"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
Effectiveness of Madre de Cacao leaves extract Gliricidia
sepium, Guava leaves extract Psidium guajava & Natural
Vinegar Dilute acetic acid in eradicating mosquito wrigglers
Culicidae diptera
John Michael Sasan
Stephanie T. Gamboa
Jermaine L. Gadiano
Anthony John C. Edoloverio
Clint MJ O. Camangyan
PAU Excellencia Global Academy Foundation, Inc., Cebu, Philippines
Abstract: Given the high cost, health, and environmental dangers associated
with currently available synthetic pesticides, finding and developing economical,
safe, and effective alternatives is critical. The goal of this research is to develop an
organic mosquito repellent that will assist to reduce the number of mosquitos in the
environment. The experiment lasted a total of 24 hours. Each treatment included
three replicates with six larvae grouped in Parallel Group Design, and each treatment
had varied doses of Madre de Cacao leaf extract, guava leaf extract, and natural
vinegar. The level of efficiency of Madre de Cacao leaves extract, Guava leaves
extract, and natural vinegar as a larvicide was determined using the Mean. At the 0.05
level of significance, the One-Way Analysis of Variance (ANOVA) was performed
to assess the significant difference in the degree of efficacy of Madre de Cacao leaf
extract, guava leaf extract, and natural vinegar at various doses. The presence of
bioactive substances such as alkaloids, saponins, tannins, flavonoids, and coumarins
on mosquitoes suggests that Madre de cacao leaf extract, guava leaf extract, and
natural vinegar might be a possible natural larvicide and an alternative to commercial
mosquito pesticide. To minimize the undesirable side effects of conventional
pesticides, the researchers recommend using Madre de cacao leaf extract as a natural
larvicide on home insects, especially in locations where the plant grows abundantly
and in moist areas where mosquitoes are widespread. Other portions of the Madre de
Cacao and Guava should be used instead of the leaves, according to further study.
Keywords: Biopesticide, Gliricidia sepium, Acetic acid, Culicidae diptera
INTRODUCTION
Mosquitoes are one of the most common insects that harm people by delivering
illnesses through their bites. Mosquitoes are responsible for millions of fatalities
worldwide each year. Female mosquitoes require blood to nourish their eggs,
www.openscience.uz 21
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
reproduce, and spread illnesses by sucking blood. Dengue and hemorrhagic fever is a
terrible disease spread by mosquitoes that is caused by a virus. Mosquito
management is necessary to prevent the spread of mosquito-borne illnesses and to
enhance the environment and public health. (Gosh, etc., 2012). To reduce the quantity
of mosquitos, a range of mosquito control measures can be employed. Commercial
larvicides (pesticides that kill mosquito larvae) may be used by local governments to
prevent mosquitos from growing into adults.
To aid the government in battling the illness and eradicating mosquito carriers,
the researchers looked at the effects of Madre de Cacao (Gliricidia sepium), guava
leaves extract (Psidium guajava), and natural vinegar (Dilute acetic acid) on mosquito
larvae. According to research, the herb Madre de Cacao serves as an insect repellent
and has the capacity to eradicate a wide range of insects that might spread illnesses.
Commercial and man-made poisons are still routinely utilized to kill pests that are
potential transmitters, but they are costly. The researchers got interested in using
natural pesticides generated from plants as a form of pest management as a result of
the hazards of high-toxic chemical compounds, which led to the production of this
study. The study's major goal was to see if Madre de Cacao (Gliricidia sepium),
Guava leaves extract (Psidium guajava), and natural vinegar (Dilute acetic acid) were
successful in killing mosquito wrigglers (Culicidae diptera).
THE PROBLEM
Statement of the Problem
The study aims to determine if the solution of Madre de Cacao leaves extract,
guava leaves extract, and natural vinegar is effective in eradicating mosquito
larvicide. Its goal is to provide answers to the following questions:
1. What are the insecticidal properties of Madre de cacao leaves extract, guava
leaves extract, and natural vinegar against mosquito larvae?
2. What is the level of effectiveness of Madre de cacao leaves extract, guava
leaves extract, and natural vinegar as a mosquito larvicide?
3. Is there a significant difference in the level of effectiveness between the
Madre de cacao leaves extract, guava leaves extract, and natural vinegar as a
mosquito larvicide solution at different concentrations?
Null Hypothesis
H01: Tannins and coumarins, a toxic chemical found in Madre de cacao leaves,
may kill practically any pest or bug. Furthermore, the presence of various bioactive
compounds such as alkaloids, saponins, tannins, and flavonoids may contribute to the
mosquito larvae's susceptibility to guava leaf extract as a lethal agent. Finally, the
vinegar's acidity is strong enough to kill a variety of pests.
www.openscience.uz 22
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
H02: The 100% concentration of Madre de cacao leaves extract, guava leaves
extract, and natural vinegar has a higher level of effectiveness as a mosquito
larvicide.
H03: There is no significant difference in the level of effectiveness of Madre de
cacao leaves extract, guava leaves extract, and natural vinegar as a mosquito larvicide
at different concentrations.
SIGNIFICANCE OF STUDY
The goal of this research is to see if Madre de Cacao leaf extract, guava leaf
extract, and natural vinegar solution can be used to kill mosquito larvae and might be
a viable alternative to commercial larvicides. The following people, according to the
researcher, will profit from this study:
Environmental health workers. The outcomes of the study will give basic
knowledge and information on the components of Madre de Cacao leaf extract, guava
leaf extract, and natural vinegar solution as a possible mosquito larvicide. It will
assist environmental health professionals in identifying alternate methods for
reducing mosquito populations in the neighborhood and preventing mosquito-borne
illnesses.
Government. The findings of this study will aid in reducing government
spending on mosquito eradication.
Community. This research will give individuals with a natural and affordable
mosquito larvicide made from Madre de Cacao leaf extract, guava leaf extract, and
natural vinegar solution. This method will help reduce the number of people infected
with the Zika virus, dengue fever, and malaria in the community, especially in
mosquito-infested regions.
Researchers. The findings of this study will serve as a starting point for
individuals interested in undertaking more research into the topic.
SCOPE AND LIMITATION
The efficiency of Madre de Cacao leaf extract, guava leaf extract, and natural
vinegar on mosquito wrigglers was investigated in this study. The experiment lasted a
total of 24 hours. Each treatment included three replicates with six larvae grouped in
Parallel Group Design, and each treatment had varied doses of Madre de Cacao leaf
extract, guava leaf extract, and natural vinegar.
At the 0.05 level of significance, the One-Way Analysis of Variance (ANOVA)
was performed to assess the significant difference in the degree of efficacy of Madre
de Cacao leaf extract, guava leaf extract, and natural vinegar at various doses.
MATERIALS AND METHODS
Instrumentation
The materials and equipment used were:
www.openscience.uz 23
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
• Graduated Cylinder - used to calculate the amount of extract and its %
concentration.
• Madre de Cacao leaves - were cut, mashed, and removed with a sieve.
• Guava leaves - the leaves were chopped, pounded, and extracted using a
strainer.
• Mortar and Pestle - used in pounding the Madre de Cacao leaves and guava
leaves to obtain the extract.
• Mosquito larvae - are wormlike eating forms that develop from domestic
mosquito eggs.
• Petri Dish - utilized as a container for setting up and treatments.
• Strainer - used in separating the extract from the blended Madre de Cacao
leaves and guava leaves.
Collection and Processing of Madre de Cacao Plant
The Madre de Cacao leaves were gathered from mountain areas like Matab-ang,
Toledo City, Cebu, Philippines. The leaves were washed and pounded using mortar
and pestle then the extracts were collected using a strainer. The extract was diluted to
various quantities by adding distilled water.
Procedure:
Culturing of Mosquito Larvae
For a period of five to seven days, mosquito larvae were cultivated. The
container was filled with water, and handfuls of dung were added to feed the
mosquitos. The mosquito eggs were tested regularly. If there were little eggs on the
surface of the water, they were transferred from the bucket to a container at room
temperature with freshwater resources and provided with fishnet for easy capture
until they reached their larval stage of development. The larvae appeared like lengthy
insects that wiggled when disturbed at this stage. After seven days, the larvae were
removed from the jar with a fishnet and employed as research subjects.
Treatment
The concentration was varied to twenty-five percent (25%), fifty percent (50%),
seventy-five percent (75%), and one hundred percent (100%) by volume respectively.
There were five treatments used. Treatment (1) is the control with zero percent (0%)
concentration of the extract. Treatment (2) uses a 25% concentration containing 10
ml of extract and 30 ml of distilled water. Treatment (3) uses a 50% concentration
containing 20ml of extract and 20 ml of distilled water. Treatment (4) uses a 75%
concentration containing 30 ml of extract and 10 ml of distilled water and lastly is
treatment (5) 100% containing 40 ml of an extract with no distilled water. Three (3)
replicates of the prepared Madre de Cacao extract were placed in a petri dish for each
treatment, for a total of fifteen (15) Petri dishes. Six (6) larvae were added to each
petri dish and were observed by the researchers for a duration of twenty-four (24)
www.openscience.uz 24
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
hours. The researchers counted and recorded the number of dead larvae in each petri
dish.
RESULTS AND DISCUSSION
Trial 1
Treatment 5 is the most successful, as seen in the table. The mosquito wrigglers
are reduced by the 100% concentration of Madre de Cacao leaves extract, Guava
leaves extract, and Natural Vinegar because of their potency as insect repellents. This
is also effective because of their characteristics, which make mosquito wrigglers
feeble. The higher the concentration of extract, the lower the survival rate of
mosquito wrigglers.
Treatment Percentage No. of wrigglers Time Survival
1 0% 6 30 min. 5
2 25% 6 30 min. 4
3 50% 6 30 min. 2
4 75% 6 30 min. 0
5 100% 6 30 min. 0
Trial 2
Treatment Percentage No. of Time Survival
Wrigglers
1 0% 6 30 min. 6
2 25% 6 30 min 6
3 50% 6 30 min. 4
4 75% 6 30 min. 2
5 100% 6 30 min. 0
According to trial 2, treatment (1) employed a 0 percent concentration of the
extract with a survival rate of 6 out of 6 wrigglers. Treatment #2 used a 25%
concentration containing ml of extract and 30 ml of distilled water, with 6 out of 6
wrigglers surviving. Treatment (3) employed a 50 percent concentration with 20 ml
of extract and 20 ml of distilled water, with 4 out of 6 wrigglers surviving. Treatment
(4) employed 75 percent of the concentration, with 30 ml of extract and 10 ml of
distilled water, and two out of six wrigglers survived. Treatment (5) employed a
100% concentration of extract in 40 mL of distilled water, with a survival rate of 0
out of 6 no. of wrigglers. Treatment (5) used 100% concentration containing 40 ml of
an extract with no distilled water with a survival of 0 out of 6 no. of wrigglers. The
more extract used, similar to treatments 1 and 2, the lower the survival rate of
mosquito wrigglers.
CONCLUSION
The study's findings led to the following conclusions:
1. From the results of the study, Madre de cacao leaf extract, guava leaf extract,
and natural vinegar can be potential natural larvicide with the presence of bioactive
www.openscience.uz 25
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
compounds such as alkaloids, saponins, tannins, flavonoids, and coumarins on
mosquitoes and can be an alternative substitute to commercial mosquito pesticide.
2. Environmental protection is now seen as critical. To be suitable, a pesticide
should not inflict excessive death on target creatures but should be environmentally
beneficial. Phytochemicals, which are reasonably safe, cheap, and easily obtainable
in many regions around the world, maybe useful. The study on Madre de cacao leaf
extract, guava leaf extract, and natural vinegar has proven success in the management
of mosquito larvae, and this technique is simple, affordable, and does not affect the
man or the ecosystem in general, and as such should be used in the control of
mosquito larvae.
3. The maximum concentration of Madre de Cacao leaves extract, Guava leaves
extract, and pure vinegar has the highest level of efficiency in killing mosquitos and
protecting our surroundings from the risks and diseases usually caused by mosquitos.
4. Because of its parricidal qualities, which are excellent in eliminating
mosquito wrigglers, the combination of Madre de Cacao leaves extract, Guava leaves
extract, and Natural vinegar has a higher level of efficiency than the other
concentrations.
RECOMMENDATION
We would like to propose this study to those who are interested in using Madre
de Cacao, Guava leaves, and Natural vinegar as mosquito repellents, as well as
finding other insects for whom the solution is relevant and conducting more research
on the solution.
The following suggestions are made:
1. For a less pressing procedure, use the bark of Madre de Cacao instead of the
leaves.
2. Instead of crushing, use a boiling procedure to ensure that the extract is as
effective as possible.
3. Add more Guava extract to the mixture for efficacy.
ACKNOWLEDGEMENTS
First of all, we would like to express our sincere gratitude to our Practical
Research adviser, Mr. John Michael V. Sasan for the endless support of our
experimental research, his patience, motivation, and immense knowledge. His
guidance helped us throughout the journey. We could not have imagined having a
better adviser and mentor for this research.
Besides our Research adviser, we would also like to thank our Principal, Dr.
Francisca T. Uy. For encouraging and reminding us to do our best. To our family for
supporting us spiritually, emotionally, and financially. We also thank our fellow
group mates through thick and thin. And most of all, we are very thankful to our
Almighty Father for giving us strength, knowledge, courage, and guidance to fulfill
www.openscience.uz 26
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
this research. Without them, we can't accomplish this. Thank you very much. We
love you from the bottom of our hearts.
Reference
de Guzman, De Ungria, Dominguez,Francisco, Maddatu, Madrid, Ortega,
Andres, Ulic-Ulic, Yim. (2018, March). KAKAWATE LEAVES (GLIRICIDIA
SEPIUM)AS MOSQUITO COIL. Coursehero.
https://www.coursehero.com/file/44661168/devdocx/?fbclid=IwAR2qN
Borreli, L. (2015, September 15). 6 Health Benefits Of Guava Leaves, From
Treating Diarrhea To Cancer. Medical Daily. https://www.medicaldaily.com/6-
health-benefits-guava-leaves-treating-diarrhea-cancer-
353666?fbclid=IwAR25f1ZlFgSPpOXhZigsDRMlMtUq7K6v6lVIuVMCW0BDeqw
nmdHoPdFtkbU
Review of Literature on Medical Properties of Gliricidia Sepium (kakawate).
(2020, July 14). Grades Fixer. https://gradesfixer.com/free-essay-examples/review-
of-literature-on-medical-properties-of-gliricidia-
sepium/?fbclid=IwAR2wQP7DKtoW5MoKcH9RtGZhcaBWHPgq8uRdlnYFS51x7B
AENd-EO6ylBTs
Faluyi, O. (2020, February 16). many ways guava leaves are beneficial to your
health. Course Hero. https://punchng.com/many-ways-guava-leaves-are-beneficial-
to-your-health/
Antonio, Antoy, Lumogda. (2014). Larvicidal Effect of Madre de Cacao Leaf
Extract (Gliricidia Sepium) on Mosquito Larvae (Culex Pipiens). Blogspot.
http://chzarmyn.blogspot.com/2014/
Lindsay Falvey, J. (2012, April 5). GLIRICIDIA MACULATA—A REVIEW.
Taylor&Francis Online.
https://www.tandfonline.com/doi/abs/10.1080/01435698.1982.9752735
Torres, C. (2020, January 8). What Makes Guavas So Healthy and How to Reap
Their Benefits. EMediHealth. https://www.emedihealth.com/nutrition/guava-benefits
Choudhary, T. (2019, March 14). 17 Best Benefits Of Guava Leaves For Skin
And Health. StyleCraze. https://www.stylecraze.com/articles/benefits-of-guava-
leaves-for-skin-hair-and-health/
Guava – Inexpensive Healthiest Fruit from Nature. (2021, August 27).
NatureLoc. https://healthyliving.natureloc.com/guava-inexpensive-healthiest-fruit-
from-
nature/#:~:text=One%20guava%20fruit%20contains%2045,heart%20disease%20and
%20high%20cholesterol.
Blogger. (2018, June 12). WHAT’S WITH THE GUAVA LEAVES? Tsena.Co.
November 2, 2021, from https://tsena.co.bw/blogs/whats-guava-leaves
www.openscience.uz 27
"Science and Education" Scientific Journal / ISSN 2181-0842 January 2022 / Volume 3 Issue 1
Guava – Health Benefits You Didn’t Know. (2016, August 3). Thingsguyana.
https://www.thingsguyana.com/guava-health-benefits-you-didnt-know/
Saep ,Domino,Arranguez. (n.d.). The Antifungal Property of Madre de Cacao
(Gliricidia sepium) Leaves Extract: Its Effects on Skin Diseases. Academia.
https://www.academia.edu/8147468/The_Antifungal_Property_of_Madre_de_Cacao
_Gliricidia_sepium_Leaves_Extract_Its_Effects_on_Skin_Diseases
Cadsawan , A.D., Olayta, J. (2020, March 13). Biopesticidal Effects of Madre
De Cacao (Gliricidia sepium) and Makabuhay (Tinospora rumpii) Extracts on
Arugula (Eruca sativa). Omicsonline. https://www.omicsonline.org/open-
access/biopesticidal-effects-of-madre-de-cacao-gliricidia-sepium-and-
makabuhaytinospora-rumpii-extracts-on-arugula-eruca-sativa.pdf
Kayedee. (2015, August 13). Nature Knows Best. PaperCramp.
https://www.papercamp.com/essay/169514/Nature-Knows-Best
Alingayao, C., Parado, M. (2008, July 22). Larvicidal Activity of Aqueous
Gliricidia sepium Madre de Cacao) Extract Against Mosquito Larvae. Blogspot.
https://www.papercamp.com/essay/169514/Nature-Knows-Best
Coimbra, J. Soares, A. Garrido, M. Sousa, C. Ribeiro, F. (2006, July). Toxicit of
plant extracts to Scutellonema bradys. Researchgate.
https://www.researchgate.net/publication/262704784_Toxicity_of_plant_extracts_to_
Scutellonema_bradys
V, Santos , E, Caixeta , E, De Campos Jr., B, Pereira. (2017, January 17).
Ecotoxicological effects of larvicide used in the control of Aedes aegypti on
nontarget organisms: Redefining the use of pyriproxyfen. J Toxicol Environ Health A
https://pubmed.ncbi.nlm.nih.gov/28095184/
Wick, K.; Bond, C.; Buhl, K.; Stone, D. 2012. Methoprene General Fact Sheet;
National Pesticide Information Center, Oregon State University Extension Services.
http://npic.orst.edu/factsheets/methogen.html.
www.openscience.uz 28
