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  1. EFFICACY OF ORIGANUM VULGARE EXTRACTS AS MOSQUITO REPELLENT A Research Paper In Partial Fulfillment of the Requirements in Practical Research I (Experimental Research) By: John Alvin Molina Jona Mae Banquillo Grade 11 HUMMS December 2019 Effecacy of Origanum Vulgare Extracts as Mosquito Repellent
  2. Chapter 1 INTRODUCTION OF THE STUDY Background and Theoretical Framework of the Study Mosquitoes are the vectors of many pathogens that continue to make a large impact on human health. Current control measures for vector-borne infectious diseases are under threat and new strategies, including genetic control, are urgently needed. Genetic strategies to control mosquito-borne diseases include population suppression and population replacement. Mosquito bites lead to a variety of mild, serious, and, rarely, life-threatening allergic reactions. These include ordinary wheal and flare reactions and mosquito bite allergies (MBA). The MBA, also termed hypersensitivity to mosquito bites (HMB), are excessive reactions to mosquito bites that are not caused by any toxin or pathogen in the saliva injected by a mosquito at the time it takes its blood- meal. Rather, they are allergic hypersensitivity reactions caused by the non-toxic allergenic proteins contained in the mosquito's saliva. Studies have shown or suggest that numerous species of mosquitoes can trigger ordinary reactions as well as MBA. These include Aedes aegypti, Aedes vexans, Aedes albopictus, Anopheles sinensis, Culex pipiens, Aedes communis, Anopheles stephensi, Culex quinquefasciatus,
  3. Ochlerotatus triseriatus, and Culex tritaeniorhynchus.[123] Furthermore, there is considerable cross-reactivity between the salivary proteins of mosquitoes in the same family and, to a lesser extent, different families. It is therefore assumed that these allergic responses may be caused by virtually any mosquito species (or other biting insect). The mosquito bite allergies are informally classified as 1) the Skeeter syndrome, i.e. severe local skin reactions sometimes associated with low-grade fever; 2) systemic reactions that range from high-grade fever, lymphadenopathy, abdominal pain, and/or diarrhea to, very rarely, life-threatening symptoms of anaphylaxis; and 3) severe and often systemic reactions occurring in individuals that have an Epstein-Barr virus- associated lymphoproliferative disease, Epstein-Barr virus-negative lymphoid malignancy,or another predisposing condition such as Eosinophilic cellulitis or chronic lymphocytic leukemia. Aglalana Integrated School is a newly established school at Dumarao, Capiz. Fact is that even though new, there are several cases of Dengue at the said school. According to recent studies, organic repellents have higher and longer chances of repelling mosquitoes. Though need some chemical compositions, the main ingredient which is the extracted oil of Origanum Vulgare also known as Oregano, is considered as an excellent antiseptic and insect repellent. It has some active ingredients such as
  4. carvacrol, thymol, and a-terpinene reported being highly effective in repelling mosquitoes. Statement of the Problem and Hypotheses Generally, this study aimed to explore the efficacy of extracted oil of Origanum Vulgare in repelling mosquitoes. Specifically, it sought to answer the following questions: 1. How long will the repellency of the extracted oil from Origanum Vulgare last? 2. Is there a difference in repellency between the organic repellent and synthetic repellents? 3. On the scale of 1-10, How effective is the repellent? In view of the aforementioned problems, the following hypotheses were tested:
  5. 1. The repellency lasts for 30 minutes up to 1 hour depending on how much extracted oil was used. 2. The organic repellent has higher and longer chance of repelling mosquitoes when extracted oil from Oregano is at high amount compared to synthetic repellents. 3. The efficacy of the repellent made from extracted oil of Origanum Vulgare on the scale of 1-10 is at 8 depending on the amount of oil used. Besides, there are still room for improvement when added with a certain oil to help the repellency last longer.
  6. Significance of the Study This study is essential to students, and other persons because it provides ideas and better actions in learning much effective and safe ways on repelling mosquitoes. This will help the students and other persons involve the community to find and discover much effective ways on repelling mosquitoes in a safe and organic way. The results of this study may be beneficial to the following: Students. The result of the study provides adequate information for the students to create an organic repellent that has higher chances of repelling mosquitoes. It also provides safety for students against blood-sucking mosquitoes. Parents. The result of the study provides trust for the parents that they and their children will be safe on mosquitoes and the effects of the repellent. The community. The result of this study provides better understanding for the people in the community that organic repellents provides not just safe but longer effect and protection against mosquitoes.
  7. Future Researchers. The result of this study will serve as a reference for future researchers in order to conduct further studies regarding the efficacy of Oreganum Vulgare as larvicide for mosquitoes and clarify the facts about it. Definition of Terms The following terms were given their conceptual and operational definitions for the purpose of clarity and precision. Effects - refer to a change that results when something done or happens; an event, condition, or state of affairs that is produces by a cause. (Merriam-Webster Dictionary, 2019) In this study, “effects” referred to the efficacy of extracted oil from Oregano as repellent for mosquitoes. Mosquitoes – a small flying insect that sucks blood of animals and people via small channel that serves as their mouth. (Merriam-Webster Dictionary, 2019)
  8. In this study, “mosquitoes” referred to the target of the organic repellent made from Oreganum Vulgare extracts. calculator as an independent variable being used. Students - refer to a person who attends a school, college, or university; a person who studies something. (Merriam-Webster Dictionary, 2019) In this study, “students” referred to the persons who are the respondents of the study. Oregano – a culinary and medicinal herb containing active ingredients such as carvacrol, thymol, and a-terpinene reported being highly effective in repelling mosquitoes. In this study, “Oregano” referred to the plant hthat was extracted to make the organic repellent. Repellent – a material with chemical compositions use to keep or drive something away such as flies, mosquitoes, etc. (Merriam-Webster Dictionary, 2019)
  9. In this study, “repellent” referred to the substance made out of Oregano extract use to repel mosquitoes. Extracts – to withdraw something, such as juice,by physical or chemical process. (Merriam-Webster Dictionary, 2019) In this study, “extracts” referred to the soluble substance removed from the Oregano. Chemical Composition – refers to the arrangement and ratio of atoms in molecules of a substance. (Merriam-Webster Dictionary, 2019) In this study, “Chemical Composition” referred to the composition of the extract of Oregano used in making the repellent.
  10. Delimitation of the Study This study aimed to contrast the efficacy of the extract of Oreganum Vulgare as a repellent for mosquitoes. This study aimed to take place at the home of the researcher located at Barangay Aglalana, Passi City, Iloilo during the second semester of the S.Y 2019-2020. The data gathering and experiment time targets the amount of days at a minimum of 10 days and a maximum of 15 days. This amount of time is required for the researcher to gather as much as information as he can to improve the efficacy of the said repellent. Addition on materials and methods are allowed to have better results for the experimental research that would be conducted by the researcher. Any result either pro or con shall be recorded to allow further clarifications and improvements on the study.
  11. CHAPTER 2 REVIEW OF RELATED LITERATURE Introduction Mosquitoes are carriers of diseases causing serious health problems in tropical regions. A variety of mosquito species transmit many diseases to humans and animals in a species-dependent manner; for example, Aedes aegypti is the vector for dengue fever, Zika fever and Chikungunya, Anopheles dirus B is a vector for malaria in Asian forested zones and Culex quinquefasciatus is a vector for zoonotic diseases that affect humans and wild and domestic animals. Almost all these diseases are severe problems in many countries, which makes it necessary to control mosquito populations. Although there are many mosquito pesticides available on the market currently, mosquitoes can develop resistance to pesticides, especially synthetic pesticides, and thus, it becomes necessary to increase the dosage used to control mosquitoes, introducing additional risks to human health and to the environment. From the past, the discovery and development of pesticides with mosquito larval toxicity derived from plant extracts have become a topic of interest to many researchers. At present, plant extract products are attractive choices for use in
  12. integrated pest management programmes to reduce the residues in the environment as it has biodegradable efficiency. Many studies have used plant extracts to control mosquitoes. This present study evaluated the use of rhizome Alpinia galanga, a herb in the Zingiberaceae family that is commonly used in cooking, especially in Thai cuisines, to control a certain species of mosquito larvae. Repellency Effects of Plant Extracts Pest management techniques are many and varied: mechanical, cultural, biological, and chemical. Treatment of adult mosquitoes – adulticiding – is achieved entirely via pesticide applications targeted to adult mosquitoes. The process of adulticiding is a step wise process that often is considered the method of last resort in an Integrated Pest Management (IPM) approach to mosquito control (Connelly 2012). Adult female of certain mosquito species have the ability to transmit diseases from an infected individual to a susceptible person. Vector control measures have, therefore, been established to control the transmission of the disease by targeting the carriers. Over the past few decades, the vector has however developed the ability to evade intervention measures, which target adult mosquitoes, thus, exacerbating the problem for vector control programmes (Sharp 1983). The current vector control technique involves the
  13. use of residual repellents which are sprayed onto walls and roofs of houses. With concern for the quality and safety of life and the environment, the emphasis on controlling mosquito vectors has shifted steadily from the use of conventional chemicals toward alternative repellents that are target-specific, biodegradable, and environmentally safe, and these are generally botanicals in origin. Although plants and their derivatives were used for controlling and eradicating mosquitoes and other domestic pests before the advent of synthetic organic chemicals, only few insecticides of plant origin have been found commercially available. Plant-derived bioproducts, however, still have encouraging results in the control of mosquito vectors if they are adequately effective and harmless to beneficial non-target organisms and the environment. Furthermore, the insect resistance to mosquitocidal botanical agents has not been documented (Shaalan et al. 2005). A study of monoterpenoids showed that for topical acute toxicity on larvae the most potent were thymol and carvacrol. The most profound synergistic effect was found in the mixture p-cymene, thymol, carvacrol. (Roman Pavela, J Biopest, 2010, 3, 573-78). The same author in another study found in laboratory conditions that the sublethal application of thyme oil to the adults of the housefly significantly reduced their longevity and the natality decreased by 80%, And even larvae which emerged from eggs from female flyies which had been treated, were non vital. In a third study (R Pavela, Phytother Research 2008, 22, 274-8) the same author studied the insecticidal activity of 34 essential oils extracted from plants and found those from Origanum vulgare,
  14. majorana, compactum, Ocimum basilicum, Thymus vulgaris were lethal in doses in the microgram range. The advantage of these plants is that they are easy to grow in many countries and have well known nutritional values. Some of the terpenes, like carvacrol, also have strong antiplasmodial properties. Oregano oils are rich in thymol (38.8%), carvacrol (32.9%), p-cymene (7,9%). They have strong toxicities against Rhizoperta dominica by contact or by fumigation (O Khalfi et al., J Sc Food and Agricul, 2008, 9, 1562-66). Efficient also against drosophila (I Karpouhtsis et al., J Agricult and Food Chem, 1998, 46, 1111-15). Thymol contributes to the acaricidal properties of Ocimum gratissimum. Repellent Bioassay Repellents make humans unattractive to a mosquito so that it will avoid areas of the body that have been treated with the products and they do not kill mosquitoes. Quality repellents will provide protection from bites for a long period of time from just one application. The use of repellents to protect humans and his animals from bites of mosquitoes already has been accepted as part of an overall integrated mosquito-borne disease control program (Chavasse and Yap 1997; Yap et al. 2000). The use of natural products such as plant extracts or oils was not common until now due to many reasons. Thus, it is now rather easy to find chemical compounds that have repellency properties
  15. and to use them as active ingredients in a repellent product (Amer and Mehlhorn 2006). Freeborn (1928) and Dover (1930) cited some insect repellent formulations consisting of a number of essential oils such as citronella, camphor, tar, pennyroyal, and castor oils that provided a long-lasting protection from insect bites. Repellents have an important place in protecting man from the bites of insect pests. An effective repellent will be useful in reducing man-vector contact and in the interruption of disease transmission. Repellent compounds should be non-toxic, nonirritating and long lasting (Kalyanasundaram 1991). Majority of commercial repellents are prepared by using chemicals like allethrin, N-N-diehyl-m-toluamide (DEET), dimethyl phthalate (DMP) and N, N-diethyl mendelic acid amide (DEM). It has been reported that these chemical repellents are not safe for public use (Zadikoff 1979; Ronald et al. 1985). In the last few years, with the increase of public concern on the safety of many chemical products that were used previously as insecticides or insect repellents, several institutes and researchers started development of natural active ingredients especially from plant sources. Synthesis Mosquitoes are the most important single group of insects in terms of public health importance. Mosquitoes not only cause nuisance by their bites but also transmit deadly diseases. Among the approximately 4000 known mosquito species, less than 10% are
  16. regarded as efficient vectors of pathogenic agents of infectious diseases having high impact, both direct and indirect, on human welfare and health. Mosquito-transmitted diseases remain a major cause of the loss of human life worldwide with more than 700 million people suffering from these diseases annually (Taubes 1997). Therefore, repellency of mosquitoes is essential to help prevent serious health problems. Using the extracted oil of Origanum Vilgare (Oregano) shows effective results of repellency effects on mosquitoes as itshows 30 minutes up to an hour repellency on mosquitoes. Preferring natural repellents to repel mosquitoes rather than synthetic enables the host to be safe not just from host-seeking mosquitoes but also to health safety concerns.
  17. Chapter 3 Methodology Research Design Experimentalresearchdesigniscentrallyconcernedwithconstructingresearchthatishighincau sal(internal)validity.Randomizedexperimentaldesignsprovidethehighestlevelsofcausalvali dity.Quasi‐experimentaldesignshaveanumberofpotentialthreatstotheircausalvalidity.Yet,n ewquasi‐experimentaldesignsadoptedfromfieldsoutsideofcriminologyofferlevelsofcausalv aliditythatrivalexperimentaldesigns. Thedesignofresearchisfraughtwithcomplicatedandcrucialdecisions.Researchersmustdecid ewhichresearchquestionstoaddress,whichtheoreticalperspectivewillguidetheresearch,how tomeasurekeyconstructsreliablyandaccurately,whoorwhattosampleandobserve,howmany people/places/thingsneedtobesampledinordertoachieveadequatestatisticalpower,andwhi chdataanalytictechniqueswillbeemployed.Theseissuesaregermanetoresearchofalltypes(ex ploratory,explanatory,descriptive,evaluationresearch).However,theterm“researchdesign” typicallydoesnotrefertotheissuesdiscussedabove.
  18. Theterm“experimentalresearchdesign”iscentrallyconcernedwithconstructingresearchthati shighincausal(orinternal)validity.Causalvalidityconcernstheaccuracyofstatementsregardin gcauseandeffectrelationships.Forexample,doesvariable1causevariationinvariable2?Ordoe svariable2causevariationinvariable1?Ordoesvariable3causevariationinbothvariables1and2 ?Andwhatisthemagnitudeofthecausalrelationshipsamongthevariables?Thus,researchdesig nasusedhereinisaconcernofexplanatoryandevaluationresearchbutgenerallydoesnotapplyt oexploratoryordescriptiveresearch. Materials/Apparatuses and Utensils These are the lists of materials, apparatuses and utensils used for the experiment. Materials and Apparatuses. Quantity. Unit Gardening gloves. 2. Pieces Pillowcaseor any cloth. 1. Pieces Hammer or Rolling Pin. 1. Pieces
  19. Bucket or large bowl. 1. Pieces Surgical-style face mask. 1. Pieces Goggles. 1 Pieces Dish soap. 100 Milliliters Strainer. 1. Pieces Spray bottle. 1. Pieces Oregano. 50. Grams Water. ½. Litters Procedures and Methods
  20. 1.Put on a pair of gardening gloves and harvest about 4 to 5 cups of oregano leaves and stems. Put the plant material in an old pillowcase. Harvest oregano after the buds have formed, but before the flowers open, because this is when its oils are strongest. 2.Place the pillowcase on the floor, and smash it several times with a mallet, or run a rolling pin over it to break up the leaves and stems and release the plant juices. Turn the pillowcase over and continue breaking up the plant material with the bat or rolling pin. 3.Open the pillowcase, turn it inside out and dump the plant material into a container, such as a bucket or large bowl. Place the empty pillowcase in the container as well. Then pour just enough water over the pillowcase and the crushed leaves and stems so that they're fully submerged. 4.Place the container in a sunny area and let the mixture steep. Stir it several times throughout the day and let it sit overnight. 5.Put on gloves, a surgical-style face mask, goggles, and clothes that cover your arms and legs, and then, stir 1 teaspoon of liquid dish soap into the mixture so the plant oils
  21. distribute throughout the water. Pour the mixture in a strainer that's placed on a clean container. This separates the plant material from the water. Thoroughly wring out the pillowcase over the container. 6.Fill the liquid in a labeled spray bottle and apply it on an insect invested plant to see if it works. The dish soap in the liquid will act as glue that helps the mixture stick to the plants. When the insects run for cover than you know the spray works
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