Cacao Leaves and Garlic as Mosquito Repelants

INTRODUCTION
“If you think you are too small to be effective, you have never been in bed with a mosquito.”

-Betty Reese

Mosquitoes are small but deadly. Everywhere you go, they are willing to suck your blood anytime. With their simple bites, they can cause you a great damage.

Garlic has not just been considered a special food for the unique flavor it imparts, but also for its medicinal value. It is thought that garlic may be an alternative mosquito repellent for humans as well. In a field study conducted in India, a preparation made of one percent garlic oil, petroleum jelly and beeswax that was rubbed on the arms and legs of the study subjects was found to be effective in preventing mosquito bites for up to eight hours. In addition, there is some evidence that heavy consumption of garlic through supplements or well-flavored foods may help ward off mosquitoes.
( http://www.news.colostate.edu/Release/390)

Nowadays, cacao is known for the production of cacao chocolate and cacao butter. It has shown its anti-HIV properties and its cacao fats have been used in many skin care products. But it can be also made as a mosquito repellant spray by its properties that are unappealing to mosquitoes.

The researchers came up with the idea of mosquito repellent spray to help our community lessen the number of dengue victims nationwide at the same time to make an eco-friendly repellent spray because it is made from natural products or it is made from organic substances which biodegrades in the environment. With this, we could help lessen the number of increasing dengue incidents, at the same time we could help people to breathe air safely.

BACKGROUND OF THE STUDY

There are a number of harmful side effects to most insect repelling products on the market today. The active ingredient in most of these products is called DEET (diethyl-meta-toluamide) which is so poisonous that even the Environment Protection Agency recommends the users to wash off their skin when they return indoors. Furthermore, the agency suggests to avoid breathing it in and not to spray it directly on face. Though DEET is used in hundreds of brands in varying concentrations, It is still a pesticide. Many consider DEET insect repellent to be harmless or even a beneficial mosquito repellent. Yet, it is a synthetic product that could harm mosquitoes it could also do damage to other living things.
(hhtp://www.mercola.com/article/pesticides/deet.html)

The mosquitoes are a family of small, midge-like flies: the Culicidae. Although a few species are harmless or even useful to humanity, most are considered a nuisance because they consume blood from living vertebrates, including humans. Mosquitoes have a battery of sensors designed to track their prey, including chemical, visual, and heat sensors. Typically, both male and female mosquitoes feed on nectar and plant juices, but in many species the mouthparts of the females are adapted for piercing the skin of animal hosts and sucking their blood as ectoparasites. In many species, the female needs to obtain nutrients from a blood meal before she can produce eggs, whereas in many other species, she can produce more eggs after a blood meal. Mosquitoes can act as vectors for many disease-causing viruses and parasites. Infected mosquitoes carry these organisms from person to person without exhibiting symptoms themselves. So all of us needs to be careful. (http://en.wikipedia.org/wiki/Mosquito)

Allium sativum (garlic) is a well known condiment or material in the kitchen to flavor the food. According to Tone Rico (http://wiki.answers.com/Q/What_do_mosquitoes_hate), garlic is a very good substance to odorize a person’s sweat that will deter mosquitoes away from you for it will secrete through the sweat glands and blanket the sweet smell of blood in the body. Hence, the researchers opted this project because a mosquito repellent spray made up from Theobroma cacao (cacao leaves) and Allium sativum (garlic) will remove the health problems caused by commercial mosquito repellents such as skin hives, redness and irritation (due to allergic reaction), a burning sensation leading to blistering and permanent scarring of the skin, nausea and vomiting, stomach irritation, mood swings, insomnia, disorientation or confusion, numbness, clumsiness and seizures (particularly in smaller children). Moreover, Theobroma cacao and Allium sativum are easy to find.

CONCEPTUAL FRAMEWORK

Theobroma cacao Allium Sativum (bulb

Fig. 1 Conceptual framework

STATEMENT OF THE PROBLEM The purpose of this study is to examine the effectiveness of Theobroma cacao leaves (cacao) and Allium sativum (garlic) extracts as a mosquito repellant. Specifically, the study seeks to answer the following questions. 1. What plant decoction can repel more mosquitoes? a. 150 grams of Theobroma cacao leaves boiled to 500 mL of water. b. 150 grams of Allium sativum (garlic) boiled to 500 mL of water. c. 150 grams of Theobroma cacao leaves and150 grams of Allium sativum boiled to 500 mL of water. 2. How effective is the decoction to repel mosquitoes based on the number of mosquitoes transferred to the other sides of the container in comparison to the commercial repellant?
STATEMENT OF THE HYPOTHESIS Ho: There is no significant difference in the repellant activity of Theobroma cacao leaves (cacao) and Allium sativum (garlic) bulb at the same proportions.
SIGNIFICANCE OF THE STUDY A mosquito repellant spray from plants do not brings out harmful substances that can risk the health of the people because it is an eco-friendly product which is safe to use. This investigation aims to make a mosquito repellent out of Theobroma cacao leaves (cacao) and Allium sativum (garlic) and to come up with a solution of the increasing number of dengue cases which is abound in Cebu. Families. Who want every member of it, safe from the mosquitoes carrying viruses (examples: dengue and malaria) who wish to buy a mosquito repellent spray which is cheaper, effective and safe to use, that they can assure that they would not inhale harmful substances instead of fresh, clean air; an air which is free from synthetic chemicals. Entrepreneurs. It is the pleasure of the entrepreneurs to have a cheaper, good quality repellent spray that is trend to the public which they can sell it at affordable price. Mother Earth. Mother Earth is the center of all beneficiaries in the study for it is an eco-friendly product. It helps save Mother Earth from the air pollutant brought by commercial mosquito repellent sprays by using natural products which biodegrades in the environment. Researchers. This study helps the researcher to know more about Theobroma cacao leaves (cacao) and Allium sativum (garlic). Another thing, they could also benefit in a way of expanding their knowledge about the mosquitoes and repellent sprays.
SCOPE AND LIMITATIONS This study dealt with the production of a mosquito repellent which is made from non-synthetic, organic chemicals which are environmentally friendly. It was focused on the decoction of Theobroma cacao leaves and the bulb of Allium sativum at various proportions to determine the concentration which can repel the most number of mosquitoes. This project was limited to two varieties of plants only namely Theobroma cacao and Allium sativum. Only the leaves of the former were gathered since they were easy to collect and were just given for free. Allium sativum, on the other hand, is used as condiment in every household in the Philippines. As such, it abounds in the market and is inexpensive. The subjects of this research were mosquitoes. They were reared from wigglers in each of the 3 mosquito cages inside a beaker which contained 500mL of plain water. They were fed with dry leaves. When the adult emerged, they were fed with sugar solutions since it is the main food of the adult mosquitoes (Foster, 1995). Identification of the mosquitoes was beyond the scope of the study since the researchers only wanted to determine the effectiveness of the aforementioned plants as repellent. The study was conducted from December 5, 2013– January 17, 2014 in Talamban National High School since Barangay Talamban ranks second in the dengue fever cases from January-June 2013 (Dengue Fever Surveillance, 2013).
DEFINITION OF TERMS For the purpose of this project and to facilitate the comprehension of this work, the terms hereafter mentioned are hereby defined. Decoction. It is the process in which the leaves of Theobroma cacao and Allium Sativum were heated to extract the substances which can repel the mosquitoes. Proportions. It refers to the various weights of Theobroma cacao and Allium Sativum. Repellant. It is behavior exhibited by the mosquitoes which are treated with the product to transfer to the untreated half of the cage.

METHODOLOGY An experimental method of research was used to test the effectiveness of the decoction of Theobroma cacao leaves and Allium sativum bulb. As mosquito repellant, decoction was done to the plants and was applied to adult mosquitoes.
MAKING OF THE CAGE A cubic form cage measured 15”x15” was prepared. A mosquito net which measured 15”x15” was attached on the cubic form cage so that the adult mosquitoes cannot escape from it. In attaching the mosquito net, a ¾ nail was used. Then, the cut mosquito net was fastened also at the middle part of the cage. This would separate the area in the cage where the application of the product would be done. A hole was made in it where a tissue tube from the tissue papers with a diameter of 2.7 cm had served as the passage way of the mosquitoes to the other side of the cage as they were repelled. A small opening which measured 5”x5” at the top of the cage was made. It served as the opening for putting the collected mosquitoes, for feeding of the adult mosquitoes and for the testing of the product. And the three cages were labeled set-ups A,B and C.
PREPARATION OF MATERIALS Fresh cacao leaves were weighed scale at 150 grams, the garlic bulb were also weighed into 150 grams and another 150 grams of cacao leaves and 150 grams of garlic were weighed for the experiment. Each weighed garlic bulbs were peeled and crushed using the mortar and pestle separately.
DECOCTIONPROCEDURE
The 150 grams washed cacao leaves and 150 grams of skinless, crushed garlic were boiled in a 500 mL of water within 15-20 minutes on a medium fire separately. Another 150 grams of cacao leaves and 150 grams of skinless, crushed garlic were also boiled together in a 500 mL of water within 15-20 minutes on a medium fire. The products were filtered using the filter paper and were place inside a container. The cacao decoction was labeled as set A, garlic decoction was labeled set B and the mixture of cacao and garlic decoction was labeled set C.
REARING OF MOSQUITOES The researchers were able to collect sixty (60) mosquitoes in which twenty (20) mosquitoes for each cage by rearing them. Four black plastic cups of equal sizes were filled with 100mL of clean tap water. Decaying leaves were placed in each of the cups. They were placed inside the 4 classrooms of Talamban National High School to collect the eggs. After a week, the mosquito traps were collected. They were placed inside the school laboratory for hatching which took 2-3 days. When hatched, the wrigglers were poured into a basin and were counted. A 100mL of clean, tap water with dry leaves was placed in each of the three beakers and twenty wrigglers were transferred in each of the set-ups using a small fish net. The three beakers were placed inside the three cages and left there for 2-3 days. They also prepared a sugar solution. 25 grams of sugar was added to a 100mL of water (Foster, 1995). It was stirred until dissolved in a beaker and transferred the solution in a container. A three (3) ball of cottons were moistened with the sugar solution in which each cotton ball was placed on a small plastic container. These containers were placed inside each cage in order to feed the adult mosquitoes.
PRODUCT TESTING Three (3) cotton balls were prepared for each of the set-ups and were placed inside each of the cages with mosquitoes ( Bernard, n.d) . The first cotton was moistened with twenty (20) sprays of repellant A (150 grams of cacao leaves) while the second cotton ball was also sprayed twenty (20) times with repellant B (150 grams of garlic) and lastly the third cotton ball was sprayed twenty (20) times with repellant C (150 grams of cacao leaves and 150 grams of garlic) . They were left for four minutes inside the mosquito cages. After which, the number of mosquitoes which transferred to the other side of the cage was counted, tallied and recorded. The same was also done for the mosquitoes which were left in the cage. The same procedure was done with the other two cages. Statistically, is of data was also done.
TREATMENT OF DATA The data gathered from actual observation were recorded, statistical treated, analyzed and interpreted.
The formulas for statistical treatment used were:
TSS=∑X² ˉ(∑X²)/N Where
TSS= total sum of square
X= individual values in each column
N= total sample size

SSB=(∑Xc)²n-(∑1X1)²/N Where
SSB= sum of squares within columns
∑xc= sum of the individual values per column n= size of the sample per column
SSW=TSS-SSB
Where
SSW= sum of squares within column.

REVIEW OF RELATED LITERATURE Garlic can be used to repel a variety of crawling and flying insects, including mosquitoes. According to Patrick Parker, SavATree Plant Health Care Program Director, one treatment with garlic is effective for 2 weeks and can repel insects for up to one month. Moreover, garlic is easy on the environments. Parker states, “It is a natural extract and does not affect insects that are beneficial to suburban landscapes.” Garlic bulbs contain an amino acid that converts to a substance called allicin when crushed, blended or chopped. The characteristic odor released as a result of this process has powerful properties. : When garlic extract is absorbed by a plant, biochemical changes take place in its foliage which cause it to actively repel insects,” says Parker, “ In short, plants are provided with a long-lasting case of “garlic breathe” that causes insects to move elsewhere.” However, the treatment is odorless to humans within minutes of application. Insects are naturally repulsed by the presence of the botanical extract. Therefore, they do not build up a resistance to garlic treatments. Garlic can also be applied to many sensitive trees, shrubs and turf that traditional treatments would harm. Parker added, “I get excited whenever a natural solution produces results that are effective enough to compete on a large scale with synthetic products.” (http://www.savatree.com/garlic-insect-repellent.html) Garlic, a common staple found in countless kitchens and recipes worldwide, long has been considered a special food - not just for the unique flavor it imparts, but also for its medicinal value. Most of its therapeutic value, as well as its flavor and odor, can be attributed to sulfur compounds contained within the garlic clove. In recent years, garlic has been widely studied for its role in promoting health. There is good evidence that garlic possesses antibacterial, antiviral, antifungal, antiprotozoan and even insect-repellent properties. Given the recent surge of the West Nile virus spread by infected mosquitoes, it is interesting to explore the value of garlic as one more method for avoiding mosquito bites. A number of studies have shown that the oil fraction of garlic destroys certain species of mosquito larvae. Garlic sprays (made primarily with garlic extract) are available on the market for us on plants as an alternative botanical pesticide to chemical pesticides. The sulfurs contained within the garlic extract have been shown to be effective against a wide range of insects, including mosquitoes, and the lingering odor can deter mosquitoes from the area for weeks. It is thought that garlic may be an alternative mosquito repellent for humans as well. In a field study conducted in India, a preparation made of 1 percent garlic oil, petroleum jelly and beeswax that was rubbed on the arms and legs of study subjects was found to be effective in preventing mosquito bites for up to eight hours. In addition, there is some evidence that heavy consumption of garlic through supplements or well-flavored foods may help ward off mosquitoes. When garlic is eaten and its components are metabolized, compounds are released from the body through the skin and the breath. (http://www.straightdope.com/columns/read/2177/does-garlic-repel-mosquitoes)
REVIEW OF RELATED STUDIES Allium sativum yields allicin, an antibiotic and antifungal compound (phytoncide). It has been claimed that it can be used as a home remedy to help speed recovery from strep throat or other minor ailments because of its antibiotic properties. It also contains the sulfur-containing compounds alliin, ajoene, diallylsulfide, dithiin, S-allylcysteine, and enzymes, B vitamins, proteins, minerals, saponins, flavonoids, and Maillard reaction products, which are not sulfur-containing compounds. Furthermore, a phytoalexin (allixin) was found, inhibiting skin tumor formation. Herin, allixin and/or its analogs may be expected useful compounds for cancer prevention or chemotherapy agents for other diseases (Ejaz et al.,2003) The composition of the bulbs is approximately 84.09% water, 13.38% organic matter, and 1.53% inorganic matter, while the leaves are 87.14% water, 11.27% organic matter, and 1.59% inorganic matter. Fresh C. citratus grass contains approximately 0.4% volatile oil. The oil contains 65% to 85% citral, a mixture of 2 geometric isomers, geraniol and neral. Related compounds geraniol, geranic acid, and nerolic acid have also been identified. More than a dozen other minor fragrant components were also found. Research has shown geraniol to be an effective plant based mosquito repellant (Rumor Kiddo). A study in Kilimanjaro region, north eastern of Tanzania found that the essential oils from of Ocimum Suaveand Ocimum Kilimandscharicum were effective against biting mosquitoes in terms of feeding inhibition, knockdown effect and mortality (Kweka, 2008). A review of plant based repellents showed that there were several plants which repelled biting arthropod. Yet, 30 species of the plants reviewed was found to have some harmful effects to humans like skin irritations and cancer causing (Maia and Moore, 2011).

RESULTS AND DISCUSSION After the data were collected, an analysis of variance (ANOVA) was performed. Table 1: The number of mosquitoes repelled in the three (3) treatments.

A
B
C
A

B

C

STEP 1:
N

T

T²

SSB=
STEP 2:
SS=
SS=
STEP 3:
C= T²/N
C=
C=
STEP 4:
SST= SS-C
SST=
SST=
STEP 5:
SSB= [T1²/N + T2²/N + T3²/N]
SSB=
STEP 6:
SSW= SST-SSB
SSB=
SSB=
STEP 7:

Sum of Square
DF
Mean Square
Source of Variation

Within Groups

RATIO
T.V

Findings: CV ( ) > TV ( )

Decision: Reject Ho

Analysis:

The proportion of Theobroma cacao leaves and Allium Sativum bulb has a significant difference

in the repellent activity.

The data on repelling the mosquitoes were subjected by one way analysis of variance. It was

found out that the different proportions of Theobroma cacao and Allium Sativum affect the number of

mosquitoes being repelled.

CONCLUSIONS Based on the results, the researchers concluded that Theobroma cacao (cacao) leaves and Allium \
Sativum (garlic) bulb decoction is effective in repelling in terms of number of mosquitoes transferred to the other side of the container.
Moreover, the decoction coming from 300 g of Theobroma cacao (cacao) leaves and Allium Sativum
(garlic) bulb can repel the mosquitoes effectively than the other two.

RECOMMENDATIONS The researchers recommend, increasing the population of the subjects so that they are not well dispersed inside the 15”x15” cage. In so doing, they can immediately transfer to the untreated area. Lastly, it is endorsed that the time in minutes in testing treatment C be varied to determine if the repellency of the product is long.

BIBLIOGRAPHIES
Books
McClung, Robert M. Aquatic Insects and How They Live. U.S.A.: William Morrow and Company Inc., 1971.
Publications.
Barnard, Donald R. et al. “Standard Method for Testing Mosquito Repellant.” Journal of the American
Mosquito Control Association, 21 (4) (2005).
Ejaz, S., L.C. Woong and A. Ejaz. “Extract of Garlic (Allium Sativum) In Cancer Chemoprevention.
Experimental Oncology. 25, 93-97 (June 2003).
Foster, Woodbridge A. “Mosquito Sugar Feeding and Reproductive Energetics.”
Kweka, Eliningaya J. et al. “Ethnobotanical study of some of mosquito repellant plants in north-eastern Tanzania.” Malaria Journal. 7 (152) (2008).
Maia, Marta Ferreira and Sarah J. Moore. “Plant-based insect repellants: a review of their efficacy, Development and testing.” Malaria Journal(2001).
Unpublished Materials
City Epidemiology Statistics and Surveillance Unit. “Dengue Surveillance 2013,” MW #26.
Internet Sources http://www.annualreviews.org/doi/abs/10.1146/annurev.en.40.010195.002303?journalCode=ento http://www.mercola.com/article/pesticides/deet.html http://en.wikipedia.org/wiki/Coumarin http://wiki.answers.com/Q/What_do_mosquitoes_hate http://www.savatree.com/garlic-insect-repellent.html http://en.wikipedia.org/wiki/Mosquito http://www.straightdope.com/columns/read/2177/does-garlic-repel-mosquitoes APPENDICES

APPENDIX A
REQUEST LETTER

TALAMBAN NATIONAL HIGH SCHOOL
TALAMBAN CEBU CITY
JANUARY 12, 2013

RHODA S. CARIEGO
Research 1
Talamban National High School
Talamban Cebu, City

Madam:

We, the undersigned students of Research 1, are conducting a research entitled Theobroma cacao leaves (cacao leaves) and Allium Sativum (garlic) as mosquito repellent spray. And we hereby ask you to be our research adviser.

We are hoping for your positive response to this matter.

Thank you and God bless.

Sincerely yours,

QUEENIBEL S. ARRIESGADO

KRISTINE JANE A. BORCES

DOMINIQUE FATIMA G. CABANSAY

JOBELYN B. COGTAS

JOSE PATERNO MONTESCLAROS

MARY ROSE A. TELAMO

Noted:

CELIA C. GEPITULAN
Research Teacher

APPENDIX B

TRANSMITTAL LETTER

TALAMBAN NATIONAL HIGH SCHOOL
TALAMBAN CEBU CITY
JANUARY 12, 2013

DR. MA. JESSICA N. ABAYON
PRINCIPAL I
Talamban National High School
Talamban Cebu, City

Madam:

As a requirement for our Research II subject we are asking your permission that we, the researchers would be allowed to use the equipment in Science Laboratory for our Investigatory Projects since the school has all the necessary materials for this study.

Your approval would be highly appreciated. We are hoping for a positive response.

Thank you and God bless.

Sincerely Yours,

QUEENIBEL S. ARRIESGADO

KRISTINE JANE A. BORCES

DOMINIQUE FATIMA G. CABANSAY

JOBELYN B. COGTAS

JOSE PATERNO MONTESCLAROS

MARY ROSE A. TELAMO

Noted:

RHODA S. CARIEGO
Research Adviser

APPENDIX C

TRANSMITTAL LETTER

TALAMBAN NATIONAL HIGH SCHOOL
TALAMBAN CEBU CITY
JANUARY 12, 2013

DR. ZENAIDA C. GRAPA
Master Teacher II
Talamban National High School
Talamban Cebu, City

Madam:

As a requirement for our Research II subject we are asking your permission that we, the researchers would be allowed to use the equipment in Science Laboratory for our Investigatory Projects since the school has all the necessary materials for this study. Moreover, we are asking from your good office for financial assistance to make this project successful.

Your approval would be highly appreciated. We are hoping for a positive response.

Thank you and God bless.

Sincerely Yours,

QUEENIBEL S. ARRIESGADO

KRISTINE JANE A. BORCES

DOMINIQUE FATIMA G. CABANSAY

JOBELYN B. COGTAS

JOSE PATERNO MONTESCLAROS

MARY ROSE A. TELAMO

Noted:

RHODA S. CARIEGO
Research Adviser

APPENDIX D
TRANSMITTAL LETTER
TALAMBAN NATIONAL HIGH SCHOOL
TALAMBAN,CEBU CITY
JANUARY 13,2014

CELIA GEPITULAN
Research I
Talamban National High School
Talamban Cebu, City

Madam:

We, the undersigned students of Research I, are conducting a research entitled “Theobroma cacao (cacao leaves) and Allium sativum (garlic) as mosquito repellent spray. And we hereby ask you to borrow the three mosquito cages in the science laboratory. Whatever damage that may happen it will be our responsibility. Your approval will contribute a great help in our research.

Thank you and God bless.

Sincerely yours,

QUEENIBEL S. ARRIESGADO

KRISTINE JANE A. BORCES

DOMINIQUE FATIMA G. CABANSAY

JOBELYN B. COGTAS

JOSE PATERNO MONTESCLAROS

MARY ROSE A. TELAMO

Noted:

CELIA GEPITULAN
RESEARCH I