AGUSAN DEL SUR STATE COLLEGE OF AGRICULTURE AND

TECHNOLOGY

MOLLUCIDAL EFFECT OF GMELINA (Gmelina arborea) SEED

EXTRACT AGAINST GOLDEN APPLE SNAIL (Pomacea
canaliculata, Lamarck.) INSIDE AT AGUSAN DEL
SUR STATE COLLEGE OF AGRICULTURE AND
TECHNOLOGY (ASSCAT), BUNAWAN,
AGUSAN DEL SUR, PHILIPPINES

JEROME MONDEJAR ROCERO

JOSHUA PUJIDA DURANGO

BACHELOR OF SCIENCE IN AGROFORESTRY

JANUARY 2022
 INTRODUCTION

The Pomacea canaliculata (Lamarck) known for golden apple snail

at a recent time is considered a rice pest in some country of Asia during

recent decades which farmers are affected (Litsinger and Estano, 1993).

The golden apple snail was first found in Malaysia in the 1990s and since

then it has become one of the major pests in the paddy industry (Nur

Suraya et al., 2017). P. canaliculata attacks and destroys young stems

and leaves of plants and could eat 7–24 rice seedlings per day (Cagauan

and Joshi, 2002).

The damage done by the snails is sufficiently serious and is a

major concern. The problem becomes more serious as P. canaliculata’s

populations can increase rapidly under favourable conditions (Ding et al.,

2018). In controlling the infestation of P. canaliculata, various methods

such as cultural control, biological control, chemical control, salt

compounds, organic compounds, and molluscicides have been used

(Liang et al., 2013). Handpicking and using ducks are among cultural

and biological controls used. These methods have been proven to control

the snails at a low cost. However, they require a large labor force and

causes damage to the rice seedlings (duck control) (Liang et al., 2013).

Chemicals are also widely used in controlling P. canaliculata as

they are very effective. The principal chemicals used effectively against

golden apple snails are metaldehyde, formulated as bait pallets, wettable

powders or flowable suspensions, and niclosamide, formulated as either

an emulsifiable concentrate, suspension concentrate or wettable powder.

Even though they are effective, the use of chemicals is discouraged as

they have long-term toxicity effects on humans and the environment.

Musman (2010) reported that the usage of chemical fertilisers such as

metaldehyde and niclosamide can pollute water sources and thus affect

the ecosystem. Furthermore, the cost of chemical molluscicides is

unaffordable to many farmers.

The P. canaliculata the snail can grow up to 80-mm shell height

(Carlsson et al,. 2004). According by Food and Organization of the United

Nation (1998), golden apple snail was introduced by the farmers in order

to export the snail to Europe and have huge profits early in the 1980s

from Florida and Latin America to the Taiwan including the Philippines.

Gmelina arborea is known for gmelina belongs to the family

Verbinaceae found in 11 countries of Asia in tropical and subtropical

region. Almost 700,000 gmelina plantations, small woodlots and

agroforestry at west-central and eastern Africa, Southeast Asia, the

South Pacific, and northern Latin America. Gmelina has major role

benefits in agroforestry systems. Intensive breeding and improved clones

will bring better growth, more uniform wood, better disease resistance,

and products of higher quality to both local and international markets

(Dvorak,2004).
 On the contrary, the molluscidal property of commercially-bought

molluscicide is manifested by looking for sufficient amounts of the active

ingredient, metaldehyde and niclosamide (Li & Wang, 2017).

These active ingredients are known to cause disruption in the

whole ecosystem as it is highly corrosive and poisonous to other

organisms not just the snails. Today the damage that the golden apple

snails caused to the Philippine rice fields gravely affects the rice

production which then cause large loses to our Filipino farmers. The use

of commercial molluscicides is one of the major solutions that they use to

decimate the snails, but the risk in their health since there are high

concentrations of toxic chemicals in the molluscicide is extremely notable

(Rejesus & Punzalan, 2009).

In this study, we aim to verify the molluscicidal potential of

(gmelina arborea) hoping the possibility to the development of plant

derived molluscicide that would efficiently decimate the snails as well

become a safer option for the farmers to use. And the ecosystem around

it. Moreover, this study could be used as baseline information for future

researchers that will experiment the same plant for other research. This

may help them in understanding the topic and have insights on the topic

they would work on.

 THE OBJECTIVES OF THE STUDY

The general objective of the study is to perform the molluscicidal

potential of gmelina arborea seed extract against golden apple snail

(Pomacea canaliculata, lamarck.).

Specific objective

Specifically, this study aims to:

1. Evaluate the significant difference between the performance of

plant derived molluscicides and a commercial molluscicide?

2. To determine the molluscicidal performance of (gmelina arborea).

3. To determine the significant difference or effect of plant-derived

molluscicide?
 SCOPE AND DELIMINATION OF THE STUDY

The study covers the determination of the molluscicidal

performance of Gmelina (gmelina arborea) seed extract against the golden

apple snail (Pomacea canaliculata L.). The study is limited to the

molluscicidal properties of certain parts of G.arborea. It also limits the

snail species to be tested which is the golden apple snail (Pomacea

canaliculata L.). Along with its limitations is the source of the snails to

be used, the source of Gmelina (gmelina arborea) seeds.

 TIME AND PLACE OF THE STUDY

The study will be conducted at agusan del sur state college of

agriculture and technology (ASSCAT), Bunawan Agusan del from

December 2021 to february 2022.

OPERATIONAL DEFINITION OF TERMS

Agroforestry refers to a collective name of land use system and

technology where woody perennials (trees, shrubs, palms, bamboos and

etc.) Are deliberately use in the same land management units as

agricultural crops or animals.

Gastropods commonly known as snails of slugs ,belong to a large

taxonomic class of invertebrates withinthe phylum Mollusca called

Gastropads ,This class comprisessnails of slugs from softwater from

freshwater and from the land .

Gmelina (gmelina arborea), beechwood,gmelina,goomer teak ,Kashmir

tree ,malay beechwood ,white teak, Yamane is a fast growing decidouos

tree in the family lamiaceae . and widely grown for timber and pulp

Golden apple snail (pomacea canaliculata ), commonly known as as the

channeled aplle snail ,is a species of large freshwater snail with gills amd

an operculum ,an aquatic gastropod mulluse in the family

ampullariidae .

Invasive species is an introduced organismthat becomes overpopulated

and negatively alters its new environment .althuogh their spread can

have benefitial asfects invasive species adversely affect the invaded

habitats and bioregions , causing ecological, environmental , and /or

economic damage

Laboratory condtion , the physical environment under which an

expeirement or procedure in a laboratory is conducted which may

introduce a specified , frequently optimized ,temperature, pressure ,

types of apparatus ,etc ) frequently as under laboratory conditions .”

Metaldehyde a crystalline compound (CH3CHO)4 that kills polymer of

acetaldehyde effect based on two principles : firstly its irritant effect

causes the slugs tp secrete large amounts of mucus which results in

desiccation and death :secondly , its toxicity as a nerve poison prevents

the slug crawling away from the poison

Mulluscicides are pesticides which kill mullosks an animal phylum of

tens of thousand of invertebrate creatures mollusks include octopi and

squid as well as snails and slugs , which are usually targeted by

mulluscicides .

Native species is indigenous to a given region or ecosystem if its

presence in that region is the results of only local natural evolution

though often popularized as without human intervention.

Niclosamide is an orally bioavailable chlorinated salicylanilide with

anthelmintic and potential antineoplastic activity. Upon oral

administration , it is also a secondary carboxamide resulting from the

formal condensation of the carboxy group of s-chlorosalicylic acid with

the amino group of z-chloro-4-nitroaniline .it is an oral anthelmintic drug

approved for use against tapeworm infections is has a role as a

piscicide,a mulluscicide an anti parasitic agent ,an anticoronaviral agent

,an anthelmintic drug.

 CONCEPTUAL FRAMEWORK OF THE STUDY

Independent Variables Dependent Variables

Treatment  Test molluscicidal performance

T1-50ml/L gmelina through mortality count. Cumulative

seed extract
mortality rate and post-mortem
T2-100ml/L gmelina
seed extract observations.
T3-150ml/L gmelina
seed extract
 Significant difference between gmelina
T4-commercial
mollucide arborea seed extract and commercial

mollucide.

 The mortality rate will be observed

every 24hrs for 3days.

Figure 1. Conceptual
 Mucus secretion
Framework of the

study  Change in the shell colour and failure

This study is of the flesh portion to withdraw into

anchored in the the shell.

concept of evaluating the mortality rate and post-mortem analysis of test

subjects to compare the molluscicidal performance of plant derived

molluscicide and commercial mollucicide.

 Figure 1 entails the flow of the study. The seed extract of Gmelina

(gmelina arborea) against golden apple snails will be the treatment of the

study. The parameters that we are going to use to measure the

molluscicidal performance of the plant-base molluscicide is the mortality

rate, cumulative mortality rate and post-mortem observations of the

P.canaliculata, L. after harnessing the parameters of the dependent

variable, the researchers then looking for the significant difference

between the plant molluscicide and commercial molluscicide.

REVIEW OF RELATED LITERATURE

BOTANICAL DESCRIPTION G. arborea

Gmelina arborea is an unarmed, moderately sized to large

deciduous tree with a straight trunk. It is wide spreading with numerous

branches forming a large shady crown, attains a height of 30 m or more

and a diameter of up to 4.5 m. Bark smooth, pale ashy-grey or grey to

yellow with black patches and conspicuous corky circular lenticels.

Inside surface of bark rapidly turns brown on exposure and exfoliates

into thick woody plates or scurfy flakes. Blaze pale orange and mottled

with a darker orange colour. Leaves opposite-decussate, mostly rather

soft and limp; petioles cylindrical, 5-15 cm long, puberulent or glabrous;

leaf blades broadly ovate, 10-25 cm x 7-20 cm wide, apically long

acuminate or caudate, entire on mature plants but strongly toothed or

lobed on young plants, usually cordate or trunkate basally, with a short

cuneate attenuation into the petiole, densely tomentose above when

young, becoming glabrous above when mature, permanently densely

fulvulous-tomentellous with stellate hairs beneath, glanduliferous just

above the petiole on the basal attenuation (Katende AB et al. 1995).

DISTRIBUTION

The species occurs in a variety of forest habitats, including tropical

semi-evergreen, sub-montane, very moist teak forests, deciduous, sal and

dry teak forests. It also occurs in Syzygium parkland and low alluvial

savannah woodland.

The tree is a light demander, although it can stand some shade. It

is moderately frost hardy and recovers quickly from frost injuries. G.

arborea occurs in the western Himalayas. Its choice of site is wide, but it

shows a preference for moist fertile valleys with sandy loam soil; in west

Bengal, this species grows best on high silt deposits near rivers. It does

not thrive where the drainage is poor, while on dry, sandy or otherwise

poor soil it remains stunted and is apt to assume little more than a

shrubby form because of repeated dying back through drought (Brose J

et. al.).

USES

Fodder: Leaves are regarded as good fodder and cattle eat the fruit.

Fuel: G. arborea is planted mostly for firewood, which has a calorific

value of 4800 kcal/kg. For firewood, a spacing of 2 x 2 m is

recommended. Plantations of G. arborea have been established for

tobacco curing. Apiculture: Flowers produce abundant nectar, which

produces high-quality honey.

Fibre: The wood produces good-quality pulp. Unmixed semi-

chemical pulp is suitable only for carton board or low-grade writing

paper, but kraft pulp of yemane wood is suitable for higher grades of

writing paper. It is also utilized for particle board.

 Timber: When 1st cut, the wood is yellowish- to reddish-white,

turning light russet or yellowish-brown with a density of 400-560

kg/cubic m. The wood seasons well without degrading, but it is slow to

dry both in the open and in a kiln. Where it is indigenous, it is regarded

as a valuable general-purpose wood because of its dimensional stability.

The natural durability of the wood is about 15 years. Uses include the

manufacture of furniture, plywood core stock, mine props, matches and

timber for light construction. Tannin or dyestuff: Both wood ash and fruit

yield a very persistent yellow dye.

Medicine: Bark, leaves and roots contain traces of alkaloids and

are used medicinally in its native range, such as in Hindu medicine. For

example, both fruit and bark have medicinal properties against bilious

fever. Other products: Recommended for silkworm culture. SERVICES

Reclamation: G. arborea is a highly light-demanding species and

regenerates naturally only in the open and on the edge of forests. It is an

ideal choice for large-scale afforestation programs. Ornamental: G.

arborea is sometimes planted as an avenue tree (Albrecht J. ed. 1993).

GOLDEN APPLE SNAIL

There are more than 100 species of apple snail that exists. Two

species, Pomacea canaliculata and Pomacea maculata, commonly

known as Golden Apple Snails, are highly invasive and cause damage to
rice crops. They were introduced to Asia, from South America, in the

1980s as potential food for people, but it unfortunately became a major

pest of rice. Golden apple snails eat young and emerging rice

plants. They cut the rice stem at the base, destroying the whole plant.

Snails are able to spread through irrigation canals, natural water

distribution pathways, and during flooding events. When water is absent,

apple snails are able to bury themselves in the mud and hibernate for up

to six months. When water is re-applied to fields, snails may emerge.

They damage direct wet-seeded rice and transplanted rice up to 30 days

old. Once the rice plant reaches 30−40 days, it will become thick enough

to resist the snail.

To distinguish golden apple snails from native snails, check its

color and size. Golden apple snails have muddy brown shell and golden

pinkish or orange-yellow flesh. They are bigger and lighter in color

compared to native snails. Its eggs are bright pink in color. To confirm

snail damage, check for missing hills, cut leaves, and cut stems.

The golden apple snail is considered a major problem of rice. If no

control measure is taken, they can completely destroy 1 m 2 of field

overnight. This damage could lead to more than 50% yield loss.

The critical time to manage golden apple snails is during land

preparation and crop establishment or planting; specifically, first 10 days

after transplanting (DAT), and during the first 21 days after direct wet-

seeding. After this, the crop is generally resistant to snail damage and

snails are actually beneficial by feeding on weeds.

Community-based snail management to best control the snail,

communities should work together to reduce snail numbers in their area.

Conduct mass snail and egg collection campaigns, involving the whole

community, during land preparation and planting or crop establishment.

Keep fields drained as much as possible during the vulnerable stages of

the rice plant (below 30 days) or transplant 25−30 day old seedlings from

low density nursery beds.

Biological control Encourage natural predators. Red ants feed on the

snail eggs while ducks (and sometimes rats) will eat young snails. Several

wild bird species have also adapted to feed on golden apple snails and

domestic ducks can be put into fields during final land preparation or

after crop establishment when plants are big enough (e.g., 30−35 DAT).

Snails can also be harvested, cooked and eaten or sold as animal feed.

However, it is important to take extra care and cook the snails

thoroughly, as they are known carriers of the rat lungworm.

Cultural control Handpick snails and crush egg masses. This is best

done in the morning and afternoon when snails are most active. Place

bamboo stakes to provide sites for egg laying that allows easy collection

of snail eggs for destruction. You can also use attractants or plants that

attract snails, such as papaya and cassava leaves, to make hand picking

easier.

Manage water Apple snails have difficulty moving in less than 2 cm of

water. Keep water level below 2 cm during the vulnerable stages of the

rice plant. Construct small canals or cannulates (e.g., 15−25 cm wide

and 5 cm deep) after the final land preparation. Pull a sack containing a

heavy object around the edges of rice paddies or at 10−15 m intervals.

Cannulates facilitate drainage and act as focal points for snails making

manual collection or killing easier.

Use toxic plants- Place toxic plants, such as tobacco leaves, heartleaf

false pickerelweed, and citrus leaves in strips across the field or in

cannulates. Prevent field entry Know your water flow. Snails can invade

fields from canals, rivers, and reservoirs. Place a barrier where water

enters and exits the field. Place a wire or woven bamboo screen or mesh

bag on the main irrigation water inlet and outlet to prevent snail entry.

Transplant-Transplanted rice is less vulnerable than direct seeded rice.

Plant healthy and vigorous seedlings. Raise seedlings in low density

nursery beds, i.e., less than 100 g seeds per m 2 and delay transplanting

(e.g., transplant 25−30 day-old seedlings). To reduce missing hills from

snail damage, multiple seedlings per hill can be planted.

Chemical control Sometimes chemical control may be needed if other

practices fail. Check locally available products that have low toxicity to

humans and the environment.  Following normal fertilizer application

rate and schedule, apply fertilizer in 2 cm of water to maximize negative

effects on apple snails. Apply products only to low spots and cannulates

rather than to the whole field. Always ensure safe application. If used,

molluscicides should only be used immediately after transplanting or

during the seedling establishment phase in direct seeded rice; and only

for rice younger than 30 days old.

 MATERIALS AND METHODS

Experimental Design

This study used true experimental design. The presences of control

and manipulation group made it ideal for the chosen design. The study

focuse on determining the molluscicidal effect of Gmelina (Gmelina

arborea) seed extract against the invasive species, Pomacea

canaliculata, Lamarck. Specifically, the design that were going to use in

the study is post-test only control group design. With this design, the

effect of experimental treatment yielded the significant difference

between the experimental group that is the three plant derived

molluscicide namely: Gmelina (Gmelina arborea) seed extract which is

the treatment group and the synthetic molluscicide that is

metaldehyde which is the positive control group in killing the golden

apple snail (Pomacea canaliculata,L.).

Subjects of the Study

Minimum of 600 samples of Golden apple snail (Pomacea

canaliculata, L.) will be utilize in the study. The samples will be

gathered from rice fields of ASSCAT Agusan del sur State College of

Agriculture and Technology where the said species are abundant. Upon

collecting the Golden apple snail, random selection was done as we are

going to expose to the plant-derived molluscicides in order to determine it

molluscicidal performance. This study used 58 sets of adult Pomacea

canaliculata, L. samples, with each individual set including 10 snails as

the subject of the study.

Preparation of Snails

The subjects, Pomacea canaliculata, Lamarck, will be collected in

the rice field of ASSCAT Agusan del sur State College of Agriculture and

Technology. The snails collected however, will be adult snails, with shell

diameter of 25 mm ± 35 mm and a weight requirement of ≥ 5 grams.

Prior to the application of treatment, the snails will be checked,

ensuring they are all alive and moving.

After collecting the snails, quarantine and acclimation will be

done to ensure that the test subjects are able to adapt to such laboratory

condition wherein the continuation of the experiment is anchored.

During the acclimation process, the snails are allowed to adapt in a

controlled environment. This is to ensure that the snails have

enough time to adapt on the newly-introduce environment. The

acclimation process is adapted from the methods of Joshi et al. (2005)

wherein 10 snails will be place in 500 ml containers, specifically

transparent containers. Each container was filled with 350 mL

dechlorinated water at 26°C (Dai et al., 2011; He et al., 2017). The

acclimatization process lasted for 24 hours. The snails then had a

three-day quarantine period where they will be expose to the typical

fresh water for two days and to the negative control, dechlorinated

water, for one day. The snails were fed as well since it could affect the

data gathered in this study. Additionally, only the ones that passed the
height and weight requirements and are actively moving (i.e. rasping

using the radula, movement of inhalant siphon, tentacles and visceral

foot outside the lip) were the ones that proceed in the actual experiment.

The snails that didn’t show active state or/and didn’t pass the shell

diameter and weight requirement will be separated crushed and

disposed as instructed by the standard protocol for disposing gastropods

(WHO, 1983).

Sampling Technique

Probability sampling, specifically simple random sampling, is use

in selecting the six hundred (600) samples of Pomacea canaliculata,

Lamarck upon collecting it from the rice fields of ASSCAT Agusan del

sur State College of Agriculture and Technology, Bunawan Agusan del

sur. The samples underwent equal distribution when it came to

exposing them to the treatments which are the plant-derived

molluscicides and the control groups. This is done to determine if

Gmelina (Gmelina arborea) seed extract have the potential

molluscicidal effect against the golden apple snail (Pomacea

canaliculata L.).

Collection of Gmelina (Gmelina arborea) seeds

Among the many possible plant molluscicide is the plant that

is chosen by the researchers which is Gmelina (Gmelina arborea) which

are available inside of ASSCAT Agusan del sur State College of

Agriculture and Technology, Bunawan Agusan del sur. We also select

and carefully inspect the condition of Gmelina (Gmelina arborea) seeds.

The amount of the plant materials to be gathered is no less than 5 kg

since sufficient amount of extract is needed for the experiments

which is done in 3 replications for the range-finding test and 5

replications for the actual experimentation.

Preparation of equipment and tools.

Since the experiment will be conducted under laboratory

conditions, laboratory tools and equipment are use. Volumetric flask,

100 mL to 500 mL beakers, and other tools must be sanitize before

usage. Distilled water is use to rinse the tools and equipment to

lessen the presence of contaminants, thus deducting other factors

that could affect the experiment. It is also a necessary precaution to

ensure the accurateness of data to be collected.

Mortality count.

In partial experimentation (3 replicates per treatment) prior to the

actual experimentation. There are three concentrations for Gmelina

(Gmelina arborea) seed extract (50 ml/L, 100 ml/L, 150 ml/L) tested

and applied for the actual experimentation. There are also a total of 330

Golden Apple Snail samples used for the partial experiment. For the

actual experiment, 25 transparent containers (5 replicates per

treatment) were labeled (e.g. Treatment 1- 50 ml/L Gmelina seed extract)

(trial 1). Ten golden apple snails were distributed randomly to every

container, with a total of 250 golden apple snails used during the
actual experimentation. The mortality of the snails are continuously

observed for the succeeding 72 hours. The test subjects were first

exposed to the negative control, dechlorinated water. The treatment

groups will be applied to the snails by preparing each amount of plant

extract solution. The positive control, metaldehyde, is also prepared to

its designated sample. Following the first 24 hours of application of

the treatments, the mortality of snails will be checked through the

use of a metal needle. By poking, it will reveal if the snail is still alive

since a live snail should quickly respond and show muscle contractions.

The number of confirmed mortalities will be removed and recorded

on the laboratory sheet. Once the number of mortalities are recorded,

the dead snails will be separated, crushed and disposed as instructed

by the standard protocol for disposing gastropods (WHO, 1983). The

data gathering will be repeated in 24 h intervals.

DATA ANALYSIS

Data obtained were analyzed by one way analysis of variance

(ANOVA) and means were compared by Duncan’s New Multiple Range

test using SPSS 15.0 version and means were separated by least

significant differences (P≤0.05), (Adzitey, et al. 2015).

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