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UNIVERSITY OF SOUTHERN PHILIPPINES FOUNDATIONCollege of PharmacyThe Effect of Pounded Allium sativum (Garlic) Clovesin Water as Mosquito RepellentA Thesis Presented to the Faculty of the College of Pharmacy in partial fulfillment of the course Bachelor of Science in PharmacyBy:Name of researchersMay...

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UNIVERSITY OF SOUTHERN PHILIPPINES FOUNDATION College of Pharmacy

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The Effect of Pounded Allium sativum (Garlic) Cloves in Water as Mosquito Repellent

A Thesis Presented to the Faculty of the College of Pharmacy in partial fulfillment of the course Bachelor of Science in Pharmacy

By:

Name of researchers

May, 2021

Acknowledgement This research was supported by University of Southern Philippines Foundation, Lahug Cebu City. We would like to express our deepest gratitude to Mrs. Althea R. Arenajo, Pharmacy Research and Thesis Writing who provided valuable feedbacks and constant encouragement throughout the duration of the study. Her valuable suggestions and insights were of immense help. Her perceptive criticism kept us working to make this research study in a much better way. We also like to give our sincere gratitude to our family who provided financial support and motivation without which this research would be incomplete.

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Abstract The main purpose of this study was to determine the effects of pounded Allium

sativum(garlic)clove in water as mosquito repellent. In this study, the effects of pounded garlic clove in water was also compared to the commercially available mosquito repellent in the market which was the Baygon spray. A quasi-experimental design was selected in order to identify the repellent effects of pounded garlic clove in water in comparison to the Baygon spray. The researchers prepared three set-ups. Each set-up has twenty mosquitoes, a spoiled milk and a container having - a pounded garlic clove in waterplaced in the first set-up, baygon sprayed on the containerin the second set-up and tap water alone in the third set-up. The mean number of mosquitoes and the distance travelled as they repelled were identified under an 8-hour observational period.The data collected were analyzed using a one-way ANOVA. The results found out that pounded garlic clove in water can repel mosquitoes. However, its repellent effects were lesser and is not comparable to the effects of Baygon spray. There is a significant difference that exist in the mean number of mosquitoes and the distance travelled between the variables being studied.

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Table of Contents Page Number Acknowledgement

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Abstract

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Chapter I – The Problem and Its Setting Introduction 5 Review of Related Literature and Studies Related Literatures

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Related Studies

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Conceptual Framework

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Statement of the Problem

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Null Hypothesis 25 Significance of the Study 26 Definition of Terms 28 Chapter II – Methodology Research Design 31 Research Subject31 Research Locale 31 Ethical Consideration 32 Data Gathering Procedure 32 Statistical Treatment 33 Chapter III – Results and Discussion 34

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Chapter IV – Summary of Findings, Conclusion and Recommendation Summary of Findings

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Conclusion40 Recommendation 41 References

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Appendices Appendix A49 Appendix B

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Appendix C

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Chapter I The Problem and Its Setting Introduction Mosquitoes are tiny but terrible creatures. Many people acquire illnesses due to mosquito bites. Mosquitoes transmit diseases caused by virus, bacteria and parasites. Vector-borne diseases account for more than 17% of all infectious diseases, causing more than seven-hundred thousand deaths annually (World Health Organization, 2020). Dengue and malaria have the most number of infected people. According to the latest World malaria report, released on 30 November 2020, there were 229 million cases of malaria in 2019 compared to 228 million cases in 2018. The estimated number of malaria deaths stood at 409,000 in 2019, compared with 411,000 deaths in 2018. In 2019, nearly half of the world’s population was at risk of malaria. In the same year, the Philippines recorded 5,730 cases of malaria, which is caused mainly by plasmodium parasites carried by mosquitoes and spread through bites (Maru, 2020). From January 1 to November 28, 2020, there have been a total of 79,218 dengue cases with 306 deaths in the Philippines (World Health Organization, 2020). The Department of HealthEastern Visayas announces last July 20, 2020 a total of 4,221 dengue cases in the region with nine (9) deaths covering morbidity weeks 1-29 (Roca, 2020). Leyte province has the majority reported cases with a total of 2,160 cases (51.17%), 279 are from Tacloban City, 228 from Baybay City, and 158 from Ormoc City. There are 571 cases (13.53%) with 113 cases recorded in Maasin City, Southern Leyte province. Last 2019, Santa Fe, a town of Leyte was placed under a state of calamity due to rising dengue 5

cases (Lubiano, 2019). There are 101 cases that were recorder from May 15 to July 8 as reported by the municipal health office. Other viral diseases such as Zika virus, yellow fever and Chikungunya are transmitted by Aedes aegypti mosquitoes. The Eastern Visayas Department of Health also reported Chikungunya virus that afflicted hundreds of residents in San Jorge, Tarangnan, and Calbayog City in Samar, San Roque, Northern Samar; and Maripipi, Biliran (Amazona, 2018). The rising emergence of these diseases and the increasing number of infected people is very alarming to many people across the world. For many years, garlic is widely used as a flavoring agent for most people, but folks had also used garlic to repel many forms of insects either crawling or flying like mosquitoes. Garlic is used as a companion plant to deter pests (Charbonneau, 2017). Garlic can be used to treat toothache, reduce blood pressure and cholesterol. Apart from this, several studies have investigated the powerful natural repelling action of garlic. The pungent scent of garlic is known to repel mosquitoes. They do not like the smell of garlic. The amino acid in garlic is converted to allicin when it is blended or crushed. The resulting odor repulsed insects (Patterson, 2019). Dezeil (2020) stated that allicin is fatal to the larvae of some mosquito. Rubbing garlic on the skin or eating it are some traditional ways of using it to repel mosquito. In Matalom, Leyte garlic clovesare used to repel mosquitoes. Some household merely place garlic around their home while others crushed a garlic clove and placed it above a container filled with water. Most residents of Matalom, Leyte believed with the repelling action of garlic while others still doubt about it. 6

In the Philippines, the use garlic as mosquito repellent is gaining popularity. There are lots of bloggers who share their experience with the homemade garlic mosquito repellent. Filipinos protect their home from disease carrier mosquitoes and save money by making a do-it-yourself anti-mosquito measure. They do it by peeling the cloves, putting it in a spray bottle and adding isopropyl alcohol. Garlic produce allicin and the alcohol help disperse this compound. They spray it around the house or directly to their skin (Gonzales, 2014). In Singapore, mosquito-related illnesses are prevalent due to their hot and humid climate. To protect their home from harmful mosquitoes they put garlic on their flower beds or around the house. They cut and scatter garlic cloves around their yard or balcony area (Lim and Liew, 2020). In South Africa, sirinol (garlic emulsion) the repellent action of the emulsion against the insects was evaluated by the three methods including petri-dish bioassay technique, Y-shape olfactometer tube bioassay technique and the glass bioassay technique. At all level tested, it was found out that garlic emulsion had effective repellent and could be considered for integration with other effective control options in the management of L. serricorne and T. castaneum (Jahromi, et al., 2012). Recently, people are looking for natural alternative repellent for a lot a reason. One of these is that some skin is sensitive to the chemicals added to commercial mosquito repellents. N, N-Diethyl-meta-toluamide, also called as DEET is commonly found in mosquito repellents, though not everyone is keen to slather it on their skin. 7

People with damaged skin should not have DEET applied to it, because it may exacerbate problems like skin irritation (Heid, 2018). The formulation of mosquito repellent includes a list of harmful chemicals that could bring danger to human health if use for a long period of time. Several studies have investigated the toxic effects of mosquito repellent. In a study conducted by Ayorinde et al. (2014), toxic effects on the hematological parameters (higher in females) and histopathology of the lungs occurs due to prolong exposure to mosquito repellent. Mosquito coil on the other hand also causes difficulty breathing, headache, bronchial irritation, skin rashes and eye problems (Chavan, 2016). Another reason as to why people look for natural mosquito repellent is that a lot of families especially those from informal settler’s do not have enough money to buy mosquito repellent.

Vector-borne diseases is prevalent mostly in low-income

communitiesdue to poor housing conditions and weak community infrastructures. Not having enough money to buy mosquito repellent like baygon sprays and mosquito coils also contribute to increasing cases among financially challenged families. Mosquito-borne diseases continue to arise. People are prone to having mosquito diseases because of lack of financial and poor sanitation that fail to make their environment free from mosquitoes. The researchers’ objective in this study is to find out the effect of

garlic in driving away mosquitoes. It is assumed that pounding of garlic

placed on top of the water will keep away the mosquitoes. In this manner, residents in the rural areas can benefit from the findings of this study.

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Review of Relate Related d Literature and Studies Related Literature Mosquito repellent activity of garlic bulb in water is reviewed due to high cost and various reports on allergic reactions caused by commercially available mosquito repellent in the market. In this study, the researchers compare the difference and similarities on the potential effects of garlic bulb in water with the already available mosquito repellent in the market. Mosquito-borne diseases. The occurrence of mosquito-borne diseases is mainly seen and experienced in tropical and subtropical environments. All the country's regions tend to be vulnerable to the mosquito-borne illnesses transmission. The Zika virus, West Nile virus, Chikungunya virus, dengue, and malaria are diseases that are spread to people by mosquitoes. The mosquitoes that transmit these diseases are females. The most common viral infection transmitted by Aedes mosquitoes in the Philippines is dengue. More than 3.9 billion people are at risk of contracting dengue in more than 129 countries, with an estimated 96 million symptomatic cases and an estimated 40,000 deaths per year (WHO, 2020). After 3 to 7 days of being bitten by mosquitoes, a person may experience symptoms such as joint pain, rashes, high fever, headache, nausea, fatigue and vomiting (Mosquito Outbreak Manila: 5 Facts About The Mosquito Scare, 2021). To let people, know how to protect themselves and their communities from mosquitoes, it is necessary to provide information and increase public awareness.

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People who have knowledge about mosquito-borne diseases are less likely to have an infection. Mosquitoes Mosquitoes. Mosquitoes are among the deadliest insects around the world in terms of the number of people it kills annually. They are the most irritating and infectious species that humans encounter. They come in over 2,500 different species and can be found in any part of the world except Antarctica (Mosquitoes: World’s Deadliest Animal, 2017). According to the Environmental Protection Agency (EPA) (2018), there are about 200 different species of mosquitoes in the United States. The invasive mosquito, C. coronator is the primary vector of Saint Louis encephalitis and West Nile viruses, both were considered as public health concerns in America according to Wilke (2020). This mosquito has spread over the different places in the United States where it is commonly found in Mississippi, Alabama, Georgia, and Florida. There are 26,825 female C. coronator collected in Miami-Dade County (Florida) and were also detected in Tennessee and Virginia (Wilke, 2020). Mosquitoes are great burden and cause of diseases even in western countries. A lot of people’s lives are in danger due to continuous presence of invasive mosquitoes. Mosquitoes are quick learners; they can easily adapt to new environments where they are. However according to the World Health Organization (WHO) (2020), tropical and subtropical areas have the highest burden of vector-borne diseases where poor families are most affected. According to Khlystov (n.d.), tropical regions like Thailand, Brazil, Indonesia, Australia and Philippines, have the largest population of mosquitoes. Mosquito-borne disease is a current concern in the Philippines. In Metro Manila, a 10

survey of mosquito species in public schools resulted in the abundance of Aedes aegypti and Aedes albopictus in both elementary and high school (Mistica, etal., 2014). Mosquitoes are spread throughout the islands of the Philippines. Aedes aegypti, Aedes albopictus, Culex pipiens pipiens and Culex pipiens quinquefasciatus are the following species of mosquito which have been described in the province of Cebu (Greif etal., 2017). In Eastern Visayas, during August of the year 2020, the province of Leyte experienced the burden of disease caused by mosquitoes. It has the most reported dengue cases having 2,194 which is 50.9% but this was 73% lower than the recorded dengue cases from the same period of previous year. (Reyes, 2020). Mosquitoes prefer to breed and lay their eggs on stagnant water. Undrained water or unclosed water tanks and rainwater puddles are few of the places where they can survive. They can also thrive in any location even if it is not their natural habitat as long as it is an environment with a hot and humid atmosphere. There are also other species of mosquitoes living in freezing temperatures, such as mosquitoes belonging to the Anopheles, Culex and Culiseta genera. For Aedes aegypti, the condition that favored their reproduction is during dry season (March to May, inclusive) if a few heavy rains occur at intervals of 15–20 days; and during wet season (June to September) according to Siler et al. (1926). Having knowledge on when mosquitoes are most active can help us protect ourselves from its dangerous bite. The American Mosquito Control Association (AMCA) recommends following the 3D’s for mosquito protection. This 3D’s include the draining of empty water containers at least once per week as this will avoid any conditions that allow will breeding grounds for mosquitoes, dress or clothes that 11

have long sleeves, long pants, and light-colored, loose-fitting clothing should be worn to reduce the risk of mosquito bites and lastly is to defend humans from mosquito bites by applying approved repellent. Mosquitoes use a lot of clues in their search for a human target, it is either through human sweat or body heat or the carbon dioxide in human breath. Mosquitoes have heat sensors. Via their heat sensors, mosquitoes can detect their victims. When the environment's humidity is very high, the performance of their sensors increases by a factor of 3. This means that from 30 feet (or 6 meters) away, they can still sense humans (Mosquito Outbreak Manila: 5 Facts About The Mosquito Scare, 2021). Mosquitoes do not use their eyesight for preying, but instead, they can sense their target through carbon dioxide that people breath out. According to Kripena (2020), approximately 4–5% of the gas that humans exhale is CO2. The more CO2 dioxide that humans exhale, the more they can be recognized easily by mosquitoes. The olfactory receptor present in the antennae of mosquitoes serves as a detector of humans. This enables mosquitoes to smell chemicals present in human sweat. Kripena (2020) states that the lactic acid released by humans when they sweat is very attractive for mosquitoes. Humans who love to eat salty foods or potassium-rich foods are more likely to produce more lactic acid. Lactic acid as mosquito attrac attractant. tant. Milk contains lactose, which is a natural sugar that humans use for energy. When milk begins to spoil, the sweetness of it is changed to a sour flavor. Milk spoils due to an overgrowth of bacteria that compromises its 12

quality. According to Hill (2019), E. coli, Salmonella, and Listeria are one of the most harmful bacteria on milk that are killed by the pasteurization process. However, pasteurization is not enough to kill all bacteria. When a carton of milk is opened, it becomes contaminated with the bacteria from the environment. These bacteria will multiply and will cause the milk to sour, have a rancid odor and get spoiled. Aside from its taste and odor, the physical appearance of milk will also change into a slimy texture with somewhat yellow color. In spoiled milk, lactococci and lactobacilli bacteria thrive in due to constant and rapid temperature changes. Lactose is converted to glucose and galactose by bacteria, resulting in lactic acid. According to Rupp (2016), the pH of milk increases until such point where Streptococci cannot grow and Lactobacilli, a second class of bacteria, appears. The rising acidity literally sours the milk and causes the milk proteins to coagulate. The increased acidity in milk causes the milk proteins to coagulate. This is not the same to unpasteurized milk or what is known as raw milk. This raw milk is somewhat different from pasteurized milk because of its natural sour taste. Raw milk can naturally ferment. Lactococcus lactis, a lactic acid bacterium, ferments milk sugar (lactose) to lactic acid in (Todar, 2010). Lactococcus lactis is found naturally is raw milk and that is the reason why raw milk is sour. Lactococcus lactis uses enzymes to extract energy (ATP) from lactose when it is added to milk. Lactic acid is produced as a byproduct of ATP synthesis. In a study conducted in Western Kenya, milk products were used as bait to trap mosquitoes. African traditional milk cream has trapped a total of 1,806 mosquitoes that includes An. gambiae s.l, An. funestus and culicine mosquitoes (Owino, 2010).

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Garlic. Garlic (Allium sativum ) is a perennial herb in the amaryllis family (Amaryllidaceae) that is grown for its flavorful bulbs. It is plant that is native to central Asia, but it also grows naturally in countries like southern France and Italy. The bulbs of garlic have an onionlike aroma and pungent taste which explains why it is not eaten raw. Garlic is used in many ways by humans in their day-to-day lives has been known among people across the world for centuries. It has a distinctive aroma is caused primarily by organosulfur compounds such as allicin found in fresh garlic cloves and ajoene formed when garlic bulbs are crushed or chopped. It is simple to grow garlic and it can be grown all year in mild climates. Garlic thrives in absolute, direct sunlight and loose, fertile, well-drained soil (Hansen, n.d.). In planting garlic, it is very important to know first the soil pH. Hansen also stated that the ideal soil pH for garlic plant is in the near-neutral range of 6.0 to 7.0.1. Garlic grows from isolated cloves which have been broken off a whole bulb. Each clove can propagate in the soil, establishing a new bulb with 5-10 cloves. Despite of its easy production, the production of garlic in the Philippines has been declining. The largest garlic-producing region is the Ilocos Region but because of the low-yielding varieties and the decline in areas for plantation, the supply of garlic in the Philippines became insufficient. The Southeast Asian Regional Center for Graduate Study and Resear...