1

VARIETIES OF PULSES FOR PRELIMINARY SURFACE WATER

TREATMENT: A COMPARATIVE STUDY

A RESEARCH SUBMITTED TO THE FACULTY OF BENGUET STATE

UNIVERSITY – SECONDARY LABORATORY SCHOOL, IN PARTIAL
FULFILLMENT TO THE REQUIREMENTS OF SENIOR HIGH
SCHOOL – SCIENCE TECHNOLOGY ENGINEERING
AND MATHEMATICS TRACK

KAWI, Dusty T.
BAS-ILAN, Aubrey T.
BOMOGAO, Islay
CHUA, Maria Francesca Nicole R.
GARCIA, Zaña Lei C.

April 2019
 ACKNOWLEDGMENT

This paper would not be successful if not for the people who supported the

researchers.

Appreciation is extended to their adviser, Ms. Alicia B. Balongyad, who

extended her firm supervision equipping the researchers with knowledge and

values. They gratefully acknowledge Ms. Vanderlee M. Batalier for her concern

and suggestions that made them endure the hardships of this research.

The authors would also like to express their gratitude to the generous

contributions of Dr. Olga B. Betudio, Mrs. Rosemarie P. Daytec and Mr. Ariston

B. Canayon for their valuable comments, suggestions and advices for the

improvement of the study.

Salutation is also extended to Engr. Rhea M. Contada for her astounding

kindness and for sharing her knowledge; Mr. Jason M. Leung for his heart

warming patience and clever ideas; and to Mrs. Elizabeth Dom-ogen for patiently

and compassionately helping in polishing the methodology and for her useful

instructions.

Appreciation is extended to CAS-Biology Department for unselfishly

lending their laboratory rooms and tools. Special thanks to Mrs. Cecilia Samonte

for letting the authors use the SLS-Chemistry Laboratory Room, as well.

The authors would also like to thank their friends who have colored this

journey with laughter, joys, nuisance and food.

 This paper is dedicated to the authors’ families who have been their

greatest inspiration, especially to their parents, for their incredible support and to

their siblings for their amazing love and understanding especially during the

tough times.

Most importantly, the authors are forever grateful to the Almighty God for

his unending love and protection all throughout this journey.

Aubrey T. Bas-ilan

Islay Erika A. Bomogao

Maria Francesca Nicole R. Chua

Zaña Lei C. Garcia

Dusty T. Kawi
 TABLE OF CONTENTS

Page

Title Page . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii
List of Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iv
List of Appendix Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . viii
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Background of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Conceptual Framework. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Statement of the Problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
MATERIALS AND 15

METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
RESULTS AND DISCUSSION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

CONCLUSIONS AND RECOMMENDATIONS. . . . . . . . . . . . . . . . . 22

LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

APPENDICES

Letter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

.
Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Pictures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
LIST OF TABLES

Table No. Page

1 Summary of weighted means as 18

 comparison among methods. . . . . . . . . . . . . . . . . . . . . . . . . . .
Summary of weighted means as
2
comparison among pulses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Summary of pH level as
3
comparison among pulses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Microbial analysis of

4 treated

wastewater. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

LIST OF APPENDIX TABLES

Table No. Page

1 Weighted mean for salt extraction in testing color. . . . . . . . . . . . 31

2 Weighted mean for salt extraction in testing odor. . . . . . . . . . . . . 31
3 Weighted mean for direct method in testing color. . . . . . . . . . . . 31
4 Weighted mean for direct method in testing odor. . . . . . . . . . . . 32
5 Weighted mean for activation method in testing color. . . . . . . . . 32
6 Weighted mean for activation method in testing odor. . . . . . . . . 32
7 Weighted mean for mung beans in testing color. . . . . . . . . . . . . . 32
 8 Weighted mean for mung beans in testing odor. . . . . . . . . . . . . . 33
9 Weighted mean for cardis beans in testing color. . . . . . . . . . . . . . 33
10 Weighted mean for cardis beans in testing odor. . . . . . . . . . . . . . 33
11 Weighted mean for navy beans in testing color. . . . . . . . . . . . . . 33
12 Weighted mean for navy beans in testing odor. . . . . . . . . . . . . . 34

ABSTRACT

An experiment was conducted to evaluate and compare three varieties of

pulses namely Cajannus cajan (Cardis beans) Phaseolus vulgaris (Navy beans)

and Vigna radiata (Mung beans). Specifically, it aimed to determine the physical

and chemical characteristics of wastewater once treated using three different

methods. These methods are activation method, direct application and salt

extraction. This study was conducted at La Trinidad, Benguet.

Using the jar test, wastewater from a swamp in La Trinidad Strawberry

Farm was treated with the three different varieties and three different methods.

The physical characteristics specifically, color and odor, was evaluated by

respondents as a survey. The chemical characteristics were tested in the laboratory

using pH paper and Gram staining.

 It was concluded that varieties of pulses possess potential natural

coagulant for surface water treatment due to its ability to remove turbidity.

However, it has effects on the basicity and acidity of the water sample.

Furthermore, it has no observable antibacterial property on water. In comparison,

Direct method was the most efficient in removing undesirable color and odor

while Cardis beans exhibit the best results in physical characteristics of treated

wastewater.
 1

INTRODUCTION

Background of the Study

Clean Water is an important resource for life. However, equality and

accessibility to it has become a major problem today. According to the World

Health Organization (WHO) and United Nations International Children’s Fund

(UNICEF) as of 2017, three out of 10 people or 2.1 billion lack access to safe,

readily available water at home. Also, six in 10 people or 4.5 billion lack

sanitation. Said alarming information will the basis of this study in solution-

finding as to natural remedies using common bean pods extract as an effective

natural water coagulant.

In the provision of clean drinking water, it must undergo several water

treatment processes which typically consist of several steps in the treatment

process according to the journal of Water Resource and Protection (2016). These

include: (1) Collection; (2) Screening and Straining; (3) Chemical Addition; (4)

Coagulation and Flocculation; (5) Sedimentation and Clarification; (6) Filtration;

(7) Disinfection; (8) Storage; and finally (9) Distribution. With the different ways

of treatment, coagulation and flocculation are classified as part of the procedure.

According to Akinbile (2018), coagulation – flocculation is regarded as one of the

most used and effective means of water treatment. It is a chemical water treatment

technique typically applied prior to sedimentation and filtration (e.g. rapid sand

filtration) to enhance the ability of a treatment process to remove particles.

Coagulation plays a major role in eliminating turbidity in the form of suspended

and colloidal material via addition of chemical coagulants such as Ferric Chloride,

Polyaluminum Chloride, Ferric Sulfate and Aluminum Sulfate which are some of

the commonly used salts. Aluminum is strongly neurotoxic and may be involved

in the development of Alzheimer’s Disease (Dwarapureddi, 2016). In addition,

disadvantages are observed such as relatively high procurement costs as well as

negative effects on the environment. Asrafuzzaman, Fakhruddin, and Hossain

(2011) stated that is therefore desirable that these chemical coagulants be replaced

with cost-effective natural coagulants to counteract the mentioned disadvantages

such as Moringa seeds, dragon fruit and cactus.

Based in their study, Asrafuzzaman et al. (2011) stated that using some

locally available natural coagulants, for example, Moringa oleifera, Cicer

arietinum, Dolichos lablab was able to significantly improve in removing

turbidity and total coliforms from synthetic raw water. Also, maximum turbidity

reduction was found for highly turbid waters after dosing water-soluble extract of

Moringa oleifera, Cicer arietinum, and Dolichos lablab. It was also found that

these natural coagulants reduced about 89–96% of total coliforms. In Tuguegarao

City, Nozaleda (2017) observed the decrease in the turbidity of water treated with

Terminalia catappa (Talisay) leaves. He also concluded that talisay leaves have no

significant effect on the pH and coliform count of the water. With the presented

results, this study aimed to use locally available natural coagulant such as the
pulses as a suitable, easier, and environment friendly option for water treatment.

A study conducted by Gayomba (2014) mentioned that pulses are a type of

crops that are intended to be harvested at a specific time. Failure to do so, leads to

an inedible waste that is often thrown away. Instead of adding up to the

community’s solid waste, it would be better to put it to other beneficial use.

Three different varieties of pulse crops were used – Phaseolus vulgaris

(Navy beans), Vigna radiata (Mung beans), and Cajannus cajan (Cardis beans).

These are the most common pulse crops in the Philippines. All these contain

globulin proteins which are responsible for its coagulative property (Fitzgerald,

Shuai & Zhu, 2018, and Kristianto, 2017). These different varieties undergo three

treatments. These treatments are the methods of extracting said proteins.

According to Johns et al. (n.d.) and Rahma et al. (2000) as cited by Fitzgerald et

al. (2018), salt extraction involves using salt as a catalyst to extract the protein.

Another method is through activating the proteins by vigorous shaking of the bean

paste (The Refreshing Point, 2017). Lastly, direct application of the bean pastes

for coagulating.

Significance of the Study

This study is an eye-opener of what people can use in the first phase of

water treatment. It is a way for people to consider organic means of coagulating

wastewater turbidity. It offers an inexpensive and environment-friendly water

coagulant for households and communities. It also encourages what extracting

method is best used for organic coagulants for future researches on other plant-

based materials. This study also helps people to determine which to use for

different and more specific types of wastewater such as household and industrial.
PROCESS
Conceptual Framework
1.Experimentation
INPUT Salt
Extraction OUTPUT
1. Common Beans Activation
Navy Beans Direct 1. Comparison
Mung Beans Application among common
Cardis Beans Survey Evaluation beans as a
2. Water Samples Microbial Analysis water coagulant
3. Distilled Water pH Testing
4. Stirrer Gram
5.Gram Staining Staining
Agents

Figure 1. Paradigm of the study

Water coagulation is the addition of positively charged polymers into a

water source in order to neutralize the negative charges of the water (Michigan

Tech, 2007). The pulse crops were used as water coagulant in exchange of

inorganic water coagulants that can harm users of such product. The process was

divided into three: experimentation, survey evaluation, and microbial analysis. By

experimentation the researchers produced treated water which was further

analyzed by the researcher. Survey evaluation and microbial analysis were

utilized for the physical and chemical characteristics of the treated water,

respectively. Hence, the results from the experiment and survey were used to

compare the different varieties of pulse crops as water coagulant.

Pulse Crops

Pulses are a leguminous crop that are harvested solely for the dry seed.

Dried beans, chickpeas, lentils and peas are the most commonly known and

consumed types of pulses (Montana Department of Agriculture, 2019).

Pulses are very high in protein and fiber, and are low in fat. Pulses are

also nitrogen-fixing crops that improve the environmental sustainability of

annual cropping systems. The term “pulse” only refers to the dried seed (Rupp,

J., 2016).

Plants’ seeds hold a promising future as alternative sources of water

purification. Most of the plants’ seeds reported to have bio-coagulative potentials

belong to the family Fabaceae, commonly known as legumes which are rich in

protein.

The suggested mechanism of coagulation property of pulse crops protein

is supposed to be that positively charged proteins bind to part of the surface of

negatively charged particles through electrostatic interactions (Bodlund, I., 2013).

Since conventional methods of assuring potable water in developing

countries are unsustainable, there is a need to consider the application of

sustainable technologies using locally available materials in treating surface water

(Pritchard et al., 2009). One area that holds a lot of prospect for the future is the

plant kingdom and particularly the uses of seeds in water treatment (Dalen et al.,

2009; Subramanium et al., 2011; Yongabi,2009).

Water Treatment Processes

To ensure cleanliness and the safety of the water we drink, it undergoes

several water treatment processes. Water is treated to remove sediment, bacteria,

and other impurities (Charleston Water System, 2013).

Hunter Water Corporation (2011) stated that water treatment process may

vary slightly at different locations, however, the basic principles are largely the

same.

The first process water goes through is Coagulation. The dirt and other

impurities in untreated water are negatively charged. Positively charged chemicals

are added to the water which will bind the particles and form larger particles

called floc which will be easier to be collected (Centers for Disease Control and

Prevention, 2015).

Water coagulation is the addition of positively charged polymers into a

water source in order to neutralize the negative charges of the water (Michigan

Tech, 2007).

Natural waters, polluted either by man or by nature, are likely to contain

dissolved inorganic and organic substances, biological forms such as bacteria and

plankton, and suspended inorganic material. To remove these substances, the

usual unit process includes sedimentation, coagulation and filtration. Coagulation,

generally followed by filtration, is far the most widely used process to remove the

substance producing turbidity in water. These substances normally producing

turbidity consist largely of clay materials and microscopic organisms and occur in

widely varying sizes, ranging from those large enough to settle readily to those

small enough to remain suspended for very long times (The American Water

Works Association, 1971)

Inorganic Coagulants

According to Ramavandi (2014) as cited by Janna (2016), many

flocculants and coagulants are commonly used in conventional water treatment

processes. These materials can be classified into inorganic coagulants. It shows

that these compounds are very efficient in removing foreign substances from

turbid water. Inorganic coagulants are used because they are cost-effective and

applicable for a broad variety of water treatment. Inorganic coagulants are

particularly effective on raw water with low turbidity (ChemTreat, 2019).

The most common inorganic coagulants in water treatment is the

aluminum sulfate (Al2(SO4)3 or alum (SNF, 2017). However, inorganic coagulants

are hazardous because of their negative effects to people. Aluminum Sulfate is a

type of salt that can cause skin irritation and mucous membranes. The primary

concern in alum is long-term exposure to low levels of the chemical. Aluminum,

from your diet or healthcare product, can cause degeneration of nervous system

tissue. It is possible that exposure to aluminum could lead to an increased risk of

certain cancers, brain plaques or Alzheimer's Disease (Abreo, V., 2009). The work

of Sies and Jones shows that high aluminum concentration causes oxidative stress.

Evidence of an oxidative stress status has been found in association with most

neurodegenerative disorder in which aluminum is present in relative high amount.

Aluminum negatively impacts neurotransmission, either by directly inhibiting the

enzymes responsible or by affecting the physical properties of synaptic

membranes

The second most used inorganic coagulant in water treatment is the Ferric

Sulfate. High content of residual iron in drinking water may be responsible for (i)

the neutralization of the disinfectants used to kill microorganisms, (ii) the

coloration of water, (iii) metallic tastes of water. Excessive ingested iron can

cause excessive levels of iron in the blood because high iron levels can damage

the cells of the gastrointestinal tract, preventing them from regulating iron

absorption. Humans experience iron toxicity above 20 milligrams of iron for

every kilogram of mass, and 60 milligrams per kilogram is a lethal dose. Ong and

Halliwell suggest that iron can be involved in Alzheimer’s disease. The important

mechanism is the interaction of iron and cholesterol in promoting oxidative

damage, causing both atherosclerosis and neurodegeneration. Increasing evidence

indicates that excessive iron in selective regions of the brain may be involved in

the neurodegenerative disorders (J.M. Siéliéchi, 2016).

Natural Coagulants

As commercialized coagulants pose threats to our environment and health,

natural coagulants are being developed and studied by many researchers

(Niquette, Patrick & Monette, Frédéric & Azzouz, Abdelkrim & Hausler, Robert.,

2004). Ali et.al. (2010), Alfugara and Muyibi (2003), Baes (1999), and

Ndabigengesere (1998) as cited by Birima et.al. (2013) developed natural

coagulants from Moringa seeds. It has been shown that Moringa seed is one of the

most effective as natural coagulant for water treatment and when compared to

conventional chemical coagulant (Amagloh and Benang, 2009).

A natural coagulant is suggested as a substitute or aid for alum. Natural

coagulants have been used for domestic household for centuries in traditional

water treatment in tropical rural areas. Some reports describe natural coagulants

from Nirmali seed and maize, mesquite bean and Cactus latifaria, and Moringa

oleifera seeds. The main advantages of using natural plant-based coagulants as

water treatment material are apparent; they are cost-effective, unlikely to produce

treated water with extreme pH and highly biodegradable. Naturally occurring

coagulants are usually presumed safe for human health (A H Birima et al, 2013).

The history of the use of natural coagulants is long. Natural organic

polymers have been used for more than 2000 years in India, Africa, and China as

effective coagulants and coagulant aids at high water turbidities. They may be

manufactured from plant seeds, leaves, and roots. Natural coagulants have bright
future and are concerned by many researchers because of their abundant source,

low price, environment friendly, multifunction, and biodegradable nature in water

purification. Using some locally available natural coagulants, for

example, Moringa oleifera, Cicer arietinum, Dolichos lablab, significant

improvement in removing turbidity and total coliforms from synthetic raw water

was found. Maximum turbidity reduction was found for highly turbid waters

(Asrafuzzaman et al., 2011).

Salt extraction

Salt extraction is a process where globulin proteins are separated from

albumins on the basis of solubility. Broadly speaking, sodium chloride (NaCl) is

one of the most commonly used salts for extraction (Karaca et al., 2016).

The improvement of coagulation efficiency by NaCl is apparently due to

the salting-in mechanism in proteins wherein a salt increases protein-protein

dissociations leading to increasing protein solubility as the salt ionic strength

increases (Mitsumasa Okada, 1999).

"Salting in" refers to the observation that at solutions of low salt

concentrations, the solubility of a protein increases with the addition of salt. As

the solubility of the salt is higher than that of the protein, it is more likely dissolve

and take up space in the solution; therefore, proteins aggregate and precipitate. By

contrast, "salting out" requires high salt concentration for the precipitation of the

protein. There are two ways of "salting out". In one method, proteins are exposed
to high concentrations of salt solutions, and in the other, the proteins are exposed

to a series of low concentrated solutions (Berg, J., 2007)

In addition, ‘salting-in’ of proteins typically occurs at low salt levels,

where the ions act to increase order of the protein's hydration layers and promote

protein-water interactions. Salts formed between cations and anions with lower

precipitation ability in the series weaken the hydrophobic interactions and result

in increasing solubility of non-polar amino acids, thus favoring the ‘salting-in’

process (Karaca et al., 2016)

Johns et al. (n.d.) used an extraction method using a saturated ammonium

sulfate ((NH4)2SO4) solution. He reported a maximum total protein dissolution

of 87.5% in mung bean proteins extracted in 5% NaCl solutions. On the other

hand, Rahma et al. (2000) used a different extraction method involving extraction,

centrifugation, filtration and micellization of mung bean proteins.

Gram Staining

Gram stain is a staining procedure developed by Hans Christian Gram in

1884 (Tortora, Funke & Case, 1995). It is one of the most used technique to

distinguish one group of bacteria from another (Nester, Anderson & Robert, 2001)

(Tortora et al., 1995). The differences in the structure of their cell walls cause the

different kinds of bacteria to stain unequally (Madigan, Martinko, & Parker, 2003)

(Nester, Anderson & Robert, 2001) (Tortora et al., 1995).

 Staining is the process of coloring microorganisms with a dye that

emphasizes certain structures. Before staining, the microorganism is smeared or

spread over the microscope slide and air dried. Some uses the flame of a Bunsen

burner to dry the smear. After that, the microorganism is flooded with a purple

dye such as crystal violet and washed off afterwards. Then, the smear is covered

with iodine (a mordant) and washed off. Afterwards, the slide is washed with

ethanol (a decolorizer) and again washed off. Lastly, the smear is counterstained

with safranin, washed off and dried. The smear is then observed under a

microscope. The bacteria that retain the color of the purple dye are Gram-positive

and the ones that do not are Gram-negative (Madigan, Martinko, & Parker, 2003)

(Nester, Anderson & Robert, 2001) (Tortora et al., 1995).

The Gram staining method is important in medical microbiology.

Although, it is not universally applicable, it provides valuable information for the

treatment of disease. For example, gram-positive bacteria tend to be killed easily

by penicillin and sulfonamide drugs. On the other hand, gram-negative bacteria

can resist these but are killed by streptomycin, chloramphenicol, and tetracycline

(Tortora et al., 1995). The Gram-positive bacteria include staphylococci ("staph"),

streptococci ("strep"), pneumococci, and the bacterium responsible for diphtheria

(Cornynebacterium diphtheriae) and anthrax (Bacillus anthracis) (MedicineNet,

2018).
Statement of the Problem

This study aimed to compare three different varieties of pulse crops for its

water coagulative ability. It specifically aimed to answer the following questions:

1. Which extraction method is most effective in coagulation of wastewater

turbidity?

a. Salt extraction

b. Direct application

c. Activation

2. Which among the three varieties of pulses is most effective in treating

wastewater in terms of its physical characteristics:

a. Color

b. Odor

3. Which among the three varieties of pulses is most effective in treating

wastewater in terms of its chemical characteristics:

a. pH

b. Microbial content

Hypotheses

1. There is an effective extraction method in coagulation of wastewater turbidity.

2. There is an effective variety of pulse in treating wastewater in terms of its

physical characteristics.
3. There is an effective variety of pulse in treating wastewater in terms of its

chemical characteristics.

Scope and Delimitation

This study focused on determining which variety of pulse crop is most

effective in treating wastewater in terms of different aspects. It is also helpful to

determine which method is most effective in extracting beans in producing a

coagulant. This study used a general source of wastewater from the Strawberry

Farm Swamp due to the presence of industrial, agricultural and household

wastewater in that area.

The experiment was done in Balili, La Trinidad which took about six hours for

coagulating and an hour in testing and evaluations. The entirety of the study took

place from January to April 2019. The final experiment was done in the month of

April.
 METHODOLOGY

This chapter discusses the research design used by the researchers. It

shows the tools used and steps done by the researchers in order to obtain

information. It presents the statistical tool used to interpret and analyze the

gathered data.

Research Design

This research used a qualitative-comparative design. It focused on the

characteristics of wastewater when treated using different varieties of common

beans. It compared the three varieties for its physical and chemical properties

which reflect its effectiveness as a water coagulant.

A comparative design essentially compares in an attempt to make a

conclusion about them. Researchers attempt to identify and analyze similarities

and differences between groups. It is still considered as experimental when the

variables are manipulated to measure how it influences what qualities are being

compared. It is also used to better understand which treatments are applicable

depending on the similarities and differences (Richardson, 2018).

Research Instrument

In order to test the coagulative properties of the beans, the researchers

conducted an experiment by using a stirrer in applying the different treatments.

The evaluation of treated water using common beans as a coagulant was done

through evaluation sheets given as a survey. The respondents were able to

distinguish which among the three common beans and the three methods are most

effective in treating color and odor. The chemical properties, pH and microbial

content, was observed in the laboratory using a pH meter and a microscope.

CONTROL SAMPLE A SAMPLE B SAMPLE C

TREATMENT 1
Color
Odor
TREATMENT 2
Color
Odor
TREATMENT 3
Color
Odor

Data Collection Process

Water samples were taken from the swamp located at the Strawberry farm

in La Trinidad. The beans were prepared by removing the seeds from the pods and

then sun-dried and lastly, crushed. After these preliminary steps, three methods

were used to extract and apply the beans. First, the activation method was done by

mixing the bean powder with distilled water and vigorously shaken for a minute

before being mixed into the water. Second, the salt extraction was done by mixing

bean powder with a brine solution before mixing it into the water. Lastly, the

direct application of the bean powder to the water was done. All three methods

were finished by letting it sit for two hours and then decantation of the treated

water was done. Once these treated waters have been freed from the coagulated

turbidity, it was tested for pH and microbial content as well as color and odor.
Treatment of Data

The tallied results from the evaluation sheets gathered through survey

were used to compute their weighted mean. All methods were compared as well

as the three varieties of beans when it come to their color and odor. The weighted

mean is a method by which a mean is used to discover a quality based on the

number that represents these qualities. It made use of a Likert scale in order to set

said standards for the qualities.

 CHAPTER III

RESULTS AND DISCUSSIONS

This chapter presents the analysis and interpretation of data collected. It

involves tabular data of tallied scores from evaluations and also a qualitative

interpretation of these scores. It also includes a comparison of treatments used in

the study.

Table 1: Summary of weighted means as comparison among methods

COLOR ODOR
Salt extraction 1.76 2.18
Direct method 2.42 2.51
Activation method 2.87 1.62

From the table above, it is shown that Activation method was the most

effective in producing a more desirable color for the treated water. Direct method

however, was most effective in treating the odor of the treated wastewater. This

shows us that different pulses are effective in successfully treating wastewater for

its undesirable color and odor. Between the two, Direct was still the best method

since it has a closely desirable color and also desirable odor. A study conducted by

Binayke and Jadhav (2013) uses the activation method in applying Moringa

olifera, Okra gum and dry flowers of Calotropis procera. It showed as high as

98% turbidity removal efficieny. This lead to a clear and colorless water

treatment. Saravanan and his team of researchers (2017) used the direct method in

treating industrial wastewater and found out Azadirachta Indica had as high as
63.01% turbidity removal efficiency. Sun and Hall (1975) had a different concept.

They believed globulins from common beans are isolated with the use of salt. A

salt commonly used was NaCl. This may be because globulins are further

classified as either euglobulins and pseudoglobulins. Euglobulins are only isolated

through the use of saline solutions however pseudoglobulins are isolated even in

plain water but still exhibit the same characterstics (Encyclopaedia Britannica

2019).

Table 2: Summary of weighted means as comparison among pulses

COLOR ODOR
Mung Beans 2.27 1.93
Cardis Beans 2.67 2.04
Navy Beans 2.11 2.33

Table 2 shows the overall scores of the three different varieties of pulses as

a water coagulant in terms of the change in color and in odor. Cardis beans which

obtained the highest weighted mean is most efficient in making the color more

desirable. Navy beans on the other hand is best at removing undesirable odor of

the wastewater. Navy beans have a globulin content of 40-50% (Tiwari&

Singh,2012), Cardis beans have 50-72% (Abrol& Shankar, 2016), and Mung

beans have 20-28%. Cardis with the highest globulin protein percentage exhibit

best performance in the treated water's color. However, odor did not reflect the

globulin content. Navy beans have 25-46% Oligosaccharides out of its total sugar

conent (Ruperez, 1998) while Cardis beans have 47-50% (Cazetta et al., 2009)
and Mung beans have 31-76% (Tajoddin et al., 2012). These Oligosaccharides are

medium chain carbohydrates that include digestible sugars such as maltodextrin

and fermentable sugars such as raffinose. Bacterial breakdown of these sugars

result to fermentation (Szczygiel, 2016). This may be a lead as to what might have

caused the change in the odor of the wastewater. Navy beans which have the

lowest percentage of Oligosaccharides was observed to have the best effect on the

odor of the wastewater when treated.

Table 3. Summary of pH level as comparison among pulses

DIRECT ACTIVATION SALT

METHOD METHOD EXTRACTION
Mung Beans 6 5 6
Cardis Beans 6 5 7
Navy Beans 6 5 8
Average 6 5 7

Table 3 shows pH analysis using pH paper. The variety of pulses do not

directly affect the pH of the water. The methods on the other hand, show

otherwise. Direct method showed a consistent pH of 6 while activation exhibits a

pH of 5. Salt extraction showed the biggest changes in pH. The addition of salt of

strong acid and strong base like NaCl to water does nothing to pH because the

species formed OH- and H+ balance each other out (Chat, 2013). All three

varieties of pulse crops have a pH range of 5-6. The water in both direct and

activation method took the pH of the pulses while those that underwent salt

extraction exhibit change in the pH of the water. Knowing the water’s pH is

important because it gives an idea of how it can be used. Only a specific water pH

can be taken by the human body without disrupting metabolic processes in the

body.

Table 4: Microbial Analysis of treated wastewater

SALT
DIRECT ACTIVATIO
EXTRACTIO AVERAGE
METHOD N METHOD
N
Mung beans Positive Positive Negative Positive
Cardis beans Negative Positive Positive Positive
Navy beans Positive Positive Negative Positive
Average Positive Positive Negative
The table shows that all three varieties of pulses have Gram-positive

bacteria. Salt extraction treatment shows mostly Gram-negative bacteria. This

may be caused by the addition of salt. A study conducted by Brown and Turner

(1963) on the membrane stability and salt tolerance of Gram-negative bacteria

showed that higher concentrations of salt had higher Gram-negative bacteria

count. Bacteria that stain purple with the Gram staining procedure are termed

Gram-positive; those that stain pink are said to be Gram-negative. Gram-positive

and Gram-negative bacteria stain differently because of fundamental differences

in the structure of their cell walls. The bacterial cell wall serves to give the

organism its size and shape as well as to prevent osmotic lysis. The material in the

bacterial cell wall which confers rigidity is peptidoglycan (Kaiser, 2019). Overall,

Gram-positive bacteria were found more evident. Although specific bacteria are

not named, gram staining gives an idea of how much of it is present. In addition,

both cocci and rod-shaped bacteria were observed.

 CHAPTER IV

CONCLUSIONS AND RECOMMENDATIONS

This chapter shows the summary of findings from the study. It also

includes several recommendations for future studies not met by this study.

Conclusions

With the results, the researchers came up with the conclusion that varieties

of pulses possess potential natural coagulant for surface water treatment due to its

ability to remove turbidity, specifically,

1. Direct method was the most efficient in removing undesirable

color and odor.

2. Cajannus cajan exhibit the best results in color while Phaseolus

vulgaris for the odor of treated wastewater.

3. Pulse crops have effects on the basicity and acidity of the water

sample.

4. It has no observable antibacterial property on water.

Recommendations

For future studies and further research, the researchers recommend:

1. A further analysis of different concentrations of pulse varieties is

recommended.

2. Testing the synergic effect of pulse crops in reducing the turbidity

of wastewater
3. Future study on other pulses locally available as a water coagulant

may also be made.

4. The use of an actual jar test to simulate coagulation-flocculation

process and a digital turbidimeter to measure turbidity in order to

increase the reliability of results.

5. Conduction of a study using combination treatment of alum and

pulse varieties in different proportions in order to investigate

effectiveness in treating raw water and further to investigate the

antibacterial property of pulses to water.

6. Treatment of different water sources such as industrial wastewater,

agricultural wastewater and household wastewater.

7. Further studies on other methods of extracting globulins from

pulses may also be done.

8. A more in-depth and accurate testing of water turbidity can be

made.

9. Future researches can also focus on microbial analysis of presend

microbes and possible pathogens.

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 APPENDIX A. LETTER

Comparative study: Varieties of Pulses as Water Coagulant

Physical Characteristics: Survey

Madam/Sir,
We, the Benguet State University – Senior High School students are
conducting a research on water coagulation. This research is intended to create a
water coagulant to treat sewage water using different varieties of beans. We then
humbly ask for your participation in filling out this questionnaire.
Thank you for your utmost support.

Sincerely yours,

Kawi, Dusty A. Bas-ilan, Aubrey T. Garcia, Zaña Lei C.

Researcher Researcher Researcher

Bomogao, Islay Erika A. Chua, Maria Francesca Nicole R.

Researcher Researcher
Noted by:
Vanderlee Batalier Alicia Balongyad
Instructor SHS coordinator

Using the scale given below, rate each sample by writing the
corresponding number for describing the change done by each treatment
compared to the control (untreated water).
Scale: 5 VERY DESIRABLE 2 UNDESIRABLE
4 DESIRABLE 1 VERY UNDESIRABLE
3 NEUTRAL
Control Sample A Sample B Sample C
Treatment 1
Color
Odor
Treatment 2
Color
Odor
Treatment 3
Color
Odor
 APPENDICES B. TABLES

Table 1: Weighted mean for Salt Extraction in testing Color

5 4 3 2 1
Mung Beans 0 0 26 510 88
Cardis Beans 0 0 26 918 44
Navy Beans 0 0 0 1224 33
Weighted Σ 0 0 4 17.33 5
Weighted μ 1.76

Table 2: Weighted mean for Salt Extraction in testing Odor

5 4 3 2 1
Mung Beans 0 0 39 816 44
Cardis Beans 0 0 0 48 1111
Navy Beans 0 832 412 36 0
Weighted Σ 0 10.67 7 10 5
Weighted μ 2.18

Table 3: Weighted mean for Direct method in testing Color

5 4 3 2 1
Mung Beans 0 0 26 1020 33
Cardis Beans 0 832 515 36 0
Navy Beans 0 0 26 918 33
Weighted Σ 0 10.67 9 14.67 2
Weighted μ 2.42

Table 4: Weighted mean for Direct method in testing Odor

 5 4 3 2 1
Mung Beans 0 0 721 510 33
Cardis Beans 0 936 412 24 0
Navy Beans 0 0 13 1020 44
Weighted Σ 0 12 12 11.33 2.33
Weighted μ 2.51
Table 5: Weighted mean for Activation method in testing Color

5 4 3 2 1
Mung Beans 15 28 1236 0 0
Cardis Beans 0 0 1030 48 11
Navy Beans 0 28 824 36 33
Weighted Σ 1.67 5.33 30 4.67 1.33
Weighted μ 2.87

Table 6: Weighted mean for Activation method in testing Odor

5 4 3 2 1
Mung Beans 0 0 26 510 88
Cardis Beans 0 0 0 612 99
Navy Beans 0 0 39 714 55
Weighted Σ 0 0 5 12 7.33
Weighted μ 1.62

Table 7: Weighted mean for Mung Beans in testing Color

5 4 3 2 1
Salt Extraction 0 0 26 510 88
Direct Method 0 0 26 1020 33
Activation 15 28 1236 0 0
Method
Weighted Σ 1.67 2.67 16 10 3.67
Weighted μ 2.27

Table 8: Weighted mean for Mung Beans in testing Odor

5 4 3 2 1
 Salt Extraction 0 0 39 816 44
Direct Method 0 0 721 510 33
Activation 0 0 26 510 88
Method
Weighted Σ 0 0 12 12 5
Weighted μ 1.93

Table 9: Weighted mean for Cardis Beans in testing Color

5 4 3 2 1
Salt Extraction 0 0 26 918 44
Direct Method 0 832 515 36 0
Activation 0 0 1030 48 11
Method
Weighted Σ 0 10.67 17 10.67 1.67
Weighted μ 2.67

Table 10: Weighted mean for Cardis Beans in testing Odor

5 4 3 2 1
Salt Extraction 0 0 0 48 1111
Direct Method 0 936 412 24 0
Activation 0 0 0 612 99
Method
Weighted Σ 0 12 4 8 6.67
Weighted μ 2.04

Table 11: Weighted mean for Navy Beans in testing Color

5 4 3 2 1
Salt Extraction 0 0 0 1224 33
Direct Method 0 0 26 918 33
Activation 0 28 824 36 33
Method
Weighted Σ 0 2.67 10 16 3
Weighted μ 2.11
Table 12: Weighted mean for Navy Beans in testing Odor

5 4 3 2 1
Salt Extraction 0 832 412 36 0
Direct Method 0 0 13 1020 44
Activation 0 0 39 714 55
Method
Weighted Σ 0 10.67 8 13.33 3

Weighted μ 2.33

APPENDICES C. FIGURES
Figure 1: Microbial analysis of Figure 3: Microbial analysis of
Mung Beans. Direct Application R2 Mung Beans. Direct Application R1

Figure 2: Microbial analysis of Figure 4: Microbial analysis of

Mung Beans. Direct Application R2 Mung Beans. Direct Application R3
Figure 5: Microbial analysis of Figure 7: Microbial analysis of
Mung Beans. Activation R2 Mung Beans. Salt Extraction R1

Figure 6: Microbial analysis of Figure 8: Microbial analysis of

Mung Beans. Activation R3 Mung Beans. Salt Extraction R2.
Figure 9: Microbial analysis of Figure 11: Microbial analysis of
Mung Beans. Salt Extraction R3. Cardis Beans. Direct Application R2

Figure 10: Microbial analysis of Figure 12: Microbial analysis of

Cardis Beans. Direct Application R1 Cardis Beans. Direct Application R3
Figure 13: Microbial analysis of Figure 15: Microbial analysis of
Cardis Beans. Activation R1. Cardis Beans. Activation R3.

Figure 14: Microbial analysis of Figure 16: Microbial analysis of

Cardis Beans. Activation R2. Cardis Beans. Salt Extraction R1.
Figure 17: Microbial analysis of Figure 19: Microbial analysis of
Cardis Beans. Salt Extraction R2. Navy Beans. Direct Application R1.

Figure 18: Microbial analysis of Figure 20: Microbial analysis of

Cardis Beans. Salt Extraction R3. Navy Beans. Direct Application R2.
Figure 21: Microbial analysis of Figure 23: Microbial analysis of
Navy Beans. Direct Application R3. Navy Beans. Activation R2.

Figure 22: Microbial analysis of Figure 24: Microbial analysis of

Navy Beans. Activation R1. Navy Beans. Activation R2.
Figure 25: Microbial analysis of
Navy Beans. Salt Extraction R1. Figure 26: Laboratory procedure for
gram staining.

Figure 27: Microbial analysis of Figure 27: Source of Water Samples.

Navy Beans. Salt Extraction R3.
Figure 28: Gram staining procedure Figure 30: Powdered Mung Beans.

Figure 29: Gram staining apparatus. Figure 31: Powdered White Beans.
Figure 32: Powdered Navy Beans

Figure 33: Water sample collection.