CHAPTER I

THE PROBLEM AND ITS SCOPE

RATIONALE OF THE STUDY

As our country continues to develop, the threat of inflation emerges as a prominent factor

affecting various aspects of our economy. Rice, being one of the most significant staple crops in

the Philippines, is vital to the country's agriculture. Disquietingly, data from the National

Economic and Development Authority (NEDA) shows a sharp rise in rice prices, rising from 1% in

January 2022 to a worrying 4.2% in July 2023. The re-entry of pests into crops, causing

consistently low yields, are serious problems which are commonly faced by farmers in the

Philippines. This perennial issue manifests itself as pests causing colossal losses to the farmers.

Agricultural workers face an ongoing threat from pests, which concludes the

importance of implementing countermeasures. Pesticides are emerging as an important

tool in farmers’ arsenal. These are designed to eliminate various pests from insects,

moths, and undesirable plants to protect crops from pests. The Golden Apple Snail,

scientifically known as Pomacea Canaliculata, poses a serious threat to rice cultivation

in the Philippines. These snails cause substantial harm by attacking and destroying young

plant stems and leaves of rice crops, consuming 7–24 rice seedlings daily.

Various control methods, including cultural, biological, and chemical approaches

have been employed, including cultural methods like handpicking, and using ducks

which are practical, but needs a ton of effort. Chemicals such as metaldehyde and

niclosamide are effective, however discouraged due to their long-term toxicity to humans
and the environment, along with affordability issues for many agricultural workers.

Fortunately, the search for plant-derived molluscicides that are target-specific,

environment and toxicologically safe, and inexpensive have gained an interest amongst

the agricultural industry.

Phyllantus Urinaria, a tropical and subtropical herb of the family Phyllanthaceae

are native plants that can be found in wastelands and dry fields. It is commonly known as

chamber bitter. Ethanolic extracts from Phyllanthus Urinaria contains phytochemicals

which has phenolic compounds such as alkaloids, flavonoids, saponins, tannins, et al.,

which possess a bitter taste, as the name indicates, due to the presence of such. It was also

asserted that plants containing bitter substances are generally disliked by snails, and

plants containing alkaloids can be extremely useful in the fight against problematic

species of snails and can cause lower harm to the environment than synthetic

molluscicides, which concludes that Phyllantus Urinaria has the potential to have

molluscicidal activities against Pomacea Canaliculata.

In the line with that, the Researchers will study the molluscicidal activities of

Phyllanthus Urinaria against Pomacea Canaliculata to improve growth control.

Phyllanthus Urinaria being organic, cost-effective, environmentally friendly, provides

protection to crops that aids Filipino farmers affected by snail pests, most specifically

Pomacea Canaliculata. Considering these various factors, the aim of this study is to

assess the molluscicidal effectiveness of the ethanolic crude extract of Phyllanthus

Urinaria to address any existing gaps in the current body of literature and explore the

potential of these plant-derived materials as a viable alternative to synthetic

molluscicides.
STATEMENT OF THE PROBLEM

This study seeks to determine the molluscicidal efficacy of the oil extract of Chamber

Bitter (Phyllanthus Urinaria) against Golden Apple Snails (Pomacea Canaliculata) to

aid in pest control, specifically Golden Apple Snails in Philippine rice cultivation.

Specifically, it will seek answers to the following questions:

1. How does the molluscicidal efficacy of Phyllanthus urinaria extracts, and the

commercial molluscicide (positive control) differ in terms of:

a. LC50 values?

b. LC90 values?

1. How does the commercial molluscicide (positive control) and Chamber Bitter

(Phyllanthus Urinaria) based molluscicide differ in terms of the following factors:

a. Accessibility

b. Duration of Action

c. Environmental Impact

d. Mortality rate

e. Target Spectrum

2. What is the lowest concentration of B. balsamifera leaf extracts that would

yield the mortality rates in the snail populations equal to:

 a. Fifty percent (LC50)?

b. Ninety percent (LC90)?

3. How well does the probit model fit the concentration-mortality relationship

among the treatments derived from the molluscicidal bioassay?

4. Based on the findings, what intervention can be done to improve the

effectiveness of Chamber bitter (Phyllanthus Urinaria) derived molluscicide

against Golden Apple Snails (Pomacea Canaliculata)?

RESEARCH HYPOTHESES

The study will use a parallel-group experimental design. To determine the

statistical significance of the findings, the researchers will use One-way ANOVA (f-test).

The use of the probit model is justified using the Chi-square Goodness of Fit test. An

alpha level of α = 0.05 will be used for all statistical tests. The following hypotheses will

be raised in the study to determine the effects of the factors involved, in the molluscicidal

effects against Pomacea Canaliculata:

Chi-square Goodness of Fit Test

Ho: The probit model adequately fits the mortality rates observed from the

molluscicidal bioassay.

Ha: The probit model does not adequately fit the mortality rates observed from the

molluscicidal bioassay.
One-way ANOVA Tests

Ho: There is no significant difference between the mortality of Golden Apple

Snails (Pomacea Canaliculata) in treatments with the varying concentrations of

the ethanolic crude extracts of Chamber Bitter (Phyllanthus Urinaria), and the

commercial molluscicide.

Ha: There is a significant difference between the mortality of Golden Apple

Snails (Pomacea Canaliculata) in treatments with the varying concentrations of

the ethanolic crude extracts of Chamber Bitter (Phyllanthus Urinaria), and the

commercial molluscicide.

PARADIGM OF THE STUDY

INPUT PROCESS OUTPUT

 Collection and

preparation of
Chamber Bitter
materials
 Chamber bitter extract (Phyllanthus Urinaria)
 Extraction of
625 mL Derived Molluscicide for
Chamber bitter
 Distilled Water 5,625 Golden Apple Snails
 Preparation for
L (Pomacea Canaliculata)
experiment

 Testing

 Data Collection

SCOPE & DELIMITATIONS OF THE STUDY

 The objective of the study will be to determine the molluscicidal efficacy of the

ethanolic crude extract of P. Urinaria against Pomacea Canaliculata to create a cost-

effective and environment-friendly product. The constituent in making the molluscicide

will be limited to the fresh Chamber bitter (Phyllanthus Urinaria). Specifically, the study

will determine the mortality rates from each treatment with varying concentrations to

calculate the LC50 and LC90 values for each of the samples. The study will look at the

effect of the various treatments in the study that will have on the mortality of the target

organisms and how changes in concentration would modify these relationships. The study

will utilize a parallel-group experimental design that uses the molluscicidal bioassay

method as recommended by the World Health Organization (2019). Moreover, statistical

methods such as the chi-square goodness of fit test, probit analysis, and the One-way

ANOVA test will be used by the researchers to analyze the data.

This study may not cover all regions affected by snail infestations worldwide,

specifically focusing only on the Golden Apple Snail (Pomacea canaliculata) and may

not extensively explore the molluscicidal effects of P. Urinaria on other snail species or

pests. Moreover, the researchers will not conduct qualitative or quantitative

phytochemical screenings for the molluscicide candidates due to financial and

instrumental limitations. The researchers will not also investigate the toxicity of these

samples to non-target organisms.

SIGNIFICANCE OF THE STUDY

This study will pave way to the discovery of a new molluscicide which prevents

harm on rice farming caused by snails, including making use of indigenous plant

materials which is much cheaper than the other synthetic materials sold in the market,

that could be utilized by agricultural industries in the production of cheaper and

environment friendly molluscicidal products. It will serve as a guide for the chemical

industries to formulate molluscicides that aren’t dangerous to man’s health and the

environment. In turn, the farmers will benefit financially from this production of a cost-

effective and environment friendly molluscicidal product.

DEFINITION OF TERMS

To ensure clarity and definiteness to their usage, the following terms were hereby

defining in accordance with how they were used in the study.

Chamber bitter (Phyllanthus Urinaria). A tropical and subtropical herb native in Asia,

it contains ethanolic compounds which contains phytochemicals which has phenolic

compounds such as which possess a bitter taste that can be used to make molluscicide.

This refers to the plant that have been extracted that isto be used to conduct this research.

Ethanolic extracts that have been found in Phyllantus Urinaria contains phytochemicals

which has phenolic compounds such as alkaloids, flavonoids, saponins, tannins, et al.,

which possess a bitter taste, as the name indicates, due to the presence of such.study.
Golden Apple Snail (Pomacea Canaliculata). Are highly invasive snails that cause

significant damage to rice crops. This refers to the sample snail used for the investigation.

Pesticides. Chemical compounds that are used to kill pests, including insects, rodents,

fungi, and unwanted plants (weeds).

Metaldehyde. a pesticide used to control snails and slugs and is approved for use in a

variety of vegetable and ornamental crops in the field or greenhouse, on fruit trees, small-

fruit plants,

Molluscicide. A substance or agent; type of pesticide used to kill harmful mollusks. This

refers to the Chamber bitter extract that will be used to kill Golden Apple Snails.

in avocado and citrus orchards, berry plants, banana plants and in limited residential

areas.

Morality. The death of large numbers (as of people or animals). The number of deaths in

a particular time or place. This refers to the death of Golden Apple Snails that will be

used for the study.

Niclosamide. is an antihelminthic medication that is taken orally and is used to treat

parasite infections.

School. This refers to Bantayan Science High School of Ticad, Bantayan, Cebu where the

investigation will be conducted.

Treatment. The techniques or actions customarily applied in a specified situation. A

substance or technique used in treating an experimental condition.

 CHAPTER III

METHODOLOGY

The study will be conducted to determine the molluscicidal efficacy of

Phyllanthus urinaria, utilizing a quantitative research design as the primary data

collection method. The subsequent sections will provide detailed insights into the

research design, outlining the orderly organization of procedures by the researchers. This

encompasses the selection of research materials, acquisition of raw materials,

identification of necessary equipment, establishment of the experimental design,

presentation of the research procedure, specification of statistical parameters, and the

environment where the study will be conducted.

Research Instrument

Materials Quantity Unit

Chamber bitter 625 mL

Golden apple snails 60 peices

Commercial molluscicide

Dechlorinated water 5,625 liter

grade 0905 crepe filter

paper

Table 2: Research Materials

Tools and Equipment Quantity Unit

Container 3 Pcs.
 Measuring cup

Incubator 1 Pcs.

Weighing scale 1 Pcs.

Tray 4 Pcs.

Mortar and Pestle 2 Pcs.

Blender 1 Pcs.

Table 3: Research Equipment

Research Environment

The researchers conducted the study at the biology laboratory of Bantayan Science High

School in Barangay Ticad, Bantayan, Cebu (11°10'20''N, 123°43'16''E). researcher’s

residence, Apatan, located at Ticad, Bantayan, Cebu. (11° 10' 7.6296"S, 123° 43'

20.5248") The chamber bitter will be collected in the wastelands of Bantayan Island.
 Research Design

The research will employ an experimental design, featuring three experimental groups

and two control groups, with the primary aim of assessing the molluscicidal efficacy of

varying concentrations of the ethanolic crude extract of Phyllanthus Urinaria. The two
control groups include a commercial molluscicide (acting as the positive control) and an

untreated dechlorinated water (serving as the negative control for observation only). The

methodology is adapted from a modified version of the methodologies outlined by the

World Health Organization (2019) and Alibo et al. (2021). Statistical analysis of the data

will involve the use of the chi-square goodness of fit test and the probit analysis method.

Additionally, the One-way ANOVA test will be applied to determine the significance of

the observed results.

Research Procedure

The study’s methodology is based on the World Health Organization’s (2019)

recommendations for molluscicide field and laboratory testing. The experiment will be

carried out in the biology laboratory of Bantayan Science High School in Barangay

Ticad, Bantayan, Cebu (11°10'20''N, 123°43'16''E). All throughout the investigation, the

laboratory protocols will be followed properly.

Collection and Preparation of Materials

First, the researchers will collect P. Urinaria one week before the

experiment. They will be collected at the dry fields and wastelands located within

Bantayan Island. For the P. Canaliculata, the researchers will purchase them from

Araño Kuyajing and will collect 60 of them or more if possible. Other materials

and instrument needed for the study are already available at the school’s

laboratory while some will be purchased or borrowed. After collecting P.

Urinaria, the plant will be washed thoroughly with distilled water.

Extraction of the Ethanolic Crude Extract

Firstly, the P. Urinaria will be left to dry at a room temperature for 24

hours. Then, they will be placed in the drying oven for 3 hours with the

temperature set at 70 oC. Then the dried plant will be powdered using a blender.

Materials for the extraction process will be prepared and assembled at the

laboratory. For this, absolute ethanol will be used in a 4:1 ratio with the powdered

P. Urinaria. The samples will then be left for 24 hours with a frequent stirring for

the first 5 hours. The solid parts of the suspension will be filtered out using a

grade 0905 crepe filter paper.

Making of Stock Solution

For the molluscicidal bioassay, stock solutions will be created. In creating

the stock solution for the plant-based molluscicide, 5 mL of the extract will be

mixed with 95 mL of water to create a 50000 PPM solution. Then, 15 mL of the

extract will also be mixed with 85 mL of water to create a 150000 PPM solution.

Lastly, 30 mL of the extract will be mixed with 70 mL of water to create a 300000

PPM solution.

Similarly, a stock solution will then be made for the commercial

molluscicide by dissolving 1.1 grams of the Niclosamide with 500 mL of water

according to its instructions.

Molluscicidal Bioassay

A molluscicidal bioassay will then be set up that follows the guidelines set

by the World Health Organization (2019) with modifications. The P. canaliculata

snails will be randomly divided into five groups: two control groups and three test

groups.

For the two control groups, one control group will be the negative control

which is only up for observation while the other control group will be the positive

control which is treated with a commercial molluscicide (Niclosamide). For the

three test groups, the proper aliquot amount from the stock solutions, as specified

in Annex 2 of the World Health Organization (2019) guidelines for laboratory and

field testing of molluscicides, will then be added to each container to yield a

specific concentration. The molluscicidal bioassay will be conducted in the

Bantayan Science High School Biology Laboratory on a photoperiod of 36 hours

in light and 36 hours in darkness.

Data Collection and Disposal of Waste

At the conclusion of the 24-hour exposure period, snails will be taken

from treated or controlled water, cleaned, and transferred to containers containing

standard snail water. Following a 24-hour recovery period, mortality will be

calculated. A snail is assumed to be dead when it doesn't move and stays still

inside its shell.

 Snails that are still alive after 24 hours will be placed in freshwater with

food and watched for an additional 48 hours (a total of 72 hours). Snails suspected

of being dead will be moved to different containers since, if they were, their

decomposition may impair the survival of other snails. Suspected dead snails will

have their deaths verified at the conclusion of the recovery time.

The waste generated from the molluscicidal bioassay will then be stored

in sealed containers and disposed of in the laboratory disposal facility for

biohazardous waste.

Statistical Parameter

Mortality of the molluscicidal bioassay will be recorded after 24 hours of

administering the treatment. The snail will be counted as dead or moribund if it does not

show signs of locomotion or reaction after the researcher touches them. The mortality

rate will then be calculated using the following equation.

number of dead ∨moribund snails

mortality rate = × 100
number of initial snails

In cases when there is observed mortality in the control group, the mortality rates

will be adjusted using a formula from Abbott (1925).

%mortality on treatment−%mortality on control

mortality (%) = × 100%
1−% mortality on control

The mortality rates for each test concentration of the treatments will be illustrated

using bar graphs. The LC50 and LC90 values with their respective 95% confidence

intervals for each of the treatments will then be calculated by probit analysis using
Microsoft Excel® for Microsoft 365 software with a toxicology calculator created by Lei

and Sun (2018). The statistical significance between the respective LC50 and LC90

values of the treatments is then determined using a One-way ANOVA test.

CHAPTER II

The researcher will conduct a study on the molluscicidal efficacy of ethanol crude extract

from chamber bitter (Phyllanthus urinaria) against golden apple snail (pomacea

canaliculata). The following sections will summarize the relevant literature that will
establish the significance of conducting research in this field, what has been done in the

field and the gap in the literature that it can potentially fill. However, it should be noted

that there is a scarcity of literature and studies surroundin the chamber biter (Phyllanthus

urinaria) extract and anything related to its molluscicidal efficacy.

Plant Based Molluscicide

According to Geethangili,M. & Ding,S. (2018), this review composition summarizes the

traditional uses of phytochemistry and pharmacology of Phyllanthus urinaria.The genus

Phyllanthus(L.) is one of the most important groups of plants belonging to the

Phyllantaceae family. Phyllanthus urinaria(L.) is an annual perennial herbal set up in

tropical Asia, China, America, and the Indian Ocean islands. P. urinaria is used in folk

medicine for the treatment of various health complications including diabetes, liver

conditions, and malaria. Phytochemical examinations reveals that 93 naturally being

composites have been insulated from P. urinaria, including tannins, lignans, terpenoids,

phenolics, flavonoids and other secondary metabolites. Pharmacological conditioning

include antimicrobial, hepatoprotective, antidiabetic, anticancer, and cardioprotective

effects. In recent times, there's interest in naturally cardioprotective agents that may

warrant side effect. Herbal products are extensively used among cases with

cardiovascular(CV) conditions, and cases freque combine herbal products with CV

specifics. This review provides traditional knowledge, phytochemistry, and natural

conditioning of Phyllanthus urinaria. This review summarizes the phytochemical

constituents and their biological activities including natural studies on various crude

excerpts and fragments both in vitro and in vivo.

According to Martin S, et al. (2009) Pomacea canaliculata snails pose a severe problem to

direct seeded rice cultivated in Southern Brazil. Snails were collected in rice fields close

to Porto Alegre (state of Rio Grande do Sul) and in Brusque (state of Santa Catarina) In

Santa Catarina the product was really effective, while in Porto Alegre it had no effect.

This unexpected behavior was probably due to the respiratory habits of the snails under
different contents of dissolved oxygen in the water. Near Porto Alegre the water used in

rice fields is heavily polluted, with dissolved oxygen levels of 1-2pm, and the snails rely

primarily on ther siphon and lungs to breathe. Since saponin control is probably due to an

interaction between saponins with the sterols present in the cell walls in the gills, no

control was observed. Control of this snail is nowadays performed with toxic chemicals

such as copper sulfate and fungicides such as fentin. By contrast, in Santa Catarina the

dissolved oxygen level of the water is a 5-6ppm, and the snails remain mostly

underwater, breathing with their gills. In this case, the snails died within 24 h at a dose of

20 and 30 ppm of product. To this observation, snails grown in polluted waters were

forced to remain underwater in saponin solutions and water(control) preventing the use of

their siphon to breathe. The snails exposef to saponin solutions died, while the controlled

snails survived, indicating that they were still able to use their gills to breathe. In

conclusion, the results indicate that Pomacea snails develop respiratory habits in relation

to the dissolved oxygen level of the water. The saponin product was only effective in

areas with high levels of dissolved oxygen, where the snails use primarily their gills to

respire. In waters with low levels of dissolved oxygen, no snail control was attained,

indicating that the saponin product affects only the gills and not the foot-sole epithelium

or internal organs, as has been reported for Bulinus africanus snails when contacted with

saponin solutions. This unexpected behavior indicates that before using the saponin-

based molluscicide, it is important to observe the dissolved oxygen levels of the water

used for irrigation and the respiratory habits of the snails.

According to Islam, S and Morshed A. 2013 Homemade bio-pesticides are prepared by

household members using local resources without having any scientific study or research.

Is is usually old aged trustworthy practices by inherently. In chemical farming, a specific

insecticide may be applied to quickly kill off a particular insect pest (animal). Chemical

controls can dramatically reduce pest populations for the short term, yet by unavoidably

killing (or starving) natural predator insects and animals, cause an ultimate increase in the

pest population. Repeated use of insecticides and herbicides and other pesticides also

encourages rapid natural selection of resistant insects, plants and other organisms,

necessitating increased use, or requiring new, more powerful controls. There are also

some general challenges with use of bio-pesticides. They tend to be more slow-acting and

may be very specific to the life cycle of the pest. There are almost CXXII biochemical

pesticide active ingredients registered with the EPA, which include XVIII floral

attractants, XX plant growth regulators, Six insect growth regulators, XIX repellents, and

XXXVI pheromones. Neem materials can affect insects, mites, nematodes, fungi,

bacteria, and even some viruses. Bio-pesticides fall on a continuum of breadth of

specificity: some active ingredients are highly specific to a particular organism at a

particular window of opportunity; others have a broader mode of action.Bio-pesticides

are certain types of pesticides derived from such natural materials as animals, plants,

bacteria, and certain minerals. Some essential oils work as repellents, and their mode of

action would be as a fragrance. In the end of my study I can say that organic pest

management system is an environmental friendly, low cost, and healthy method of pest

control system. Gradually farmers getting aware of this method and this pest management

system are getting popular day by day because of its beneficial features. Bio-pesticides
are a set of tools and applications that will help our farmer’s transition away from highly

toxic conventional chemical pesticides into an era of truly sustainable agriculture. Of

course bio-pesticides are only a part of a larger solution; sustainable agriculture is a broad

and deep field. But helping farmers move from their current chemical dependency to

organic agriculture and beyond requires tools for the transition and tools for a new era.

Bio- pesticides can and will play a significant role in this process. There remain,

however, serious questions about the safety of these products from both a human and

ecosystem health standpoint. Current regulations do not go nearly far enough in

evaluating systemic broader impacts of bio-pesticides. By definition, green chemistry is

about continuous improvements aimed at reducing or eliminating hazard. We must

encourage pest management solutions and regulations to continuously evolve and ensure

that multi-disciplinary teams, including green chemists, environmental health sciences

and other sciences, approach these products systemically to both discover and refine

them. Bio-pesticides offer powerful tools to create a new generation of sustainable

agriculture products. They are the most likely source for alternatives to some of the most

problematic chemical pesticides currently in use that are under ever-increasing scrutiny.

According to Updahay A. et.al July-August,2015 The molluscicidal activity of the leaf

powder of Moringa oleifera and lyophilized fruit powder of Momordica charantia against

the snail Lymnana aruminata was time and concentration dependent. M. oleifera leaf

powder (96 h LC 197.59 ppm) was mote toxic than M. charanzia lyophilized fruit powder

(96 h LC, 318.29 ppm). The ethanolic extracts of M. oleifera leaf powder and Momordica
charantia lyophilized fruit powder were more toxic than other organic solvent extracts.

The 96 h LC of the column purified fraction of M. oleifera lest powder was 22.52 ppm,

while that of M. charantia lyophilized fruit powder was 6.21 ppm. Column, thin layer and

high performance liquid chromatography analysis show that the active molluscicidal

components in M. oleifera leaf powder and lyophilized frait of M. charantia me

benzylamine (96 h LC 2.3 ppm) and momordicine (96 h LC 1.2 ppm), respectively

Benzylamine and momordicine significantly inhibited, d, in vivo and in ving, the

acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the

nervous tissues of L. acuminate, Inhibition of AChE, ACP and ALP activity in the

nervous tissues of L. acuminata by benzylamine and momordicine may be respransible

for the molhuscicidal activity of M. oleifera and M. charantia fruits, respectively. In

conclusion, the results of this study indicate that the ethanolic extract of Momordica

charantia leaf powder has significant antibacterial activity against a variety of foodborne

pathogens. The authors found that the leaf extracts of Mallotus le viched and Momordica

charantia had significant AChE inhibitory activity. This suggests that these plants may

have potential as therapeutic agents for Alzheimer's disease and other neurological

conditions associated with AChE deficiency.

According to Radwan, Mohamed A. and Gad, Radwan Amira F. 2023 Despite the great

benefits of using synthetic molluscicides as mollusc control agents, they can destroy the

entire ecosystems, causing detrimental effects on human health and non-target organisms.

Essential oils and/or their constituents are gaining increasing interest for using as safe

alternatives to pesticides for controlling various pests including gastropods. The present
review was conceived to overview on essential oils and/or their components as novel

mollusc control agents that can be used in sustainable agriculture and medical public

health sectors. We also intended to find approaches for new essential oils based on

pesticidal products as a promising alternative for mollusc control. Indeed, this review

covers their mollusci- cidal, antifeedant and repellent properties and discusses their

structare-activity relationships. Phytochemical analysis of aromatic plants using GC and

GC-MS to elucidate the possible secondary metabolites present which could be

responsible for their molluscicidal efficacy and to know of their mechanism of action is

also addressed. Furthermore, the performance of component blends explored and efforts

evolved for maximizing the molluscicidal activity of essential oil components by either

synthesized new derivatives or mixing with synergists are also discussed. We also looked

at additional assessment of various issues before adopting these products as general

molluscicides. All the data in this review supported the promising uses of essential oils

and/or their components in harmful gastropod pest control. In conclusion, essential oils

(EOs) are promising molluscicidal agents against terrestrial and aquatic gastropods. They

have a wide range of molluscicidal activities, including contact toxicity, repellency, and

antifeedant properties. EOs are also relatively safe for non-target organisms and the

environment. The study by Radwan et al. (2023) provides a comprehensive overview of

the molluscicidal potential of EOs and their active constituents. The authors conclude that

EOs are promising new molluscicide agents that could be used to develop safe and

effective pest management strategies for gastropods.

According to Geethangili,M. & Ding,S. (2018), this review composition summarizes the

traditional uses of phytochemistry and pharmacology of Phyllanthus urinaria.The genus

Phyllanthus(L.) is one of the most important groups of plants belonging to the

Phyllantaceae family. Phyllanthus urinaria(L.) is an annual perennial herbal set up in

tropical Asia, China, America, and the Indian Ocean islands. P. urinaria is used in folk

medicine for the treatment of various health complications including diabetes, liver

conditions, and malaria. Phytochemical examinations reveals that 93 naturally being

composites have been insulated from P. urinaria, including tannins, lignans, terpenoids,

phenolics, flavonoids and other secondary metabolites. Pharmacological conditioning

include antimicrobial, hepatoprotective, antidiabetic, anticancer, and cardioprotective

effects. In recent times, there's interest in naturally cardioprotective agents that may

warrant side effect. Herbal products are extensively used among cases with

cardiovascular(CV) conditions, and cases freque combine herbal products with CV

specifics. This review provides traditional knowledge, phytochemistry, and natural

conditioning of Phyllanthus urinaria. This review summarizes the phytochemical

constituents and their biological activities including natural studies on various crude

excerpts and fragments both in vitro and in vivo.

According to Barrion, M. A,. et al. (2019) The study is intended to test the molluscicidal

exertion of Mangifera indica dinghy depending on the following concentrations: 15%,

25%, 50% and 75% within 15, 30 and 45 minutes based on the mortality rates of the

Pomacea canaliculata Lamarck (Golden Apple Snails). The concentrations were attained

using the process of decoction in which 15 grams of Mangifera indica bark was
subordinated into 100 ml distilled water and the same goes with the other concentrations.

The Philippines is one of the most rice consuming countries. Rice is Philippine's staple

food and most fibers grained nutrient. With this, the population in the Philippines is

continuously growing and this threatens the production of rice. But one of the major

problems that a Filipino farmer faces is a lower harvest beacause of crop losses due to

pest irruption. In the field of agriculture, farmed crops are vulnerable from the infection

of pests which cause damages leading to a significant reduction of crop(The Food Journal

and Food Nutrition & Science, 2015). Pesticides are categorized based on how it affects

its target which is called "modes of action". Other than that, they can also be classified

according to their chemical composition for example, organic pesticides. Organic

pesticides are produced with active ingredients derived from living matter such as

plants.The natural ingredients which are not only cost-effective have a minimal effect on

the soil as well on humans.The results indicate that M. indica extracts have potential as

natural molluscicides and could provide an alternative to synthetic pesticides for

controlling golden apple.

According to QUI Jian-Wen (2008) Wetland macrophyte diversity, trophic status and

phytoplankton: a case study of the cascading effects of an invasive herbivore preference

in the apple snail;design and some brief preliminary results. cormmon local macrophytes

to the apple snail in following 12 species of macrophytes are being tested: To standardize

consumption rate for different species, determined for cach plant species, as well as the

apple plant in treatment with snails, and Co and Cfare the The consumption rate on

different macrophyte species Preliminary results: This apple snail is a voracious This task
tests the feeding of apple snails in the the apple snail habitats were selected as the test

changes in biomass for different species of changes in biomass for different species

ofnorphology and toughness, but different nutrition andeducation, water quality

improvement and wild lifegbobs snails to test whether plant structure i important in the

plants that apple snails do not cat as whole tissue or as agar together with the extract of a

plant that the apple species that the apple snail likes, contains only 1% Assessment of

impact of the apple snail on macrophyte species composition and coverage. and plant

species diversity, and water qualitys snail grazing on macrophyte biomass, nutrient levels

Each species is tested similar biomass of the same species of macrophytes and 4 different

densities of apple snails. The experiment with each plant species lasts one different plant

species to grazing, and whether nutrient Part 2: Two species of macrophytes have been

tested. Snail density has great influence on the biomass of High snail density results in a

great Invading herbivory: The golden apple snail alters.

According to Alampayan M. (2019) Molluscicidal Activityon Golden Kuhol Applied

With Ipil-Ipil Leaves (Leucaena glauca L.),Makahiya (Mimosa pudica)and Guava

(Psidium guajava L.) Marivic Alampayan Keywords: Golden Apple Snail, Tannins,

Saponins ABSTRACT INTRODUCTION The golden apple snail, popularly known as

"golden kuhol" [Pomacea canaliculata Lamarck], was However, a few years after its

introduction, the golden apple snail became a major pest of It is known to be one of the

farmer's worst enemy in the rice field. This pest can be very destructive. METHODS by

using a plant molluscicide in the form of ipil-ipil, guava and Makahiya leaves extract

applied to The plant materials of ipil-ipil, guava and makahiya were gathered in a farm
lot and were about 100 grams of each botanical plants (makahiya, ipil-ipil, guava).

botanical plants in the Erlenmeyer flask. extract in the cylinder and then place it in the

hot plate. Using the hot plate let the extract be heated After 30 minutes, allow the extract

to cool doWn. Then label and store for application on the mortality rate of snails.

RESULTS The study shows the effectivity of botanical plants as molluscicide with the

golden kuhol applied With different time interval of 10 minutes. among the three

botanical plant is the makahiya leaves. DISCUSSIONS Guava is the most potent hence

tannins and saponins were abundantly present, ipil-ipil is abundantly present in saponins

and moderately abundant with tannins. moderately presence saponins and slightly present

in tannins. between plants and phytophagous insects are multiple particularly those

focused on toxicity.

According to Yang. Chen-Min, et al. (2005).Acetone, ethanol and methanol extracts of

Phyllanthus urinaria inhibit HSV-2 infection in vitro. In this study the anti-HSV-1 and

HSV-2 activities of different solvent extracted from P. urinaria were investigated in vitro

by plaque reduction assay.The plant is traditionally believed that it can reduce heat,

remove food stagnancy, improve eyesight, relieve inflammation, calm the liver, suppress

yang hyperactivity of liver(subdue the excessive syndrome and heat syndrome cause by

liver for example hypertension, neurasthenia etc), detoxify poison from body and

increase the flow of urine (Committee on Chinese Medicine and pharmacy, 2003), in

Taiwan, the whole plant of P. urinaria is habitually used as the therapy for diarrhea,

dysentery, contagious hepatitis, edema, infantile malnutrition, acute conjunctivitis,

aphthae and unreasonable pyogenic infections(Committee on Chinese medicine and

pharmacy, 2003).The traditionally usage of P. urinaria as therapy for virus caused-

hepatitis hinted that it is an antiviral agent.However, the anti-HSV activity of P. urinaria

has not been reported in literature, in this study, a series of experiments were conducted

to investigate the anti-HSV-1 and HSV-2 activity of P. urinariain vitro. Results showed

that acetone, ethanol and methanol extracts of P. urinaria Inhibited HSV-2 but not HSV-1

infection.The 50% inhibitory concentration against HSV-2 infection(IC50) of acetone,

ethanol and methanol extracts was 4.3±0.5, 5.0±0.4 and 4.0±0.9mcg/ml, respectively. All

three extracts showed no cytotoxic effect against Vero cells at concentrations of

10.0mcg/ml or below.Therefore, the acetone, ethanol and methanol extracts of P. urinaria

were concluded to likely inhibit HSV-2 infection through disturbing the early stage of

virus infection and through diminishing the virus Infectivity.

According to Cabrera, Z. W.& Manlatican, R. (2020). Molluscicidal Activities of

Leucaena leucocephala and Moringa oleifera seed extracts against achatina fulica. The

molluscicidal activities of Moringa oleifera and Leucaena leucocephala seed were

evaluated using the invasive and farm pest giant African land snail (Achatina fulica). This

study employed Complete Randomized Design (n=360), involving two solvent

concentrations (95% ethanol and 75% ethanol) with three extract concentrations each

(200 ppm, 600 ppl, and 1000 ppm) and two control groups (Positive Control =

Niclosamide and Negative Control = distilled water).This study examined the

molluscicidal activities of two plant species that are environment-friendly and locally

available to Filipino farmers.The molluscicidal properties from the seeds of Leucaena

leucocephala and Moringa oleifera through ethanol extraction were tested by

administering each ethanolic seed extract to adult Achatina fulica using different product

concentrations. The molluscicidal assay of both Moringa oleifera and Leucaena

leucocephala showed significant differences in mortalities between the different seed

extract concentrations in 75% and 95% ethanol (n=360). Each snail demonstrated a

normal activity (no observer effects on hyperactivity or escape behavior and food intake)

throughout the 24-hour period with three follow-up observations in 5-hour interval after

the application of distilled water, indicating that the factors that contribute to the

mortality of the other other groups were entirely attributed to the applied

treatment.Meanwhile, the positive control group exposed to Niclosamide demonstrated

100% mortalities (Appendix G), within the 24-hour observation period after the

application of Niclosamide, snails exhibited hyperactivity (Ebenso, 2003).Among the

treatments, only the 600 ppm and 1000 ppm of M. oleifera seed extract in 95% showed

result statistically similar to the positive control (Niclosamide).

According to Schneiker, J., et al. (2016), Golden apple snails (GAS) have been a

significant invasive pest in rice crops. Reduced government support for training is

attributed to the perceived knowledge of farmers. Only 23% of farmers had undergone

prior training in GAS management. The training demonstrated no significant impact on

the adoption of sustainable methods or avoidance of molluscicides. Notably, 74% of

farmers continue to use molluscicides despite recommendations for sustainable

alternatives. There is an emphasis on the need for ongoing education, revisiting training

strategies, and exploring community-driven solutions for sustainable GAS management.

According to Balmores, M.G.,et al. (2020), There are various kinds of pesticides, and

molluscide is one of them. Molluscide is a type of pesticide specifically used to kill

mollusks, including snails and slugs. Ampalaya (Momordica Charantia) contains

alkaloids and other phenolic compounds, responsible for killing Golden Apple Snails.

The experiment was conducted in a plastic tray with water and Ampalaya leaf extract.

The concentration of Ampalaya leaf extract and the weight of Golden Apple Snails

influence their mortality rates. Concluding that the difference in Ampalaya leaf extract

concentration determines effectiveness in killing Golden Apple Snails, and the weight of

the snails plays a significant role in their mortality rates.

According to Cabrera, Z. W.& Manlatican, R. (2020). Molluscicidal Activities of

Leucaena leucocephala and Moringa oleifera seed extracts against achatina fulica. The

molluscicidal activities of Moringa oleifera and Leucaena leucocephala seed were

evaluated using the invasive and farm pest giant African land snail (Achatina fulica). This

study employed Complete Randomized Design (n=360), involving two solvent

concentrations (95% ethanol and 75% ethanol) with three extract concentrations each

(200 ppm, 600 ppm, and 1000 ppm) and two control groups (Positive Control =

Niclosamide and Negative Control = distilled water).This study examined the

molluscicidal activities of two plant species that are environment-friendly and locally

available to Filipino farmers.The molluscicidal properties from the seeds of Leucaena

leucocephala and Moringa oleifera through ethanol extraction were tested by

administering each ethanolic seed extract to adult Achatina fulica using different product

concentrations. The molluscicidal assay of both Moringa oleifera and Leucaena

leucocephala showed significant differences in mortalities between the different seed

extract concentrations in 75% and 95% ethanol (n=360). Each snail demonstrated a

normal activity (no observer effects on hyperactivity or escape behavior and food intake)
throughout the 24-hour period with three follow-up observations in 5-hour interval after

the application of distilled water, indicating that the factors that contribute to the

mortality of the other other groups were entirely attributed to the applied

treatment.Meanwhile, the positive control group exposed to Niclosamide demonstrated

100% mortalities (Appendix G), within the 24-hour observation period after the

application of Niclosamide, snails exhibited hyperactivity (Ebenso, 2003).Among the

treatments, only the 600 ppm and 1000 ppm of M. oleifera seed extract in 95% showed

result statistically similar to the positive control (Niclosamide).

Based on the study of Silva Yrr., et al. (2020), the crude ethanolic extracts of the Persea

americana stem bark which showed a presence of flavonoids, anthraquinone heterosides,

comarins and tannins showed molluscicidal activity against all life cycle stages of B.

glabrataembryos. It somehow showed how the early developmental stages of B.

glabrataembryos are more sensitive to the toxic effects of extracts when compared to

adults. Thus, the crude is a potential candidate for molluscicide, contributing to the

control of host snails and reducing the transmission of schistosomiasis.