serratia
Escherichia coli (E. coli), a rod-shaped member of the coliform group,
can be distinguished from most other coliforms by its ability to ferment
lactose at 44°C in the fecal coliform test, and by its growth and color
reaction on certain types of culture media. When cultured on an EMB
plate, a positive result for E. coli is metallic green colonies on a dark
purple media. Unlike the general coliform group, E. coli are almost
exclusively of fecal origin and their presence is thus an effective
confirmation of fecal contamination. Typically, E. coli are about 11% of
the coliforms in human feces

COLIFORM INDEX
The coliform index is a rating of the purity of water based on a count of
fecal bacteria. Coliform bacteria are microorganisms that primarily
originate in the intestines of warm-blooded animals. By testing for
coliforms, especially the well known E.Coli, which is a thermo tolerant
coliform, one can determine if the water has probably been exposed to
fecal contamination; that is, whether it has come in contact with human
or animal feces. It is important to know this because many disease-
causing organisms are transferred from human and animal feces to
water, from where they can be ingested by people and infect them.
Water that has been contaminated by feces usually contains pathogenic
bacteria, which can cause disease. Some types of coliforms cause
disease, but the coliform index is primarily used to judge if other types
of pathogenic bacteria are likely to be present in the water. The coliform
index is used because it is difficult to test for pathogenic bacteria
directly. There are many different types of disease-causing bacteria, and
they are usually present in low numbers which do not always show up in
tests. Thermotolerant coliforms are present in higher numbers than
individual types of pathogenic bacteria and they can be tested for
relatively easily. However, the coliform index is far from perfect.
Thermotolerant coliforms can survive in water on their own, especially in
tropical regions, so they do not always indicate fecal contamination.
Furthermore, they do not give a good indication of how many pathogenic
bacteria are present in the water, and they give no idea at all of whether
there are pathogenic viruses or protozoa which also cause diseases and
are rarely tested for. Therefore, it does not always give accurate or
useful results regarding the purity of water.
INDICATOR ORGANISM
Indicator organisms are used to measure potential fecal
contamination of environmental samples. The presence of coliform
bacteria, such as E. coli, in surface water is a common indicator of fecal
contamination. Coliform bacteria in water samples may be quantified
using the most probable number (MPN) method, a probabilistic test
which assumes cultivable bacteria meet certain growth and biochemical
criteria. If preliminary tests suggest that coliform bacteria are present at
numbers in excess of an established cut-off (the Coliform Index), fecal
contamination is suspected and confirmatory assays such as the
Eijckman test are conducted. Coliform bacteria selected as indicators of
fecal contamination must not persist in the environment for long periods
of time following efflux from the intestine, and their presence must be
closely correlated with contamination by other fecal organisms.
Indicator organisms need not be pathogenic. Non-coliform bacteria,
such as Streptococcus bovis and certain clostridia may also be used as
an index of fecal contamination.

PLAN OF WORK

Standard Method for Testing Microbiological

Safety of Drinking Water
Background

Bacteria originate from faecal matter and pollute streams and

groundwater. Most bacteria are only 1 micrometer in diameter, but some
bacteria range in size from 0.1 to 10 micrometers. Bacteria of the
coliform group are considered the primary indicators of fecal
contamination and are often used to assess water quality. Coliform is
used to describe a group of Gram-negative, facultative, anaerobic, rod-
shaped, non-spore-forming bacteria, capable of growing in bile salts and
other surface active agents and able to ferment lactose to produce acid
and gas within 48 hours at 35’C. Coliform bacteria, having the same
properties at 44’C, are referred to as thermo-tolerant coliforms or E. coli
(Escherichia coli).This is nothing but faecal coliform bacteria, found in
the intestines of warm-blooded animals, including humans.Coliform
bacteria generated from the waste of warm-blooded animals, may get
into the river or groundwater through surface runoff, especially after a
heavy rainfall. Water contaminated with faecal matter can seep into
bigger bodies of water, which may ultimately affect drinking water. A
spring, hand-dug well, buried well, cistern, etc. are all very likely to be
contaminated with coliform bacteria. Thus, contamination of water
sources by the spread of such bacteria is the leading cause of water
quality impairment in rivers. These bacteria may cause a serious health
threat to humans, causing diseases like typhoid fever, hepatitis,
gastroenteritis, dysentery, etc. Even a very small percentage of faecal
coliform bacteria may cause intestinal distress and in more severe cases
nausea, vomiting and even death.

It is often difficult to detect specific disease-causing organisms in water

that we drink everyday. Therefore, indicator organisms are detected in
drinking water, which are of faecal origin. Smoke is an indicator of fire.
Similarly, bacteria of faecal origin (coliforms / E. coli) are indicators of
pollution from faecal sources, thereby warning that there may be more
dangerous organisms like Cholerae, Salmonella, Hepatitis virus etc,
present in water. In this exercise, this principle is employed to ensure
microbiological safety of our drinking water.

Objective
To detect the presence of coliform bacteria in drinking water with the
help of multiple tube method.

Materials / Equipment Required

1. Mac Conkey broth double strength 10 ml in 5 tubes.

2. Mac Conkey broth single strength 5 ml in 10 tubes.
3. Dropper to measure water in little quantities.
4. Incubator
5. Water samples
6. Binocular microscope or powerful magnifying lens

Methodology
1. First sterilize all the 15 tubes in hot, boiling water.
2. After cooling, add –
3. 1. 5 ml of water to 5 tubes of double strength medium
4. 2. 1 ml of water to 5 tubes of single strength medium
5. 3. 0.1 ml of water to 5 tubes of single strength medium
6. Keep all the 15 tubes in the incubator for 48 hours and at a
temperature of 35-37’C. If you do not have an incubator a closed
 chamber with a light bulb and thermometer can be a poor
substitute. Periodic manual checking of temperature along with
switching off / on of the bulb will be needed.
7. Now look for some change of colour and gas produced in the
tubes.
8. Plot the number of tubes, which has changed colour and match it
with Mc Crady table given below. This table gives you the number
of bacteria present in 100 ml of water.
9. Incubate the tubes showing colour sub culture in fresh medium at
a temperature of 44’C.
10. The positive result is then matched in the same table, which
shows thermo tolerant bacteria in 100 ml of water.

How Safe should be our Water?

1. Piped and treated (urban)water supply: No coliforms and E. coli in
100 ml of water
2. Untreated water entering the system: E. coli – nil / 100 ml,
Coliforms – 3/100 ml
3. Rural unpiped supply: E. coli – nil / 100 ml, Coliforms 10 / 100 ml

Note
Measurement of the number of coliform bacteria per 100 ml is often
used as an important criterion in determining the degree of pollution and
the sanitary quality of a simple of water. The allowable number limit per
100 ml of surface water sample is less than 100 organisms. The absence
of E. Coli in drinking water is an absolute indispensability

Further Activity
The presence of faecal coliform bacteria in sources other than drinking
water can also pose health hazards. For instance, when irrigating crops
with faecal coliform contaminated water (such as wastewater), chances
are high that disease may spread after consumption of the grains and
vegetables grown with such water. Crops that are eaten uncooked are
especially dangerous for the transmission of such diseases. One method
for controlling the
spread of such bacterial diseases is by delaying the consumption of the
crop, as bacteria dissipates and perishes once exposed to air.
Think….
• Think of some new and innovative solutions to kill faecal coliform
bacteria and the disease causing bacteria found in
irrigation/municipal water.
• Is there any quick and easy method of killing the bacteria source in
the vegetables you buy before you put it on the table?
• Do you think disinfectant chemicals such as chloride can
effectively kill the bacteria without disturbing the balance of the
aquatic environment? Show your teacher with the help of some
experiment the possible negative or positive results of using
disinfectants to kill bacteria.
• Imagine yourself the head municipal authority in your village or
town. Now think of the measures that you would be taking to stem
or control microbiological pollution of water sources in your
locality? Make a list of all possible activities / regulations /
enforcements that you can think of.