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Pesticidal Action of Certain Plant Extracts Against Mosquito Vectors

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Pesticidal Action of Certain Plant Extracts Against Mosquito Vectors PESTICIDAL ACTION OF CERTAIN PLANT EXTRACTS AGAINST MOSQUITO VECTORS (CULICIDAE: DIPTERA) Thesis submitted in fulfillment of the requirements for the degree of Doctor of Philosophy in Zoology by Lalrotluanga Ph.D. Registration No: MZU/Ph.D/228 of 28.11.2008 Under the Supervision of Dr. G. Gurusubramanian Associate Professor Department of Zoology & Joint- Supervision of Dr. N. Senthil Kumar Professor Department of Biotechnology Department of Zoology School of Life Sciences Mizoram University, Aizawl, Mizoram, India 2012 CERTIFICATE I certify that the thesis entitled “Pesticidal action of certain plant extracts against mosquito vectors (Culicidae: Diptera)” sumitted to the Mizoram University for the award of the degree of Doctor of Philosophy in Zoology by LALROTLUANGA is a record of research work carried out by him during the period from 2008 to 2012 under my guidance and supervision, and that this work has not formed the basis for the award of any degree, diploma, associateship, fellowship or other similars titles in this university or any other university or institution of higher learning. Signature of the Supervisor (G. GURUSUBRAMANIAN) Signature of the Co-Supervisor (N. SENTHIL KUMAR) DECLARATION I declare that the thesis entitled “Pesticidal action of certain plant extracts against mosquito vectors (Culicidae: Diptera)” submitted to the Mizoram University for the award of degree of Doctor of Philosophy in Biotechnology is a bonafide record of work carried out by me during the period from 2008 to 2012 under the guidance of Dr. G. Gurusubramanian (Supervisor), Department of Zoology and Prof. N. Senthil Kumar (Co- Supervisor), Department of Biotechnology and has not formed the basis for the award of any degree, diploma, associateship, fellowship or other similar titles in this University or other University or institution of higher learning. Signature of the Candidate (LALROTLUANGA) ACKNOWLEDGEMENT I express my heartfelt gratitude to my respected supervisor Dr. G. Gurusubramanian for his immaculate guidance throughout my research. I express my sincere thanks to Professor N. Senthil Kumar, my co-supervisor and Head, Department of Biotechnology, Mizoram University for his intense involvement and assistance throughout my studies. My heartfelt thanks to Miss Esther Lalhmingmawii, Dr. HT Lalremsanga for their assistance and encouragement. I thank Prof G.C. Jagetia and Dr. N. Solanki for providing necessary facilities to carry out my Ph.D research work successfully. I also would like to thank all the non-teaching staff of Department of Zoology. I express my gratitude to my laboratory members- Rita Zomuanpuii, Zothansangi, Lalhmachhuani, Laltanpuii Catherine Vanlalruati, Vanlalsawmi, Zothansanga, Zothansanga, K Vanlalhruaia, Vabeihrulai, Lalrinzuali, Zothansiama and C. Lalrinchhana. I thank UGC, New Delhi for the Meritorius fellowship . Words can not express my gratitude to my parents, my sister and my nieces for their everlasting love, encouragement and support. Above all, I thank God for his guidance and blessing throughout my life. (LALROTLUANGA) LIST OF CONTENTS 1. Introduction and review of literature 1 - 46 2. Objectives of the study 47 3. Materials and methods 48 - 63 3.1 Mosquito culture 3.2 Preparation of plant extracts 3.3 Larvicidal bioassay 3.3.1 Preliminary screening of larvicidal bioassay 3.3.2 Dose–response larvicidal bioassay 3.3.3 Time response Larvicidal Bioassay (LT50) 3.3.4 Estimation of nutrient reserves 3.3.4.1 Sugar analysis 3.3.4.2 Glycogen analysis 3.3.4.3 Lipid analysis 3.3.4.4 Protein analysis 3.3.4.5 Standard curves 3.4 Insect Growth Regulators 3.5 Adulticidal Bioassay 3.6 Repellent Bioassay 3.7 Oviposition Deterrent Test 3.8 RAPD-PCR profiling of treated mosquito larvae 3.8.1 DNA Extraction 3.8.2 RAPD-PCR amplification 3.8.3 Agarose gel Electrophoresis 3.9 Statistical analysis 4. Results 64 - 97 5. Discussion 98 - 114 6. Summary 115 - 120 7.References 121 -157 LIST OF TABLES Table Titles of the table No. Efficacy of petroleum ether extracts of plants against mosquito 1 vectors Efficacy of chloroform extracts of plants against mosquito 2 vectors 3 Efficacy of acetone extracts of plants against mosquito vectors 4 Efficacy of Methanol extracts of plants against mosquito vectors 5 Efficacy of aqueous extracts of plants against mosquito vectors Ethnomedicinal plants tested for their insecticidal actions 6 against mosquitoes and their properties 7 Traditional plants used for mosquito repellent 8 List of random primers used for RAPD analysis 9 RAPD-PCR reaction conditions Screening of larvicidal effecfs of ethnomedicinal plants against 10 Ae. Albopictus at 500 ppm concentration in different solvents Screening of larvicidal effects of ethnomedicinal plants against 11 An. barbirostris at 500 ppm concentration in different solvents Screening of larvicidal effecfs of ethnomedicinal plants against 12 Cx. quinquefasciatus at 500 ppm concentration in different solvents Mortality, log probit and regression analysis of third larval 13 instars of A. albopictus, A. barbirostris and C. quinquefascitus in different concentrations of crude root extract of H. benghalensis Completely randomized three-way factorial ANOVA using mosquito species, period of bioassay and different 14 concentrations as variables Lethal time (LT50) of acetone extract of H. benghalensis (root) 15 against the three mosquito vector species Mortality, log probit and regression analysis of third larval instars of A. albopictus, A. barbirostris and C. quinquefascitus in 16 different concentrations of petroleum ether extract of Curcuma longa rhizome 17 Log probit and regression analysis of larvicidal activity of petroleum ether extract of Elsholtzia communis (leaf) against different mosquito species Log probit and regression analysis of larvicidal activity of extract 18 of Elsholtzia communis leaf (chloroform) against different mosquito species Log probit and regression analysis of larvicidal activity of acetone 19 extract of Elsholtzia communis leaf (Acetone) against different mosquito species Mortality, log probit and regression analysis of third larval instars of Ae. albopictus, An. barbirostris and Cx. quinquefascitus 20 in different concentrations of Petroleum ether extract of Homalomena aromatica rhizome Larvicidal action of Syzygium aromaticum leaves against 3rd 21 instar larvae of Culex quinquefaciatus (Culicidae:Diptera) Biochemical profile of nutrient reserves and primary metabolites 22 in freshly emerged III instar larvae treated with sublethal concentration (5 ppm) of acetone extract of H. benghalensis root Consequence of different concentrations of petroleum ether 23 extract of Melia azedarach fruit on the development of Aedes albopictus treated as the first instar larvae . Preliminary effect of acetone extract of H. benghalensis root on Ae. albopictus, An.barbirostris, and Cx. quinquefasciatus adults 24 exposed continuously for 3 hours at different concentrations and mortality after 24 hour exposure Table 25. Repellent activity of Gmelina arborea leaf extracts 25 against blood-starved female Ae. albopictus Repellent activity of Curcuma longa rhizome extracts against 26 blood-starved female Ae.albopictus Repellent activity of Homalomena aromatica rhizome extracts 27 against blood-starved female Ae.albopictus Repellent activity of Gmelina arborea leaf extracts against blood- 28 starved female An. barbirostris Oviposition deterrency of different plant extracts against Ae. 29 albopictus female Alteration in number of total bands in treated genomic DNA from 30 larvae of C. quinquefasciatus List of Plates Plate No. Title of the Plate 1. Mosquito vectors used in the present study - Aedes albopictus - Anopheles barbirostris - Culex quinquefasciatus 2. Mosquito culture 3. Plants tested for mosquitocidal properties 4. Effect of petroleum extract of Melia azedarach on the development of Cx. quinquefasciatus larvae List of Figures Fig. No. Title of the Figures 1. Global distribution of human Plasmodium spp. and lymphatic filarial species A) malaria) and (B) lymphatic filariasis 2. Distribution of Dengue disease throughout the world 3. Study design for repellent test 4. Comparison of RAPD-PCR profiles of III instar larvae of Culex quinquefasciatus treated with plant extracts 1 1. INTRODUCTION AND REVIEW OF LITERATURE Mosquitoes are the most important single group of insects in terms of public health importance. Mosquitoes not only cause nuisance by their bites but also transmit deadly diseases. Among the approximately 4000 known mosquito species, less than 10% are regarded as efficient vectors of pathogenic agents of infectious diseases having high impact, both direct and indirect, on human welfare and health. Mosquito-transmitted diseases remain a major cause of the loss of human life worldwide with more than 700 million people suffering from these diseases annually (Taubes 1997). Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases (Fradin and Day 2002). Mosquitoes transmit a number of diseases, such as malaria, filariasis, dengue, Japanese encephalitis, etc. Dengue fever incidence has increased fourfold since 1970 and nearly half the world’s population is now at risk. In 1990, about 1.5 billion people lived in regions where the estimated risk of dengue transmission was greater than fifty percent (Hales et al. 2002). An outbreak of chikungunya virus infection emerged in the
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    ACTA ENTOMOLOGICA J'viUSEI NATIONALIS PRAGAE 1948, XXVI, 367 367. A BIBLIOGRAPHY OF THE ANOPLURA OR SUCKING LICE. L. FREUND Zool. Institute of the Martin-Luther-University, Halle-S. (Accepted for publication December 31, 1949.) This is a further collection of the literature concerning the sucking lice or Anoplura, comprehending the publications from 1927 til1948. The publications prior to 1927 has the author published in the Zentralblatt fiir Bakteriologie und Parasitenkunde, Abt. I, Ref. 1927, p. 343-384. Therewith is the bibliography completed up to date, including a couple of titles omitted in the former publication. Regarding the importance of the sucking lice as transporters of dieases especially of the spotted fever is the utility of such a collection for further investigations without doubt. ADLER, S. & AsBEL, R., The behavior of Spirochaeta persica in Pediculus humanus. Ann. trop. Med. Paras. 36. 1942, 83-96. AKINSOHIN, TH., Parl:J,sitic lice on animalis and their destruction. The Agriculturist St. Petersburg, 7. 1914, 287. - Rev. Appl. Ent. B. 2. 1914, ll8. ALESS.ANDRINI, G., e MrssmoLI, A., Sulla struttura dell'apparato respira­ torio dmle ,Anopheles". Riv. Malariol. 5. 1926, 35-43. (Stigma of Anopl.) ALVERDES, F. & BrELING, R., Untersuchungen iiber bakterielle Infektio­ nen bei Kleiderlausen (Pediculus vestimenti). Ztschr. Natfschg. 4 b, 1949, 150-157. ALPATov, V. V., The louse of man, its development and survival at diffe­ rent temperatures. Adv. Mod. Biol. Moscow, 15. 1942, 190-207. ANGEVINE, W. K., Pyrethrum ointment. A report on its use in the control ofliceandscabies.Soap, 17.1941, ll7-ll9. ANIGSTEIN, L. & LAWKOWIOZ, W., Researches of strains of Rickettsia and Proteus cultivated from experimental typhus of the murine type.
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