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New Era University

College of Arts and Sciences

Biology Department

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS IN

BIOLOGY RESEARCH 2 (THESIS WRITING) UNDER DR. MYLENE O. ANDAYA

The Utilization of powdered Janitor fish bones from

Marikina river as soil acidity reducing agent

Researchers:

de Asis, Dorothy Rica G.

Bunda, Wising John B.

Juanir, Timmy Art S.

Napay, Arman P.
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TABLE OF CONTENTS
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ABSTRACT
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CHAPTER I

BACKGROUND OF THE STUDY

The Janitor fish (Pterygoplichthys pardalis) is one of the many species of fish that

has been considered by the Bureau of Fisheries and Aquatic Resources (BFAR) as a

pest in the Philippines due to its uncontrollable and ballooning population. Sometimes

used as an ornamental fish, it eats not only the algae in aquariums but also those fishes

that are with it. The Janitor fish already occupying much of the freshwater, especially in

the Marikina River, Pasig River, and Laguna de Bay, has raised awareness to the

government and the community as it eats and outnumbers the carps, tilapia and other

fishes that are present in those bodies of water. Many innovative ways and researches

have been conducted to lessen their population; one of this is the extraction of biodiesel

by a Marikina High School student in 2006.

Many problems also caused by Janitor fish are soil erosion in riverbanks,

competition in the source of food by our local fishes, resulting in lack of food source for

the other fishes and thus, not maintaining ecological balance.

In this research study, the researchers aim to create a solution in the growing

number of janitor fish, not only regulating their population from bodies of water but most

importantly, to innovate and make a new product out of the janitor fish through its

chemical components.

A study from the University of the Philippines Los Baños in 2013, proved that the

bones of the Janitor fish contain calcium phosphate and calcium carbonate. These

substances could be of great use to the growing problem of our agricultural sector in the
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acidity of soil in the Philippines due to the use of commercial fertilizers and pesticides,

and acid rain.

STATEMENT OF THE PROBLEM

1. Are the Janitor fish bones effective as a soil acidity reducing agent?

2. Is there any significant difference among the effect of different formulations in

Janitor fish bones in reducing the soil acidity?

SIGNIFICANCE OF THE STUDY

The Philippines is known to be one of the countries that is relied upon on the

agricultural sector. But because of the continuous use of pesticides and commercial

fertilizers and pollution the soil becomes acidic. In addition to that, acid rain also

contributes to the acidity of the soil. The Janitor fish bones, if proven to be effective as a

soil acidity reducing agent will be a great help to maintain the normal pH of the soil.

SCOPE AND LIMITATIONS

This study only covers the use of the Janitor fish (Pterygoplichthys pardalis)

taken from the Marikina river in reducing soil acidity.

HYPOTHESIS

1. The Janitor fish bones will not be effective as a soil acidity-reducing agent.

2. There is no significant difference in the changes made by the formulations.

DEFINITION OF TERMS

1. Acid – a compound usually having a sour taste and capable of neutralizing

alkalis and reddening blue litmus paper, containing hydrogen that can be

replaced by a metal or an electropositive group to form a salt, or containing

an atom that can accept a pair of electrons from a base. Acids are proton

donors that yield hydronium ions in water solution, or electron-pair donors or

bases.

2. Acid rain – precipitation, as rain, snow, or sleet, containing relatively high

concentrations of acid-forming chemicals, as the pollutants from coal smoke,

chemical manufacturing, and smelting, that have been released into the

atmosphere and combined with water vapor: harmful to the environment

3. Base – a compound that reacts with acid to form a salt, ammonia, calcium

hydroxide, or certain nitrogen-containing organic compounds.

4. Calcium Phosphate – (Ca 3O8P2) any of several phosphates of calcium

occurring naturally in some rocks and animal bones, used as a fertilizer and

food additive as well as in baking powder and dentifrices.

5. Calcium Carbonate – a white, crystalline, water-insoluble, tasteless powder,

CaCo3, occurring in nature in various forms, as calcite, chalk, and limestone:

used chiefly in dentifrices and polishes and in the manufacture of lime and

cement.

6. Fertilizers – any substance used to fertilize the soil, especially a commercial

or chemical manure.

7. Janitor Fish – a member of the Genus Pterygoplichthys that preys on the local

fishes in some areas of Metro Manila

8. Pest – an insect or other small animal that harms or destroys garden plants,

trees, etc.

9. pH (power of Hydrogen) – the symbol for the logarithm of the reciprocal of

hydrogen ion concentration in gram atoms per liter, used to express the

acidity or alkalinity of a solution on a scale of 0 to 14, where less than 7

represents acidity, 7 neutrality, and more than 7 alkalinity.

10. Predator – any organism that exists by preying upon other organisms

11. Soil – the portion of the earth’s surface consisting of disintegrated rock and

humus.

12. Teleosts – belonging or pertaining to the Teleostei, a group of bony fishes

including most living species

CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

Janitor fish
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Scientific classification

Kingdom: Animalia

Phylum: Chordata
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Class: Actinopterygii

Order: Siluriformes

Family: Loricariidae

Subfamily: Hypostominae

Genus: Pterygoplichthys

The Janitor fish is one of the armored catfishes that belong to the genus

Pterygoplichthys. This species is native to Brazil, Peru and other South American

nations. It was imported to the Philippines as a helpful fish to help clean algae in

aquariums. In time, however, these creatures quickly multiplied in number, not in their

supposed habitat under the supervision of aquaculturists, but instead have plagues

freshwater lakes and rivers in vast proportions. Soon enough, the population has

skyrocketed and threatened the livelihood of local fishermen in areas where the fish

have proliferated, as this creature is also a predator of catfish, tilapia, milkfish, and their

eggs. In addition to that, digestive wastes produced by janitor fish results to water

turbidity, and males of this species burrow into mud banks and cause soil to erode.

Researchers from Pililla National High School (2011) have conducted a

comprehensive study regarding this matter and found out that Pterygoplichthys pardalis

is found in the Marikina River and Lake Paitan in Cuyapo, Nueva Ecija, and the species

Pterygoplichthys disjunctivus is found in the Laguna de Bay. Also, they have discovered

that the Pterygoplichthys from the Laguna de bay have successfully invaded the

Agusan Marsh in Agusan del Sur, which results to a competition for food between the
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janitor fish and other local fishes. The said researchers stated that this current state of

janitor fish overpopulation in local waters should come as a wakeup call for a more

drastic method extinguishing these creatures to be devised, as the Agusan del Sur is

Asia’s largest marsh land and have yet to be fully explored by scientists.

Local government units have initiated projects here and there, at times even

encouraging citizens to participate, in order to help reduce, if not completely eradicate

the threat that janitor fish poses to the delicate ecological balance of local freshwater

lakes and rivers.

Janitor fish bone as a source of calcium-rich compounds

A number of studies have investigated and proven the existence of calcium-

containing materials in certain animals such as hydroxyapatite, a type of a calcium

phosphate compound. Venkatesan and Kim (2010, as cited by Piencenaves in 2013)

successfully obtained HAp from tuna bones, by thermal calcination method, stating that

“the production of HAp from natural sources is inexpensive and uncomplicated.”

In the extension of Amurao’s research (2005), he studied the use of janitor fish

bones as carbon filters. It involved extracting calcium and activated carbon from the

bones, which, as he concluded has the potentiality to be used in water purifiers.

Another project undertaken by Agusan National High School’s John Michael D.

Piencenaves (2013) investigated the production of calcium-based bioceramic powder

from P. disjunctivus. Janitor fish bones calcined at 1000°C for four hours resulted in the
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formation of the calcium phosphate bioceramic through Fourier Transform Infra-Red

Spectroscopy.

Ideal Soil pH

To measure the acidity or alkalinity of soil, soil pH is used as the logarithmic

scale where 0 represents strongest acid, 14 represents strongest alkali or base, and 7

represents neutrality. Slightly acid is considered to be 5.2 to 6.0, moderately acid 5.6 to

6.0, strongly acid 5.1 to 5.5, very strongly acid 4.5 to 5.0, and extremely acid below 4.5.

(Halcomb, 2009).

Brady, 1990, suggested that 6.0-6.8 pH range is “the most ideal soil pH for most

crops because it coincides with optimum solubility of the most important plant nutrients.”

Contribution of commercial fertilizers to soil acidity

Soil acidity is a measure of the concentration of H+ ions in the soil solution. As

discussed in an article published in AG Professionals (2013), the greatest injury to crop

growth from low pH soils comes not from the H+ ions, but from the release of aluminum

into the soil solution at low pH levels. As the pH decreases below 5.5, the availability of

aluminum and manganese increase and may reach a point of toxicity to the plant.

Excess Al in the soil solution interferes with root growth and function, as well as

restricting plant uptake of certain nutrients, namely, Calcium and Magnesium. Liming

acid soils reduces the activity of Aluminum and Manganese.

Of all the major fertilizer nutrients, nitrogen is the main nutrient affecting soil pH,

and soils can become more acidic or more alkaline depending on the type of nitrogen

fertilizer used (Mosaic Fertilizer Technology Research Centre, 2013). The primary

reason for soils becoming more acidic over time is the use of nitrogen (N) fertilizers

containing ammonium-N, including mono and diammonium phosphates, 11-52-0 and

18-46-0. As the ammonium-N in fertilizers nitrifies, acidity is released (Kansas State

University, 2013).

Factors, including soil type and organic matter, affect the amount of materials

needed to change soil pH (Crouse & Denny, 2015). Fertilizers, rain, organic matter and

soil microorganisms affect soil pH. Agricultural limestone normally is used to increase

the soil’s pH, making it alkali, while sulfur is used to lower (acidify) it.

Environmental factors affecting soil acidity

It should be remembered that soil acidification is a natural process in high rainfall

environments and is not just a man-made condition. Intensive agriculture can speed up

acidification due to removal of produce from the land, enhanced leaching of bicarbonate

and soluble organic matter in soil percolates, and accumulation of organic matter in soil

over time (MFTRC, 2013).

According to Halcomb, 2009, some of the factors which affect soil acidity are: (1)

Application of acid forming fertilizers which produce large amounts of H ions, such as

urea, ammonium and sulfate fertilizers, (2) Removal of base forming ions by crops. (3)
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Removal of base forming ions by leaching and erosion and (4) Decomposition of

organic matter which produces carbonic acids.

Soil Degradation

Soil plays a very important role in agriculture for this is where the plant grows and

gets its nutrients; it is also used for storage of water and detoxifies pollutants and also

regulates the Earth’s temperature (Blum, 2007). The Philippines have a significant

percentage of eroded lands in Luzon, Visayas and Mindanao. Soil degradation includes

the loss of nutrients and organic matter, salinization, acidification, compaction, and

subsidence (Asio, et al, 2009). Soil degradation could affect plant growth and thus

decrease the quality of plants that are produced.

Nutrients affected by pH

Soil pH is vital for the growth of the plant. For this reason, the ideal soil pH must

be known for this greatly affects the nutrient uptake of plants. In Halcomb’s study in

2009 in the University of Tennessee, “Manganese availability increases as pH drops

with toxic symptoms occurring on some plants at pH 5.0 or below. In some soils in

Tennessee this is an important factor, as manganese is concentrated heavily enough to

exist as “pellets”. These look like shot gun pellets on the soil surface after a rain; the
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availability of phosphorus, one of the major plant nutrients, decreases as pH rises or

falls from about 6.3; the availability of copper and zinc is greatest between 5.0 and 6.0.

Calcium, magnesium, and molybdenum become less and less available as pH drops

from 7.0 to 4.0; the activity of soil microorganisms affecting organic matter

decomposition and nutrient availability (especially nitrogen) may be greatly reduced as

soils become more and more acid.” It is therefore recommended that the soil pH be in

between 5.5 and 6.5 for most plants.

Ideal soil pH should be determined for too much acidity or alkalinity could cause

insolubility of nutrients and unavailability to the plants. Various macronutrients and

micronutrients cannot support the growth of plants. If the soil’s pH is too low, nitrogen,

phosphorus, calcium, magnesium and potassium may not be available to plants. In

addition to that, aluminum, iron, and manganese toxicity will also increase but in soils

with proper pH, sulfur, calcium, magnesium, and molybdate are non-toxic and can be

available for plant uptake. (Halcomb, 2009).

Furthermore, there are plants that prefer acidic soil like ericaceous plants to

which heath, heather, azalea, rhododendron, mountain laurel (Kalmia), leucothoe,

huckleberry, bearberry, pernettya, andromeda, blueberry, etc. belongs. They do well in

soils with pH of 4.0-6.0, and with preferred range of 5.0-5.5. Lilac and mockorange

thrive in alkaline soils with somewhere in 7.0-7.5 pH (Halcomb, 2009).

Microbial and plant growth also depends on ion solubility which is affected by the

soil pH. At lower pH, heavy metals are more soluble (Brady, 1990).
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CHAPTER III

METHODOLOGY AND RESEARCH MATERIALS

This chapter includes the experimental approach will be applied in the study to

prove the effectivity of Pterygoplichthys pardalis (Janitor fish) bones as soil buffer and

as soil remediates.

1. Research Design
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The researchers will use experimental method of research. The trials on the

application of the P. pardalis’ bone powder on the agricultural soil with low pH (6 and

below) is an experimental type of research.

2. Research Materials

2.1. P. pardalis

2.2. Preparation of P. pardalis’ bone powder

2.2.1. P. pardalis bones

2.2.2. Distilled water

2.2.3. Laboratory oven

2.2.4. Mortar and pestle

2.2.5. Analytical balance

2.3. P. pardalis bone powder application on agricultural soil

2.3.1. Sample agricultural soil

2.3.2. P. pardalis bone powder

2.3.3. Beaker

2.3.4

2.3.5. pH meter
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2.3.6. Distilled water

2.3.7. Spatula

3. Research Setting

3.1. P. pardalis bone powder as acidity-reducing agent

The experimental trials will be conducted in a sample acidic soil (pH 5.5).

Changes in pH will be monitored every 3-4 days to see if the desired results are

accomplished.

4. Research procedure

4.1. Preparation of P. pardalis bone powder

The bones of P. pardalis will be removed and cleaned with tap water. The fish

bones will be scraped and washed with distilled water until no meat can be found

sticking to them. Then, the fish bones will be oven dried for 5 hours at 150°C to

eliminate moisture and for the bones to be brittle. By using mortar and pestle, the dried

samples will be crushed and powdered. The dried samples collected then will be

weighed using analytical balance.

4.2 Testing for the Reduction of Soil Acidity

There will be four setups and fours trials in this experiment. The first set will have

three pots, each with 150g of soil and 100g of powdered Janitor fish bones; the second

set will also have three pots, but this time, each will have 150g of soil and 75g of

powdered Janitor fish; the third set will also have three pots, each with 150g of soil and
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50g of Janitor fish bones. The set-up will be monitored every three to four days to see if

there is any difference from the previous soil pH. In testing for the soil pH, first, a

sample of the mixture is put on a beaker and mixed with 10mL of distilled water, enough

to test it using the pH meter. The pH meter is then put on the mixture and the pH is

observed and recorded.

CHAPTER IV

DATA ANALYSIS AND INTERPRETATION

Table 1.1 The Formulations Used for the Study

Soil Bones

Set 1 A, B, C 150g 100g

Set 2 A, B, C 150g 75g

Set 3 A, B, C 150g 50g

Control 150g None

In this table, we can see the different formulations used in the different setups.

The first set-ups (Set 1 A,B,C) contained 150g of soil and 100g of powdered Janitor fish
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bones. The second setups (Set 2 A,B,C) contained 150g of soil and 75g of powdered

Janitor fish bones, The third setups (Set 3 A,B,C) contained 160g of soil and 50g of

powdered Janitor fish. The control only contained 150g of soil with no powdered Janitor

fish bones.

Table 2.1 pH observations

Observations

Sets Trial 1 Trial 2 Trial 3 Trial 4

A. 100g 7.3 7.3 7.6 7.6

B. 100g 7.3 7.4 7.4 7.4

C. 100g 7.3 7.4 7.6 7.6

A. 75g 6.9 7.2 7.2 7.4

B. 75g 7.1 7.3 7.2 7.3

C. 75g 6.9 7.2 7.1 7.2

A. 50g 6.9 6.8 7.0 7.0

B. 50g 6.5 7.0 7.0 7.0

C. 50g 6.6 6.9 7.0 7.0

Control 5.5

It can be observed based on the data above that the biggest change in pH was in

the 100g setup with 7.6 as the highest recorded pH when the powdered bones were

applied. The least change was in the 50g setup, with 6.5 as the lowest pH, from 5.5

which is the pH of the soil. In most of the setups, there was a constant increase in pH or

reduction of the soil acidity. In the 50g setup, it met the ideal soil pH which should be

around 6.0-6.8.

Table 2.2 Average pH based on the data collected

Set Average Difference from control

A.100g 7.45 1.95

B.100g 7.38 1.88

C.100g 7.48 1.98

A.75g 7.18 1.68

B.75g 7.22 1.72

C.75g 7.1 1.6

A.50g 6.92 1.42

B.50g 6.88 1.38

C.50g 6.88 1.38

Set A, 100g had the highest change in pH when compared to the control with

1.95. The least pH change was in Sets B and C, 50g with 1.38.

CHAPTER V.
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Summary
The utilization of different materials from our environment always plays an

important role in maintaining the balance in our ecology. In this study, the utilization of

the janitor fish bones to be one of the ways to reduce their population in the Marikina

river plays a significant role, as it is used to reduce soil acidity. Through this, the

researchers not only contribute an idea to help the government control the janitor fish

population, but also help the agricultural sector in reducing the acidity of their soil.

The preparation of the janitor fish bones may be a cumbersome task especially

when separating the bones from the meat, but the desirable results await. Based on the

data observed and provided by the researchers, the bones were instrumental in

reducing the acidity of the soil. There has been a significant difference based on the

average pH that resulted from the application of the bones on the acidic soil.

Conclusion
In this study, the researchers can conclude that the powdered Janitor fish bones

were effective in reducing the acidity of the soil. There was also a significant difference

among the different formulations in the different setups.

Recommendation
It is recommended that in 150g of soil, 50g of powdered Janitor fish bones must

be used to achieve the most desirable soil pH (6.0-6.8). It is also recommended for

aquaculturists to suggest this idea to farmers as their alternative to liming to get the

ideal soil pH.

BIBLIOGRAPHY
Asio, V., et al. (2009). A Review of Soil Degradation In The Philippines. Annals of

Tropical Research. 31(2), 69-94. Leyte, Philippines: Visayas State University.

Brady, N.C. (1990). The Nature and Properties of Soils. Macmillan Publishing

Company, New York, New York.

Gutierrez, R. and Barraquio, W. (2010). Acid -Tolerant Rhizobia of Phaseolus vulgaris

L. from the Intensively Cropped Soils of La Trinidad, Benguet, Philippines.

Philippine Journal of Science. 139 (1), 79-90. ISSN 0031 - 7683.

Halcomb, M., et al. (2009). Soil pH Explained. University of Tennessee Extension.

Iligan, L., et al. (2014). Soil Fertility Evaluation For Rice Production In Catanduanes

Province, Philippines. International Journal Of Scientific & Technology Research.

3(12). ISSN 2277-8616.

McCauley, A., et al (2003). Soil pH and Organic Matter

U.S.D.A. (2014). Soil Physical and Chemical Properties. Retrieved from

[Link]
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APPENDICES