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The Feasibility Of Makahiya Edited

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Dengue is the most common arthropod-borne viral (arboviral) illness. It is transmitted by the mosquitoes of the genus Aedes which are widely distributed in tropical or sub-tropical countries particularly in the Philippines. The AedesAegypti Mosquitoes are the vectors of Dengue fever and the eggs have the ability to survive desiccation for long periods of time, allowing eggs to be easily spread to new locations thus contributing to the rampant spread of the disease when left unchecked. Death tolls in the Philippines over Dengue matters reached at least 42,207 dengue case, 193 of them fatal, had been recorded from January to the first week of June 2013 (Department of Health, 2013). This study uses the benefit we can get from the Makahiya (mimosa pudica) roots extract to prevent the propagation of mosquito eggs.

This study will serve as the basis for future plans of action by the school administrators, community officials and the society with regards to the necessary actions for exterminating the mosquito eggs and larvae. Throughout the history, many plants have been used as an alternative insecticide, ovicide, larvicide, and repellants. The increasing number of dengue cases not only in our country but also to other countries all over the world. Experiencing the abnormal change in weather is not of recent origin to us. In that case, they are harder to intercept in flight that is why it is more advisable to focus on the places where they lay their eggs and exterminate them as early as possible.

Statement of the problem
This study aims with the investigation of Makahiya as an alternative larvicide for mosquito larvae. Specifically, it seeks to answer the following questions:
1. What are the effects of Makahiya extract to the mosquito larvae after having been exposed to: a. 80% makahiya extract and 20% water within 5 mins., 15 mins., 30 mins., and 45 mins. b. 60% makahiya extract and 40% water within 5 mins., 15 mins., 30 mins., and 45 mins. c. 100% makahiya extract within 5 mins., 15 mins., 30 mins., and 45 mins. 2. In which of the following time duration will the solution still be affective?

a. 5 days
b. 1 week
c. 10 days
d. 2 weeks
H0: The Makahiya (Mimosa Pudica) extract is effective as a larvicide for mosquito larvae. H1: The Makahiya (Mimosa Pudica) extract is not effective as a larvicide for mosquito larvae.

Significance of the Study
Among the people who will benefit are the following:
To the Students:
This study will create a milieu of awareness about the harmful disease brought by mosquitoes and the knowledge they can get on how to prevent such dengue cases in their respective communities. They can use this study to further put this into action in order to help subside the cases. To the Parents:

The parents of the children will be aware that an alternative solution that cannot harm both their health and their children’s is available and possible. Therefore, this study can assist them in helping their own community free from possibilities of mosquito habitat. To the School Faculty:

In providing a safe environment for the students, this study can provide awareness for the school faculty that an alternative larvicide will enable them to use this full knowledge and safety assurance in the student’s health. To the Future Researchers:

This study will serve as a theoretical model for future studies of the same nature if ever the existing problem has penetrated in this case will exist in the future. This will provide them the facts needed to compare their study during respective time and usability.

Scope and Limitations
This study aims to explore the capability of Makahiya root extract as an larvicide for mosquito larvae. It is focused on the effectiveness of the solutions in having the ability to kill a larva. The study will be conducted in Ormoc City, Leyte. It involves an experimentation using the said solutions, involving the use of the roots of the Makahiya plant. This study is focused in providing people with a less expensive and more resourceful way of killing mosquito larvae.

The variables involved in this study making the Makahiya root extract are stopwatch, mosquito larvae, water, and plastic container. The said solution is directly applied to the mosquito larvae samples.

Conceptual Framework

Definition of Terms
Makahiya – also called sensitive plant, is a creeping annual or perennial herb; heavily armed with recurved thorns and having sensitive soft grey-green leaflets that fold and droop at night or when touched or cooled (Filipino Herbs, 2007) (Encarta Dictionaries, 2010) Ovicide – an agent that kills eggs; an insecticide effective against the egg stage (Merriam Webster Dictionary, 2010) Commercial – a paid advertisement on television or radio (Merriam Webster Dictionary, 2010) Extract – a concentrated preparation of the essential constituents of substances (Merriam Webster Dictionary, 2010) Longevity – long duration of an animal’s life (Encarta Dictionaries, 2010) Dengue – a tropical disease caused by a virus that is transmitted by mosquitoes and marked by high fever and severe muscle and joint pains (Microsoft Encarta, 2009) Alternative – able to serve as a substitute for something else (Microsoft Encarta, 2009) Larvicide – a chemical used to kill larvae (Microsoft Encarta, 2009) Capability – the power or practical ability necessary for doing something (Encarta Dictionaries, 2010)

Review of Related Studies and Literature

The following is summary of the previous researches made in Makahiya (M. Pudica) and mosquito ovicide and larvicide. This review allows the reader to be brought up to date regarding the state of research or familiarizes the reader with contrasting perspectives and viewpoints on the topic. Makahiya found throughout the Philippines is found wild everywhere on Mt. Banahaw and other tropical areas. It is considered a weed with beneficial uses to both animals and humans. Having the name Mimosa Pudica is derived from the meaning “.sensitive” given the fact that its leaves contract when exposed to human touch. Makahiya is known as diuretic and is considered alterant and antiasthmatic. It is used for urinary complaints and is useful in diseases arising from corrupt blood and bile. It has also been used as a functional food. All of its parts from its thorns to its leaves and its stems and roots, all play an essential role in Herbal Medication.

Not only Makahiya have the genes to cure but also together with its relatives as sensitive plants. Sensitive plant contains a lot of biological compounds which are useful for human health (Filipino Herbs Healing Wonders, 2004-2014). One of them is mimosine. Mimosine can inhibit the proliferation of a number of lung and liver cancer cells. Besides, mimosine can inhibit the development of bacteria and be used as skin bactericide. Mimosine, β-[N-(3-hydroxypyridone-4)]-α-aminopropionic acid, occurs in the shoots and stem of Mimosa pudica. L. Mimosine has a number of interesting pharmacological properties, including the capacity to block the cell cycle before the G1/S transition, inhibit cardiac fibrosis, and prevent neuronal death and acts as antimicrobial agent. The mechanism through which mimosine blocks the progression of the cell cycle are not fully understood, but there was evidence that the inhibition of DNA replication may be caused by chelation of iron from ribonucleotidereductased by mimosine (Thu et al., n.d.).

It has an apoptopic effect, the same attribute that is provoked during cell disintegration. Mimosine possesses antimitotic activity that blocks the cell cycle in the large G1 phase and inhibits DNA synthesis, which prevents the formation of the replication fork by altering deoxyribonucleotide metabolism (Tran Dang Xuan, ShinkichiTawata &Tran Dang Khanh, n.d.). At high concentrations, mimosine reversibly arrests the progression of the cell cycle at G1 before the onset of DNA replication, and this arrest differs from the cell cycle arrest caused by other chemical agents, such as aphidicolinor hydroxyurea, that inhibit DNA polymerases or ribonucleotidereductase, respectively. While this property of mimosine makes a valuable tool for cell cycle-related studies, the exact point of cell cycle arrest and the mechanisms contributing to this effect are presently unknown (Soon-Young Park, n.d.).

Cell cycle arrest in mimosine response to environmental effects can lead to DNA breaks. Inhibition of DNA replication during the initiation step can lead to DNA damage and characterized a cell-cycle-arrest point at the replication initiation step before the establishment of active replication forks. This arrest can be elicited by mimosine, iron chelators, ciclopiroxolamine or 2,2′-bipyridyl, and can be reversed by the removal of the drugs or the addition of excess iron. While the mimosine penetrates deeper and deeper, its chemicals block the cells into entering the next development stage. While it blocks off the cells, the latter is disintegrated resulting to the death of the cells. Not an exception is the roots of the Makahiya plant. They are found to have a great medicinal value.

The findings of Pancle and Pathack (2010) were based on the results of the pharmacognistic and phytochemical studies including qualitative chemical examinations on the roots of the plant. Results of the chemical tests indicated presence of flavonoids, phytostenol, amino acids, tannins, glycosine and fatty acids. Another study that can prove the effectiveness ofmakahiya plant is the study of the effect of mimosa pudica root extract on vaginal estrous and serum hormones for screening of antifertility activity in albino mice. Mimosa pudica is one of the folk medicinal plants commonly used as antifertility agent in some places in India. The present work was carried out to evaluate the claimed antifertility effect of the plant by carrying out pharmacological studies with the root extract of the plant.The root extract of M. pudica has antifertility effect as it prolongs the estrous cycle and disturbs the secretion of gonadotropin hormones in albino mice (Nong Maikaen, 2007).

One of the characteristics of the Makahiya plant is its antifertility that is effective to delay on-going reproduction stages. The plant also is used by the local people to treat snakebites. It was effective to neutralize the lethality and toxic enzymes of NajaKarouthia venom. The unusual C-gylcosidic flavonoid, 4-hydroxymaysine isolated from the touch sensitive legume M. Pudica could contribute to the resistance of this plant to generalist insect herbivores (Lobstein et al., 2012). It contains an alkaloid mimosine which has been found to have potent antiproliferative and apoptopic effects. In the present study, the synthesized AgNPs from M. Pudica were tested for larvicidal activity against the larvae of Anopheles Subpictus and R. Microplus which showed increased activity from the synthesized AgNPs and it was also tested against bacteria and fungi (Springer-Verlag, 2010). Ovicide means a chemical agent that kills eggs, especially the eggs of insects and larvicide means an insecticide designed to kill larval pests. It is found to be sensitive and efficient to detect the population of Aedes, even when the population densities were low. In addition proved that it is safe, economical and environmental friendly.

It can monitor and detect Aedes mosquito population acting as an early warning signal to pre-empt any impending dengue out outbreaks. It can be lethal to eggs once a larvicide is incorporated on the oviposition substrate (Pelayo, 2013). Vasuki (1990), Ali et al. (1995), Su and Mulla (1998) and Suman et al. (2010) have suggested that larvicidal and ovicidal efficacies are governed by the type and concentration of different classes of insect growth regulators. Vasuki (1990) and Su and Mulla (1998) reported diflubenzuron, pyriproxyfen and azadirachtin to be less toxic to Cx. Quinquefasciatus eggs at a WHO recommended concentrations for larviciding (WHO, 2006). In order to manage mosquito populations at egg stages, the eggs need to be exposed at higher concentrations of insect growth regulators for shorter rather than long durations.

Eggs exposed for longer durations at low concentrations will defeat the idea of mosquito control as eggs would hatch unaffected since there is always a short duration between when the eggs are laid (susceptible stage) and embryonic development (resistant stage) begins. Freshly laid eggs are more vulnerable to the toxicity of insect growth regulators than embryonated eggs. Similar observations were made by Miura et al. (1976) and Vasuki (1990). Miura et al. (1976) suggested that freshly laid and 2–14 h old eggs were more vulnerable to diflubenzuron exposure. While, Vasuki (1990) demonstrated species specific variation for ovicidal action of chitin synthesis inhibitor and juvenile hormone analogagainst eggs of Anopheles stephensi (Liston), Cx. quinquefasciatus and Ae. aegypti.

These differences may be attributed to the inability of insect growth regulators to disrupt hormone actions during egg development and the loss of shell permeability due to endochorion tanning and wax layer formation (Clements, 1992). Such variations associated with the egg structure, egg morphology and physiological adaptation of eggs to a particular habitat bear more significance in the light of the observations made by Linley and Craig (1994), Suman et al. (2008, 2011) and Jagadeeshan and Singh (2007). They concluded that variations in a habitat bring changes to egg structures that may result into decreased ovicidal efficacy.

According to M. Govindarajan (2013) all leaf and seed extracts of Pithecellobium dulce (Roxb.) Benth. (Fabaceae) showed moderate larvicidal and ovicidal effects. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Zero hatchability was observed at 400 mg/l for leaf methanol extract and 625 mg/l for seed methanol extract of P. dulce against An. stephensi and Ae. aegypti, respectively. Compared to leaf extracts, seed extracts have low potency against the two mosquitoes. The plant world comprises a rich untapped pool of phytochemicals that may be widely used in place of synthetic insecticides in mosquito control programme (Anupam Ghosh, 2011). Aqueous and organic solvent extracts of plants/plant parts were effective in killing the mosquito larvae (K. Raghavendra, 2008).

Phytochemicals are botanicals which are naturally occurring insecticides obtained from floral resources. Applications of phytochemicals in mosquito control were in use since the 1920s, but the discovery of synthetic insecticides such as DDT in 1939 side tracked the application of phytochemicals in mosquito control programme. After facing several problems due to injudicious and over application of synthetic insecticides in nature, re-focus on phytochemicals that are easily biodegradable and have no ill-effects on non-target organisms was appreciated. Since then, the search for new bioactive compounds from the plant kingdom and an effort to determine its structure and commercial production has been initiated. At present phytochemicals make up to 1 per cent of world’s pesticide market. Plants produce numerous chemicals, many of which have medicinal and pesticidal properties. More than 2000 plant species have been known to produce chemical factors and metabolites of value in pest control programmes.

Members of the plant families- Solanaceae, Asteraceae, Cladophoraceae, Labiatae, Miliaceae, Oocystaceae and Rutaceae have various types of larval, adulticidal or repellent activities against different species of mosquitoes (Anupam Ghosh N. C., 2012). Phytochemicals may serve as suitable alternatives to synthetic insecticides in future as these are relatively safe, inexpensive, and are readily available in many areas of the world. Different parts of plants contain a complex of chemicals with unique biological activity which is thought to be due to toxins and secondary metabolites, which act as mosquitocidal agents. Furthermore, the crude extracts may be more effective compared to the individual active compounds, due to natural synergism that discourages the development of resistance in the vectors. There has been an increasing interest in anti-mosquito products derived from natural origin because the continued applications of synthetic compounds have some drawbacks, including the widespread development of insecticide resistance.

Another drawback with the use of chemical insecticides is that these are non-selective and could be harmful to other organisms in the environment. The toxicity problem, together with the growing incidence of insect resistance, has called attention to the need for novel insecticides, and for more detailed studies of naturally-occurring insecticides. It is, therefore, necessary to develop new materials for controlling mosquitoes in an environmentally safe way, using biodegradable and target-specific insecticides against them (M. Govindarajan, M. Rajeswary, & R. Sivakumar, 2013). Human beings have used plant parts, products and secondary metabolites of plant origin in pest control since early historical times. Vector control has been practiced since the early 20th century. During the pre-DDT era, reduction of vector mosquitoes mainly depended on environmental management of breeding habitats, i.e., source reduction.

The efficacy of phytochemicals against mosquito larvae can vary significantly depending on plant species, plant parts used, age of plant parts (young, mature or senescent), solvent used during extraction as well as upon the available vector species. Sukumar et al.(n.d.) have described the existence of variations in the level of effectiveness of phytochemical compounds on target mosquito species vis-à-vis plant parts from which thesewere extracted, responses in species and their developmental stages against the specified extract, solvent of extraction, geographical  origin of the plant, photosensitivity of some of the compounds in the extract, effect on growth and reproduction. Today, environmental safety is considered to be of paramount importance. An insecticide does not need to cause high mortality on target organisms in order to be acceptable but should be eco-friendly in nature.

Phytochemicals may serve as these are relatively safe, inexpensive and readily available in many parts of the world. Several plants are used in traditional medicines for the mosquito larvicidal activities in many parts of the world. According to Bowers et al. (n.d.), the screening of locally available medicinal plants for mosquito control would generate local employment, reduce dependence on expensive and imported products, and stimulate local efforts to enhance the public health system. The ethno-pharmacological approaches used in the search of new bioactive toxins from plants appear to be predictive compared to the random screening approach. The recently developed new isolation techniques and chemical characterization through different types of spectroscopy and chromatography together with new pharmacological testing have led to an interest in plants as the source of new larvicidal compounds. Synergistic approaches such as application of mosquito predators with botanical blends and microbial pesticides will provide a better effect in reducing the vector population and the magnitude of epidemiology (Anupam Ghosh N. C., 2012). Chapter 3

This chapter introduces the research strategy and the empirical techniques applied in the research. This also presents what samples are being used in the study and how the researchers choose the specific samples. This includes the procedures that the researchers did in order for them to come up with the desired product or mixture. Research Design

The experimental method of research was used in this study. Experimental method involves manipulating one variable. This is a method or procedure involving the control or manipulation of conditions for the purpose of studying relative effects of various treatments applied to members of a sample or of the same treatment applied to members of different samples. The experimental method involves manipulating one variable to determine if changes in one variable cause changes in another variable. This method relies on controlled methods, random assignment and the manipulation of variables to test a hypothesis (Cherry, 2014). Research Locale of the Study

The research locale of this study is St. Paul’s School of Ormoc Foundation, Inc. The researchers conducted the study in the said institution and gathered the mosquito larvae at the house of one of our researchers. The researchers conducted the study in school because the school provided us with apparatuses that we can use in our experiment.

Samples of the Study
The researchers chose the appropriate samples in order to create the product that could be effective enough as an alternative larvicide for mosquito larvae. The researchers used the Makahiya plant as the main ingredient. A specific amount was supplied as the researchers conducted the experiment for the desired result with the help of other materials such as blender, beaker, timer, dropper, magnifying glass, graduated cylinder, and a stirring rod. Sampling Design

The appropriate sampling design for our experimental research regarding the feasibility of Mimosa Pudica plant extract as an alternative larvicide is purposive sampling wherein the samples obtained are chosen based on the researchers’ judgment. The samples obtained are chosen because these are the sole ingredients in proving our research. Validation of Instrument

Research Procedure
Specifically, the researchers used the following:
Medicine Dropper
Magnifying Glass
Graduated Cylinder
Mosquito Larvae
Makahiya Plant
Stirring rodss

The set-ups had different numbers of the following ratios: 8ml makahiya extract: 2ml water; 6ml makahiya extract: 4ml water; 10ml of makahiya extract. Before you start:
1. Blend the makahiya plant.
2. Squeeze the makahiya plant after blending.
3. Separate three set-ups according to the desired measurement of the solution. Application of the Product:
1. In set-up A with the use of a medicine dropper, apply 8ml makahiya extract and 2ml water in the mosquito larvae samples and see the reaction of the mosquito larvae from the makahiya extract solution for 5 to 45 minutes. 2. In set-up B with the use of a medicine dropper, apply 6ml of makahiya extract and 4ml water in the mosquito larvae samples and see the reaction of. 3. In set-up C with the use of a medicine dropper, apply 10 ml of makahiya extract in the mosquito larvae.

(2007). Retrieved from Filipino Herbs.
Anupam Ghosh, N. C. (2011, April 13). Retrieved May 2012, from http://icmr.nic.in/ijmr/2012/may/0502.pdf Anupam Ghosh, N. C. (2012, May). Plant extracts as potential mosquito larvicides. Retrieved from NCBI: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401688/ Cherry, K. (2014). Retrieved from About.com: http://psychology.about.com/od/eindex/g/experimental.htm Encarta Dictionaries. (2010).

K. Raghavendra, S. S. (2008, February 4). Retrieved July 2009, from http://icmr.nic.in/ijmr/2009/july/0711.pdf Merriam Webster Dictionary. (2010).
Microsoft Encarta. (2009).
Nong Maikaen, A. P. (2007). The effect of Mimosa Pudica root extract on vaginal estrous and serum hormones for screening of antifertiliy activity in
albino mice. In Contraception (pp. 482-485). Bangkok, Thailand: Springer Netherlands. Pelayo, R. (2013, August 31). Retrieved from FILIPINO DISCOVERIES, INVENTIONS, INNOVATIONS and PRODUCTS: http://devcomconvergence.wordpress.com/2013/08/31/dost-region-6s-mosquito-ovicidallarvicidal-ol-trap-as-an-alternative-mosquito-trap-in-reducing-dengue-mosquito/ Soon-Young Park, J.-S. I.-R.-E.-J.-K. (n.d.). Retrieved from Researcher’s Group: http://www.researcher.com/plantsandalternatives

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