BARTOLOME SANGALANG NATIONAL HIGH SCHOOL

BARAWID ST., SAINT JOHN DIST., GUIMBA, NUEVA ECIJA

Potential of Nile Tilapia (Oreochromis niloticus) Scale as an Adsorption of Heavy metals

Lead (Pb) in Soil Environment

A Research Proposal Presented to the Panel of Oral Examiners

In Partial Fulfilment of the Requirements in Practical Research II

Dedan Kaye Suplito

Diana E. Belarminno
Paul Vincent Sabaddo
Proponents

Jobelle Ann B. Nacino

Research Adviser

Address: Barawid St., Saint John District, Guimba, Nueva Ecija

Logo 300809
Tel. No.:(044) 611 0617
Email Address:bartolomesangalang102179@gmail.com SCHOOL ID
0.76in
RESEARCH PLAN/ PROJECT SUMMARY

A. Rationale

Soil environment became contaminated by the accumulation of heavy metals and

metalloids due to emissions from rapidly expanding industrial areas, mining tailings, disposal of

high metal waste, leaded gasoline and paint, fertilizer application, animal manure, sewage

sludge, pesticides, wastewater irrigation, coal combustion residues, petrochemical spillage and

atmospheric deposition. Heavy metals form an ill-defined category of inorganic chemical risk

like lead (Pb), chromium (Cr), arsenic (As), zinc (Zn), cadmium (Cd), copper (Cu), mercury

(Hg), and nickel (Ni) are the most frequently identified at polluted sites ( Wuana,2011) .

Even though these metals have no biological role, their toxic effects exist in some form

that is harmful to the human body and its proper functioning. They can act as a phantom part of

the body at times, and they can even interfere with metabolic processes. So on, it became an

extremely risk for both animals and humans that can cause several threats to one’s health.

Heavy metals such as lead (Pb) tend to stay in the upper soil layers and can be

metabolized into plant tissues , especially because it heavy metals have a strong attraction to

certain organic materials. Lead (Pb ) is consumed by plants from their roots because heavy

metals attacks leaves and the root’s outer layer. Once infected plants are ingested, a part of Pb

that enters the body is absorbed into the bloodstream and accumulates in body tissues and other

organs can cause death.

 As stated by Ambika et al. (2016) Lead (Pb) is one of the ubiquitously distributed most

abundant toxic elements in the soil. It exerts adverse effect on morphology, growth and

photosynthetic processes of plants. Lead is known to inhibit seed germination of Spartiana

alterniflora, Pinus helipensis . Inhibition of germination may result from the interference of lead

with important enzymes. In maize (Zea mays) reduction in germination percentage; suppressed

growth; reduced plant biomass; decrease in plant protein content has been noticed. Whereas in

Portia tree (Thespesia populnea) Reduction in a number of leaves and leaf area; reduced plant

height; decrease in plant biomass and in Oat (Avena sativa) Inhibition of enzyme activity which

affected CO2 fixation.

Nowadays, soil pollution spread in our environment due to heavy metals contamination

consequently the agricultural field, industrial field, and other livelihood resources that demand

soil are highly at risk. Environmentalists developed a low-cost way of solving this crisis which is

soil remediation. Soil remediation is a way to purify and revitalize the soil. It is a process to

remove contaminants to protect both the health of the population and the environment. So on, the

process aims to restore the soil to its natural, pollution-free state.[ CITATION Sav13 \l 1033 ]

According to Abdbulla (2014) experimental studies on the removal by adsorption

technique of lead (Pb) pollution from contaminated water using local fish scales, i.e. Tilapia and

Merah (Oreochromos and Lutjanus). Adsorbent fish scales balanced almost 100% of the Lead

uptake across the spectrum of concentrations and pH tested. The efficiency of fish scales as an

adsorbent is stronger compared to commercially available activated carbon.

 Nile tilapia (Oreochromos niloticus) often seen in the wet market particularly in those

areas far from the sea. It is a freshwater fish that has been cultured into farming for marketing.

Its scales has been a problem waste to markets because of improper waste disposal not knowing

it has a benefit to adsorption of heavy metals (Pb) in the contaminated water.

This study will use fish scales as an alternative adsorbent for reducing heavy metals in a

soil sample which will let them formulate a solution to the widely known problem of soil

contamination. Okra (Abelmoschus esculentus) will be used to test the amount of heavy metal

present in the soil sample

Okra (Abelmoschus esculentus) is a flowering plant whose seed pods are edible. It's

grown in warm, tropical climates like those found in Africa and South Asia. Biologically

classified as a fruit and it is generally utilized like a vegetable in cooking. It also contains many

health benefits to humans but most of the crops nowadays were contaminated with heavy metals

that are highly risk for human life.

Furthermore, this study will be conducted to determine the potential of Nile tilapia

(Oreochromos niloticus) as an adsorbent of heavy metals in the soil environment

B. Hypotheses / Research Questions / Goals / Expected Outcomes

B1. Hypothesis of the Study

 1. There is no potential of Nile tilapia (Oreochromos niloticus) scale in the adsorption of

lead (Pb)

2. There is no significance of Nile tilapia (Oreochromos niloticus) scale to the growth of

variables.

3. There is no other potential of Nile tilapia (Oreochromos niloticus) scale in a soil

environment.

B2. Research Questions

1. Does the Nile tilapia (Oreochromis niloticus) scale adsorb the lead (Pb)?

2. Is there any significance of the Nile tilapia (Oreochromis niloticus) scale to the growth of

variables?

3. Is there any potential that can the Nile tilapia (Oreochromis niloticus) scale for soil

remediation?

B3. Agricultural Goals

This study aims to remove heavy metals in soil from the scale of Nile Tilapia

(Oreochromis niloticus). This will be helpful to farmers to provide a low-cost for soil

remediation.

B4. Expected Outcomes

 The Nile tilapia (O. niloticus) scale will expected to adsorb the heavy metals in the

environment. At the end of this study, it will provide a new-found knowledge and a new low-

cost product for removing soil pollutant. Also, the success of the study will benefits the

agricultural field to have a healthy environment for their crops.

METHODOLOGY

In this section of the study, comprising the step by step methods and discusses the proper

analysis used for evidence of data.

Collection of Materials

Preparation of Adsorbent, Soil and other Variables

 Analysis of Chemical Properties of Soil

Planting of Abelmoschus esculentus seedlings on soil

with Heavy metals and Nile tilapia(O. niloticus) scale

Observation of Abelmoschus esculentus plant growth

Collection of Abelmoschus esculentus plant

Analysis for soil with Heavy metals content

Analysis for Abelmoschus esculentus plant

Statistical Analysis

Figure 1. Methodological Framework of the Study

 The researchers will use ten scientific methods in this study. These methods will be

discussed in the following:

Collection of Materials

The source of the scale of Nile tilapia (Oreochromos niloticus) that will be used in the

study will be obtained by collecting from the public wet market of Guimba, Nueva Ecija. The

soil will be acquired at the rice field of Maturanoc,Guimba, Nueva Ecija.

Preparation of Materials

Clay is a wet soil and only dries in summer, therefore it will be air-dried for one week to

assure that it is dry enough to be crushed. O. niloticus scale will be used as the main substance to

lessen the amount of heavy metal. O. niloticus was wash with water to remove the excess

contents and boiled for cleansing. Scales will be store in a room temperature and air-dried. When

the scales were fully dried, it will be crushed and pulverized. The seedlings of A. esculentus will

be soaked in the water for one day to germinate faster (I. E. Agbozu, 2014).

Preparation of Heavy Metals

After all the analysis in soil and O. niloticus scale, the solution will be prepared. The

different levels of heavy metal and will be added in small pots with and without the O. niloticus

scale (Santiago et al., 2020).

Analysis of Chemical Properties of Soil

pH Level

It will analyze the nutrient state of the air-dried and sieved soil. Twelve (12) solutions of

Chlorophenol Red (CPR) will be applied to the sample for five minutes. The pH level of the

treated soil will be defined by comparing the color of the solution to the color of the indicator pH

map. However, if the outcome is greater than, the predicted result was between these two, "the

greater than to and the less than or equal to 5" it must be measured using Bromothymol blue

(BTB) and if it is less and equal, it will be used (BCG) again in the testing (Kimberly Frye,et al.,

2020).

Nitrogen-Phosphorus-Potassium (NPK) Analysis

Three test tubes will be prepared for Nitrogen-Phosphorus-Potassium (NPK) Analysis.

Test tubes will partially filled with soil samples added with heavy metal solution were settled

for 30 minutes and results showed the availability of Nitrogen in the soil sample. Another test

tube filled with soil sample will be added with 24 drops of Phosphorus (P) solution and 4

drops of Phosphorus 2 (P2), results will show the availability of Phosphorus in the soil

sample. Other test tubes filled with soil sample will be added with 24 drops of Potassium (K)

solution and 8 drops of Potassium 2 (K 2) , the results will show the availability of Potassium
 on the soil sample. The appearance of the solution will be observed to determine the

availability of Nitrogen-Phosphorus-Potassium (NPK) on the soil sample. (Vigyanashram,

2020)

Planting of Abelmoschus esculentus seedlings on soil with Heavy metals and Nile tilapia (O.

niloticus) scale

The air-dried and sieved soil will measure 600 grams and then added to 6 grams O.

niloticus scale. Thirty (30) mL of distilled water will be poured into the plant with 0 ppm and

100 ppm of heavy metals until it sinks within the soil. In each pot, 250 mL of distilled water was

added for 15 minutes of absorption. Subsequently, 3 seedlings of A.esculents will be planted in

each pot

( Santiago et al., 2020).

Observation of A.esculentus plant growth

The A.esculentus will be observed in two concentrations with different time of exposure

to observe the changes in the growth of A.esculentus. The soil with 0ppm of heavy metal will be

planted with A.esculentus and observes for 10 days while the soil with 100ppm will be observes

for 30 days ( Santiago et al., 2020).

Collection of A.esculentus plant

The A.esculentus will be harvest and wash with distilled water to cleanse it. The length of

the roots and stem will be measure as well as the number of leaves and weight of plants. The

sample will subjected to an air drying procedure at 70C (Santiago et al., 2020).

Analysis for Soil with Heavy Metals Content

The Nile tilapia (O.niloticus) scale will be exposed to heavy metals for 15 days and 40

days. The soil with heavy metals and adsorbent (O. niloticus ) scale content will be examined

after the days of exposure. By the Atomic Emission Spectrometer (MP-AES), the effects of Nile

tilapia (O. niluticus) as an adsorbent will determined ( Santiago et al., 2020).

Analysis for A. esculentus plant

As the plant A. esculentus was exposed to heavy metals and Nile tilapia(O. niloticus)

scale, it will analyze by Atomic absorption spectroscopy (AAS) to determine the heavy metal

content that absorb in the plant and if the Nile tilapia (O. niloticus) also has significant to it

(Aquisman et al., 2019).

Statistical Analysis

The data from the correlational analysis will be need to see the effect of Nile Tilapia ( O.

niloticus) scales to the soil and to the ( A. esculentus)

 BACKGROUND OF THE STUDY

INTRODUCTION

Soil environment became contaminated by the accumulation of heavy metals and

metalloids due to emissions from rapidly expanding industrial areas, mining tailings, disposal of

high metal waste, leaded gasoline and paint, fertilizer application, animal manure, sewage

sludge, pesticides, wastewater irrigation, coal combustion residues, petrochemical spillage and

atmospheric deposition. Heavy metals form an ill-defined category of inorganic chemical risk

like lead (Pb), chromium (Cr), arsenic (As), zinc (Zn), cadmium (Cd), copper (Cu), mercury

(Hg), and nickel (Ni) are the most frequently identified at polluted sites ( Wuana,2011) .

Even though these metals have no biological role, their toxic effects exist in some form

that is harmful to the human body and its proper functioning. They can act as a phantom part of

the body at times, and they can even interfere with metabolic processes. So on, it became an

extremely risk for both animals and humans that can cause several threats to one’s health.

Heavy metals such as lead (Pb) tend to stay in the upper soil layers and can be

metabolized into plant tissues , especially because it heavy metals have a strong attraction to

certain organic materials. Lead (Pb ) is consumed by plants from their roots because heavy

metals attacks leaves and the root’s outer layer. Once infected plants are ingested, a part of Pb

that enters the body is absorbed into the bloodstream and accumulates in body tissues and other

organs can cause death.

 Nowadays, soil pollution spread in our environment due to heavy metals contamination

consequently the agricultural field, industrial field, and other livelihood resources that demand

soil are highly at risk. Environmentalists developed a low-cost way of solving this crisis which is

soil remediation. Soil remediation is a way to purify and revitalize the soil. It is a process to

remove contaminants to protect both the health of the population and the environment. So on, the

process aims to restore the soil to its natural pollution-free state[ CITATION Sav13 \l 1033 ].

According to Abdbulla (2014) experimental studies on the removal by adsorption

technique of lead (Pb) pollution from contaminated water using local fish scales, i.e. Tilapia and

Merah (Oreochromos and Lutjanus). Adsorbent fish scales balanced almost 100% of the lead

(Pb) uptake across the spectrum of concentrations and pH tested. The efficiency of fish scales as

an adsorbent is stronger compared to commercially available activated carbon.

Nile tilapia (Oreochromos niloticus) often seen in the wet market particularly in those

areas far from the sea. It is a freshwater fish that has been cultured into farming for marketing.

Its scales has been a problem waste to markets because of improper waste disposal not knowing

it has a benefit to adsorption of heavy metals in the contaminated water.

Furthermore, this study will be conducted to determine the potential of Nile tilapia

(Oreochromos niloticus) as an adsorbent of heavy metals in the soil environment

Specifically, this study aims to:

1. Determine whether the Nile Tilapia (Oreochromos niloticus) adsorbs the heavy metals;
 2. Ascertain the potential of the Nile Tilapia (Oreochromos niloticus) scale for the growth of Okra

(Abelmoschus esculentus)

3. Substantiate the Potential of Nile Tilapia (Oreochromos niloticus) scale for soil remediation.

METHODOLOGY

In this section of the study, comprising the step by step methods and discusses the proper

analysis used for evidence of data.

Collection of Materials

Preparation of Materials

Analysis of Chemical Properties of Soil

Preparation of Heavy Metals

Planting of Abelmoschus esculentus seedlings on soil

with Heavy metals and Nile tilapia(O. niloticus) scale
 Observation of Abelmoschus esculentus plant growth

Collection of Abelmoschus esculentus plant

Analysis for soil with Heavy metals content

Analysis for Abelmoschus esculentus plant

Statistical Analysis

Figure 1. Methodological Framework of the Study

 The researchers will use ten scientific methods in this study. These methods will be

discussed in the following:

Collection of Materials

The source of the scale of Nile tilapia (Oreochromos niloticus) that will be used in the

study will be obtained by collecting from the public wet market of Guimba, Nueva Ecija. The

soil will be acquired at the rice field of Maturanoc,Guimba, Nueva Ecija.

Preparation of Materials

Clay is a wet soil and only dries in summer, therefore it will be air-dried for one week to

assure that it is dry enough to be crushed. O. niloticus scale will be used as the main substance to

lessen the amount of heavy metal. O. niloticus was wash with water to remove the excess

contents and boiled for cleansing. Scales will be store in a room temperature and air-dried. When

the scales were fully dried, it will be crushed and pulverized. The seedlings of A. esculentus will

be soaked in the water for one day to germinate faster (I. E. Agbozu, 2014).

Preparation of Heavy Metals

After all the analysis in soil and O. niloticus scale, the solution will be prepared. The

different levels of heavy metal and will be added in small pots with and without the O. niloticus

scale (Santiago et al., 2020).

Analysis of Chemical Properties of Soil

pH Level

It will analyze the nutrient state of the air-dried and sieved soil. Twelve (12) solutions of

Chlorophenol Red (CPR) will be applied to the sample for five minutes. The pH level of the

treated soil will be defined by comparing the color of the solution to the color of the indicator pH

map. However, if the outcome is greater than, the predicted result was between these two, "the

greater than to and the less than or equal to 5" it must be measured using Bromothymol blue

(BTB) and if it is less and equal, it will be used (BCG) again in the testing (Kimberly Frye,et al.,

2020).

Nitrogen-Phosphorus-Potassium (NPK) Analysis

Three test tubes will be prepared for Nitrogen-Phosphorus-Potassium (NPK) Analysis.

Test tubes will partially filled with soil samples added with heavy metal solution were settled

for 30 minutes and results showed the availability of Nitrogen in the soil sample. Another test

tube filled with soil sample will be added with 24 drops of Phosphorus (P) solution and 4

drops of Phosphorus 2 (P2), results will show the availability of Phosphorus in the soil

sample. Other test tubes filled with soil sample will be added with 24 drops of Potassium (K)

solution and 8 drops of Potassium 2 (K 2) , the results will show the availability of Potassium
 on the soil sample. The appearance of the solution will be observed to determine the

availability of Nitrogen-Phosphorus-Potassium (NPK) on the soil sample (Vigyanashram,

2020).

Planting of Abelmoschus esculentus seedlings on soil with Heavy metals and Nile tilapia (O.

niloticus) scale

The air-dried and sieved soil will measure 600 grams and then added to 6 grams O.

niloticus scale. Thirty (30) mL of distilled water will be poured into the plant with 0 ppm and

100 ppm of heavy metals until it sinks within the soil. In each pot, 250 mL of distilled water was

added for 15 minutes of absorption. Subsequently, 3 seedlings of A.esculents will be planted in

each pot

( Santiago et al., 2020).

Observation of A.esculentus plant growth

The A.esculentus will be observed in two concentrations with different time of exposure

to observe the changes in the growth of A.esculentus. The soil with 0ppm of heavy metal will be

planted with A.esculentus and observes for 10 days while the soil with 100ppm will be observes

for 30 days ( Santiago et al., 2020).

Collection of A.esculentus plant

The A.esculentus will be harvest and wash with distilled water to cleanse it. The length of

the roots and stem will be measure as well as the number of leaves and weight of plants. The

sample will subjected to an air drying procedure at 70C (Santiago et al., 2020).

Analysis for Soil with Heavy Metals Content

The Nile tilapia (O.niloticus) scale will be exposed to heavy metals for 15 days and 40

days. The soil with heavy metals and adsorbent (O. niloticus ) scale content will be examined

after the days of exposure. By the Atomic Emission Spectrometer (MP-AES), the effects of Nile

tilapia (O. niluticus) as an adsorbent will determined ( Santiago et al., 2020).

Analysis for A. esculentus plant

As the plant A. esculentus was exposed to heavy metals and Nile tilapia(O. niloticus)

scale, it will analyze by Atomic absorption spectroscopy (AAS) to determine the heavy metal

content that absorb in the plant and if the Nile tilapia (O. niloticus) also has significant to it

(Aquisman et al., 2019).

Statistical Analysis

The data from the correlational analysis will be need to see the effect of Nile Tilapia ( O.

niloticus) scales to the soil and to the ( A. esculentus).

 REFERENCES

Agbozu, (2014) Academic Journals.Africa: African Journal of Environmental Science

Technology
Ambika et al. (2016). ResearchGate India: International Journal of Application or Innovation in
Engineering & Management
Aquisman et al. (2019) Scientific & Academic Publishing. Malaysia : Analytical Environmental
Chemistry Research Group
Ambreen, (2016) Journal of BioSource Management. Faisalabad Pakistan: University of Agriculture

Kimberly et al., (2020) Journal of Hazardous. Massachusetts: JoVE Science Education Database,
Environmental Science

Monichsa et al., (2014) National Center for Biotechnology Information. Toxicology

Rizzo, (2019). Healthline. Nutrition
Shahan et al., (2019) AIP. Pakistan: Institute of Environmental Engineering and Management Mehran
UET Jamshoro, Sindh, Pakistan

Wuana, (2011). Hndawi. Nigeria : Analytical Environmental Chemistry Research Group

Zayadi , (2013)ResearchGate. Malaysia : Faculty of Civil and Engineering, Universiti Tun Hussein Onn
Malaysiaing Tilapia