Exploring the Potential of Calamansi (Citrofortunella microcarpa) Fruit

Extract as a Molluscicide Against Golden Apple Snail (Pomacea

canaliculata)

Proponents:

Bañas, Giolo Steffano F.

Mangupit, April Rose P.

In Fulfillment of the Requirements in Research Capstone

Lamba National High School

Science, Technology, Engineering, and Mathematics Strand

Sitio Matlong, Barangay Lamba, Banga, South Cotabato

A.Y. 2023-2024

May 2024
Abstract

The golden apple snail, originally from Argentina and introduced to farmers in the
Philippines via Taiwan in the 1980s, has emerged as an unexpected rice pest in several
Asian countries. In the Philippines, its presence has resulted in substantial economic
losses to aquatic crops, not accounting for additional damages to human health and
natural ecosystems. Additionally, its control has led to environmental concerns, as
resource-limited farmers resort to indiscriminate and toxic agrochemicals, exacerbating
ecosystem pollution, endangering their health, and contributing to the decline of aquatic
biodiversity.

The study finds that exposure to Treatment 3, containing 6mL of C. microcarpa fruit
extract, leads to snail mortality within 24 hours, indicating its impact on snail health. As
exposure time increases, so does the number of dead snails, suggesting a correlation
between exposure duration and mortality rate. This demonstrates the potential of C.
microcarpa extract as a molluscicide for controlling Golden Apple Snail populations,
urging further research into its mechanisms and application for pest control.

The study investigates the use of C. microcarpa as an alternative molluscicide to control

Golden Apple Snail populations. Treatment 1, not involving C. microcarpa, achieved
nearly 100% snail mortality within 72 hours, indicating its effectiveness. However, as the
study focuses on C. microcarpa's potential, further research is needed to compare its
efficacy with Treatment 1 and explore any advantages or disadvantages it may offer.

This study highlights an effective treatment method for controlling Golden Apple Snail
populations, involving exposure to 6mL of C. microcarpa fruit extract over 72 hours. The
extract's efficacy is attributed to specific chemical compounds present. Researchers
suggest further tests with different extract concentrations to determine optimal
molluscicidal activity. They also recommend exploring its potential against other pest
species, emphasizing the importance of thorough scientific inquiry to understand its
efficacy and broader applications in pest management.
 Introduction

In recent decades, every administration has emphasized the importance of

achieving food self-sufficiency for the nation. This commitment has translated into a
focus for political leaders and agricultural authorities on domestically producing an ample
quantity of rice, the staple food, to meet the needs of the growing population (Arsenio,
B., 2003).

The 2008 rice crisis presented significant hurdles for Philippine policymakers.
They faced the task of securing a sufficient rice supply by importing it during periods of
escalating global prices. Concurrently, they needed to stabilize domestic rice prices.
Since rice plays a central part in the political economy, stabilizing rice prices ranks as
one of the government’s highest policy objectives (Calvo, P., 2014).

The golden apple snail, originally from Argentina and introduced to farmers in the
Philippines via Taiwan in the 1980s, has emerged as an unexpected rice pest in several
Asian countries. In the Philippines, its presence has resulted in substantial economic
losses to aquatic crops, not accounting for additional damages to human health and
natural ecosystems. Additionally, its control has led to environmental concerns, as
resource-limited farmers resort to indiscriminate and toxic agrochemicals, exacerbating
ecosystem pollution, endangering their health, and contributing to the decline of aquatic
biodiversity (R.C. Joshi, 2007).

Calamondin (Citrofortunella microcarpa) fruits or locally known as Kalamansi is

widely cultivated in the Philippines and is used as a condiment almost in every famous
dish made in the Philippines (Morte, M. Y., 2017). Citrus, a genus encompassing various
fruits like oranges and lemons, has long been celebrated for its manifold health benefits,
supported by extensive pharmacological research. Among these citrus varieties,
calamansi (Citrus microcarpa) stands out for its remarkable antibacterial properties,
which have been scientifically documented (Husni, E., 2021).

Investigating the effectiveness of calamansi fruit extract (Citrofortunella

microcarpa) as a possible molluscicide against the golden apple snail (Pomacea
canaliculata) is the main goal of this study. The project aims to investigate the feasibility
of using Calamansi extract as a technique of limiting the population of golden apple
snails in agricultural settings through extensive experimentation and analysis. By
conducting this investigation, the study hopes to advance environmentally sound and
sustainable approaches to controlling pest populations, which would ultimately boost
agricultural output and environmental preservation initiatives.
 Literature Review

This literature review provides a concise overview of research on natural

compounds as agents against agricultural pests, focusing on the golden apple snail
(Pomacea canaliculata). With a spotlight on calamansi (Citrofortunella microcarpa) fruit
extract, it examines the potential of botanical extracts for sustainable pest management
in agriculture, aiming to offer insights into their efficacy and environmental implications
for advancing sustainable agricultural practices.

Pomacea canaliculate (Golden Apple Snail)

The Golden apple snail, also known as Pomacea canaliculata, is a fascinating

creature with a rich cultural and culinary significance in Southeast Asia. Its distinct spiral-
shaped shell, ranging from light brown to golden hues, adds aesthetic charm to both
culinary dishes and freshwater habitats. Despite its small size, typically measuring
between 2-3 centimeters in diameter, the golden apple snail packs a flavorful punch,
boasting tender, succulent flesh with a delicate seafood-like taste. Found predominantly
in warm freshwater ecosystems like rice fields and ponds, these snails thrive amidst
ample vegetation, contributing to the biodiversity of their habitats (Braganza, L., 2024).

Beyond their culinary appeal, Golden apple snail offer additional health benefits.
Widely used in regional cuisines across Southeast Asia, these mollusks are not only
delectable but also nutritious. With their protein-rich meat and potential medicinal
properties, consumption of Golden apple snail presents a holistic approach to well-being,
bridging cultural heritage with nutritional value and sustainability.

Golden apple snail (GAS) is not a serious pest in its native range because of
ecological balance with population controlling mechanisms already existing in those
environments. Sriyani Wickramasinghe cited other studies stating that GAS becomes a
pest in the absence of natural enemies such as predators and parasites
(Wickramasinghe, S., 2006).

Citrofortunella microcarpa (Calamansi)

Calamansi or calamodin is very abundant and one of the sources of staple fruit
juice in the Philippines. It is grown principally for its fruit juice, since it is widely known as
good source of Vitamin C (Acero, LH., 2017).

According to Tamaray, M., et al. (2020), calamansi has the potential as a

microbial agent for aquaculture use. The presence of major compounds, limonene had
contributed immensely to the aroma, antibacterial, antifungal, and insect repellant
properties.

Citrus is a plant that has many health benefits and has been widely reported for
its pharmacological effects. One of the citruses that has been reported to have
antibacterial activity is calamansi. The study of Fitri Yeni et. al. of 2021 aims to determine
the chemical content profile of the essential oil of fruit peels and leaves of calamansi.
The study shows that the main components of essential oil in the peels of the calamansi
are D-limonene, Citronellal, Cyclopentene, and Citronellol. While the calamansi leaves
consist of Citronellal, Citronellol, Carene, and Phellandrene. Hence, the essential oil of
the fruit peels and leaves of calamansi showed antibacterial activity against MRSA.
Molluscicidal Activity

Some snail species pose a serious threat for human health, economy, and the
environment due to their widespread distribution and the transmission of dangerous
parasites causing, among others, schistosomiasis and fascioliasis. Scientists from
around the world have been studying the effects of plant extracts on snails for many
years in order to find an alternative to molluscicides of synthetic origin. Especially, the
study of Sebastian Wszelaki, et al. of 2023 main purpose was to collect the results
obtained so far on the effect of plant alkaloids on snails. Results shows that plant
alkaloids can be extremely useful in the fight against problematic species of snails and
cause much lower harm to the environment than synthetic molluscicides.

According to Christopher Boyd, 2015. Molluscicides are substances designed to

eliminate mollusks, a diverse group of invertebrate animals numbering in the tens of
thousands. Among mollusks are octopi, squid, as well as slugs and snails, which are the
primary targets of molluscicides. Slugs and snails are well-known by farmers and
gardeners as some of the most annoying and destructive pests. They can destroy the
leaves and fruit of a large variety of plants, old and young.

The study of Nur Suraya of 2020 was conducted to investigate the molluscicidal
activity of four different Ipomoea batatas leaf extracts against Pomacea canaliculata and
screen the phytochemical compounds of I. batatas leaf extracts. For all extracts, results
showed a positive relationship between the concentration and the golden apple snails.
Addition, the study concludes that the I. batata leaf extracts have potency in controlling
golden apple snails especially I. batata methanol leaf extracts that can be used as an
alternative molluscicides.
 Methodology

The methodology involves systematically evaluating Calamansi fruit extract's

effectiveness against Golden Apple Snails. Steps include extract preparation, controlled
experiments, data analysis, and discussion. This study aims to provide insights into
using Calamansi as a sustainable solution for pest management.

Extract Preparation

The C. microcarpa was gathered in Purok Sampaguita, Sitio Matlong, Lamba,

Banga at Dela Cruz’s residence on May 09, 2024. The C. microcarpa fruit was washed
and cleaned using cold water and sliced using a knife. Extracting the fruit by squeezing
the fruit by hand and strained to avoid mixing the seeds with the juice. Preparing and
extraction of C. microcarpa was conducted and observed in Sumbria’s residence on May
09, 2024.

Controlled Experiments

Golden Apple Snails are exposed to varying concentrations of Calamansi fruit

extract while being monitored for behavior and mortality rates.

Treatment 1 – No Extract (Control)

Treatment 2 – 25% Extract (3mL of C. microcarpa fruit extract)

Treatment 3 – 50% Extract (6mL of C. microcarpa fruit extract)

Treatment 4 – 75% Extract (9mL of C. microcarpa fruit extract)

This setup includes establishing control groups for comparison and repeating
trials for reliability. Statistical analysis is employed to assess the differences in mortality
rates between treatment groups and controls.

Data Gathering

The mortality rate of the P. canaliculata snail were observed and recorded every
24 hours for 3 days.

Preparation of different treatments

A volume of 50mL paddy field water was poured into containers with two tested
snails each treatment. The mortality rate was observed every 24 hours for 3 days. Each
treatment has different volume of extract poured in every container using a syringe.
 Results

This part is the presentation of the data through the results of the experiment.
Tabular presentations of the data are furthermore presented for comprehensive analysis
and interpretation.

Molluscicidal Activity

Treatment Percentage of Mortality (%)

Concentration (mL)
24hrs 48hrs 72hrs
No treatment (C. 0 0 0
microcarpa fruit extract)
3mL of C. microcarpa 0 100 100
fruit extract
6mL of C. microcarpa 100 100 100
fruit extract
9mL of C. microcarpa 0 0 100
fruit extract

Table 1. Result of Mortality Rate of P. canaliculata

This observation stems from an experiment where Golden Apple Snails (GAS)
were subjected to different treatments involving C. microcarpa fruit extract.

Firstly, after 24 hours of exposure to Treatment 3, which utilizes 6mL of C.

microcarpa fruit extract, signs of snail mortality become apparent. This indicates that the
C. microcarpa extract has a noticeable impact on the snails' health and viability, leading
to their eventual demise.

Additionally, as the duration of exposure to Treatment 3 increases beyond the

initial 24 hours, the number of deceased snails continues to rise. This suggests a
correlation between the duration of exposure to the C. microcarpa extract and the
mortality rate of the snails. Longer exposure times result in a greater number of snails
succumbing to the effects of the extract.

Overall, these observations highlight the potential of C. microcarpa fruit extract

as a molluscicide for managing Golden Apple Snail populations. It also emphasizes the
importance of further research to understand the mechanisms underlying the extract's
molluscicidal activity and to optimize its application for effective pest control.

Discussion

This statement describes a scientific study that examined the effectiveness of

using C. microcarpa, a specific plant or compound, as an alternative to traditional
molluscicides for controlling the population of P. canaliculata, also known as the Golden
Apple Snail. Molluscicides are substances used to kill or control mollusks, such as snails
and slugs, which can be agricultural pests.

The results of the study indicate that Treatment 1, which did not involve the use
of C. microcarpa, showed promising molluscicidal activity. Specifically, it took 72 hours
for the treatment to achieve an almost 100% mortality rate among the Golden Apple
Snail population.

This suggests that Treatment 1, possibly involving another molluscicide or

method, was highly effective in controlling the snail population within the given
timeframe. However, it's important to note that the study was investigating the potential
of C. microcarpa as an alternative, implying that further research is needed to compare
its effectiveness with Treatment 1 and to explore any potential advantages or
disadvantages it may have.

Conclusion

The statement highlights a specific treatment method that proved effective in

controlling the population of P. canaliculata, the Golden Apple Snail. The treatment
involved exposing the snails to 6mL of C. microcarpa fruit extract over a period of 72
hours. The effectiveness of this treatment can be attributed to the chemical compounds
present in the C. microcarpa fruit extract, including carane, cyclopentanone, cychexanol,
decanal, L-alpha-Terpineol, and D-Crvone.

The researchers suggested further investigation into the gathered data by

conducting additional tests using different concentrations of C. microcarpa fruit extract.
This would help determine the optimal concentration required to achieve maximum
molluscicidal activity against P. canaliculata. Additionally, they recommended exploring
the potential of C. microcarpa fruit extract as a molluscicide against other pest species,
indicating its broader applicability beyond just Golden Apple Snails.

This recommendation emphasizes the importance of thorough scientific inquiry to

fully understand the efficacy and potential applications of C. microcarpa fruit extract as a
molluscicide. Further experimentation and analysis can provide valuable insights into its
effectiveness, safety, and practical use in pest management strategies.
 References

Arsenio, B., 2003. The Rice Problem in the Philippines: Trends, Constraints, and Policy
Imperatives. MPRA Paper. Link: https://mpra.ub.uni-
muenchen.de/24865/?fbclid=IwAR33Fp-qMXFGYr3i4vt1R64dStOkjJTPeUp-
CoJqOnVJE6HVvO0BLt6lT5E

Calvo, P. A., et al., 2014. Trade Policies, Household Welfare, and Poverty Alleviation.
Case Study. Link: https://unctad.org/system/files/official-
document/gds2014d3_en.pdf?fbclid=IwAR0UfCb1IVpqUXMoi_mssPM6T16v
2VTj78qQYFuQ5vYJB-Ego0X6XhSE0vE#page=40

Joshi, R. C., 2007. Problems with the Management of the Golden Apple Snail Pomacea
canaliculata: An Important Exotic Pest of Rice in Asia. Conference Paper. Link:
https://link.springer.com/chapter/10.1007/978-1-4020-6059- 5_24?
fbclid=IwAR0DNvYgmHyxHUsJ4ETxf54psWNmNEcsoD5PX2qudPW4
wawMSYl37FVIhZU

Acero, L. H., and Morte, M. Y., 2017. Potential of Calamansi (Citrofortunella

microcarpa) Fruit Peels Extract in Lowering the Blood Glucose Level of
Streptozotocin in Induced Albino Rats (Rattus albus). Journal. Link:
http://www.ijfe.org/uploadfile/2017/0816/20170816113539776.pdf?fbclid=IwA
R05H_6c-hrvvMOE8n5UgtfJSASg_cnDgtQCMOaAU0F2Sk8313wLWUxuxCo

Fitri, Y., et al., 2021. Chemical Contents Profile of Essential Oil from Calamansi (Citrus
microcarpa Bunge) Peels and Leaves and Its Antibacterial Activities. Journal.
Link: https://www.atlantis-press.com/proceedings/iccscp- 21/125962517?
fbclid=IwAR30jaaFS0I9cPNLzzP8lBn1K2E- IhUwK6cMBdtKlZt3IJmZFn_UDSgYQa0

Braganza, L., 2014. Golden Apple Snail: 4 Health Benefits of Golden Kuhol, Description,
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apple-snail-4-health-benefits-of-golden-kuhol-description-and-
disadvantages/?fbclid=IwAR0_WSezBJiMxjQZYqcMJfv3u4vfd1yEItERp0aMJBfR
oyDWpzE6hUUwNP8

Wickramasinghe, S., 2006. Golden Apple Snails in the World: Introduction, Impact, and
Control Measures. Blog. Link:
https://www.researchgate.net/publication/312976059_Golden_apple_snails_in_th
e_world_introduction_impact_and_control_measures

Nur S., et al., 2020. Molluscicidal Activity of Ipomea batatas leaf extracts against
Pomacea canaliculata (Golden Apple Snail). Journal. Link:
https://pdfs.semanticscholar.org/6287/7bad83e72d9422a1c03430bc96d02e04da
1e.pdf?
fbclid=IwAR1Y0Tub0cY4wAfEOlprxP9_JnTRp3MKjBqmZwW0Bfm0XDgb a31hbV3j_Sc

Boyd, C., 2015. What is a Molluscicide? Journal. Link:

https://www.chemservice.com/news/what-is-a- molluscicide/?
fbclid=IwAR0lBvxyu3dlZejakMAUVNm0-
pcDMOp0mLr_WYvruq_H76GnzuP0WVTUcSg
A
P
P
E
N
D
I
C
E
S
 Appendix A
Gathering of Materials

Appendix B
Preparation of Fruit Extract
 Appendix C
Preparation of Different Concentration

Appendix D
Exposure of P. canaliculata to different concentrations
 Appendix E
Observation and Data Gathering