BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY:
Insect-transmitted diseases remain a major source of illness and death worldwide. Mosquitoes alone transmit disease to more than 700 million people annually. Malaria alone kills 3 million each year, including 1 child every 30 seconds. Although mosquito-borne diseases currently represent a greater health problem in tropical and subtropical climates, no part of the world is immune to this risk1.
Control of such diseases is becoming increasingly difficult because of increasing resistance of mosquitoes to pesticides. An alternative approach for mosquito control is the use of natural products of plant origin. The botanical insecticides are generally pest specific, readily biodegradable and usually lack toxicity to higher animals. One plant species may possess substances with a wide range of activities, for example extracts from the neem tree Azadirachta indica showed antifeedant, oviposition deterrent, mosquito repellent and growth regulating activity.
In recent years interest in plant based products has been revived because of the development of cross-resistance and possible toxicity hazards associated with synthetic insecticides and their rising cost. Phytochemicals obtained from the huge diversity of plant species are important source for safe and biodegradable chemicals, which can be screened for mosquito repellent and insecticidal activities and tested for mammalian toxicity2.
In addition to application as general toxicants against mosquito immature, phytochemicals may also have potential uses as growth and reproduction inhibitors, repellents, and oviposition deterrents.
The most common mosquito repellent formulations available on the market contain DEET (N,N-diethyl-3-methylbenzamide), which has shown excellent repellency against mosquitoes and other biting insects .However, human toxicity reactions after the applications of DEET vary from mild to severe. To avoid these adverse effect, research on repellents that are derived from plant essential oil to replace DEET. Botanical repellents are promising in that they are effective, safe to users, and also inexpensive1.
A large number of such plants are available in ayurveda, which if validated scientifically helps a lot to the society. Hence the present study is proposed to investigate the LARVICIDAL ACTION OF ESSENTIAL OILS FROM THE FLOWERS OF NELUMBO NUCIFERA AND TAGETES ERECTA.
6.2 REVIEW OF THE LITERATURE:
Synonym: Nelumbium nelumbo.
Description: Perennial aquatic herb, Root stock stout, cylindrical. Leaves peltate, radiately nerved. Flowers large, solitary, fragnant, ovoid, fleshy, sunk separately in cavities of receptacle, maturing into nut-like achenes.
Parts used: Root, leaves, flowers, seeds
Chemical constituents: Leaves contain alkaloids: nuciferine, roemerine, flavanoid: quercetin. The plumules yield proteins, sugars and vitamins. The receptacles contain quercetin.
Uses: Lotus is helpful in controlling the burning sensation, due to its cold potency. The leaf paste is applied to the body in case of fever and inflammatory skin conditions. The flower stalk, mixed with other herbs, is used to treat bleeding from the uterusons. The leaves are also used in treating sunstroke, diarrhea, dysentery, dizziness and vomiting of blood.
Synonym: Mexican marigold.
Description: Tagetes erecta is a bushy composite plant, having pinnate leaves and large yellow or orange flower heads. Also called Aztec marigold.
Parts used: Flowers, leaves, stem.
Chemical constituents: Tagetes erecta contains thiophenes, flavonoids, carotenoids, triterpenoids. Glucoside of Quercetagenin, phenolics, syringic acid, quercetin, thienyl and ethyl gallat.
Uses: The flowers of Tagetes erecta are the source of two food colourant products: Marigold meal and Marigold extract. The plant is most effective against the nematode species Pratylenchus penetrans. T. erecta is grown to extract lutein, a common yellow/orange food colour. The essential oil of the flower contains antioxidants.
Studies on the anti-inflammatory activity of rhizomes of Nelumbo nucifera was carried out by P K Mukharjee.et.al.3.
Anti-HIV benzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera and structure–activity correlations with related alkaloids by Y Kashiwada.et.al.4
Antioxidant activity of Nelumbo nucifera seeds by S Rai, A Wahile.et.al.5
Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats was carried out by Yuka onoa.et al.6
Antipyretic activity of Nelumbo nucifera rhizome extract by P K Mukharjee.et.al.7
Synthesis of silver nanoparticles using Nelumbo nucifera leaf extract and its larvicidal activity against malaria and filariasis vectors was carried out by T Santosh kumar.et.al.8
Hypoglycemic activity of Nelumbo nucifera gaertn.(Fam. Nymphaeaceae) rhizome (methanolic extract) in streptozotocin‐induced diabetic rats by PK Mukharjee.et.al.9
Inhibitory effects of Nelumbo nucifera leaves on rat lens aldose reductase, advanced glycation endproducts formation, and oxidative stress by H A Jung.et.al.10
Studies on essential oils chemical and biocidal investigations on Tagetes erecta leaf volatile oil by G Singh.et.al.11
Effects of Tagetes erecta extracts on glutathione S-transferase and protease activities and protein content in Tetranychus viennensis was carried out by G L Shi.et.al.12
Growth response of marigold (Tagetes erecta L.) to inoculation withGlomus mosseae, Trichoderma aureoviride andPythium ultimum in a peat-perlite mixture by C Calvet.et.al.13
Insecticidal activity of callus culture of Tagetes erecta was carried out by R Sarin et al.14
Enzyme-mediated solvent extraction of carotenoids from marigold flower by E Barzana.et.al.15
Studies on the antioxidant and analgesic activities of Aztec marigold (Tagetes erecta) flowers by S Bashir.et.al.16
Antimutagenicity of xanthophylls present in Aztec Marigold against 1-nitropyrene by E González de Mejı́a.et.al.17
Larvicidal activity of Tagetes minuta (marigold) toward Aedes aegypti by MM Green.et.al.18
Light‐dependent toxicity of the extract of plant Tagetes erecta and alpha‐terthienyl toward larvae of mosquito Culex tritaeniorhynchus was carried out by SP Singh.et.al.19
Insecticidal activity of flower of Tagetes erecta L. against Tribolium castaneum (Herbst) by F Nikkon.et.al.20
Larvicidal action of essential oils from plants against the vector mosquitoes Anopheles stephensi (Liston), Culex quinquefasciatus (Say) and Aedes aegypti (L.) was carried out by Namrata pathak.et.al.21
6.3 THE OBJECTIVE OF THE STUDY:
Collection and authentification of flowers of Nelumbo nucifera (Nelumbonaceae) and Tagetes erecta (Asteraceae)
Determination of larvicidal activity from the flowers of Nelumbo nucifera and Tagetes erecta by Steam distillation using Clevenger’s apparatus.
To establish a pharmacological profile of obtained extract for its larvicidal activity.
MATERIALS AND METHODS:
7.1 SOURCE OF DATA
Whole work is planned to generate data from laboratory studies. Experiments are performed as described in reference, experimental studies in journals and in text books available within the college, IISC library, Bengaluru, RGUHS digital library (Helinet), Text books of Pharmacognosy and Botany.
Place of work: P.E.S college of pharmacy,Bengaluru-560 050.
I.C.M.R ; National Institute Of Malarial Research.
Nirmal bhavan. Bengaluru -562 110.
Websites : www.sciencedirect.com
will be used to obtain related information regarding this research protocol.
7.2 PLAN OF WORK:
The whole study is divided as:
PLANT COLLECTION AND EXTRACTION:
The plants Nelumbo nucifera (Nelumbonaceae) and Tagetes erecta (Asteraceae) will be collected from wild source and will be authentified. They will be subjected for steam distillation by using Clevenger’s apparatus. The concentrate extract will be useful for our studies.
The larvicidal activity of medicinal plants will be determined against Aedes aegypti and Anopheles stephensi urban mosquito vector after making serial dilutions5. 2,1,0.5,0.25 and 0.125% in acetone. Later 1 ml of the dilution will be made up to 250 ml with distilled water to obtain a final concentration ranging between 200 and 6.25 ppm. Four replicates will be used in the bioassays against 3 instar larvae of the three species along with concurrent controls. Mortality will be recorded after 24 h and percent corrected mortality will be determined by Abotts formula. LC 50 and LC 90 will be calculated.
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATIONS OR
INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR
OTHER HUMANS OR ANIMALS?
– NO –
7.4 HAS ETHICAL CLEARENCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
1. Narayan DP, Purohit SS, Arun KS, A.. K. Handbook of Medicinal plants,a complete source book.
2. Kirithikar KR, BD B. Indian medicinal plants. Vol:1.
3. Pulok KM, Kakali S, Das J, Pal M, B.P. S. Studies on the Anti-Inflammatory Activity of Rhizomes of Nelumbo nucifera. Planta Med. 1997;63(4):367-9.
4. Yoshiki K, Akihiro A, Yasumasa I, Yuh P.C, Hiroshi F, Masataka I ea. Anti-HIV benzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera and structure activity correlations with related alkaloids. Bioorganic & Medicinal Chemistry. 2005 17;13(2):443–8.
5. Sujay R, Atul W, Kakali Mukherjee, Bishnu Pada Saha, Pulok KM. Antioxidant activity of Nelumbo nucifera (sacred lotus) seeds.
6. Yuka Onoa, Eri Hattoria, Yukitaka Fukayaa, Shoji Imaia, Ohizumi. Y. Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats. Journal of Ethnopharmacology. 2006 30;106 (2):238–44.
7. Mukherjee PK, Das J, Saha K, Giri SN, Pal M, BP. S. Antipyretic activity of Nelumbo nucifera rhizome extract. Indian J Exp Biol. 1996;34(3):275-6.
8. Santhoshkumar T, Rahuman AA, Rajakumar G, Marimuthu S, Bagavan A, Jayaseelan C ea. Synthesis of silver nanoparticles using Nelumbo nucifera leaf extract and its larvicidal activity against malaria andfilariasis vectors. Parasitol Res. 2011 10;108(3):693-702.
9. Pulok KM, Saroj KP, Kakali Saha, BP. S. Hypoglycemic activity of Nelumbo nucifera gaertn. (Fam. Nymphaeaceae) rhizome (methanolic extract) in streptozotocin-induced diabetic rats. Phytotherapy Research. 1995;9(7):522–4.
10. Hyun AJ, Yu JJ, Na YY, Da MJ, Hyun JB, Dong WK ea. Inhibitory effects of Nelumbo nucifera leaves on rat lens aldose reductase, advanced glycation endproducts formation, and oxidative stress. Food and Chemical Toxicology. 2008;46(12):3818–26.
11. Gurdip Singh, Om PS, M PDL, ANC. C. Studies on essential oils. Part 35: chemical and biocidal investigations onTagetes erecta leaf volatile oil. Flavour and Fragrance Journal. 2003;18(1):62-5.
12. Shi GL, Wang YN, Wang HL, Zhao LL, Liu SQ, Cao H ea. Effects of Tagetes erecta extracts on glutathione S-transferase and protease activities and protein content in Tetranychus viennensis. The Journal of Applied Ecology. 2007;18(2):400-4.
13. Calvet C, Pera J, JM. B. Growth response of marigold (Tagetes erecta L.) to inoculation withGlomus mosseae, Trichoderma aureovirideandPythium ultimum in a peat-perlite mixture. Plant and Soil. 1993;148(1):1-6.
14. Sarin. R. Insecticidal activity of callus culture of Tagetes erecta. Fitoterapia. 2004;75(1):62–4.
15. Barzana E, Rubio D, Santamaria RI, Garcia CO, Garcia F, Ridaura SVE ea. Enzyme-Mediated Solvent Extraction of Carotenoids from Marigold Flower (Tagetes erecta). J Agric Food Chem. 2002;50 (16):4491–6.
16. Samra Bashir, HG. A. Studies on the antioxidant and analgesic activities of Aztec marigold (Tagetes erecta) flowers. Phytotherapy Research. 2008;22(12):1692–4.
17. Elvira GM, Guadalupe LP, RG. M. Antimutagenicity of xanthophylls present in Aztec Marigold (Tagetes erecta) against 1-nitropyrene. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 1997 389(2-3):219–26.
18. Green MM, Singer JM, Sutherland DJ, CR. H. Larvicidal activity of Tagetes minuta (marigold) toward Aedes aegypti. J Am Mosq Control Assoc. 1991;7(2):282-6.
19. Singha SP, Pushpa Sharmab, LK. V. Light dependent toxicity of the extract of plantTagetes erecta and alpha terthienyl toward larvae of mosquito Culex tritaeniorhynchus. Toxicological & Environmental Chemistry. 1987;16(1):81-8.
20. Farjana Nikkon, Rowshanul HM, Rezaul KM, Ferdousi. Z. Insecticidal Activity of Flower of Tagetes erecta L. against Tribolium castaneum (Herbst). Research Journal of Agriculture and Biological Sciences. 2009;5(5):748-53.
21. Namrata PM, Singh K, Sagar OP, VDV. P. Larvicidal action of essential oils from plants against the vector mosquitoesAnopheles stephensi (Liston), Culex quinquefasciatus (Say) and Aedes aegypti (L.). International Pest Control. 2000;42(2):53-5.