SavithaSekharNair,VinayaShetty,andNadikereJayaShettyJournal of Insects Earlier studies have shown the e cacy of Eucalyptus oils as a larvicide against mosquitoes [ ]. In the year ,

Research ArticleRelative Toxicity of Leaf Extracts of Eucalyptus globulus andCentella asiatica against Mosquito Vectors Aedes aegypti andAnopheles stephensi

Savitha Sekhar Nair Vinaya Shetty and Nadikere Jaya Shetty

Centre for Applied Genetics Bangalore University J B Campus Bengaluru Karnataka 560056 India

Correspondence should be addressed to Nadikere Jaya Shetty shetty njyahoocoin

Received 17 July 2014 Accepted 1 October 2014 Published 22 October 2014

Academic Editor Jose A Martinez-Ibarra

Copyright copy 2014 Savitha Sekhar Nair et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The larvicidal activity of different solvent leaf extracts (hexane diethyl ether dichloromethane andmethanol) ofEucalyptus globulusand Centella asiatica against two geographically different strains of Aedes aegypti and Anopheles stephensi was investigated Theextracts were tested against the late third instar larvae of Aedes aegypti and Anopheles stephensi and larval mortality was observedafter 24 hours of treatment LC

50and LC

90were calculatedThe LC

50values of hexane extract of Eucalyptus globulus against the late

third instar larvae of the BSN and JPN strains of Aedes aegypti and the DLC and KNG strains of Anopheles stephensi were 22521677 1188 and 1928 ppm while those of the hexane extract of Centella asiaticawere 2465 2687 506 and 2435 ppm respectivelyThe LC

50values of diethyl ether extract of Centella asiatica were 3396 1345 241 and 147 ppmThe hexane extracts of both plants

and the diethyl ether extract of C asiatica presented the highest potential for the control of Aedes aegypti and Anopheles stephensiThe present findings also reveal the necessity of assaying multiple strains of a species to fully comprehend the larvicidal efficacy ofa compound

1 Introduction

Apart from being a social nuisance mosquitoes pose serioushealth threats to both men and animals considering thatthey are the principal vectors for many vector borne diseasesincludingmalaria dengue yellow fever and Chikungunya [12] inmen and equine encephalitis haemorrhagic septicaemiaof buffaloes and enzootic hepatitis in animals [3] In someindividuals mosquito bites also result in acute systemicallergic reactions defined by the presence of one or moreof the following urticaria angioedema wheezing dyspneahypotension and decrease or loss of consciousness [4]The mosquito Aedes (Stegomyia) aegypti (Linn) (DipteraCulicidae) is the primary vector of dengue yellow feverand Chikungunya [2] According to WHO over 40 ofthe worldrsquos population is now at risk of dengue and thereare 200 000 estimated cases of yellow fever causing 30 000deaths worldwide each year Malaria on the other hand alife threatening disease which caused an estimated 627 000deaths in 2012 is transmitted exclusively through the bites

of Anopheles mosquitoes [5] Anopheles stephensi Liston(Diptera Culicidae) is a major vector in India as well as insome of the West Asian countries and has been shown to bedirectly responsible for about 40ndash50 of the annual malarialincidence [6 7]

Mosquito control is an extensively researched topic con-sidering that presently we do not have vaccines for malariaand dengue [8 9] and therefore it becomes necessary tocheck the spread of these diseases Vector control methodsthus are of utmost prominenceThe incessant use of chemicalinsecticides has often led to the disruption of natural biolog-ical control system and outbreak of insect species [10] Theselective pressure of conventional insecticides is enhancingthe resistance of mosquito strains at an alarming rate [11]increasing the demand for newproducts that are eco-friendlytarget-specific and biodegradable Plant-derived insecticidesprovide an alternative to synthetic pesticides because of theirgenerally low environmental pollution low toxicity to menand other advantages [12]

Hindawi Publishing CorporationJournal of InsectsVolume 2014 Article ID 985463 7 pageshttpdxdoiorg1011552014985463

2 Journal of Insects

Earlier studies have shown the efficacy of Eucalyptus oilsas a larvicide against mosquitoes [13ndash16] In the year 2004Yang et al reported that Eucalyptus leaf oil particularly someof its components such as 18-cineole 1-120572-terpineol and (E)-pinocaveol merits further study as lead compounds for thecontrol of the human hair louse Pediculus humanus capitisDe Geer (Pediculidae) [17] Constituents of the essential oilof Eucalyptus camaldulensisDehnh and Eucalyptus urophyllaS T Blake (Myrtaceae) a-phellandrene limonene p-cymenec-terpinene terpinolene and a-terpinene have been shownto possess strong larvicidal effects against Aedes aegypti andAedes albopictus [15] The various components of Eucalyptusessential oil have been shown to act synergistically (and notadditively) to bring the overall bactericidal activity [18]

Centella asiatica is mostly studied for its medicinal prop-erties Very few studies have documented its potential as abiopesticide A study has suggested that the leaf extract ofC asiatica is promising as a larvicide and adult emergenceinhibitor against Culex quinquefasciatus Say (Culicidae) [19]Shukla et al [20] isolated and studied the growth inhibitoryeffect of 2120572 3120573 6120573 23-120572 tetrahydroxyurs-12-ene-28-oicacid a triterpenoid glycoside against larvae of SpilarctiaobliquaWalker (Arctiidae)

Numerous studies have examined the variations in lar-vicidal efficacies of plant extracts tested against differentmosquito species however we were not able to find anyreports of the larvicidal efficacies of plant extracts testedagainst different strains of mosquitoes of the same speciesSusceptibility studies involving strains from different loca-tions could be beneficial to gauge the efficacy of the drugbeing studied and as a prelude to investigations involving theinherent resistance mechanisms of the vectors to the drugs

Different solvent types can significantly affect the potencyof the extracted plant compounds and there is a differencein the chemoprofile of the plant species [21] This is becausepolar solvents extract polar constituents of the plant whilenonpolar solvents extract the nonpolar constituents of theplant Hence the best results will be observed using the plantextract of the solvent whose polarity matches that of themolecules in the plant responsible for insecticidal activity(if any) Additionally extracts or pure compounds derivedfrom specific solvents can influence the bioactivity probablybecause of the active components which are present in largequantities [22]

In the present study we have investigated the larvicidalactivity of the hexane diethyl ether dichloromethane andmethanol extracts of the leaves of Eucalyptus globulus Labill(Myrtaceae) and Centella asiatica (Linn) Urban (Apiaceae)against two geographically different strains of Aedes (Ste-gomyia) aegypti (Linn) (Diptera Culicidae) and Anophelesstephensi Liston (Diptera Culicidae)

2 Materials and Methods

21 Mosquito Culture Four strains which consisted of lab-oratory colonies of Aedes aegypti collected from J P Nagar(JPN) (129120∘N 775930∘E) and Basaveshwaranagar (BSV)(129867∘N 775386∘E) and Anopheles stephensi collectedfrom Dollars Colony (DLC) (13∘210158403010158401015840N 77∘341015840310158401015840E) and

Kengeri (KNG) (129100∘N 774800∘E) were used for thisstudy Larvae collected from the said locations of BBMPregion Karnataka India during the months March-April2012 had been reared continuously in the insectary at theCentre for Applied Genetics Bangalore University followingthe method of Shetty [23]

22 Collections of Plant Materials The leaves of Eucalyptusglobulus and Centella asiatica were collected from Bangalorecity (11∘110158404810158401015840N 77∘210158403610158401015840E elevation 399m) KarnatakaIndia in July 2013 and were authenticated at the Departmentof Botany Bangalore University Bangalore India

23 Preparation of Plant Extracts The leaves (2 kg each) wereair dried in shade for 15ndash30 days The dried leaves werethen powdered mechanically using a commercial electricalstainless steel blender One kg of powdered leaves wasextracted successively by maceration using nonpolar to polarsolvents namely hexane diethyl ether dichloromethane andmethanol In each solvent the plant material was soaked for48 h at 35∘C and filtered twice using Whatman number 1filter paper to obtain the extract and to the residue the samesolvent was added againThe procedure was repeated twice toobtain maximum extract The extracts were concentrated atreduced temperature using a rotary vacuum evaporator andstored at a temperature of 4∘C One gram of the concentratedplant extractwas dissolved in 100mLof 1 1 acetone dimethylsulfoxide (DMSO) and considered as 1 stock solution Fromthis stock solution varying concentrations of each extractwasprepared and these solutionswere used for larvicidal bioassayAll chemicals used in this study were of extra pure grade andwere obtained from Sisco Research Laboratories PVT LtdIndia

24 Larvicidal Bioassay Larvicidal activity of each extractderived from the leaves of Eucalyptus globulus and Centellaasiatica against the two strains ofAe aegypti andAn stephensiwas assessed by using a slightlymodified version of theWHOstandard method [24]

Initially the mosquito larvae were exposed to a broadrange of test concentrations to determine the activity range ofeach extract Based on the results of preliminary screeningbatches of 25 late third instar larvae were added to 300mLwide mouth disposable bowls containing serial concentra-tions (6ndash9 concentrations yielding between 10 and 95mortality in 24 hrs) of each plant extract made up to 250mLby volume using tap water The test was carried at a tem-perature of 25 plusmn 2∘C and relative humidity of 75 plusmn 5The numbers of dead larvae were counted after 24 hours ofexposure and percentage mortality was calculated for eachtest as follows

Percentage Mortality

=

Number of dead larvaepupaeNumber of larvae introduced

times 100

(1)

The final percentage mortality was calculated from theaverage of three replicates Solutions containing tapwater and1 1 acetone dimethyl sulfoxide butwithout the plant sample

Journal of Insects 3

served as controls The control mortalities were corrected byusing Abbottrsquos formula [25]

25 Statistical Analysis The average larval mortality datawere subjected to probit analysis for calculating LC

50and

LC90

and other statistics at 95 fiducial limits of upperconfidence limit (UCL) and lower confidence limit (LCL)and Chi-square values were calculated using the SPSS 120(Statistical Package of Social Sciences) software (Finney1971) The Chi-square values were considered significant at119875 lt 005 level

3 Results

The larvicidal efficacies of hexane diethyl ether dichloro-methane and methanol extracts of both Eucalyptus globu-lus and Centella asiatica were established for two differentmosquito strains of each of the above said species fromthe Bangalore Karnataka The data were recorded andstatistical data including regression equation LC

50 LC90

LCL UCL and Chi-square values were calculated andpresented (Table 1) Nomortality was observed in the control

All plant extracts showed varied level of toxic effects withspecies after 24 hours of exposure while themethanol extractdemonstrated very low larvicidal efficacy in the case of bothof the plants

31 Eucalyptus globulus Among the An stephensi strainsthe DLC strain displayed the highest susceptibility to thehexane extract (LC

501188 and LC

904504 ppm) followed by

the diethyl ether extract (LC50

4641 and LC90

8001 ppm)The KNG strain of Anopheles stephensi on the other handshowed the highest larval mortality in the dichloromethaneextract (LC

501620 and LC

905824 ppm) followed by the

hexane extract (LC501928 and LC

90827 ppm) The BSN and

JPN strains ofAe aegyptiweremost susceptible to the hexaneextract (LC

502252 1677 and LC

904236 3040 ppm resp)

The methanol extract was the least effective in all cases Ahigher level of tolerance was recorded in the BSN strain ofAe aegypti to the methanol extract (LC

5018390 and LC

90

32359 ppm)

32 Centella asiatica The hexane extract produced the high-est mortality in the case of the BSN and JPN strains of Aeaegypti (LC

502465 2687 and LC

905931 16068 ppm resp)

and also the DLC strain of An stephensi (LC50506 and LC

90

3288 ppm) the KNG strain of An stephensi however wasmost susceptible to the diethyl ether extract (LC

50147 and

LC90

1180 ppm) Again the methanol extract was the leastpotent in all cases The BSN strain of Ae aegypti displayeda higher level of tolerance (LC

507765 and LC

9038534 ppm)

upon treatment with the methanol extract

4 Discussion

The leaf extracts of Eucalyptus globulus and Centella asiaticashowed an overall moderate larvicidal effect when testedagainst different strains of Ae aegypti and An stephensi

Among the four extracts studied the hexane extracts of bothplants and the diethyl ether extract of C asiatica demon-strated higher potential for the control of strains of mosquitovectors followed by dichloromethane The methanol extractproduced a comparatively less promising result The effectof the extracts on larval mortality was dose-dependent withincreasing rates of larval mortality observed at increasingconcentrations The highest level of tolerance was observedin the BSN strain of Ae aegypti towards the methanol extractof E globulus

Although the larvicidal efficacy of the extracts is notas promising as that of synthetic insecticides commonly inuse today [26] the present results are comparable to thoseof earlier authors who worked on various plant extracts aslarvicides against different mosquito species The methanolleaf extract of Clitoria ternatea L (Leguminosae) showed adose-dependent larvicidal activity against An stephensi withan LC

50value of 5556 ppm [27] In the case of Morinda

citrifolia L (Rubiaceae) the highest larvicidal activity wasexhibited by the methanol extract of the leaf when comparedto the hexane chloroform acetone and water extracts theLC50in case of An stephensi and Ae aegypti was observed to

be 26196 and 27792 ppm respectively The chloroform leafextracts ofNyctanthes arbortristisL (Oleaceae) demonstratedan LC

50value of 7806 ppm while the LC

50value of the

methanol flower extract of the same plant was 6794 ppmwhen tested against An stephensi Studies have also beencarried out on the larvicidal potential of the essential oilextracted from the Eucalyptus species [13ndash15] and E globulusin particular [28] However this is the first time report ofthe larvicidal efficacy of serial extracts of Eucalyptus globulusand Centella asiatica leaves using multiple solvents againstdifferent geographical strains ofAe aegypti andAn stephensiIn the present study strains of both species of mosquitoeswere found to be highly susceptible to hexane leaf extractswhen compared to the other solvent extracts

Numerous authors have observed a converse relationshipbetween extract effectiveness and solvent polarity where theefficacy increases with decreasing polarity [29ndash32] Singhet al [33] reported the LC

50values of the hexane extract

of Eucalyptus citriodora Hook (Myrtaceae) against the IVthinstar larvae of An stephensi (6986 ppm) and Ae aegypti(9176 ppm) Hexane extracts of other plants have also shownsimilar satisfactory larvicidal potency LC

50of hexane leaf

extract of Citrus sinensis (L) Osbeck (Rutaceae) against earlyfourth instars of Ae aegypti was found to be 44684 ppm[34] The LC

50of hexane leaf extract ofMurraya koenigii (L)

Spreng (Rutaceae) against third instars Culex quinquefascia-tus was found to be 96353 ppm [35] In the case of Citrulluscolocynthis (L) Schrad (Cucurbitaceae) however the LC

50of

the hexane extract was found to be 145129 ppm while that ofthe diethyl ether extract was found to be 50339 ppm Thisresult is similar to that of the KNG strain of An stephensiin our studies where the diethyl ether extract of C asiaticaproduced a better result than the hexane extract Contrarilythe other An stephensi strain that we worked with (DLC)exhibited higher susceptibility for the hexane extract againstthe diethyl ether extract


Table1Eff

ectsof

relativ

etoxicity

ofsolventextracts(hexanediethyletherdichloromethaneand

methano

l)of

theleaveso

fEucalyptusg

lobu

lusa

ndCe

ntellaasiaticaagainstthe

latethird

larvalsta

geof

BSVandJPNstr

ains

ofAe

desa

egyptiandDLC

andKN

Gstr

ains

ofAn

opheles

stephensi

Botanicaln

ame

Solvents

Species

Strain

LC50(ppm

)(UCL

-LCL

)LC

90(ppm

)(UCL

-LCL

)Re

gressio

nequatio

n119903

1205942

df

Eucalyptus

globu

lus

Hexane

Aeaegypti

BSV

2252(2109ndash2635)

4236(4004ndash5989)

119910=46638119909minus59716

09364

01902

7JPN

1677

(1521ndash

1867)

304(2663ndash4

009)

119910=49559119909minus60248

0984

00238

6

Anstephensi

DLC

1188(881ndash1512

)4504(3738ndash9

533)

119910=22119119909+04106

09137

01123

7KN

G1928(1547ndash

2551)

827(6552ndash1634)

119910=20242119909+03744

09378

01134

7

Diethylether

Aeaegypti

BSV

6868(6308ndash7564)

12167(10737ndash15825)119910=51546119909minus96228

09594

00244

7JPN

3188(14

67ndash64

12)

20699(10851ndash348885)119910=15756119909+10554

09114

01811

6

Anstephensi

DLC

4641(4288ndash5242)

8001(7261ndash10507)

119910=54107119909minus9428

09382

00767

6KN

G318(2706ndash3917)

9519

(7374ndash16187)

119910=2688119909minus17265

09279

006

056

Dichlorom

ethane

Aeaegypti

BSV

3146(2761ndash

385)

8299

(6771ndash

12907)

119910=3038119909minus2588

09846

00832

6JPN

3622(1716ndash6

201)

29046(106

86ndash

139251)119910=14157119909+13773

0889

01362

6

Anstephensi

DLC

3003(2563ndash3942)

12354(8725ndash22839)

119910=20839119909minus0163

09747

01345

7KN

G162(1125ndash1994)

5824(4526ndash13234)

119910=23036119909minus00899

09623

00536

7

Methano

lAe

aegypti

BSV

18393(16958ndash20497)

32359(29302ndash40

511)

119910=52171119909minus12032

09805

00504

7JPN

14679(1281ndash17868)

35302(29117ndash56123)

119910=33587119909minus5636

096

4400634

5

Anstephensi

DLC

12633(7981ndash

18173)

36143(29883ndash127366)

119910=28038119909minus3696

08769

02807

7KN

G8896

(764

7ndash11243)

26551(21770

ndash49074

)119910=26955119909minus29496

09204

01292

6

Centellaasiatica

Hexane

Aeaegypti

BSV

2465(2131ndash

3071)

5931(5273

ndash10139)

119910=33561119909minus3027

09284

01366

5JPN

2687(1744ndash3747)

16068(10821ndash5866

2)119910=16479119909+09969

0929

00583

5

Anstephensi

DLC

506(183ndash105)

3288(19

85ndash8730)

119910=15752119909+23154

08489

02479

6KN

G2435(2211ndash

2742)

4286(3627ndash

5845)

119910=52115119909minus74372

09963

00055

5

DiethylEther

Aeaegypti

BSV

3396

(2581ndash

4723)

15993(11736ndash

3840

5)119910=19022119909+01855

08489

00844

5JPN

1345(206ndash

2041)

7708(5767ndash

31917)

119910=16885119909+14054

09362

004

635

Anstephensi

DLC

241(2034ndash

3015

)6307(540

4ndash11162)

119910=3063119909minus22959

08489

01046

5KN

G147(3ndash35)

118(637ndash17122)

119910=14175119909+33433

08862

01491

5

Dichlorom

ethane

Aeaegypti

BSV

3397

(281ndash7463)

21277(12419

ndash403246)

119910=16065119909+09337

09384

01913

5JPN

8376

(7779ndash10302)

16272(15423ndash25588)119910=44382119909minus7973

09

01722

5

Anstephensi

DLC

2368(14

55ndash3933)

33471(15767ndash279272

)119910=11127119909+23581

09338

00533

5KN

G1555(209ndash

2955)

30229(17325ndash335591

)119910=09933119909+28229

08881

00528

5

Methano

lAe

aegypti

BSV

7765(59099ndash

109078)

38534(272801ndash10164

21)119910=21701119909minus12027

08905

00966

5JPN

7215

(36954ndash

135804)

2806

(186607ndash3248971)119910=21701119909minus12027

08905

01928

5

Anstephensi

DLC

4768(46194ndash

49349)

5879

(56722ndash64399)119910=14064119909minus32668

09832

00333

8KN

G177(11928ndash22509)

5599

(48397ndash126017

)119910=25595119909minus07538

09002

0110

75

LC50lethalcon

centratio

nthatkills

50of

thee

xposed

larvaeLC 9

0lethalconcentrationthatkills

90of

thee

xposed

larvaeU

CLupp

erconfi

dencelim

itLC

Llower

confi

dencelim

it119903correlationcoeffi

cient

1205942C

hi-squ

ares

ignificantat119875lt005

levelanddfdegreeo

ffreedom


Even though our studies show methanol extracts toproduce the least mortality other plants such as Morindacitrifolia L (Rubiaceae) and Erythrina indica Lam (Legumi-nosae) have been shown to produce greater susceptibilityfor their methanol extract when compared to their hexaneextract [36 37] Therefore the present study suggests that inthe case of E globulus andC asiatica the compoundsmajorlyresponsible for the insecticidal activity are most probably inthe nonpolar and intermediate range and are not polar

There is probably also the result of the synergistic activityof the active compounds that influences the efficacy of theextract against the physiological characteristics of each straintested For instance dichloromethane and diethyl ether aresolvents with very similar polarity and should typically elutecompounds of the same class and the results show thatin some instances dichloromethane extracts prove that theyhave higher efficacy while diethyl ether extracts show greaterpotency Further studies involving larger number of strainsand purified isolated constituents should reveal a clearerpicture of interaction between the bioactive molecules andtarget species

In the present investigation it was also interesting tonote that the variation in the response of different strains ofmosquito of the same species to a specific plant extract isalmost the same as the variation in the response of differentstrains of mosquito of different species to the same extractFor instance the LC

50values for diethyl ether extract of

C asiatica against the An stephensi strains KGR and DLCare 147 ppm and 241 ppm while the LC

50values for the

same extract against the Ae aegypti strains JPN and BSV are1345 and 3396 ppm From this study it is revealed that it isimperative to assay the extract against different geographicalstrains of the same species to ascertain the efficacy of acompound

Earlier studies on different geographical strains of Anstephensi and Ae Aegypti have reported varied levels ofsusceptibility to various insecticides commonly used inmosquito vector control [26 38ndash43] Unlike conventionalinsecticides which are based on a single active ingredientplant derived insecticides comprise botanical blends of chem-ical compounds which act concertedly on both behavioraland physiological processes Thus there is very little chanceof pests developing resistance to such substances Identifyingbioinsecticides that are efficient as well as being suitable andadaptive to ecological conditions is imperative for continuedeffective vector control management [21] In order to developcost effective pesticides from the plant leaves used in thepresent study it will be necessary to carry out further studiesto determine the active compounds in these plants theirlarvicidal efficacies their individual and synergistic modesof action the feasibility of large scale use and stability ofthese active compounds under field conditions Most studiesusing biological extracts have been tested against a singlestrain of a species This is probably because one would notexpect to observe large variations in the susceptibility of onestrain to another Unlike the varied resistance status observedin the case of synthetic insecticides the biological extractsused in this study have not been used by municipal bodiesas insecticides Therefore the observed variation in response

between different strains does not appear to have been causedby such a selection event but rather by the intervention ofbiological and genetic factors resulting in the change in thesusceptibility to different plant extracts

5 Conclusion

In the present study the hexane extracts of both plantsand the diethyl ether extract of C asiatica demonstratedthe highest potential for the control of strains of mosquitovectors The results indicate that the compounds responsiblefor the insecticidal activity of both plants are most probablyin the nonpolar and intermediate range Also the presentfindings show that it is necessary to assay multiple strainsof a species to fully comprehend the larvicidal efficacy of acompound

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] World Health Organization ldquoVector control for malaria andother mosquito-borne diseasesrdquo WHO Technical Report 8571995

[2] A C Morrison E Zielinski-Gutierrez T W Scott and RRosenberg ldquoDefining challenges and proposing solutions forcontrol of the virus vector Aedes aegyptirdquo PLoS Medicine vol5 no 3 article e68 2008

[3] S Christophers Aedes Aegypti (L) The Yellow Fever MosquitoIts Life History Bionomics and Structure Cambridge UniversityPress London UK 1960

[4] Z Peng A N Beckett R J Engler D R Hoffman N L Ottand F E R Simons ldquoImmune responses to mosquito saliva in14 individuals with acute systemic allergic reactions tomosquitobitesrdquo Journal of Allergy and Clinical Immunology vol 114 no5 pp 1189ndash1194 2004

[5] World Health Organization ldquoMalaria factsheet94rdquo 2014httpwwwwhointmediacentrefactsheetsfs094en

[6] C F Curtis ldquoShould DDT continue to be recommended formalaria vector controlrdquo Medical and Veterinary Entomologyvol 8 no 2 pp 107ndash112 1994

[7] F H Collins and S M Paskewitz ldquoMalaria current and futureprospects for controlrdquoAnnual Review of Entomology vol 40 pp195ndash219 1995

[8] World Health Organization Malaria Vaccine Rainbow TablesWorld Health Organization Geneva Switzerland 2013 httpwwwwhointimmunizationresearchdevelopmentRainbowtablesen

[9] World Health Organization ldquoDengue and dengue hemorrhagicfever factsheet117rdquo 2014 httpwwwwhointmediacentrefactsheetsfs117en

[10] U Chaithong W Choochote K Kamsuk et al ldquoLarvicidaleffect of pepper plants on Aedes aegypti (L) (Diptera Culici-dae)rdquo Journal of Vector Ecology vol 31 no 1 pp 138ndash144 2006

[11] AW Brown ldquoInsecticide resistance inmosquitoes a pragmaticreviewrdquo Journal of the American Mosquito Control Associationvol 2 no 2 pp 123ndash140 1986


[12] S Liu G Shi H Cao F Jia and X Liu ldquoSurvey of pesticidalcomponent in plantrdquo in Entomology in China in 21st CenturyProceedings of the Conference of Chinese Entomological Soci-ety pp 1098ndash1104 Science amp Technique Press 2000

[13] S Senthil Nathan ldquoThe use of Eucalyptus tereticornis Sm(Myrtaceae) oil (leaf extract) as a natural larvicidal agentagainst the malaria vector Anopheles stephensi Liston (DipteraCulicidae)rdquoBioresource Technology vol 98 no 9 pp 1856ndash18602007

[14] A Lucia L W Juan E N Zerba L Harrand M Marco andH M Masuh ldquoValidation of models to estimate the fumigantand larvicidal activity of Eucalyptus essential oils against Aedesaegypti (Diptera Culicidae)rdquo Parasitology Research vol 110 no5 pp 1675ndash1686 2012

[15] S-S Cheng C-G Huang Y-J Chen J-J Yu W-J Chen andS-T Chang ldquoChemical compositions and larvicidal activitiesof leaf essential oils from two eucalyptus speciesrdquo BioresourceTechnology vol 100 no 1 pp 452ndash456 2009

[16] S M Medhi S Reza K Mahnaz et al ldquoPhytochemistry andlarvicidal activity of Eucalyptus camaldulensis against malariavector Anopheles stephensirdquo Asian Pacific Journal of TropicalMedicine vol 3 no 11 pp 841ndash845 2010

[17] Y-C Yang H-Y Choi W-S Choi J M Clark and Y-J AhnldquoOvicidal and adulticidal activity of Eucalyptus globulus leafOil Terpenoids against Pediculus humanus capitis (AnopluraPediculidae)rdquo Journal of Agricultural and Food Chemistry vol52 no 9 pp 2507ndash2511 2004

[18] K Cimanga K Kambu L Tona et al ldquoCorrelation betweenchemical composition and antibacterial activity of essential oilsof some aromatic medicinal plants growing in the DemocraticRepublic of Congordquo Journal of Ethnopharmacology vol 79 no2 pp 213ndash220 2002

[19] S Rajkumar and A Jebanesan ldquoLarvicidal and adult emergenceinhibition effect of Centella asiatica Brahmi (Umbelliferae)against mosquito Culex quinquefasciatus say (Diptera Culici-dae)rdquo African Journal of Biomedical Research vol 8 no 1 pp31ndash33 2006

[20] Y N Shukla R Srivastava A K Tripathi and V PrajapatildquoCharacterization of an ursane triterpenoid from Centella asi-atica with growth inhibitory activity against Spilarctia obliquardquoPharmaceutical Biology vol 38 no 4 pp 262ndash267 2000

[21] A Ghosh N Chowdhury and G Chandra ldquoPlant extractsas potential mosquito larvicidesrdquo Indian Journal of MedicalResearch vol 135 no 5 pp 581ndash598 2012

[22] P V Oliveira J C Ferreira Jr F S Moura et al ldquoLarvicidalactivity of 94 extracts from ten plant species of northeastern ofBrazil against Aedes aegypti L (Diptera Culicidae)rdquo Parasitol-ogy Research vol 107 no 2 pp 403ndash407 2010

[23] N J Shetty ldquoChromosomal translocations and semisterility inthe malaria vectorAnopheles fluviatilis Jamesrdquo Indian Journal ofMalariology vol 20 pp 45ndash48 1983

[24] World Health Organization ldquoInstructions for determining thesusceptibility or resistance of mosquito larvae to insecticidesrdquoTech Rep WHOVBC81807 World Health OrganizationGeneva Switzerland 1981

[25] W S Abbott ldquoA Method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 no 2 pp265ndash267 1925

[26] V Shetty D Sanil and N J Shetty ldquoInsecticide susceptibilitystatus in three medically important species of mosquitoesAnopheles stephensi Aedes aegypti and Culex quinquefasciatus

from Bruhat Bengaluru Mahanagara Palike Karnataka IndiardquoPest Management Science vol 69 no 2 pp 257ndash267 2013

[27] N Mathew M G Anitha T S L Bala S M Sivakumar RNarmadha and M Kalyanasundaram ldquoLarvicidal activity ofSaraca indica Nyctanthes arbor-tristis and Clitoria ternateaextracts against three mosquito vector speciesrdquo ParasitologyResearch vol 104 no 5 pp 1017ndash1025 2009

[28] B Selamawit and R Nagappan ldquoEvaluation of water andethanol extract of Eucalyptus globulus labillardiere (Myrtaceae)leaves against immature stages of filarial vector Culex quinque-fasciatus say (Diptera Culicidae)rdquo Current Research Journal ofBiological Sciences p 4 2012

[29] A-A Aivazi and V A Vijayan ldquoLarvicidal activity of oakQuer-cus infectoria Oliv (Fagaceae) gall extracts against Anophelesstephensi Listonrdquo Parasitology Research vol 104 no 6 pp 1289ndash1293 2009

[30] G Sharma H Kapoor M Chopra K Kumar and V AgrawalldquoStrong larvicidal potential of Artemisia annua leaf extractagainst malaria (Anopheles stephensi Liston) and dengue (Aedesaegypti L) vectors and bioassay-driven isolation of the markercompoundsrdquo Parasitology Research vol 113 no 1 pp 197ndash2092014

[31] M S Mulla and T Su ldquoActivity and biological effects of neemproducts against arthropods of medical and veterinary impor-tancerdquo Journal of the American Mosquito Control Associationvol 15 no 2 pp 133ndash152 1999

[32] K P Prathibha B S Raghavendra andV A Vijaya ldquoEvaluationof larvicidal effect of Euodia ridleyi Hochr Leaf extract againstthreemosquito species atMysorerdquoResearch Journal of BiologicalSciences vol 5 no 6 pp 452ndash455 2010

[33] R K Singh R C Dhiman and P K Mittal ldquoStudies onmosquito larvicidal properties of Eucalyptus citriodora Hook(Family Myrtaceae)rdquo Journal of Communicable Diseases vol39 no 4 pp 233ndash236 2007

[34] R Warikoo A Ray J K Sandhu R Samal N Wahab and SKumar ldquoLarvicidal and irritant activities of hexane leaf extractsof Citrus sinensis against dengue vector Aedes aegypti Lrdquo AsianPacific Journal of Tropical Biomedicine vol 2 no 2 pp 152ndash1552012

[35] KKovendan S Arivoli RMaheshwaran K Baskar and SVin-cent ldquoLarvicidal efficacy of Sphaeranthus indicus Cleistanthuscollinus andMurraya koenigii leaf extracts against filarial vectorCulex quinquefasciatus say (Diptera Culicidae)rdquo ParasitologyResearch vol 111 no 3 pp 1025ndash1035 2012

[36] K Kovendan K Murugan S P Shanthakumar S Vincentand J-S Hwang ldquoLarvicidal activity of Morinda citrifoliaL (Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012

[37] M Govindarajan and R Sivakumar ldquoLarvicidal ovicidaland adulticidal efficacy of Erythrina indica (Lam) (FamilyFabaceae) against Anopheles stephensi Aedes aegypti and Culexquinquefasciatus (Diptera Culicidae)rdquo Parasitology Researchvol 113 no 2 pp 777ndash791 2014

[38] A Ponlawat J G Scott and L C Harrington ldquoInsecticidesusceptibility of Aedes aegypti and Aedes albopictus acrossThailandrdquo Journal of Medical Entomology vol 42 no 5 pp 821ndash825 2005

[39] S C Rawlins ldquoSpatial distribution of insecticide resistance inCaribbean populations of Aedes aegypti and its significancerdquoRevista Panamericana de Salud Publica vol 4 no 4 pp 243ndash251 1998


[40] G Albrieu Llinas E Seccacini C N Gardenal and S LicastroldquoCurrent resistance status to temephos in Aedes aegypti fromdifferent regions of ArgentinardquoMemorias do Instituto OswaldoCruz vol 105 no 1 pp 113ndash116 2010

[41] P Baskar and N J Shetty ldquoSusceptibility status of Anophelesstephensi Liston to insecticidesrdquo Journal of CommunicableDiseases vol 24 no 3 pp 188ndash190 1992

[42] C Ghosh B Rajashree B Priyalakshmi and N J ShettyldquoSusceptibility status of different strains of Anopheles stephensiListon to fenitrothion deltamethrin and cypermethrinrdquo Pestol-ogy vol 26 no 4 pp 47ndash52 2002

[43] N J Shetty T Zin T P N Hariprasad and M Z MinnldquoInsecticide susceptibility studies in thirty strains of Anophelesstephensi Listonmdasha malaria vector to alphamethrin bifenthrin(synthetic pyrethroids) and neem (a botanical insecticide)rdquoPestology vol 30 no 10 pp 21ndash28 2006

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


Earlier studies have shown the efficacy of Eucalyptus oilsas a larvicide against mosquitoes [13ndash16] In the year 2004Yang et al reported that Eucalyptus leaf oil particularly someof its components such as 18-cineole 1-120572-terpineol and (E)-pinocaveol merits further study as lead compounds for thecontrol of the human hair louse Pediculus humanus capitisDe Geer (Pediculidae) [17] Constituents of the essential oilof Eucalyptus camaldulensisDehnh and Eucalyptus urophyllaS T Blake (Myrtaceae) a-phellandrene limonene p-cymenec-terpinene terpinolene and a-terpinene have been shownto possess strong larvicidal effects against Aedes aegypti andAedes albopictus [15] The various components of Eucalyptusessential oil have been shown to act synergistically (and notadditively) to bring the overall bactericidal activity [18]

Centella asiatica is mostly studied for its medicinal prop-erties Very few studies have documented its potential as abiopesticide A study has suggested that the leaf extract ofC asiatica is promising as a larvicide and adult emergenceinhibitor against Culex quinquefasciatus Say (Culicidae) [19]Shukla et al [20] isolated and studied the growth inhibitoryeffect of 2120572 3120573 6120573 23-120572 tetrahydroxyurs-12-ene-28-oicacid a triterpenoid glycoside against larvae of SpilarctiaobliquaWalker (Arctiidae)

Numerous studies have examined the variations in lar-vicidal efficacies of plant extracts tested against differentmosquito species however we were not able to find anyreports of the larvicidal efficacies of plant extracts testedagainst different strains of mosquitoes of the same speciesSusceptibility studies involving strains from different loca-tions could be beneficial to gauge the efficacy of the drugbeing studied and as a prelude to investigations involving theinherent resistance mechanisms of the vectors to the drugs

Different solvent types can significantly affect the potencyof the extracted plant compounds and there is a differencein the chemoprofile of the plant species [21] This is becausepolar solvents extract polar constituents of the plant whilenonpolar solvents extract the nonpolar constituents of theplant Hence the best results will be observed using the plantextract of the solvent whose polarity matches that of themolecules in the plant responsible for insecticidal activity(if any) Additionally extracts or pure compounds derivedfrom specific solvents can influence the bioactivity probablybecause of the active components which are present in largequantities [22]

In the present study we have investigated the larvicidalactivity of the hexane diethyl ether dichloromethane andmethanol extracts of the leaves of Eucalyptus globulus Labill(Myrtaceae) and Centella asiatica (Linn) Urban (Apiaceae)against two geographically different strains of Aedes (Ste-gomyia) aegypti (Linn) (Diptera Culicidae) and Anophelesstephensi Liston (Diptera Culicidae)

2 Materials and Methods

21 Mosquito Culture Four strains which consisted of lab-oratory colonies of Aedes aegypti collected from J P Nagar(JPN) (129120∘N 775930∘E) and Basaveshwaranagar (BSV)(129867∘N 775386∘E) and Anopheles stephensi collectedfrom Dollars Colony (DLC) (13∘210158403010158401015840N 77∘341015840310158401015840E) and

Kengeri (KNG) (129100∘N 774800∘E) were used for thisstudy Larvae collected from the said locations of BBMPregion Karnataka India during the months March-April2012 had been reared continuously in the insectary at theCentre for Applied Genetics Bangalore University followingthe method of Shetty [23]

22 Collections of Plant Materials The leaves of Eucalyptusglobulus and Centella asiatica were collected from Bangalorecity (11∘110158404810158401015840N 77∘210158403610158401015840E elevation 399m) KarnatakaIndia in July 2013 and were authenticated at the Departmentof Botany Bangalore University Bangalore India

23 Preparation of Plant Extracts The leaves (2 kg each) wereair dried in shade for 15ndash30 days The dried leaves werethen powdered mechanically using a commercial electricalstainless steel blender One kg of powdered leaves wasextracted successively by maceration using nonpolar to polarsolvents namely hexane diethyl ether dichloromethane andmethanol In each solvent the plant material was soaked for48 h at 35∘C and filtered twice using Whatman number 1filter paper to obtain the extract and to the residue the samesolvent was added againThe procedure was repeated twice toobtain maximum extract The extracts were concentrated atreduced temperature using a rotary vacuum evaporator andstored at a temperature of 4∘C One gram of the concentratedplant extractwas dissolved in 100mLof 1 1 acetone dimethylsulfoxide (DMSO) and considered as 1 stock solution Fromthis stock solution varying concentrations of each extractwasprepared and these solutionswere used for larvicidal bioassayAll chemicals used in this study were of extra pure grade andwere obtained from Sisco Research Laboratories PVT LtdIndia

24 Larvicidal Bioassay Larvicidal activity of each extractderived from the leaves of Eucalyptus globulus and Centellaasiatica against the two strains ofAe aegypti andAn stephensiwas assessed by using a slightlymodified version of theWHOstandard method [24]

Initially the mosquito larvae were exposed to a broadrange of test concentrations to determine the activity range ofeach extract Based on the results of preliminary screeningbatches of 25 late third instar larvae were added to 300mLwide mouth disposable bowls containing serial concentra-tions (6ndash9 concentrations yielding between 10 and 95mortality in 24 hrs) of each plant extract made up to 250mLby volume using tap water The test was carried at a tem-perature of 25 plusmn 2∘C and relative humidity of 75 plusmn 5The numbers of dead larvae were counted after 24 hours ofexposure and percentage mortality was calculated for eachtest as follows

Percentage Mortality

=

Number of dead larvaepupaeNumber of larvae introduced

times 100

(1)

The final percentage mortality was calculated from theaverage of three replicates Solutions containing tapwater and1 1 acetone dimethyl sulfoxide butwithout the plant sample




50and

LC90


3 Results


50 LC90




501188 and LC



4641 and LC90


501620 and LC





502252 1677 and LC

904236 3040 ppm resp)


5018390 and LC

90

32359 ppm)


502465 2687 and LC

905931 16068 ppm resp)


90


50147 and

LC90


507765 and LC

9038534 ppm)


4 Discussion








50value of the





50of hexane leaf




50of



Table1Eff

ectsof

relativ

etoxicity


methano

l)of

theleaveso

fEucalyptusg

lobu

lusa

ndCe


latethird

larvalsta

geof

BSVandJPNstr

ains

ofAe

desa

egyptiandDLC

andKN

Gstr

ains

ofAn

opheles

stephensi

Botanicaln

ame

Solvents

Species

Strain

LC50(ppm

)(UCL

-LCL

)LC

90(ppm

)(UCL

-LCL

)Re

gressio

nequatio

n119903

1205942

df

Eucalyptus

globu

lus

Hexane

Aeaegypti

BSV

2252(2109ndash2635)

4236(4004ndash5989)

119910=46638119909minus59716

09364

01902

7JPN

1677

(1521ndash

1867)

304(2663ndash4

009)

119910=49559119909minus60248

0984

00238

6

Anstephensi

DLC

1188(881ndash1512

)4504(3738ndash9

533)

119910=22119119909+04106

09137

01123

7KN

G1928(1547ndash

2551)

827(6552ndash1634)

119910=20242119909+03744

09378

01134

7

Diethylether

Aeaegypti

BSV

6868(6308ndash7564)

12167(10737ndash15825)119910=51546119909minus96228

09594

00244

7JPN

3188(14

67ndash64

12)

20699(10851ndash348885)119910=15756119909+10554

09114

01811

6

Anstephensi

DLC

4641(4288ndash5242)

8001(7261ndash10507)

119910=54107119909minus9428

09382

00767

6KN

G318(2706ndash3917)

9519

(7374ndash16187)

119910=2688119909minus17265

09279

006

056

Dichlorom

ethane

Aeaegypti

BSV

3146(2761ndash

385)

8299

(6771ndash

12907)

119910=3038119909minus2588

09846

00832

6JPN

3622(1716ndash6

201)

29046(106

86ndash

139251)119910=14157119909+13773

0889

01362

6

Anstephensi

DLC

3003(2563ndash3942)

12354(8725ndash22839)

119910=20839119909minus0163

09747

01345

7KN

G162(1125ndash1994)

5824(4526ndash13234)

119910=23036119909minus00899

09623

00536

7

Methano

lAe

aegypti

BSV

18393(16958ndash20497)

32359(29302ndash40

511)

119910=52171119909minus12032

09805

00504

7JPN

14679(1281ndash17868)

35302(29117ndash56123)

119910=33587119909minus5636

096

4400634

5

Anstephensi

DLC

12633(7981ndash

18173)

36143(29883ndash127366)

119910=28038119909minus3696

08769

02807

7KN

G8896

(764

7ndash11243)

26551(21770

ndash49074

)119910=26955119909minus29496

09204

01292

6

Centellaasiatica

Hexane

Aeaegypti

BSV

2465(2131ndash

3071)

5931(5273

ndash10139)

119910=33561119909minus3027

09284

01366

5JPN

2687(1744ndash3747)

16068(10821ndash5866

2)119910=16479119909+09969

0929

00583

5

Anstephensi

DLC

506(183ndash105)

3288(19

85ndash8730)

119910=15752119909+23154

08489

02479

6KN

G2435(2211ndash

2742)

4286(3627ndash

5845)

119910=52115119909minus74372

09963

00055

5

DiethylEther

Aeaegypti

BSV

3396

(2581ndash

4723)

15993(11736ndash

3840

5)119910=19022119909+01855

08489

00844

5JPN

1345(206ndash

2041)

7708(5767ndash

31917)

119910=16885119909+14054

09362

004

635

Anstephensi

DLC

241(2034ndash

3015

)6307(540

4ndash11162)

119910=3063119909minus22959

08489

01046

5KN

G147(3ndash35)

118(637ndash17122)

119910=14175119909+33433

08862

01491

5

Dichlorom

ethane

Aeaegypti

BSV

3397

(281ndash7463)

21277(12419

ndash403246)

119910=16065119909+09337

09384

01913

5JPN

8376

(7779ndash10302)

16272(15423ndash25588)119910=44382119909minus7973

09

01722

5

Anstephensi

DLC

2368(14

55ndash3933)

33471(15767ndash279272

)119910=11127119909+23581

09338

00533

5KN

G1555(209ndash

2955)

30229(17325ndash335591

)119910=09933119909+28229

08881

00528

5

Methano

lAe

aegypti

BSV

7765(59099ndash

109078)

38534(272801ndash10164

21)119910=21701119909minus12027

08905

00966

5JPN

7215

(36954ndash

135804)

2806

(186607ndash3248971)119910=21701119909minus12027

08905

01928

5

Anstephensi

DLC

4768(46194ndash

49349)

5879

(56722ndash64399)119910=14064119909minus32668

09832

00333

8KN

G177(11928ndash22509)

5599

(48397ndash126017

)119910=25595119909minus07538

09002

0110

75

LC50lethalcon

centratio

nthatkills

50of

thee

xposed

larvaeLC 9


90of

thee

xposed

larvaeU

CLupp

erconfi

dencelim

itLC

Llower

confi

dencelim


cient

1205942C

hi-squ

ares


levelanddfdegreeo

ffreedom







50values for the




5 Conclusion




References






















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




50and

LC90


3 Results


50 LC90




501188 and LC



4641 and LC90


501620 and LC





502252 1677 and LC

904236 3040 ppm resp)


5018390 and LC

90

32359 ppm)


502465 2687 and LC

905931 16068 ppm resp)


90


50147 and

LC90


507765 and LC

9038534 ppm)


4 Discussion








50value of the





50of hexane leaf




50of



Table1Eff

ectsof

relativ

etoxicity


methano

l)of

theleaveso

fEucalyptusg

lobu

lusa

ndCe


latethird

larvalsta

geof

BSVandJPNstr

ains

ofAe

desa

egyptiandDLC

andKN

Gstr

ains

ofAn

opheles

stephensi

Botanicaln

ame

Solvents

Species

Strain

LC50(ppm

)(UCL

-LCL

)LC

90(ppm

)(UCL

-LCL

)Re

gressio

nequatio

n119903

1205942

df

Eucalyptus

globu

lus

Hexane

Aeaegypti

BSV

2252(2109ndash2635)

4236(4004ndash5989)

119910=46638119909minus59716

09364

01902

7JPN

1677

(1521ndash

1867)

304(2663ndash4

009)

119910=49559119909minus60248

0984

00238

6

Anstephensi

DLC

1188(881ndash1512

)4504(3738ndash9

533)

119910=22119119909+04106

09137

01123

7KN

G1928(1547ndash

2551)

827(6552ndash1634)

119910=20242119909+03744

09378

01134

7

Diethylether

Aeaegypti

BSV

6868(6308ndash7564)

12167(10737ndash15825)119910=51546119909minus96228

09594

00244

7JPN

3188(14

67ndash64

12)

20699(10851ndash348885)119910=15756119909+10554

09114

01811

6

Anstephensi

DLC

4641(4288ndash5242)

8001(7261ndash10507)

119910=54107119909minus9428

09382

00767

6KN

G318(2706ndash3917)

9519

(7374ndash16187)

119910=2688119909minus17265

09279

006

056

Dichlorom

ethane

Aeaegypti

BSV

3146(2761ndash

385)

8299

(6771ndash

12907)

119910=3038119909minus2588

09846

00832

6JPN

3622(1716ndash6

201)

29046(106

86ndash

139251)119910=14157119909+13773

0889

01362

6

Anstephensi

DLC

3003(2563ndash3942)

12354(8725ndash22839)

119910=20839119909minus0163

09747

01345

7KN

G162(1125ndash1994)

5824(4526ndash13234)

119910=23036119909minus00899

09623

00536

7

Methano

lAe

aegypti

BSV

18393(16958ndash20497)

32359(29302ndash40

511)

119910=52171119909minus12032

09805

00504

7JPN

14679(1281ndash17868)

35302(29117ndash56123)

119910=33587119909minus5636

096

4400634

5

Anstephensi

DLC

12633(7981ndash

18173)

36143(29883ndash127366)

119910=28038119909minus3696

08769

02807

7KN

G8896

(764

7ndash11243)

26551(21770

ndash49074

)119910=26955119909minus29496

09204

01292

6

Centellaasiatica

Hexane

Aeaegypti

BSV

2465(2131ndash

3071)

5931(5273

ndash10139)

119910=33561119909minus3027

09284

01366

5JPN

2687(1744ndash3747)

16068(10821ndash5866

2)119910=16479119909+09969

0929

00583

5

Anstephensi

DLC

506(183ndash105)

3288(19

85ndash8730)

119910=15752119909+23154

08489

02479

6KN

G2435(2211ndash

2742)

4286(3627ndash

5845)

119910=52115119909minus74372

09963

00055

5

DiethylEther

Aeaegypti

BSV

3396

(2581ndash

4723)

15993(11736ndash

3840

5)119910=19022119909+01855

08489

00844

5JPN

1345(206ndash

2041)

7708(5767ndash

31917)

119910=16885119909+14054

09362

004

635

Anstephensi

DLC

241(2034ndash

3015

)6307(540

4ndash11162)

119910=3063119909minus22959

08489

01046

5KN

G147(3ndash35)

118(637ndash17122)

119910=14175119909+33433

08862

01491

5

Dichlorom

ethane

Aeaegypti

BSV

3397

(281ndash7463)

21277(12419

ndash403246)

119910=16065119909+09337

09384

01913

5JPN

8376

(7779ndash10302)

16272(15423ndash25588)119910=44382119909minus7973

09

01722

5

Anstephensi

DLC

2368(14

55ndash3933)

33471(15767ndash279272

)119910=11127119909+23581

09338

00533

5KN

G1555(209ndash

2955)

30229(17325ndash335591

)119910=09933119909+28229

08881

00528

5

Methano

lAe

aegypti

BSV

7765(59099ndash

109078)

38534(272801ndash10164

21)119910=21701119909minus12027

08905

00966

5JPN

7215

(36954ndash

135804)

2806

(186607ndash3248971)119910=21701119909minus12027

08905

01928

5

Anstephensi

DLC

4768(46194ndash

49349)

5879

(56722ndash64399)119910=14064119909minus32668

09832

00333

8KN

G177(11928ndash22509)

5599

(48397ndash126017

)119910=25595119909minus07538

09002

0110

75

LC50lethalcon

centratio

nthatkills

50of

thee

xposed

larvaeLC 9


90of

thee

xposed

larvaeU

CLupp

erconfi

dencelim

itLC

Llower

confi

dencelim


cient

1205942C

hi-squ

ares


levelanddfdegreeo

ffreedom







50values for the




5 Conclusion




References






















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table1Eff

ectsof

relativ

etoxicity


methano

l)of

theleaveso

fEucalyptusg

lobu

lusa

ndCe


latethird

larvalsta

geof

BSVandJPNstr

ains

ofAe

desa

egyptiandDLC

andKN

Gstr

ains

ofAn

opheles

stephensi

Botanicaln

ame

Solvents

Species

Strain

LC50(ppm

)(UCL

-LCL

)LC

90(ppm

)(UCL

-LCL

)Re

gressio

nequatio

n119903

1205942

df

Eucalyptus

globu

lus

Hexane

Aeaegypti

BSV

2252(2109ndash2635)

4236(4004ndash5989)

119910=46638119909minus59716

09364

01902

7JPN

1677

(1521ndash

1867)

304(2663ndash4

009)

119910=49559119909minus60248

0984

00238

6

Anstephensi

DLC

1188(881ndash1512

)4504(3738ndash9

533)

119910=22119119909+04106

09137

01123

7KN

G1928(1547ndash

2551)

827(6552ndash1634)

119910=20242119909+03744

09378

01134

7

Diethylether

Aeaegypti

BSV

6868(6308ndash7564)

12167(10737ndash15825)119910=51546119909minus96228

09594

00244

7JPN

3188(14

67ndash64

12)

20699(10851ndash348885)119910=15756119909+10554

09114

01811

6

Anstephensi

DLC

4641(4288ndash5242)

8001(7261ndash10507)

119910=54107119909minus9428

09382

00767

6KN

G318(2706ndash3917)

9519

(7374ndash16187)

119910=2688119909minus17265

09279

006

056

Dichlorom

ethane

Aeaegypti

BSV

3146(2761ndash

385)

8299

(6771ndash

12907)

119910=3038119909minus2588

09846

00832

6JPN

3622(1716ndash6

201)

29046(106

86ndash

139251)119910=14157119909+13773

0889

01362

6

Anstephensi

DLC

3003(2563ndash3942)

12354(8725ndash22839)

119910=20839119909minus0163

09747

01345

7KN

G162(1125ndash1994)

5824(4526ndash13234)

119910=23036119909minus00899

09623

00536

7

Methano

lAe

aegypti

BSV

18393(16958ndash20497)

32359(29302ndash40

511)

119910=52171119909minus12032

09805

00504

7JPN

14679(1281ndash17868)

35302(29117ndash56123)

119910=33587119909minus5636

096

4400634

5

Anstephensi

DLC

12633(7981ndash

18173)

36143(29883ndash127366)

119910=28038119909minus3696

08769

02807

7KN

G8896

(764

7ndash11243)

26551(21770

ndash49074

)119910=26955119909minus29496

09204

01292

6

Centellaasiatica

Hexane

Aeaegypti

BSV

2465(2131ndash

3071)

5931(5273

ndash10139)

119910=33561119909minus3027

09284

01366

5JPN

2687(1744ndash3747)

16068(10821ndash5866

2)119910=16479119909+09969

0929

00583

5

Anstephensi

DLC

506(183ndash105)

3288(19

85ndash8730)

119910=15752119909+23154

08489

02479

6KN

G2435(2211ndash

2742)

4286(3627ndash

5845)

119910=52115119909minus74372

09963

00055

5

DiethylEther

Aeaegypti

BSV

3396

(2581ndash

4723)

15993(11736ndash

3840

5)119910=19022119909+01855

08489

00844

5JPN

1345(206ndash

2041)

7708(5767ndash

31917)

119910=16885119909+14054

09362

004

635

Anstephensi

DLC

241(2034ndash

3015

)6307(540

4ndash11162)

119910=3063119909minus22959

08489

01046

5KN

G147(3ndash35)

118(637ndash17122)

119910=14175119909+33433

08862

01491

5

Dichlorom

ethane

Aeaegypti

BSV

3397

(281ndash7463)

21277(12419

ndash403246)

119910=16065119909+09337

09384

01913

5JPN

8376

(7779ndash10302)

16272(15423ndash25588)119910=44382119909minus7973

09

01722

5

Anstephensi

DLC

2368(14

55ndash3933)

33471(15767ndash279272

)119910=11127119909+23581

09338

00533

5KN

G1555(209ndash

2955)

30229(17325ndash335591

)119910=09933119909+28229

08881

00528

5

Methano

lAe

aegypti

BSV

7765(59099ndash

109078)

38534(272801ndash10164

21)119910=21701119909minus12027

08905

00966

5JPN

7215

(36954ndash

135804)

2806

(186607ndash3248971)119910=21701119909minus12027

08905

01928

5

Anstephensi

DLC

4768(46194ndash

49349)

5879

(56722ndash64399)119910=14064119909minus32668

09832

00333

8KN

G177(11928ndash22509)

5599

(48397ndash126017

)119910=25595119909minus07538

09002

0110

75

LC50lethalcon

centratio

nthatkills

50of

thee

xposed

larvaeLC 9


90of

thee

xposed

larvaeU

CLupp

erconfi

dencelim

itLC

Llower

confi

dencelim


cient

1205942C

hi-squ

ares


levelanddfdegreeo

ffreedom







50values for the




5 Conclusion




References






















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







50values for the




5 Conclusion




References






















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology











































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology













Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

SavithaSekharNair,VinayaShetty,andNadikereJayaShettyJournal of Insects Earlier studies have shown the e cacy of Eucalyptus oils as a larvicide against mosquitoes [ ]. In the year ,