Physico – Chemical and Microbiological Analysis of

Water Source in Brgy. 105 San Isidro,

Tacloban City

An Investigatory Project
Presented to the Faculty of Science Department
In partial fulfillment of the requirements in Research II

Angel Rose C. Condes

Arian Jane A. Cabilos
Marvielyn B. Pedotera
Precious Mae P. Tabuloc
Sheena Ann C. Herrera
Shuntle Mae Leizbeth C. Cañezo

RESEARCHERS

February 2019
 ACKNOWLEDGEMENT

The researchers would like to extend their sincerest gratitude to those who

in one way or the other helped on the realization of this research paper.

To the DOST, Palo for testing the water samples and providing test results;

To the Faculty and Administration of Tacloban City National High School,

for the acceptance and approval of this study;

To our research adviser, Mr. Rodelito G. Quinte, for his continued support,

guidance, and encouragement;

To Ramos family, owner of the deep well water, for letting the researchers

get water samples from it;

To the respondents, for sparing their time in participating in the researchers’

study, particularly in answering the questionnaires set by the researchers;

To all our friends for their continued support and prayers;

To our love ones for their understanding, love, and inspiration;

To our beloved parents for their moral and financial support; and,

Above all, to ALMIGHTY GOD for the gift of knowledge and good health.
 ABSTRACT

This study focuses on The Physico-chemical and Microbiological Analysis

of Water Source in Brgy. 105 San Isidro, Tacloban City. This study is undertaken

to determine the analysis of the pump well in Brgy. 105 San Isidro Tacloban City,

if it is a safe and uncontaminated drinking water for the families who drink water

from it. Specifically, the study aims: to determine the physical characteristic of the

water sample as to color; to analyse physico-chemical parameters in terms of

turbidity, salinity, hardness and total suspended solids (TSS); and, to analyse

microbiological parameters in terms of Heterotrophic Plate Count, E. coli (MPN)

Count, Total Coliform.

According to the results, the physical characteristic of the water sample as

to colour passed the standard value of 5 color units; its microbiological parameters

didn’t pass the standard and absolute value of a safe drinking water while its

physico-chemical analysis passed the Philippine National Standards for Safe

Drinking Water.

Constant monitoring of the deep well water in Brgy. 105 San Isidro, Tacloban

City is suggested. Sewerage facilities and septic tanks should be constructed far

from the location of the pump well. The inclusion for test of inorganic constituents

and pesticides is recommended in researches related to the quality of water in

deep wells. The conduct of the barangay officials’ training workshops on how to

formulate policies or guidelines that will help in maintaining the deep well water

potability is also suggested.

 TABLE OF CONTENTS
Title Page
Acknowledgement
Abstract
Table of Contents
Chapter I: Introduction
Background of the Study 2
Statement of the Problem
Null Hypothesis 8
Significance of the Study 8
Conceptual Framework 10
Definition of Terms 11
Chapter II: Review of Related Literature 12
Chapter III: Methodology
Research Design 19
Research Locale 19
Research Instrument 20
Scope and Limitations 20
Procedure 21
Chapter IV: Results and Discussion 22
Chapter V: Conclusion and Recommendations

Conclusions 26
Recommendations 26
Bibliography 28
Appendices

Appendix A 32
Appendix B 33
Appendix C 35
 CHAPTER I

INTRODUCTION

Background of the Study

Groundwater is generally considered a safe source of drinking water

because it is abstracted with low microbial assessment of microbial and physico-

chemical qualities of water (Palamuleni & Akoth, 2015). On a global scale,

groundwater represents the world’s largest and most important source of fresh

potable water.

Generally, the groundwater quality varies from place to place, seasonal

changes, (Trivede, Bajpai & Thareja, 2010). Or even the types of soils, rocks and

surfaces through which it moves, Seth, Tagbor & Bernard (2014). Naturally

occurring contaminants are present in the rocks and sediments. As groundwater

flows through the sediment metals such as iron and manganese are dissolved and

may later be found in high concentrations in the water, (Moyo, 2013). In addition,

human activities can alter the natural composition of groundwater through the

disposal or dissemination of chemicals and microbial matter on the land surface

and into soils, or through injection of wastes directly into groundwater,

(Govindarajan & Senthilnathan, 2014).

A study by De Vera (2015) states that contaminated deep well water results

in problems of gastro-enteritis and other waterborne diseases. High levels of

salinity, high hardness and microbiological problems have also been reported in

groundwater. Water quality problems have partly been associated with inadequate
sanitation, Van Vuuren (2013). Watershed management programs often rely on

monitoring for a large number of quality parameters to define contaminant issues.

While coliforms have traditionally been used to identify microbial contamination,

these indicators cannot discriminate among potential contaminant sources.

Microbial source tracking (MST) can provide the missing link that implicates the

sources of contamination, Plummer et al. (2007).

According to Dileep (2015) in her study, “Physico-chemical and

Microbiological Analysis of Municipality Drinking water,” the open well water has

the highest content of bacteria and total conidial spores rather than the municipal

tap water and can water. The total plate and coliform count revealed that 82.6%

and 92.4% of drinking water samples are found to cross the WHO guideline value

for drinking water.

There is now a growing awareness on the use of safe water for

consumptions in Brgy. 105 San Isidro, Tacloban City. Some residents use

disinfection methods such as boiling and chlorination of water which comes from

wells for drinking purposes. Others prefer to buy purified or commercial mineral

water to ensure that their drinking water is safe and potable. However, there are

still residents who drink water from wells directly and is presumed to be safe.

This is one of the major problems faced every day. Thus, the researchers

come up with the study, “Physico-chemical and Microbial Analysis of Water Source

in Brgy. 105 San Isidro, Tacloban City.”

Statement of the Problem

This study is undertaken to determine the analysis of the deep well in Brgy.

105 San Isidro Tacloban City, if it is a safe and uncontaminated drinking water for

the families who drink water from it. Specifically, the study aims:

a) to determine the physical characteristics of the water sample as to color;

b.) to analyse microbiological parameters in terms of Heterotrophic Plate

Count, E. coli (MPN) Count, Total Coliform (MPN) Count and Fecal Coliform

(MPN) Count; and

c.) to analyse physico-chemical parameters in terms of turbidity, salinity,

hardness and total suspended solids (TSS).

Null Hypothesis

There is no significant difference between the purified or the commercial

mineral water and the water from the deep well. Hence, the consumers can utilize

the water from the deep well or the purified or commercial mineral water as their

source of drinking water.

Significance of the Study

This research entitled, “Physico-chemical and Microbiological Analysis of

water source in Brgy. 105 San Isidro, Tacloban City,” is beneficial to the following;
To the community

This study will help the community to ensure the safeness and potability of

the water source used as a drinking water. It can help minimize risk and therefore,

it can prevent deleterious health repercussions through the consumption of water

as a drinking water.

To the researchers

This study will serve as a tool and reference material to help improve the

other researchers’ study related to this topic. It can provide more information about

the physico-chemical properties and the microscopic forms of life present in the

water sample.

To the Industry

This study will help the industry in reproducing safe and potable drinking

water not only to in Brgy. 105 San Isidro, Tacloban City but also to various

barangays. It can be widely utilized for various rural and urban industrial purposes.
Conceptual framework of the study

This chapter focuses in discussing about Physico-chemical and

Microbial Analysis of water source in Brgy. 105 San Isidro, Tacloban City.
The researchers will conduct a The researchers will get a
survey, employing a combination water sample form the deep
of.observation and questionnaire. well.
.

All numerical values obtained The water sample will be

from the laboratory analyses of transported to the DOST
water parameters will be immediately. The sample will
compared with the values set by be analysed.
PNSDW 2007.

The figure 1: Physico-chemical and Microbiological analysis of water source.

The diagram above shows the flow of the research study. First, the

researchers will conduct a survey in Brgy. 105 San Isidro, Tacloban City. Ten

respondents will be chosen to answer the questions in the questionnaire. Then,

the researchers will collect a water sample from the pump well. After the collection,

the collected water sample will be transported immediately to the DOST. The water

sample will be analysed with the use of national standard methods of analysis and

laboratory apparatus for both the physico-chemical and microbiological analysis.

The physical characteristic, microbiological and the physico-chemical content of

the water sample will be evaluated. All numerical values obtained from laboratory

analyses of water parameters will be compared with the values set by PNSDW

2007, to evaluate if the sample will pass or fail the national standard for a good

and safe drinking water.

Definition of Terms

In order to provide the reader a better understanding about the content in

the study, the following key terms are defined literally as they were used in the

study.

1. Analysis –a careful study of something to learn about its parts, what they

do, and how they are related to each other.

2. Microbiology –a branch of biology dealing with microscopic forms of life.

3. Physico-chemical -relating to chemistry that deals with the physico-chemical

properties of substances.

4. Water sources –are sources of water that are useful or potentially useful to

humans. It is important because it is needed for life to exist.

 Chapter II

REVIEW OF RELATED LITERATURES

According to the study “Physico-chemical and Microbiological Analysis of

Underground Water in and around Gwalior City, MP, India” (K. Ajit et al., 2009),

ground water is one of the major source of drinking water in both rural and urban

areas. The essence of ground water for humans’ usage cannot be over

emphasized. Groundwater contains chemicals which made people use it as a

potable water, unaware of its various water-borne diseases. Unsafe drinking water

contributed to countless health problems in developing countries such as the one

billion or more incidents of diarrhea occur annually.

This study of K. Ajit et al. (2009) explains the fact that most consumed water

is ground water. The underground water intake is not ensured with its quality and

potability. This would help the researchers prove the fact that safe water sources

are highly essential to everyday living.

De Vera Ph.D. (2015), in her study “Microbiological and Physico-chemical

Quality of Deep well water in Selected Public Schools”, stated that contaminated

deep well water results in gastro-enteritis and other water-borne diseases.

Groundwater which is the main source of drinking water should be periodically

monitored.

This study by De Vera (2015) is connected to the researchers’ study

because nowadays, the consumers can’t easily determine if the water source is
safe, clean and microbes free. Deep well water is prone to pollutants, and thus; it

alters its purity making its microbial content percentage increase rapidly.

In the study “Physicochemical and microbiological assessment of drinking

water from different sources Junagadh City, India” (V.G Vyas, M. Hassan, S.I.

Vindhani, H.J. Parmar, V. am. Bhalani, 2014), the quality of water is generally

depending on its chemical and microbial condition. The water can easily be

contaminated during its transportation from its supply and finally to the consumer.

The condition of the water supply networks is a factor which contributes to the

water quality. Water becomes contaminated by pathogens such as coliform group

bacteria, Salmonella and dysentery causing human faecal material carried in

sewage is often dumped into the rivers and lakes that may lead to water

contamination. Ground water crisis is not a result of natural factors. Prolonged

discharge of industrial effluents, domestic sewage and solid waste dump causes

the groundwater to be polluted and create health problems.

This study of (V.G Vyas, M. Hassan, S.I. Vindhani, H.J. Parmar, V. Am.

Bhalani, 2014) proves that the quality of underground water is highly affected by

various factors such as transportation and its microbial contents. The study would

help the researchers determine the possible factors which would have an impact

to the water to be consumed by countless residents in a specific work place.

A research by AMA Dawood, A Dawood & NN Okine (2013), the United

Nation had bespoken that water is essential and to access a safe water source is

imperative to humans. The assessment of water quality became urgent globally

due to recent cases alarming the countries. A report about global development

outlook indicates that 30% of the population lack access to safe drinking water.

Thus, the research by AMA Dawood, A Dawood & NN Okine (2013)

represents the theory being highlighted by UN wherein the water being consumed

resulted to some cases of water-borne diseases. A report/outlook indicated that

most of the population lack access to safe sources causing a global alarm. It is

one of the major problems faced globally. This shows the capacity of water-borne

diseases to affect the health of its consumers where the researchers’ study is

looking forward to.

Generally, groundwater quality varies from place to place, sometimes

depending on seasonal changes, the types of soils, rocks and surfaces through

with it moves. Naturally occurring contaminants are present in the rocks and

sediment (Palamuleni & Akoth, 2015).

Palmuleni & Akoth’s study is connected to the researchers’ study because

it constantly gives emphasis to the contaminants which may affect the water

source and its quality such that it would have a greater content of microbes with

least ensured safety of certain consumers. This may help the researchers in this

research work on discovering the water's quality in Brgy. 105 San Isidro, Tacloban

City.

A work by Hunter et al., (2010), a poor water supply impacts health by

causing acute diarrhea episodes, and non-diarrheal diseases, which arise from
chemical species such as arsenic and fluoride. It can also affect health by limiting

productivity and the maintenance of personal hygiene.

In the researchers’ study “Physico-chemical and Microbiological Analyses

of water source in Brgy. 105 San Isidro, Tacloban City” the community in the said

barangay has a problem in identifying whether their water source is safe, clean

and microbes free. One of the main aims of this study is to give the community an

assuring result that would ensure the safety of the water source for them to be able

to avoid various water-borne diseases. In addition, they will also have an adequate

amount of consumable water in their daily living.

N. Rahmanian et.al, (2015) also states that contaminants in the water can

affect the water quality and consequently to human health. The potential source of

water contamination and geological conditions, industrial and agricultural activities,

and water treatment plants. These contaminants are further categorized as micro-

organisms, inorganics, organics, radionuclides and disinfectants.

The researchers’ study is connected to Rahmanian et. al, (2015) because

this research aim is to know the physico-chemical analysis and its microbiological

content to enable the consumers in the community to evaluate the potability and

safeness of their source of water used in industrial purposes.

Jensen et al., (2002) states that even if drinking water of poor rural

communities is obtained from a safe source it can become contaminated during

storage in the house.

 Jensen et al. (2002) explains that even if the source of drinking water is

safe, it can become contaminated while it is being stored in the house. This is

possible because of the microbes and other elements present in its environment

which can affect the water's quality.

According to Edema et al., (2001) “Confirmation of microbiological standard

is of special great concern because water has great potential to spread diseases

within a big population. Although the standards vary from one place to another

place, the main objective is to reduce the possibility of spreading water borne

diseases.”

The United Nations Environmental Programme Global Environment

Monitoring System (UNEP GEMS 2000) states that water is vital to the existence

of all living organisms, but this valued resource is increasingly being threatened as

human population grow and demand more water of high quality for domestic

purposes and economic activities.

This statement of UNEP GEMS (2000) clearly gives emphasis that water is

very significant to all living things, without safe water, the lives of biotic factors are

at risk. As population increases, the demand for water by consumers also

increases. Thus, the amount of safe water is already inadequate for consumers

nowadays due to human activities and environmental factors.

According to Palmer et. al, (2005), it is essential that people be informed

about goods and services provided by freshwater ecosystems. Humans utilize the

services provided by aquatic ecosystems for food crops in agriculture, skins,

medicinal products, ornamental products (such as aquarium fish), implementation

of biological control of insects and weeds of aquatic ecosystems in order to better

manage them, and increasingly for recreational purposes.

This research conducted by Palmer et al. (2005) states that consumers

should know the significance of water. Humans use water in their daily living, in

recreational purposes and industrial means.

Lack of access to safe water and adequate sanitation services especially in

developing countries often results in the death of about two million infants annually

(UNICEF, 2005; Cosgrove and Rijsberman, 2000; Gomez and Nakat, 2002).

This statement of UNICEF (2005) means that a clean and potable drinking

water is essential to the health of our global community. Improving and expanding

existing water treatment and sanitation systems is more likely to provide safe and

sustainable sources of water for the consumers.

 Chapter III

METHODOLOGY

Research Design

This study made use of the descriptive research and in particular, it utilized

a descriptive-survey and descriptive-analytic design. Descriptive survey design is

used for evaluating the physical characteristic of the deep well water, as to color.

The descriptive analytic design is used to describe the nature of an object by

separating it into parts, its purpose is to discover the nature of things, particularly,

the chemical and microbial analysis of the pump well water (Calmorin, Calmorin,

2007).

The researchers’ general procedure of describing had the chief purpose of

description of the water source’ analysis in Brgy. 105 San Isidro, Tacloban City

that might had affected by both physical and environmental factors.

Research Locale

This study is conducted in Brgy. 105 San Isidro, Tacloban City,

employing a combination of observations and questionnaire.

Research Instrument

Research materials in conducting the experiment.

1. Survey questionnaire- It is used to secure certain questions and is used to

obtain information on the Physico-chemical and Microbiological Analysis of

Water Source in Brgy. 105 San Isidro, Tacloban City.

2. Water Bottles- It is used as a container in which the water sample will be

placed.

3. Water samples- These are the primary components in experimentation.

4. Pail- It is used as storage for the unused waters.

Scope and Limitations

This study aimed to enlarge the knowledge capacity about the safeness and

potability of the available water consumed by residents from a pump well in Brgy.

105, San Isidro, Tacloban City.

It only focused on investigating the physico-chemical and microbiological

content of the water source in Brgy. 105 San Isidro, Tacloban City.
Procedure

Before the start of actual water sampling, a survey of the pump well water

as the water source in Brgy. 105 San Isidro, Tacloban City was accomplished. A

questionnaire on a physical characteristic of the pump well water as to color was

prepared. Ten respondents were chosen through purposive sampling to answer

the questions set by the researchers in the questionnaire. After the survey, the

researchers proceeded on getting water samples from the pump well. Then after

that, the researchers went to DOST-Palo and gave the water samples for it to be

investigated and analysed. The researchers waited for the go signal of the DOST-

Palo if the water’s analysis is already done before the researchers get the values

and results.

This study employed the use of standard laboratory procedures to analyse

the water quality. All values were compared with the values set by Philippine

National Standards for Safe Drinking Water 2007 (PNSDW 2007).

 Chapter IV

RESULTS AND DISCUSSIONS

Table1. Physical Characteristic of.Pump Well.Water.Sample

Sample Sample Parameter Method Results

Number Information

Color (True) Photometric 1.00 HU

MIC- Water sample
0032 placed in 1.5 L
PET..bottle
labelled as
“DEEP WELL
WATER”
collected on 18
Jan 2019 at
2:15 PM”

Table 1 shows the physical characteristic of water sample in Brgy. 105 San

Isidro, Tacloban City. It can be gleaned from the table that the color description

passed the Philippine National Standards for Safe Drinking Water 2007 (PNSDW

2007).

Colour may occur in drinking water for any one or more several reasons. It

may be due to the presence of coloured organic substances originating in the

decay or aqueous extraction of natural vegetation, such as in soil runoff; presence

of metals or highly coloured industrial waste commonly pulp and paper textile (Am,

J., 1966).
Table 2. Microbiological.Analyses.of.Deep.Well.Water.Sample

Sample Sample Parameter Method Results

Number Information

Total.Colifor Multiple-Tube
m (MPN) Fermentation ›23.0
MIC- Water Count Technique MPN/100
0032 sample mL
placed in 350 Fecal Multiple-Tube
mL Coliform Fermentation ›23.0
PET..bottle (MPN) Count Technique MPN/100
labelled as mL
“DEEP E. coli Multiple-Tube
WELL (MPN) Count Fermentation ›23.0
WATER” Technique MPN/100
collected on mL
18 Jan 2019 Heterotrophic Pour Plate Technique 28,000 or
at 2:15 PM” Plate Count 35°C/48h 28 x 10³
Plate Count Agar CFU/mL

Table 2 reveals the data on total coliform counts, fecal coliform count, E.

coli and heterotrophic plate count of the deep well water sample. It shows that the

total bacterial count in the study site did not pass the standard for drinking water

as per standard methods of detection and values for microbiological quality. E. coli

is detected in the deep well water sample. The PNSDW standard value for total

coliform is <1.1 MPN/100 mL, fecal coliform is <1.1 MPN/100 mL, E. coli is <1.1

MPN/100 mL and heterotrophic plate count is <500 CFU/mL.

The presence of coliform bacteria warns of the potential presence of

diseases causing organisms and should alert the person responsible for the water

to take precautionary actions A.L. Smith (1971). According to Edberg, et.al. (2000)

no detection of E. coli is evident in all areas because E. coli have relatively short
life span and is considered to be the most sensitive to environmental stresses, so

its survival is short live. These bacteria survive for about 4-12 weeks in water

containing a moderate amount of microflora at a temperature of 15° – 18°. When

concentrations of fecal coliform are elevated, concentrations of E. coli are elevated

too.

Table 3. Physico-Chemical Analyses of Water Sample

Sample Sample Parameter Method Results

Number Information

CHE- Water Turbidity Photometric <1.00 FAU

0014 sample
placed in 1.5
L PET..bottle Salinity Refractometer <1.00‰
labelled as
“DEEP
WELL Hardness EDTA Titrimetric 37.8 mg/L
WATER” (𝑎𝑠 𝐶𝑎𝐶𝑂3 )
collected on
18 Jan 2019
Gravimetric, Dried
at 2:15 PM”
Total Suspended @ 103-105°C < 1.00 𝑚𝑔
Solids /𝐿𝑎𝑏
(TSS)

Table 3 presents the results obtained from the physico-chemical analysis of

the deep well water sample. It can be gleaned in the table that the turbidity, salinity,

hardness and total suspended solids (TSS) of the deep well water sample is low

compared with the allowable limit of 5 NTU for turbidity, <500 mg/L for salinity, 300

mg/L (as Ca𝐶𝑜2) for hardness and <10 mg/L for total suspended solids (TSS).
 The possible sources of salinity in groundwater are from agriculture and

natural sources. Evaporation of irrigation water will remove water and leave salts

behind. More salt can be dissolved from soil as irrigation water percolates

downward. Plants can naturally increase soil salinity as they uptake water and

exclude salts. Groundwater contains naturally-occuring salts from dissolving rocks

and organic materials. Some rocks dissolve very easily; groundwater in these

areas can naturally be very high in salinity. The table reflects that the water sample

passed the allowable limit of 1 mg/L of total suspended solids. According to Taiwo

(2011) high total suspended solids in drinking water could harbour microbiological

pollutant during and after rainfall, citing in particular the groundwater in Abeokuta

which becomes turbid and laden with suspended solids.

 Chapter V

CONCLUSIONS AND RECOMMENDATIONS

Conclusions

The salient findings of the study are the following:

1. After the laboratory analysis, the researchers conclude that the deep well

is contaminated and unsafe for the families who drink water from it.

2. The physical characteristic of the water sample as to color passed the

description of a standard value.

3. The pump well water’s microbiological parameters as to Heterotrophic

Plate Count, E. coli (MPN) Count, Total Coliform (MPN) Count and Fecal

Coliform (MPN) Count did not possess the standard value for a safe drinking

water.

4. The physico-chemical parameters in terms of salinity, turbidity, hardness

and total suspended solids (TSS) passed the standard values set by the

PNSDW 2007.

Recommendations

1. Constant monitoring of the deep well water in Brgy. 105 San Isidro,

Tacloban City is suggested.

2. Sewerage facilities and septic tanks should be constructed far from the

location of the pump well.

3. The inclusion for test of inorganic constituents and pesticides is

recommended in researches related to the quality of water in deep wells.

4. The conduct of the barangay officials’ training workshops on how to

formulate policies or guidelines that will help in maintaining the deep well

water potability is also suggested.

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to 200 MLD filteration. Retrieved from

www.researchgate.net/publication228771058_Comparative_Study_of

_Seasonal_Variation_in_Physico=Chemical_Characteristics_in_Drinking_Water

Quality_of_Kanpur_IndIA_With_Reference_To_200_MLD_filteration on August 8,

2018.

UNICEF, (2005). National Rural water supply and Sanitation investment

programme. Final draft, pp: 1.

World Health Organization (2010). Guidelines fpr Drinking-water Quality.

Recommendation, Geneva, pp: 1-6. Retrieved from

http://www.who.int/water_sanitation_health/WHS_WWD2010_guidelines_2010_6

_en. On October 3, 2018.

APPENDICES
 Appendix A

TIME TABLE OF ACTIVITIES

o June 13, 2018- Submission of the statement of the problem and significance of

the study

o June 25, 2018- Deadline of chapter I

o July 3, 2018- Proposal of chapter I

o October 1, 2018- Gathering data in chapter II review of related literature

o January 18, 2018- Giving the water samples to the DOST, Palo

o January 25, 2019- Paying the analysis fee in DOST

o January 28, 2019- Presentation of results and discussion

o February 6, 2019- Checking of the final draft of the paper

o February 7, 2019- First complete draft of the paper

 Appendix B
DOCUMENTATION

Figure 1: Actual pumping of a water sample

into a 350 mL and 1.5 L bottle for Physico-
chemical and Microbiological analysis.

Figure 2: Paying for the analysis fee at the

DOST VIII counter.
Figure 3: Waiting for the
laboratory results.
 Appendix C
METHODOLOGY
These are the different methods used in acquiring the result of each

parameter.

Physico-chemical

1. Color (True)

Sample was filtered through a standard glass fiber filter. Filtered sample

was transferred to a 50 mm cell and color was read at 455nm using

Pharo 300 Spectroquant Spectrophotometer.

2. Turbidity

Sample was transferred in a 50mm rectangular cuvette and inserted in

the cell receptacle of Pharo 300 Spectroquant Spectrophotometer.

Turbidity was determined using 550 nm filter. Result displayed in the

photometer’s LCD was recorded.

3. Salinity

A drop or two of a well-mixed sample was placed unto the prism surface

of Atago Salinity Refractometer, previously adjusted to 0‰ using

distilled water. Salinity value was viewed through the eyepiece.

4. Hardness (Total)

Sample was diluted with distilled water, added with hardness buffer

solution and drops of Eriochrome Black T solution and titrated with

standardized EDTA titrant to the blue end point.

 5. Total Suspended Solids

Sample was filtered using Whatman GF/C glass fiber filter previously

washed and dried to constant weight. Filter with the solids was dried to

constant weight at 103-105°C in a drying oven.

Microbiological

1. Multiple -Tube Fermentation

Undiluted sample was inoculated to fermentation tubes with

appropriate volume of Lauryl Sulfate Broth (Merck), incubated at ( 35 ±

0.5 ) °C, ( 48 ± 3 ) h. Tubes exhibiting growth and/or gas formation were

further confirmed using Brilliant Green Lactose Bile Broth (Merck)

incubated at ( 35 ± 0.5 ) °C, ( 48 ± 3 ) h for estimation of Total Coliform,

EC medium (HiMedia) and Tryptone water medium (Merck) incubated

at ( 44.5 ± 0.2 ) °C for ( 24 ± 2 ) h for estimation of Fecal Coliform and

E. coli, respectively.

The result, reported as Most Probable Number per 100mL of sample

(MPN/100 mL), is an estimate of the mean density of coliform in the

sample based on probability formula.

2. Heterotrophic Plate Count (HPC)

The HPC, also known as Heterotrophic Plate Count, estimates the

number of live heterotrophic bacteria in the sample. Following pour

plate method, samples (undiluted and diluted) were plated with Plate
Count Agar (Merck), incubated at 35°C for 48 h. Result is reported as

Colony-Forming Units per mL of sample (CFU/mL).