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CHAPTER I
Problem and its Background
Introduction
Dengue Dengue is a pande pandemic mic concern concern and it common commonly ly affect affectss the the people peoples s health health in de!eloping countries" Although ad!anced technology and medicines are no# a!ailable$ cases of dengue are still rising up to date d ate #ith outstanding numbers"
The Philippines ha!e been a dengue fe!er hotspot for se!eral years" During the first %uarter of &'(($ The Philippine Department of Health reported that the incidence of dengue fe!er doubled doubled in )etro )anila" Compared Compared to last years statist statistical ical record record of dengue cases in the Philippines$ the ($*+, dengue case in &'(' has significantly increased to ,$-** dengue cases during the same period in &'((" .Tube/a$ Philip$ 0D1H 0D1H tags deng de ngue ue 2ho hott sp spot otss in Pa Pasi sig$ g$ 3alen en/u /uel ela4 a4$$ Apr April
(,$ (,$ &'( &'(($ ###"in%uirer"net ###"in%uirer"net$$
http566###"in%uirer" http566###" in%uirer"net6specialfeatures6denguealert6!ie#" net6specialfeatures6denguealert6!ie#"php7db8(9article8&'((',(,: php7db8(9article8&'((',(,: --((+;<< --((+;
The main cause of dengue is a female mos%uito kno#n as Aedes as Aedes aegypti #hich aegypti #hich carries the dengue !irus" 1nce the !irus enters the human body$ it puts the persons persons life at risk" risk" The Aedes The Aedes aegypti usually aegypti usually breeds on areas #ith stagnant #ater and filthy damp places such such as disc discar arded ded tire tires$ s$ flo# flo#er er !ases !ases and and unco! unco!er ered ed barr barrel els" s"
=tudi =tudies es sho# sho# that that
mos%uitoes are fast reproducing animals$ thus pre!enting the population gro#th of the lar!ae is the best #ay to lessen the alarming number of dengue cases"
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Jasminum sambac is sambac is a bushy !ine #ith dark green lea!es and has fragrant little flo#ers flo#ers used as decorations decorations that ha!e aromatherapy aromatherapy purposes" purposes" It is #ell regarded regarded as an aphrodisiac and also considered as an antidepressant$ sedati!e and antispasmodic"
It is a species of >asmine nati!e to =outh and =outheast Asia and it is the national flo#er of the Philippines" Jasminum Philippines" Jasminum sambac is sambac is abundant to the country especially in =an Pedro$ ?aguna"
Background of the =tudy
Dengue is an acute acute fe!er caused by a !irus" !irus" It occurs in t#o t#o forms forms namely$ namely$ the .(< dengue fe!er and the .&< dengue dengue haemorrhagic haemorrhagic fe!er" fe!er" Dengue spreads spreads rapidly rapidly and may affect large number of people during an epidemic resulting in reduced #ork producti!ity$ but most importantly causing the loss of li!es"
Dengue is spread spread by the bite of an infected infected female female mos%uito mos%uito (Aedes aegypti)" aegypti)" The mos%ui mos%uito to gets the !irus by biting biting the infected infected persons" persons" There There is no #ay to tell tell if a mos%uito is carrying the dengue !irus" !irus" Therefore$ people must protect themsel!es from all mos%uito bites" All efforts of control should be directed against the mos%uitoes" mos%uitoes" It is important to take control measures to eliminate the mos%uitoes and their breeding places" Ho#e!er$ the efforts should be intensified before the transmission season .during and after the rainy season< and at the time of the epidemic"
In the year (**+$ Philippines Philippines had its its #orst cases of dengue" This caused and started
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Jasminum sambac is sambac is a bushy !ine #ith dark green lea!es and has fragrant little flo#ers flo#ers used as decorations decorations that ha!e aromatherapy aromatherapy purposes" purposes" It is #ell regarded regarded as an aphrodisiac and also considered as an antidepressant$ sedati!e and antispasmodic"
It is a species of >asmine nati!e to =outh and =outheast Asia and it is the national flo#er of the Philippines" Jasminum Philippines" Jasminum sambac is sambac is abundant to the country especially in =an Pedro$ ?aguna"
Background of the =tudy
Dengue is an acute acute fe!er caused by a !irus" !irus" It occurs in t#o t#o forms forms namely$ namely$ the .(< dengue fe!er and the .&< dengue dengue haemorrhagic haemorrhagic fe!er" fe!er" Dengue spreads spreads rapidly rapidly and may affect large number of people during an epidemic resulting in reduced #ork producti!ity$ but most importantly causing the loss of li!es"
Dengue is spread spread by the bite of an infected infected female female mos%uito mos%uito (Aedes aegypti)" aegypti)" The mos%ui mos%uito to gets the !irus by biting biting the infected infected persons" persons" There There is no #ay to tell tell if a mos%uito is carrying the dengue !irus" !irus" Therefore$ people must protect themsel!es from all mos%uito bites" All efforts of control should be directed against the mos%uitoes" mos%uitoes" It is important to take control measures to eliminate the mos%uitoes and their breeding places" Ho#e!er$ the efforts should be intensified before the transmission season .during and after the rainy season< and at the time of the epidemic"
In the year (**+$ Philippines Philippines had its its #orst cases of dengue" This caused and started
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the great alarm alarm to the health health of people people li!in li!ing g in the country country"" And And until until the presen present$ t$ @ili @ilipi pinos nos are stil stilll in dange dangerr of the the endem endemic ic dise diseas asee here here in the the Phili Philippi ppine nes" s"
The The
Department of Health reported that cases of dengue fe!er doubled in )etro )anila during the first four months of the year &'((" &'((" The Philippines has been a dengue hotspot for years and the cases are most alarming5 ,$-** cases in &'(( #hile ($*+, cases in &'('" The Depart Departmen mentt of Health Health cited )anila )anilas Baranga Barangay y =an )iguel )iguel in Pasig Pasig City City and Barangays in 3alen/uela 3alen/uela as particularly high h igh risk areas"
There There is is no specific specific medicine medicine for the treatment treatment of the disease" disease" @or this reason$ reason$ this this research study is to be conducted to produce an effecti!e #ay of lessening the greater risks of dengue by using the lar!icide lar!icide produced from an organic compound compound found in the =ampaguita leaf to kill the lar!ae of mos%uitoes"
Theoretical @rame#ork
Reinfestati Reinfestation$ on$ aggra!ation aggra!ation of the dengue situation$ situation$ economic economic difficult difficulties$ ies$ and kno#ledge deficit are the threats of the urbani/ation of dengue fe!er$ thus it continues the cycle of dengue fe!er and number number of dengue cases started started to increase increase again" Based on Carl Carlo o " @inl @inlay ay and and Dr" Dr" uan uan uite uitera ras s theory theory #hic #hich h says says that that the the infe infect ctio ion n #as #as transmitted from person to person through the bite of the mos%uito$ the basis of this study is to ha!e effecti!e measures in controlling the disease first by pre!enting the population
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gro#th of Aedes aegypti #hich carries the dengue !irus by eradicating the mos%uito lar!ae using the leaf etract of =ampaguita . Jasminum sambac< as a lar!icide"
The =ampaguita or Jasminum sambac ha!e the constituents of alkaloids$ glycoside$ fla!anoid$ terpines$ tannin$ resin and salicylic acid" The =ampaguita contains alkaloids that are highly poisonous and can ha!e a pharmacological effect on animals" .)o ?i Hua" =ampaguita$ Philippine )edicinal Plants" 1ctober &'('" http566###"stuartchange"org6=ampaguita"html< Because of this$ there might be a possibility that the =ampaguita . Jasminum sambac< can help in pre!enting further gro#th of Aedes aegypti.
Conceptual @rame#ork
IPT
:ature of =ampaguita lea!es containing Alkaloid -Aedesa egypti lar!ae
PR1CE==
:Crude method
1TPT
etraction
:Testing the leaf etract on Aedes aegypti lar!ae to determine its lar!icidal property and effecti!eness"
Jasminum sambac crude leaf etract as an effecti!e lar!icide against Aedes aegypti.
@igure ("( A paradigm sho#ing the conceptual frame#ork of the study
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@irst$ the researchers #ould ha!e the =ampaguita lea!es #ith the Aedes aegypti lar!ae for the etraction and eperimentation process" In the Process table$ the researchers ha!e the crude etraction method to take out the leaf etract from the =ampaguita leaf" After#ards$ the researchers #ould look for#ard to ha!ing an effecti!e lar!icide against Aedes aegypti lar!ae" This paradigm #ould be follo#ed during the #hole study and #ill be used as the guide in continuing #ith this study"
=tatement of the Problem )ain Problem5 Does the leaf extract of Jasminum sambac contain effective larvicidal property? The follo#ing %uestions #ill be ans#ered by the end of this research5 (" Fhat are the effects of the crude leaf etract to Aedes aegypti lar!ae7 a" In terms of the mortality rate b" In terms of time of effecti!ity b"(< after (; minutes b"&< after -' minutes b"-< after G' minutes b",< after (&' mintues
c" In terms of the concentration of the etract
c"(< ( ppm
c"&< (' ppm
c"-< ('' ppm
c",< (''' ppm
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Assumptions
A solution to one of the alarming dangers in the Philippines #hich is the dengue is the production of efficacious lar!icides" In this study$ Jasminum sambac contains glycosides and alkaloid that can be used as lar!icides #hich can pre!ent the greater risks of dengue therefore$ lessening the cases regards to the loss of life" There are also specific methods to etract the re%uired organic compounds from the said plant material to produce an effecti!e lar!icide" The lar!icidal property of the Jasminum sambac #ill greatly affects the population gro#th of the mos%uito$ causing high mortality rate" Aedes aegypti$ resulting to a safer place to li!e in"
Hypothesis
Null ypothesis! The Jasminum sambac leaf etract has no significant effect on Aedes aegypti lar!ae"
Alternative ypothesis! The Jasminum sambac leaf etract has a significant effect on Aedes aegypti lar!ae"
=ignificance of the =tudy The aim of the study is to make use of the lar!icidal property of =ampaguita . Jasminum sambac< lea!es as an effecti!e lar!icide" The lar!icide #ould be pre!enting the population gro#th of Aedes aegypti lar!ae that carries Dengue" The number of
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Dengue cases according to the Department of Health is ($*+, last year to ,$-** this year and the number of deaths caused by this disease is already only in )etro )anila" This research study is efficient and is one of the best #ays in pre!enting dengue that may benefit the #hole community"
=cope and ?imitations of the =tudy The study is focused on testing the lar!icidal property of =ampaguita . Jasminum sambac< lea!es and its effect to Aedes aegypti lar!ae" The Jasminum sambac lea!es #ould be gathered from =an Pedro$ ?aguna" The =ampaguita leaf etract #ould be etracted by using an electric blender to pul!eri/e then a =ohlet etractor to take out the alkaloid from the po#der and after#ards to be placed in a petri dish" The lar!icide #ould be sprayed in different amounts in four setups containing not less than (; female mos%uito lar!ae" The Aedes aegypti #ith the lar!icide #ould be eposed for &, and ,+ hours" The study #ould last for about t#o to three months" The emphasis is not to cure Dengue fe!er but rather to help in pre!enting the population gro#th of the Aedes aegypti"
Definition of Terms
Alkaloids" It is a chemical compound that is considered most po#erful and effecti!e
among all elements found in plants" Its effecti!eness commonly includes all substances that are poisonous in the plants" Alkaloids are deri!ed from amino acids and include an
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enormous number of bitter$ nitrogenous compounds according to R"@" Raffauf .Plant Alkaloids5 A uide To Their Disco!ery and Distribution$ (**G<" Alkaloids are considered to be disintegrate substances that comprise proteins" Dengue fe!er"an acute infectious disease transmitted by Aedes aegypti mos%uitoes$ and characteri/ed by headache$ se!ere >oint pain$ and a rash" .0Dengue"4 )erriam: Febsters Collegiate Dictionary" ('th ed" &''("< Dengue Haemorrhagic fe!er"It is a se!ere$ potentially deadly infection spread by certain species of mos%uitoes . Aedes aegypti<" Bleeding and sometimes shock occurs J leading to death" .3aughn DF$ Barrett A$ =olomon T" @la!i!iruses .Kello# @e!er$ Dengue$ Dengue Hemorrhagic @e!er$ apanese Encephalitis$ Fest ile Encephalitis$ =t" ?ouis Encephalitis$ Tick:Borne Encephalitis<" In5 )andell ?$ Bennett E$ Dolin R$ eds" Principles and Practice of Infectious Diseases" th ed" Philadelphia$ Pa5 Else!ier Churchill ?i!ingstone &''*5chap (;-"< Dog Heart#orm". Dirofilaria immitis)$ a disease common in canines caused by a nematode" .0Does Kour Dog Ha!e a Heart#orm74 .Crans$ F" Does Kour Dog Ha!e A Heart#orm7Rutgers"
Retrie!ed
@ebruary
($
&'(&$
from
###:
rci"rutgers"edu6Linsects6hart#"htm< Epidemic" Affecting a disproportionally large number of indi!iduals #ithin a population or community" .0Epidemic"4 )erriam:Febsters Collegiate Dictionary" ('th ed" &''("< ?ar!ae" It is an immature$ #ingless$ and often !ermiform feeding form that hatches from the egg of a many insects$ increases in si/e$ undergoes other minor changes #hile passing through se!eral molts$ and is finally transformed into a pupa or chrysalis from
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#hich the adult ultimately emerges".0?ar!a"4 )erriam:Febsters Collegiate Dictionary" ('th ed" &''("< ?ar!icide" It is an insecticide or other pesticide used for killing lar!ae" .0lar!icide"4 )erriam:Febster"com" &'((" http566###"merriam:#ebster"com" @ebruary ($ &'((<" )urray 3alley" It is a rare disease caused by the )urray 3alley encephalitis !irus" It is spread to humans by infected mos%uitoes" .)arch &$ &'((" )urray 3alley Encephalitis$ =F
o!ernment$
Health"
Retrie!ed
@ebruary
(($
&'(&$
from
http566###"health"ns#"go!"au6factsheets6infectious6murrayM!alleyMenceph"html< Pandemic" It is occurring o!er a #ide geographic area and affecting an eceptionally high proportion of the population" .0pandemic"4 )erriam:Febsters Collegiate Dictionary" ('th ed" &''("< Ross Ri!er !irus" It is a germ that infects people$ particularly in rural areas$ sometimes causing a flu:like illness #ith >oint pains$ rash and fe!er" .anuary (G$ &''+" )urray 3alley Encephalitis$ =F o!ernment$ Health" Retrie!ed @ebruary (($ &'(&$ from http566###"health"ns#"go!"au6factsheets6infectious6rossri!er"html< =ampaguita" .scientific name Jasminum sambac< It is a species of the genus >asmine and is #idely gro#n as an ornamental plant" This species is nati!e to southern Asia$ India and =ri ?anka".=ampaguita"The @lo#er Epert" Retrie!ed @ebruary (($ &'(&$ from http566###"theflo#erepert"com6content6aboutflo#ers6tropicalflo#ers6sampaguita< 3accine" It is a biological preparation that impro!es immunity to a particular disease" A !accine typically contains an agent that resembles a disease:causing microorganism$ and is often made from #eakened or killed forms of the microbe".0!accine"4 )erriam: Febster"com" &'((" http566###"merriam:#ebster"com" @ebruary ($ &'((<"
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CHAPTER II Re!ie# of Related ?iterature and =tudies Related ?iterature @oreign ?iterature Dengue is an epidemic disease that occurs on #armer climates caused by a !irus trans: mitted by the mos%uito$ Aedes aegypti$ #hich also spreads yello# fe!er" In =outheast
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Asia$ a se!ere form of the ailment called dengue haemorrhagic fe!er has caused numerous deaths for the past fe# years" .0Dengue fe!er4$ rolier Academic Encyclopedia$ (*++<
The 0tiger mos%uito4 or Aedes aegypti$ is a species that breeds in
artificial containers found in houses" This type of insect bites on its hosts during daytime" Aedes aegypti is the !ector of urban yello# fe!er and dengue" This species are associated #ith the transmission of the !iral haemorrhagic fe!ers$ of =outheast Asia" The female Aedes aegypti has mouthparts modified for sucking blood from its hosts" The female mos%uitoes need it in order to mature its eggs" In dengue !irus infected mos%uitoes$ the !irus is present in the sali!ary glands of the mos%uito" Fhen a female Aedes aegypti bites a human for food$ she in>ects sali!a into the #ound #here the anti: coagulants contained in her sali!a facilitate feeding" .0 Aedes aegypti4$ Dengue 3irus et$ http566###"dengue!irusnet"com6aedes:aegypti"html< )ost mos%uito species spend much of their life cycle in lar!al stage #hen they are highly susceptible to both predation and control efforts" They often are concentrated #ithin defined #ater boundaries$ immobile #ith little ability to disperse$ and accessible" ?ar!ae feed on organic particulate matter in the #ater like algae" As long as the temperature is cool$ the Aedes aegypti lar!ae can remain in its lar!al stage for months" .0?ife Cycle of Aedes aegypti4$ Dengue 3irus et$ http566###"dengue!irusnet"com6life: cycle:of:aedes:aegypti"html< Adult mos%uitoes$ on the other hand$ fly in search of males$ blood meals$ or #ater sources for egg laying and are usually inaccessible$ not concentrated$ and highly distributed"
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There are se!eral theories regarding alkaloids stating that this type of element is a by: product of plant:metabolism that they are means of protection of plants against animal and insect attacks or that they are reser!oirs for protein synthesis$ regulators of gro#th and reproduction$ or detoifying agents" .0Alkaloids4$ rolier Academic Encyclopedia$ (*++< This element has a ma>or positi!e effect on the intensity of making a lar!icide" A lar!icide$ on the other hand$ is a chemical used for killing lar!ae" ?ar!icides are chemicals used to kill mos%uito lar!ae" ?ar!icides are more effecti!e in controlling mos%uitoes because they kill mos%uitoes before they become adults" .0?ar!icides4$ @air Harbor$ http566###"fairharbor"com6fhca6ca&,'*lar!icides"htm< The lar!al stage of a mos%uito #ould be its #eakest point" Therefore$ effecti!e lar!iciding can reduce the number of mos%uitoes a!ailable to disperse$ potentially spread diseases and lay eggs #hich leads to more mos%uitoes" .0?ar!icides and ?ar!iciding4$ )os%uito Information Febsite$ http566mos%uito"ifas"ufl"edu6?ar!iciding"html$ &* =eptember &'((<
?ocal ?iterature Dengue has been sei/ing headlines again in !arious local papers and publications in Asia about another outbreak in the Philippines and a predicted upcoming outbreak #hich #ould be much #orse than that of (**+$ #here there #ere more than -+$''' cases" .0&'(( Dengue 1utbreak"4$ Biotech Philippines$ http566###"biotech"ph6&'((:dengue: outbreak$ &* =eptember &'((<
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The mos%uitos bullet:shaped eggs are singly laid and are #ithout floats" The tiger mos%uito lays its eggs in any type of #ater$ mainly in polluted areas" ?ar!al de!elopment is affected critically by the presence of ade%uate moisture and surrounding !egetation$ light intensity$ salinity or alkalinity of the a%uatic medium$ and temperature" )os%uito control is largely a matter of familiarity #ith the biological and ecological re%uirement of the target species #hene!er possible" .Balta/ar$ Clare and =ala/ar$ elia 0Philippine Insects an Introduction4$ Diliman$ Nue/on City$ ni!ersity of the Philippines Press$ (**< =ampaguita . Jasminum sambac< or Oampopot$ a climbing shrub #ith #hite flo#ers$ usually #ith a double corolla$ #hich has a delightful fragrance and nati!e to southern Asia$ contains a po#erful element called alkaloid" .=teiner$ )ona ?isa$ 0Philippine 1rnamental Plants and Their Care4$ )anila$ Philippines" (**&<
Related Readings The related readings tell about the certain topics such as administrati!e orders$ health programs$ campaigns and ordinances concerning dengue pre!ention and control" Thus$ using the full kno#ledge ac%uired can help in the process of pre!enting such epidemic disease to #orsen" The alarming number of dengue cases ser!es as a #ake:up call to e!eryone that #e must pre!ent and control the spread of dengue"
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The Administrati!e order no" &+( of (**G entitled 0The ational Tepok ?amok and Dengue =apok Program4 eplains about the pre!ention of the spread of dengue !irus by a ation#ide clean:up dri!e operation and elimination of mos%uito breeding places in 1bser!ance of the @our 1clock Habit #ith the participation of the ational Agencies and ?ocal o!ernment nits particularly the Department of Health that leads this pro>ect"( Though the ordinance stated abo!e is a good #ay to pre!ent the massi!e effect of the dengue !irus$ it is still not good enough to say that #e are safe from being affected by this disease" )os%uitoes are fast reproducing insects and cleaning our surroundings #ont be enough to stop the spread of dengue" According to the Administrati!e order no" ,; of &''($ The Aedes aegypti mos%uitoes are container:breeders" Aedes aegypti could breed in any container that can hold #ater" Plastic bottles$ tin cans and plastic cans can easily be disposed but pails$ plastic drums and flo#er pots are usually used in our homes may re%uire chemical lar!iciding #hich is on a long:term basis but it ha!e the best result in situations #here the location and period at risk is indicated or specified" ?ar!icide application is more beneficial because of the fact that the mos%uitoes are destroyed before they mature and disperse to human habitation that can cause massi!e effects especially to e!ery indi!iduals health" It can also be done at home and can obtain fast results"&
1 Administrati!e 1rder no" &+( s" (**G$ '6(*6*+$ Department of Health$ '*6&*6(($ http566###"chanrobles"com6administrati!eorders6administrati!eorderno&+(&'(**G"html
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Related =tudies @oreign =tudies The researchers primarily choose this research proposal because of its significance to the common problems in the community and the increasing number of dengue cases is one of them" The researchers #ere able to find out that there are studies that are interconnected #ith the present study that #ill ser!e as the basis of this research proposal" The research proposal deri!ed its etraction process from the study in India by Oumar$ aresh et" Al about Carica Papaya leaf etract being an effecti!e lar!icide" It similar to the current study ecept that they used Papaya leaf etract as the independent !ariable" The etraction process is to use the sohlet etractor to etract the crude from the lea!es of the Carica Papaya" The plant etract sho#ed lar!icidal and pupicidal effects after &, h of eposure ho#e!er$ the highest lar!al and pupal mortality #as found in the leaf etract of methanol C" papaya against the first: to fourth:instar lar!ae and pupae of !alues ?C.;'< 8 I instar #as ;("G ppm$ II instar #as G("+ ppm$ III instar #as ,"' ppm$ and I3 instar #as +&"(+ ppm$ and pupae #as ,,'"G; ppm$ respecti!ely$ and bacterial insecticide$ spinosad against the first to fourth instar lar!ae and pupae of !alues ?C.;'< 8 I instar #as ;("G ppm$ II instar #as G("+ ppm$ III instar #as ,"' ppm$ and I3 instar #as +&"(+ ppm$ and pupae #as *-",, ppm$ respecti!ely" )oreo!er$ combined treatment of !alues of ?C.;'< 8 I instar #as ;;" ppm$ II instar #as G;" ppm$ III instar #as
2 Administrati!e 1rder no" ,; s" &''($ '+6(G6'($ Forld Health 1rgani/ation$ '*6&*6((6$http566###"doh"go!"ph6files6ao,;:'("pdf
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G"-G ppm$ and I3 instar #as *&"+ ppm$ and pupae #as ('"G& ppm$ respecti!ely" o mortality #as obser!ed in the control" The results that the lea!es etract of C" papaya and bacterial insecticide$ "pinosad is capable of being effecti!e lar!icidal and pupicidal properties of against chi#ungunya !ector$ A. aegypti" =ince the study #ill also be using the same plant part in #hich the crude #ill be etracted from$ the researchers #ill use this study as the basis for its eperimentation process"
The study of Priya Joy and Dr. D. Patric Raa entit!ed "#nti$%acteria! #cti&ity 'tudies of Jasminum grandiforum and Jasminum sambac( on the year 2008 in )ndia* +i&es ,ay in stren+thenin+ the foundation of the research -ro-osa! ecause of its -hytoche/ica! ana!ysis on Jasminum Grandiforum and Jasminum Sambac.
Phytoconstituents
Jasminum sambac
Alkaloids
Q
Tannin
Q
@la!anoid
Q
=teroid lycoside
Q Q
Terpenes
Q
Resin
Q
=alicylic acid
Q
Ta!e 3.1 Phytoche/ica! #na!ysis on Jasminum Sambac
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The ta!e ao&e sho,s that Jasminum sambac is -ositi&e in containin+ a!a!oid and +!ycoside ,hich are eecti&e in+redients in /ain+ !ar&icide. #!a!oid is said to e an eecti&e constituent for a !ar&icide accordin+ to a study y erdinand . Ta!ontsi* Jos-hat . atasyoh* et. #!. entit!ed "osuito ar&icida! #cti&ity of #!a!oids fro/ anthoy!u/ !e/airei a+ainst the a!aria ector Anopheles gambiae( ,hich ,as conducted in :er/any on ;o&e/er 2010.
The study
sho,s that the
oca! 'tudies
The researchers ased the concentrations of the etracts on the study entit!ed "The !ar&icida! eect of :uyaano = Annona muricata> !eaf etract on Aedes aegypti /osuito( y Tan++a$an* ?ie oren in eu on 1999 in ,hich used 1 --/* 10 --/* 100 --/* and 1000 --/ as its concentration that otained the fo!!o,in+ /orta!ity rates as the resu!t of the study@ 1000 --/ A 0.61B 100 --/. A 0.403B 10 --/ A
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0.203B 1 --/ A 0.10 and conc!uded that the :uyaano !eaf etract eerted !ar&icida! eect on the t,o$day o!d Aeges aegypti !ar&ae ,ith the hi+her concentration eertin+ +reater -otency.
The -eriod of ti/e of recordin+ the resu!ts of the research is ased on a study in the Phi!i--ines durin+ 1993$1994* entit!ed "The !ar&icida! eects of nee/ tree = Azadirachta indica> !eaf and seed etracts on /osuito !ar&ae ='econd year schoo! year 1993$1994>( y P. Padi!!a in ,hich the researcher used the said etracts for dierent -eriods of ti/e@ 15 /inutes* 30 /inutes* 60 /inutes and 120 /inutes. #fter three re-!icates ,ith a /ai/u/ -eriod of t,o hours* resu!ts sho,ed an increasin+ !ar&icida! eect as the concentration of the ;ee/ etract fro/ oth seed and !eaf is increased. or the -ositi&e contro!* a 100C !ar&icida! eect ,as oser&ed 15 /inutes after the ti/e of a--!ication.
In this study$ the researchers are to in!estigate if Jasminum sambac has lar!icidal property" =ubse%uent the in!estigations abo!e$ Jasminum sambac may contain lar!icidal property that can be beneficial in pre!enting diseases specifically Dengue"
ustification of the =tudy )any approaches ha!e been de!eloped to control the mos%uito threats" 1ne approach is killing the mos%uito at lar!al stage" In this study$ =ampaguita Jasminum sambac leaf
19
etract is to be tested #hether it is an effecti!e lar!icide to mos%uitoes specifically Aedes aegypti" The study is distinct #ith the other studies because it has the same sub>ect matter and it both aims to make use of an etract from a plant to be made into a lar!icide" Based on the gi!en related studies$ the phytochemical analysis of the Jasminum sambac callus etract$ it is sho#n that alkaloid is present" =tudies ha!e sho#n that some plant etracts like the ones mentioned in the related studies contain lar!icidal property" These plant etracts from different parts of the plant contain alkaloids and glycosides that can be made as a lar!icide" There is also a study sho#ing t#el!e plants tested for ha!ing lar!icidal property but Jasminum sambac is none of the listed tested plants"
?ar!icidal application is more beneficial than the other
methods in controlling Aedes aegypti because it sho#s more accurate and effecti!e results because it inhibits the gro#th of the lar!ae thus makes it unable to gro# fully into a mos%uito and spread the dengue !irus to humans" The related literature and studies pro!es that the research proposal is not a duplicate of any other study"
20
CHAPTER III Research )ethodology Research Design In order to test the effecti!eness of the =ampaguita leaf etract #hich is the treatment to be applied to the mos%uito lar!ae$ different le!els of the etract must be tested upon sets of lar!ae" =ince there is only one treatment but has different le!els$ #ithout a control group$ the completely randomi/ed design #ill be used" The amount of the lar!ae must be obser!ed first before applying the treatment" Replications of the sets of lar!ae #ill be made for each le!el of treatment to reassure the accuracy of the treatments effect" The containers #ith lar!ae are obser!ed repeatedly and #ill be recorded" If a change in the amount of the mos%uito lar!ae is seen$ then it can
21
be a strong e!idence to the lar!icides effecti!eness" Fhen the eperiment is done after se!eral obser!ations$ data #ill be collected" Random =election
(+' Aedes aegypti lar!ae
Treatment =et:up A 8(; lar!ae6( ppm .#ith - replicates< =et:up B8 (; lar!ae6(' ppm .#ith - replicates< =et:up C8 (; lar!ae6('' ppm .#ith - replicates< =et:up D8 (; lar!ae6(''' ppm .#ith - replicates<
Posttest umber of dead lar!ae In set:up A umber of dead lar!ae In =et:up B umber of dead lar!ae In =et:up C umber of dead lar!ae In =et:up D
Table -"& The table sho#s the research design to be used" In order to see the effecti!eness of the lar!icide$ data is charted" After#ards$ the differences bet#een each set of containers are obser!ed" If there is a clear distinction bet#een the amounts of lar!ae from the others$ then an inference #ould be much easier to come up on"
Determination of =ample =i/e There #ill be a number of treatments each #ith different le!els of concentration of the etract from the =ampaguita leaf" These treatments #ill be applied to containers #ith lar!ae" Each petri dish contains fifteen lar!ae" There #ill be three replicates for each container assigned to a certain le!el of treatment to determine the differences of each
22
result accurately" In total$ there #ould be (+' Aedes aegyptilar!ae" The researchers may form an unbiased conclusion" A large sample si/e #ill make the records homogeneous" This type of data #ill make accurate statistical treatments #ith small amounts of !alues"
=ampling Design and Techni%ue The researchers #ill use probability sampling techni%ue to obtain unbiased results" The researchers #ill use the method of lottery sampling to set each petri dish$ containing (; lar!ae$ to its corresponding set" After#ards$ each set #ill be paired to a different le!el of treatment" The =ub>ect The sub>ect to be used in this study is the mos%uito lar!ae that carries dengue !irus$ #hich is kno#n as the Aedes aegypti" The Aedes aegypti$ a female mos%uito is recogni/ed as a small$ dark mos%uito #ith #hite markings on their legs" This type of mos%uito breeds on stagnant #aters and filthy damp places" The mos%uito carries other diseases rather than >ust dengue" The Aedes aegypti lar!ae may be obtained through o!icidal traps$ breeding and gathering the sub>ect" The population of Aedes aegypti are decreased by the etermination of its lar!ae for further gro#th"
23
The Research Instrument The researchers #ould be using a research instrument to record and gather the data needed in the eperiment" This research instrument #ould be in the form of obser!ation sheets that #ould be used during the eperiment" It #ould contain the results of the eperiment" The researchers chose to ha!e four concentrations #ith three replicates under each concentration" The first column #ould contain the factors to be considered during the eperiment" In the net columns$ records about the four concentrations ( ppm$ (' ppm$ ('' ppm and (''' ppm #ould be sho#n" Thus$ the e!aluation of the study #ould be more organi/ed and efficient #ith the help of the research instrument"
3alidation of the Research Instrument The research instrument must be !alidated and re!ised first by the research teacher before using it during the eperiment"
The re!ised research instrument includes the
suggestions to secure the legitimacy of the instrument" This #ould gi!e a possibility for the eperiment to be more specific and accurate"
Procedures$ )aterials and E%uipments The researchers #ould be using Jasminum sambac lea!es as the independent !ariable of this study" =ampaguita lea!es #ould be a!ailable in =an Pedro$ ?aguna #herein the plant is abundantly found" The Aedes aegypti lar!ae #ould be the population of this research" The lar!ae to be tested must be approimately (+'"
The lar!ae #ould be
24
collected from stagnant #aters and after#ards #ould be brought to a laboratory and thus the lar!ae #ould be tested #hether it is Aedes aegypti or not" The study #ould last for about t#o to four months. The procedure #ould begin #ith the crude etraction method" The etraction method #ould be starting from #ashing the =ampaguita leaf #ith tap #ater" Then$ lea!e it to dry #herein the room temperature is about &, to &+ C" Fhen the leaf is dried$ place it in an electrical blender to pul!eri/e" =ohlet etractor #ould be used to etract alkaloid from the po#der" The =ohlet etractor is a flask and condenser de!ice for the continuous etraction of alcohol or either soluble materials .0=ohlet etractor4" )cgra# Hill Dictionary of =cientific and Technical Terms" &''- =ith Edition< The pul!eri/ed =ampaguita lea!es are placed inside a SthimbleS made from thick filter paper$ #hich is loaded into the main chamber of the =ohlet etractor" The =ohlet etractor is placed onto a flask containing the etraction sol!ent" The =ohlet is then e%uipped #ith a condenser" The sol!ent is heated on a traditional hotplate to reflu" The sol!ent !apour tra!els up a distillation arm$ and floods into the chamber housing the thimble of solid" The condenser ensures that any sol!ent !apour cools and drips back do#n into the chamber housing the solid material" The chamber containing the solid material slo#ly fills #ith sol!ent" =ome of the desired compound #ill then dissol!e in the sol!ent" Fhen the =ohlet chamber is almost full$ the chamber is automatically emptied by a siphon side arm$ #ith the sol!ent running back do#n to the distillation flask" During each cycle$ a portion of the non:!olatile compound dissol!es in the sol!ent" After many cycles the desired compound is concentrated in the distillation flask" The ad!antage of this system is that instead of many portions of sol!ent being
25 passed through the sample$ >ust one batch of sol!ent is recycled" After etraction the sol!ent is remo!ed$ typically by means of an e!aporator$ yielding the etracted compound" The non:soluble portion of the etracted solid remains in the thimble$ and is usually discarded" .0=ohlet etraction
procedure"4
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There #ould be four concentrations #ith three replicates of each petri dish" In each petri dish$ there #ould be (; lar!ae" After the etraction$ the lar!icide #ould be placed in the petri dish and the lar!ae #ould be eposed to different concentrations (ppm$ (' ppm$ ('' ppm and ('''ppm" Ha!ing different concentrations #ould indicate the difference and the effecti!eness of the lar!icide" The changes #ould be obser!ed e!ery (;$ -'$ G' and (&' minutes" After#ards$ the data #ould be recorded and analy/ed" Then a conclu: sion #ould be formed"
o!!ect sa/-a+uita !eaf and #edes ae+y-ti !ar&ae
Record the resu!ts ased on ho, /any !ar&ae died and !i&ed after 24 hours and 48 hours of !ar&icide e-osure
Dse the crude etract as a !ar&icide to the set$u-s.
@igure -"( A flo#chart sho#ing the procedures
Dsin+ an e!ectrica! !ender* -o,deriEe the dried 'a/-a+uita !eaf.
Ftract the 'a/-a+uita crude fro/ the -o,deriEed !eaf usin+ a 'oh!et etractor
26
Data athering Procedure The lar!icidal property of =ampaguita leaf etract re%uires eperimentation as the data gathering procedure" The researchers #ould be conducting an eperiment to test #hether the =ampaguita plant contains alkaloid #hich #ill be used as a lar!icide for mos%uitoes" The researchers #ould be collecting data through obser!ation sheets #hich are based on the eperiment conducted" Through these obser!ation sheets$ the researchers #ill be able to gather the data needed for the completion of the problem"
Data Processing )ethod After performing the procedure$ the data #ill be tabulated in a matri" The measure of the mortality rate of each container #ith mos%uito lar!ae #ill be recorded after the analy sis" After#ards$ the mean percentage lar!al mortalities of =ampaguita leaf etract is sho#n in the figure belo#" Table -"- )ean lar!al mortalities induced by leaf etract of =ampaguita . Jasminum sambac< on the late third instar lar!ae of Aedes aegypti e!ery (;$ -'$ G' and (&' minutes
27
Treatment
)ean )ortality )( after (; minutes
( ppm concentration
)( after -' minutes )( after G' minutes )( after (&' minutes )& after (; minutes
(' ppm concentration
)& after -' minutes )& after G' minutes )& after (&' minutes )- after (; minutes
('' ppm concentration
)- after -' minutes
)- after G' minutes )- after (&' minutes ), after (; minutes ), after -' minutes (''' ppm concentration ), after G' minutes ), after (&' minutes Then statistical treatments #ill be applied to the data"
=tatistical Treatment of Data The statistical techni%ue that the researchers #ill use to treat the research data for in
MS b
depth solution of problems in the study is the @:test #ith
MS w
as its main e%uation"
@irst$ the researchers #ould ha!e to #rite the hypothesis5 the null and the alternati!e" et is to find the critical !alue in #hich you #ill base on #hether to re>ect or accept the null or alternati!e hypothesis" After finding the critical !alue$ conduct the test" @or this
28
problem$ the researchers #ould be using @:test" @or the f:test$ ra# should be computed in #hich the !alues of the ra# score #ill be needed to determine the sum of for each group$ the total $ and the sum of the ra# scores" et$ compute for the sum of s%uares$ the degree of freedom and the mean of s%uares" Then determine the significance of the @: ratio" @:test is used in this problem to determine the differences of the means of the , different concentrations of Jasminum sambac leaf etract on the , set:ups of petri dishes #ith (; Aedes aegypti lar!ae" By determining this$ the researchers #ill be able to make a conclusion on #hether the null hypothesis formulated should be accepted or re>ected"
BDETARK RENIRE)ET eeds )aterials and E%uipments ?aboratory Epenses Transportation 1ther Epenses T1TA?
Cost ,$;'' G$;'' ;' ($''' (&$;'
29
=CHED?E 1@ ACTI3ITIE= =K &'((:&'(& Au g
(" athering of Aedes aegypti lar!ae and =ampaguita lea!es &" Eperimentation -" 1bser!ation ," Analysis
Sept
Oct
Nov
Dec
Jan
Feb
30
;" Presentation of the @inal Research paper G" Recommendations
BIB?I1RAPHK A" Books 0Alkaloids"4 .(*++< rolier Academic Encyclopedia" Balta/ar$ Clare and =ala/ar$ elia" .(**< $hilippine %nsects an %ntroduction" Diliman$ Nue/on City" ni!ersity of the Philippines Press" 0Dengue fe!er"4 .(*++< rolier Academic Encyclopedia" allo$ Romulo" .(*+,< The Philippine ournal of =cience" De ?a =alle ni!ersity" -G(: -'"
31
=teiner$ )ona ?isa" .(**&< $hilippine &rnamental $lants and 'heir are" )anila$ Philippines"
B" Internet =ites Tube/a$ Philip" .&'(($ April (,<" D1H tags dengue 2hot spots in Pasig$ 3alen/uela" %nuirer " @rom http566###"in%uirer"net6specialfeatures6denguealert6!ie#"php7 db8(9article8&'((',(,:--((+; &'(( Dengue 1utbreak" *iotech $hilippines" @rom http566###"biotech"ph6&'((:dengue: outbreak Administrati!e 1rder no" &+( s" (**G" .(**G< han +obles ,irtual a ibrary" Retrie!ed =eptember &*$&'((" @rom http566###"chanrobles"com6administrati!eorders6administrati!eorderno&+(&'(**G"htm l Administrati!e 1rder no" ,; s" &''(" .&''(<" Department of ealth" Retrie!ed =eptember &* &'((" @rom http566###"doh"go!"ph6files6ao,;:'("pdf )o ?i Hua" =ampaguita$ Philippine )edicinal Plants" "tuartxchange" Retrie!ed 1ctober &'('" @rom http566###"stuartchange"org6=ampaguita"html ?ar!icidal effects of atrophacurcas ?" against Anopheles arabiensis .Diptera5 Culicidea<" $etros" Retrie!ed o!ember (($ &'((" @rom http566###"mu"edu"et6me>s6pdfs6!-n(6e#dneh"pdf 0?ar!icide"4 'he /ree Dictionary" @rom http566###"thefreedictionary"com6lar!icide =teps of the =cientific )ethod" Retrie!ed o!ember &$ &'((" @rom http566###"eperiment:resources"com
32
Dundas Data 3isuali/ation$ Inc" and others" .&''(:&''*< A13A @ormula" Retrie!ed &''*" @rom http566support&"dundas" com61nlineDocumentation6FinChart&'';6Ano!a"html ?ife
Cycle
of
Aedes
Aegypti" Dengue
,irus
Net "
Retrie!ed
&'('"
@rom
http566###"dengue!irusnet"com6life:cycle:of:aedes:aegypti"html
APPEDIU A =ample Co!er ?etter MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM MMMMMMMMMMMMMM
Dear .)adam6 =ir< MMMMMMMMMM$
Research I is one of the sub>ects in )akati =cience High =chool" In this sub>ect$ students are re%uired to pass a research proposal and perform !arious eperiments after#ards"
33
The researchers are conducting a study about the 0?AR3ICIDA? PR1PERTK 1@ THE Jasminum sambac ?EA@ EUTRACT= 1 Aedes aegypti lar!ae"4 This #ould re%uire the researchers to ask your participation in this study through ans#ering the obser!ation sheets needed in the eperiment" The obser!ation sheet is needed to gather data about the ongoing eperiment" The in!estigators #ould maintain the pri!acy of your personal information" The students performing the study are hoping for your kind consideration and cooperation" Fe are grateful for your kind response to our re%uest" If you #ould be in!ol!ed$ #e #ould gladly for#ard you the results of the dissertation" 3ery truly yours$ The researchers$ lory Tolentino =amantha Bismonte )arian Boneo
APPEDIU B 1bser!ation =heet A)E5 MMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMMM DATE5 MMMMMMMMMMMMMMMM Concentration
Concentration
Concentration
Concentration
(ppm
(' ppm
('' ppm
(''' ppm
@actors to be considered Time of eposure upon application Time of effecti!ity
(
&
-
(
&
-
(
&
-
(
&
-
34
)ortality rate
CRRIC?) 3ITAE Personal Information ame5 )arian H" Boneo Birth Date5 anuary (($ (**+ Birth Place5 )akati City$ )etro )anila Parents5 =yl!ia H" Boneo Antonio D" Boneo Educational Information Kear5 &'(':Present Course5 High =chool =chool5 )akati =cience High =chool Kear5 &'',:&'(' Course5 rade =chool =chool5 Cembo Elementary =chool Fork6Research Agencies5 Kear Employed5 &'((:&'(&
35
Position5 Researcher Agency5 )akati =cience High =chool Honors6A#ards Recei!ed5 Honors5 @irst Honors .Cembo Elementary =chool< Research Conducted The ?ar!icidal Property of =ampaguita Jasminum sambac ?eaf Crude Etract on )os%uito lar!ae . Aedes aegypti) #ith lory race )" Tolentino and =amantha Ally 3" Bismonte
CRRIC?) 3ITAE Personal Information ame5 =amantha Ally 3" Bismonte Birth Date5 )arch &+$ (**+ Birth Place5 Binan$ ?aguna Parents5 )aria Alma ?ynn 3" Bismonte =am B" Bismonte Educational Information Kear5 &'(':Present Course5 High =chool =chool5 )akati =cience High =chool Kear5 &'',:&'(' Course5 rade =chool =chool5 Ann Arbor )ontessori ?earning Center Binan Fork6Research Agencies5 Kear Employed5 &'((:&'(&
36
Position5 Researcher Agency5 )akati =cience High =chool Honors6A#ards Recei!ed5 Honors5 =econd Honorable )ention .Ann Arbor )ontessori ?earning Center< Research Conducted The ?ar!icidal Property of =ampaguita Jasminum sambac ?eaf Crude Etract on )os%uito lar!ae . Aedes aegypti) #ith )arian H" Boneo and lory race )" Tolentino
CRRIC?) 3ITAE Personal Information ame5 lory race )" Tolentino Birth Date5 )arch &G$ (** Birth Place5 Nue/on City$ )etro )anila Parents5 Chonalyn )" Tolentino regory R" Tolentino Educational Information Kear5 &'(':Present Course5 High =chool =chool5 )akati =cience High =chool Kear5 &'',:&'(' Course5 rade =chool =chool5 =t" Bridget =chool NC Fork6Research Agencies5 Kear Employed5 &'((:&'(&
