Parasitology Research

, Volume 110, Issue 5, pp 1801–1813 | Cite as

A novel herbal formulation against dengue vector mosquitoes Aedes aegypti and Aedes albopictus

  • Rajan Maheswaran
  • Savarimuthu IgnacimuthuEmail author
Original Paper


The objective of this study was to develop a herbal formulation to control dengue vector mosquitoes. PONNEEM, a novel herbal formulation prepared using the oils of neem (Azadirachta indica), karanj (Pongamia glabra) and their extracts, was tested for larvicidal, ovicidal and oviposition deterrent activities against Aedes aegypti and Aedes albopictus at 1, 0.5, 0.3 and 0.1 ppm concentrations. Cent percent larvicidal and ovicidal activities were observed at 0.1 ppm in the two mosquito species under laboratory and sunlight-exposed conditions up to 12 months from the date of manufacture. Oviposition deterrent activity of 69.97% and 71.05% was observed at 1 ppm concentration of PONNEEM against A. aegypti and A. albopictus, respectively. Reduction in enzyme levels for α-esterase was 0.089 ± 0.008 and 0.099 ± 0.140 μg napthol produced/min/mg larval protein; for β-esterase, it was 0.004 ± 0.009 and 0.001 ± 0.028 μg napthol produced/min/mg larval protein; for glutathione S-transferase, it was 10.4814 ± 0.23 and 11.4811 ± 0.21 μmol/min/mg larval protein and for total protein, it was 0.177 ± 0.010 and 0.008 ± 0.005 mg/individual larva in treated groups of A. aegypti and A. albopictus, respectively. The nontarget organisms such as Gambusia affinis and Diplonychus indicus were not affected. No mortality was observed in control. PONNEEM can be used effectively for the management of human vector mosquitoes.


Azadirachtin Larvicidal Activity Herbal Formulation Ovicidal Activity Oviposition Deterrent 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors are thankful to the Entomology Research Institute, Loyola College for the financial support.


  1. Abbot WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18:265–267Google Scholar
  2. Adebayo TA, Gbolade AA, Olaifa JI (1999) Comparative study of toxicity of essential oils to larvae of three mosquito species. Nigerian J Nat Prod Med 3:74–76Google Scholar
  3. Ascher KRS (1993) Nonconventional insecticidal effects of pesticides available from the neem tree, Azadirachta indica. Arch Insect Biochem Physiol 22:433–449CrossRefGoogle Scholar
  4. Batabyal L, Sharma P, Mohan L, Maurya P, Srivastava CN (2009) Relative toxicity of neem fruit, bitter gourd, and castor seed extracts against the larvae of filaria vector, Culex quinquefasciatus (Say). Parasitol Res 105(5):1205–1210PubMedCrossRefGoogle Scholar
  5. Chaithong U, Choochote W, Kamsuk K, Jitpakdi A, Tippawangkosol P, Chaiyasit D, Champakaew D, Tuetun B, Pitasawat B (2006) Larvicidal effect of pepper plants on Aedes aegypti (L.) (Diptera: Culicidae). J Vector Ecol 31:138–144PubMedCrossRefGoogle Scholar
  6. Choochote W, Tuetun B, Kanjanapothi D, Rattanachanpichai E, Chaithong U, Chaiwong P, Jitpakdi A, Tippawangkosol P, Riyong D, Pitasawat B (2004) Potential of crude seed extract of celery, Apium graveolens L., against the mosquito Aedes aegypti (L.) (Diptera: Culicidae). J Vector Ecol 29:340–346PubMedGoogle Scholar
  7. Corbet SA, Danahar CW, King V, Chalmers CL, Tiley CF (1995) Surfactant-enhanced essential oils as mosquito larvicides. Entomol Exp Appl 75:229–236CrossRefGoogle Scholar
  8. DeBach P, Rosen D (1991) Biological control by natural enemies, 2nd edn. Cambridge University Press, CambridgeGoogle Scholar
  9. Deo PG, Hasan SB, Majumdar SK (1988) Toxicity and suitability of some insecticides for household use. Int Pest Control 30:118–129Google Scholar
  10. Dondji B, Duchan S, Diabate A, Herve JP, Corbel V, Hougard JM, Santus R, Schrevel J (2005) Assessment of laboratory and field assays of sunlight induced killing of mosquito larvae by photo sensitizers. J Med Entomol 42:652–656PubMedCrossRefGoogle Scholar
  11. Figueiredo LTM, Fonseca BAL (1996) Dengue. In: Veronesi R, Focaccio R (eds) Tratado de Infectologia. Editora Atheneu, São Paulo, pp 201–214Google Scholar
  12. Ganesh KN, Urmila J, Vijayan VA (2003) Pyrethroid susceptibility and enzyme activity in two malaria vectors, Anopheles stephensi (Liston) and Anopheles culicifacies (Giles) from Mysore, India. Indian J Med Res 117:30–38PubMedGoogle Scholar
  13. Garcez WS, Garcez FR, da Silva LMGE, Hamerski L (2009) Larvicidal activity against Aedes aegypti of some plants native to the West-Central region of Brazil. Biores Technol 100:6647–6650CrossRefGoogle Scholar
  14. Gbolade AA, Oyedele AO, Sosan MB, Adewayin FB, Soyela OL (2000) Mosquito repellent activities of essential oils from two Nigerian Ocimum species. J Trop Med Plants 1:146–148Google Scholar
  15. Ghani A (1998) Medicinal plants of Bangladesh. Asiatic Society of Bangladesh, Dhaka, p 270Google Scholar
  16. Govindarajan M, Mathivanan T, Elumalai K, Krishnappa K, Anandan A (2011) Mosquito larvicidal, ovicidal, and repellent properties of botanical extracts against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). Parasitol Res 109(2):353–367PubMedCrossRefGoogle Scholar
  17. Hafeez F, Akram W, Shaalan EA (2011) Mosquito larvicidal activity of citrus limonoids against Aedes albopictus. Parasitol Res 109(1):221–229PubMedCrossRefGoogle Scholar
  18. Halstead SB (2007) Dengue. Lancet 370:1644–1652PubMedCrossRefGoogle Scholar
  19. Hayes JB Jr, Laws ER Jr (1991) Handbook of pesticide toxicology, vol I. Academic Press, San DiegoGoogle Scholar
  20. Islam ME, Khatune NA, Wahed MII, Haque ME, Mosaddik MA (2003) Larvicidal activity of new glycoside, phenyl ethyl β-d glucopyranoside from the stem of the plant Sida rhombifolia. Pak J Biol Sci 6:73–77CrossRefGoogle Scholar
  21. Isman MB (2000) Plant essential oils for pest and disease management. Crop Prot 19:603–608CrossRefGoogle Scholar
  22. Kady GAE, Kamel NH, Mosleh YY, Bahght IM (2008) Comparative toxicity of two bio-insecticides (Spinotoram and Vertemic) compared with methomyl against Culex pipiens and Anopheles multicolor. World J Agri Sci 4:198–205Google Scholar
  23. Kalaivani K, Senthil-Nathan S, Murugesan AG (2011). Biological activity of selected Lamiaceae and Zingiberaceae plant essential oils against the dengue vector Aedes aegypti L. (Diptera: Culicidae). Parasitol Res. doi: 10.1007/s00436-011-2623-x
  24. Kanis LA, Prophiro JS, da Silva Vieira E, do Nascimento MP, Zepon KM, Kulkamp-Guerreiro IC, da Silva OS (2011). Larvicidal activity of Copaifera sp. (Leguminosae) oleoresin microcapsules against Aedes aegypti (Diptera: Culicidae) larvae. Parasitol Res. doi: 10.1007/s00436-011-2610-2
  25. Kirtikar KR, Basu BD (1994) Indian medicinal plants, 2nd edn. Dehradun Publisher Ltd., India, pp 830–832Google Scholar
  26. Locantoni L, Guisti F, Cristofaro M, Pasqualini L, Esposito F, Lupetti P, Habluetzel A (2006) Effect of neem extract on blood feeding oviposition and oocyte ultra structure in Anopheles stephensi Liston (Diptera: Culicidae). Tissue Cell 38:361–371CrossRefGoogle Scholar
  27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  28. Mohsen ZH, Jawad ALM, Al-Saadi M, Al-Naib A (1995) Anti-oviposition and insecticidal activity of Imperata cylindric (Gramineae). Med Vet Entomol 9:441–442PubMedCrossRefGoogle Scholar
  29. Mouches C, Magnin M, Berge JB, Silvestri M, Beyssatt V, Pasteur N, Georghiou GP (1987) Overproduction of detoxifying esterases in organophosphate resistant Culex mosquitoes and their presence in other insects. Proc Natl Acad Sci U S A 84(8):2113–2116PubMedCrossRefGoogle Scholar
  30. National Research Council (1992) Neem: a tree for solving global problems. Report of an Adhoc Panel of the Board on Science and Technology for International Development National Academy Press, WashingtonGoogle Scholar
  31. Nicoletti M, Serafini M, Aliboni A, D'Andrea A, Mariani S (2010) Toxic effects of neem cake extracts on Aedes albopictus (Skuse) larvae. Parasitol Res 107(1):89–94PubMedCrossRefGoogle Scholar
  32. Nishimura H (2001) Aroma constituents in plants and their repellent activities against mosquitoes. Aroma Research 2:257–267Google Scholar
  33. Parmar BS, Dutta S (1987) Evaluation of some oils as malathion synergists. Int J Trop Agric 5:223–226Google Scholar
  34. Prophiro JS, da Silva MA, Kanis LA, da Rocha LC, Duque-Luna JE, da Silva OS (2011) First report on susceptibility of wild Aedes aegypti (Diptera: Culicidae) using Carapa guianensis (Meliaceae) and Copaifera sp. (Leguminosae). Parasitol Res (in press)Google Scholar
  35. Rajkumar S, Jenanesan A (2002) Oviposition attractancy of Solunum aeriathum D. Don. leaf extract for Culex quinquefasciatus Say. J Exp Zoology India 5:221–224Google Scholar
  36. Rajkumar S, Jebanesan A (2008) Bioactivity of flavonoid compounds from Poncirus trifoliata L. (Family: Rutaceae) against the dengue vector, Aedes aegypti L. (Diptera: Culicidae). Parasitol Res 104(1):19–25PubMedCrossRefGoogle Scholar
  37. Rao GR, Dhingra S (1997) Synergistic activity of some vegetable oils in mixed formulation with cypermethrin against instars of Spodoptera litura (Fabricius). J Entomol Res 21:153–160Google Scholar
  38. Rozendaal JA (1997) Vector control Geneva. World Health Organization, SwitzerlandGoogle Scholar
  39. Schmutterer H (1990) Properties of natural pesticides from the neem tree, Azadirachta indica. Ann Rev Entomol 35:271–297CrossRefGoogle Scholar
  40. Schmutterer H (2002) The neem tree (Azadirachta indica) and other Meliceous plants. In Source of unique natural products for integrated pest management, medicine, industry and other porposes, 1st edn. Neem Foundation, MumbaiGoogle Scholar
  41. Shanmugasundaram R, Dutt MS, Arivazhagan A, Gayathri V, Jeyalakshmi T, Murthy PB (2001) Comparative study on the larvicidal property of various products against the larvae of Culex quinquefasciatus (Culicidae: Diptera). Pestology 25:33–35Google Scholar
  42. Shanmugasundaram R, Jeyalakshmi T, Dutt MS, Murthy PB (2008) Larvicidal activity of neem and karanja oil cakes against mosquito vectors, Culex quinquefasciatus (Say), Aedes aegypti (L.) and Anopheles stephensi (L.). J Environ Biol 29(1):43–45PubMedGoogle Scholar
  43. Sharma VP, Dhiman RC (1993) Neem oil as a sand fly (Diptera: Psychodidae) repellent. J Am Mosq Control Assoc 9:364–366PubMedGoogle Scholar
  44. Sivagnaname N, Kalyanasundaram M (2004) Laboratory evaluation of methanolic extract of Atlantia monophylla (Family: Rutaceae) against immature stages of mosquitoes and non-target organisms. Mem Inst Oswaldo Cruz 99:115–118PubMedCrossRefGoogle Scholar
  45. Su T, Mulla MS (1998a) Antifeedancy of neem products containing Azadirachtin against Culex tarsalis and Culex quinquefasciatus (Diptera: Culicidae). J Vector Ecol 23:114–122PubMedGoogle Scholar
  46. Su T, Mulla MS (1998b) Ovicidal activity of neem products (Azadirachtin) against Culex tarsalis and Culex quinquefasciatus (Diptera: Cilicidae). J Am Mosq Control Assoc 14:204–209PubMedGoogle Scholar
  47. Templeton W (1969) An introduction to the chemistry of terpenoids and steroids. Butterworths, LondonGoogle Scholar
  48. Tripathi AK, Prajapati V, Agarwal KK, Khanuja SPS, Kumar S (2000) Repellency and toxicity of oil from Artemisia annua to certain stored product beetles. J Econ Entomol 93:43–47PubMedCrossRefGoogle Scholar
  49. Tripathi AK, Prajapati V, Verma N, Bahl JR, Bansal RP, Khanuja SPS (2002) Bioactivities of the leaf essential oil of Curcuma longa (var. ch-66) on three species of stored-product beetles (Coleoptera). J Econ Entomol 95:183–189PubMedCrossRefGoogle Scholar
  50. Tsao R, Lee S, Rice PJ, Jensen C, Coats JR (1995) Monoterpenoids and their synthetic derivatives as leads for new insect control agents. In: Baker DR, Fenyes JG, Basarab GS (eds) Synthesis and chemistry of agrochemicals IV. American Chemical Society, Washington, pp 312–324CrossRefGoogle Scholar
  51. Warrier PK, Nambiar VPK, Ramankutty C (1995) Indian medicinal plants: a compendium of 500 species, vol 4. Universities Press, Mumbai, IndiaGoogle Scholar
  52. World Health Organization (WHO) website (2008a) Report on global surveillance of epidemic-prone infectious diseases. Dengue and dengue haemorrhagic fever (chapter 6). Accessed May 2008
  53. World Health Organization (WHO) website (2008b) Dengue and dengue haemorrhagic fever. Accessed Mar 2009
  54. World Health Organization (1996) Report of the WHO informal consultation on the “Evaluation and testing of insecticides”. CTD/WHOPES/IC/96. pp. 1–69.Google Scholar
  55. Zebitz CPW (1984) Effects of some crude and azadirachtin enrichd neem Azadirachta indica seed kernel extracts on larvae of Aedes aegypti. Entomol Exp Appl 35:11–16CrossRefGoogle Scholar
  56. Zebitz CPW (1986) Potential of neem seed kernel extracts in mosquito control. Proc. 3rd Int Neem Conf Nairobi, pp.555-573Google Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Entomology Research InstituteLoyola CollegeChennaiIndia

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