Journal of Parasitic Diseases

, Volume 39, Issue 3, pp 385–392 | Cite as

RETRACTED ARTICLE: Mosquitocidal properties of Bacillus species isolated from mangroves of Vellar estuary, Southeast coast of India

  • S. BalakrishnanEmail author
  • K. Indira
  • M. Srinivasan
Original Article


Samples collected from the mangroves of Vellar estuary yielded a mosquitocidal bacterium, whose secondary metabolites exhibited mosquito larvicidal and pupicidal activity. The bacterium was isolated using standard microbiological methods and identified using classical biochemical tests. The mosquitocidal bacterium was identified as Bacillus subtilis, Bacillus thuringiensis, Bacillus sphaericus and Bacillus cereus. Mosquitocidal metabolite(s) was separated from the culture supernatant of the bacterium and its efficacy was against the larval and pupal stages of two different species of mosquitoes and determined in terms of LC50 and LC90. Mosquito larvicidal activity in terms of LC50 against Anopheleus stephensi and Aedes aegypti was 4.374 and 7.406 μl/ml and its pupicidal activity was 4.928 and 9.865 μl/ml, respectively. The present study proved that the mosquitocidal properties of the Bacillus species isolated from mangroves of Vellar estuary was evaluated as target species of mosquito vectors. This is an ideal eco-friendly approach for the vector control programs.


Bacillus species Culture supernatant Mosquito pupicidal Mangroves Vellar estuary 



We thank the authorities of Annamalai University for providing the necessary facilities and the first author thanks to the INCOIS-SATCORE Project (G4/515/2008), Ministry of Earth Sciences (Government of India) and second author thanks to the DST-PURSE Programme, Department of Science and Technology (Government of India) for financial support during the period of study. We also thank the anonymous referees for the valuable comments, which greatly improved our manuscript.


  1. Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18:265–267CrossRefGoogle Scholar
  2. Achs J, Malaney P (2002) The economic and social burden of malaria. Nature 15:680–685Google Scholar
  3. Ali A, Nayar JK, Xue RD (1995) Comparative toxicity of selected larvicides and insect growth regulators to a Florida laboratory, population of Aedes albopictus. J Am Mosq Control Assoc 11(1):72–76PubMedGoogle Scholar
  4. Armengol G, Hernandez J, Velez JG, Orduz S (2006) Long-lasting effects of a Bacillus thuringiensis serovar israelensis experimental tablet formulation for Aedes aegypti (Diptera: Culicidae) control. J Econ Entomol 99:1590–1595CrossRefGoogle Scholar
  5. Aronson AI, Beckman W, Dunn P (1986) Bacillus thuringiensis and related insect pathogens. Microbiol Rev 50:1–24PubMedPubMedCentralGoogle Scholar
  6. Balaraman K (1995) Mosquito control potential of Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus. ICMR Bull 25:45–51Google Scholar
  7. Beatty ME, Letson W, Edgil DM (2007) Estimating the total world population at risk for locally acquired dengue infection. In: Proceedings of 56th annual meeting of American society of tropical medicine and hygiene, Philadelphia, Pennsylvania, USA, pp 4–8Google Scholar
  8. Blackwood KS, Turenne CY, Harmsen D, Kabani AM (2004) Reassessment of sequence-based targets for identification of Bacillus species. J Clin Microbiol 42:1626–1630CrossRefGoogle Scholar
  9. Chandra G, Bhattacharjee I, Chatterjee SN (2008) Mosquito control by larvivorous fish. Indian J Med Res 127:13–27PubMedGoogle Scholar
  10. Chatterjee S, Subhra Ghosh T, Das S (2010) Virulence of Bacillus cereus as natural facultative pathogen of Anopheles subpictus Grassi (Diptera: Culicidae) larvae in submerged rice-fields and shallow ponds. Afr J Biotechnol 9(41):6983–6987Google Scholar
  11. Cooping LG, Menn JJ (2001) Biopesticides: a review of their action, applications and efficacy. Pest Manag Sci 56:651–676CrossRefGoogle Scholar
  12. Darriet F, Hougard JM (2002) An isolate of Bacillus circulans toxic to mosquito larvae. J Am Mosq Control Assoc 18:65–67PubMedGoogle Scholar
  13. Das PK, Amalraj DD (1997) Biological control of malarial vectors. Indian J Med 106:174–197Google Scholar
  14. Das K, Mukherjee AK (2006) Assessment of mosquito larvicidal potency of cyclic lipopeptides produced by Bacillus subtilis strains. Acta Trop 97:168–173CrossRefGoogle Scholar
  15. Das NG, Goswami D, Rabha B (2007) Preliminary evaluation of mosquito larvicidal efficacy of plant extracts. J Vector Borne Dis 44:145–148PubMedGoogle Scholar
  16. de Barjac H, Sebald M, Charles JF, Cheong WH, Lee HL (1990) Clostridium bifermentans serovar Malaysia, a new anaerobic bacterium pathogen to mosquito and blackfly larvae. CR Acad Sci III 310:383–387Google Scholar
  17. Federici BA, Park HW, Sakano Y (2006) Insecticidal protein crystals of Bacillus thuringiensis. In: Shively JM (ed) Microbiology monographs series, inclusions in prokaryotes, vol 1. Springer, Berlin, pp 195–236CrossRefGoogle Scholar
  18. Finney DJ (1971) Probit analysis. Cambridge University Press, CambridgeGoogle Scholar
  19. Geetha I, Manonmani AM (2008) Mosquito pupicidal toxin production by Bacillus subtilis subsp. subtilis. Biol Control 44(2):242–247CrossRefGoogle Scholar
  20. Geetha I, Prabakaran G, Paily KP, Manonmani AM, Balaraman K (2007) Characterisation of three mosquitocidal Bacillus strains isolated from mangrove forest. Biol Control 42:34–40CrossRefGoogle Scholar
  21. Geetha I, Manonmani AM, Paily KP (2010) Identification and characterization of a mosquito pupicidal metabolite of a Bacillus subtilis subsp. subtilis strain. Appl Microbiol Biotechnol 86(6):1737–1744CrossRefGoogle Scholar
  22. Georghiou GP, Wirth MC (1997) Influence of exposure to single versus multiple toxins of Bacillus thuringiensis subsp. israelensis on development of resistance in the mosquito Culex quinquefasciatus (Diptera: Culicidae). Appl Environ Microbiol 63:1095–1101PubMedPubMedCentralGoogle Scholar
  23. Gupta DK, Vyas M (1989) Efficacy of Bacillus subtilis against mosquito larvae Anopheles culicifacies. Zeitschrift fuer Angewandte Zoologie 76:85–91Google Scholar
  24. Khyami-Horani H, Katbeh-Bader A, Mohsen ZH (1999) Isolation of endospore forming bacilli toxic to Culiseta longiareolata (Diptera: Culicidae) in Jordan. Lett Appl Microbiol 128:57–60CrossRefGoogle Scholar
  25. Kovendan K, Murugan K, Vincent S, Kamalakannan S (2011) Larvicidal efficacy of Jatropha curcas and bacterial insecticide, Bacillus thuringiensis, against lymphatic filarial vector, Culex quinquefasciatus Say. (Diptera: Culicidae). Parasitol Res 109:1251–1257CrossRefGoogle Scholar
  26. Kovendan K, Murugan M, Vincent S, Barnard DR (2012) Studies on larvicidal and pupicidal activity of Leucas aspera Willd. (Lamiaceae) and bacterial insecticide, Bacillus sphaericus, against malarial vector, Anopheles stephensi Liston. (Diptera: Culicidae). Parasitol Res 110:195–203CrossRefGoogle Scholar
  27. Krattiger F (1997) Insect resistant crops: a case study of Bacillus thuringiensis (BT) and its transfer to developing countries. ISAAA Briefs 2:42Google Scholar
  28. Kumar A, Valecha N, Jain T, Dash AP (2007) Burden of malaria in India: retrospective and prospective view. Am J Trop Med Hyg 77:69–78CrossRefGoogle Scholar
  29. Lee YW, Zairi J (2005) Laboratory evaluation of Bacillus thuringiensis H-14 against Aedes aegypti. Trop Biomed 22:5–10PubMedGoogle Scholar
  30. Lingenfelser A, Rydzanicz K, Kaiser A (2010) Mosquito fauna and perspectives for integrated control of urban vector-mosquito populations in Southern Benin (West Africa). Ann Agric Environ Med 17(1):49–57PubMedGoogle Scholar
  31. Maeda M, Mizuki E, Hara M, Tanaka R, Akao T, Yamashita S, Ohba M (2001) Isolation of Bacillus thuringiensis from intertidal brackish sediments in mangroves. Microbiol Res 156:195–198CrossRefGoogle Scholar
  32. Mahesh Kumar P, Murugan K, Kovendan K, Subramaniam J, Amaresan D (2012) Mosquito larvicidal and pupicidal efficacy of Solanum xanthocarpum (Family: Solanaceae) leaf extract and bacterial insecticide, Bacillus thuringiensis, against Culex quinquefasciatus Say (Diptera: Culicidae). Parasitol Res. doi: 10.1007/s00436-011-2797-2 CrossRefGoogle Scholar
  33. Medeiros FP, Santos MA, Regis L, Rios EM, Rolim Neto PJ (2005) Development of a Bacillus sphaericus tablet formulation and its evaluation as a larvicide in the biological control of Culex quinquefasciatus. Mem Inst Oswaldo Cruz 100:431–434CrossRefGoogle Scholar
  34. Murugan K, Thangamathi P, Jeyabalan D (2002) Interactive effect of botanical and Bacillus thuringiensis subsp. israelensis on Culex quinquefasciatus Say. J Sci Ind Res 61:1068–1076Google Scholar
  35. Nielsen-Leroux C, Charles JF, Thiery I, Georghiou GP (1995) Resistance in a laboratory population of Culex quinquefasciatus (Diptera: Culicidae) to Bacillus sphaericus binary toxin is due to a change in the receptor on midgut brush-border membranes. Eur J Biochem 228:206–210CrossRefGoogle Scholar
  36. Ohba M, Wasano N, Mizuki E (2000) Bacillus thuringiensis soil populations naturally occurring in Ryukyus, a subtropic region of Japan. Microbiol Res 155:17–22CrossRefGoogle Scholar
  37. Pandey V, Agrawal V, Raghavendra K, Dash AP (2007) Strong larvicidal activity of three species of Spilanthes (Akarkara) against malaria (Anopheles stephensi Liston, Anopheles culicifacies, species C) and filaria vector (Culex quinquefasciatus Say). Parasitol Res 102:171–174CrossRefGoogle Scholar
  38. Park HW, Federici BA (2009) Genetic engineering of bacteria to improve efficacy using the insecticidal proteins of Bacillus species. In: Stock SP (ed) Insect pathogens: molecular approach and techniques. CABI International, Cambridge, pp 275–305CrossRefGoogle Scholar
  39. Park HW, Bideshi DK, Federici BA (2010) Properties and applied use of the mosquitocidal bacterium, Bacillus sphaericus. J Asia Pac Entomol 13:159–168CrossRefGoogle Scholar
  40. Plearnpis L, Haruyuki A, Sakol P (2001) Isolation of bacterial strains colonizable in mosquito larval guts as novel host cells for mosquito control. J Biosci Bioeng 92(4):342–345CrossRefGoogle Scholar
  41. Poopathi S, Tyagi BK (2006) The challenge of mosquito control strategies; from primordial to molecular approaches. Biotechnol Mol Biol Rev 1(2):51–65Google Scholar
  42. Poopathi S, Mani TR, Rao DR, Baskaran G, Kabilan L (1999) Cross-resistance to Bacillus sphaericus strains in Culex quinquefasciatus resistant to B. sphaericus 1593M. Southeast Asian J Trop Med Publ Health 30:477–481Google Scholar
  43. Rao DR, Mani TR, Rajendran R (1995) Development of a high level of resistance to Bacillus sphaericus in a field population of Culex quinquefasciatus from Kochi, India. J Am Mosq Control Assoc 11:1–5PubMedGoogle Scholar
  44. Rodriguez MM, Bisset J, de Fernandez DM, Lauzan L, Soca A (2001) Detection of insecticide resistance in A. aegypti (Diptera: Culicidae) from Cuba and Veninzula. J Med Entomol 38:623–628CrossRefGoogle Scholar
  45. Sharma SK, Upadhyay AK, Haque MA, Tyagi PK, Raghavendra K, Dash AP (2010) Wash-resistance and field evaluation of alphacypermethrin treated long-lasting insecticidal net (Interceptor) against malaria vectors Anopheles culicifacies and Anopheles fluviatilis in a tribal area of Orissa, India. Acta Trop 116(1):24–30CrossRefGoogle Scholar
  46. Shida O, Takagi H, Kadowaki K, Yano H, Komagata K (1996) Differentiation of species in the Bacillus brevis group and the Bacillus aneurinolyticus group based on the electrophoretic whole-cell protein pattern. Antonie Van Leeuwenhoek 70:31–39CrossRefGoogle Scholar
  47. Silva-Filha MH, Regis L, Nielson-LeRoux C (1995) Low-level resistance to Bacillus sphaericus in a field-treated population of Culex quinquefasciatus (Diptera: Culicidae). J Econ Entomol 88:525–530CrossRefGoogle Scholar
  48. Sneath PHA (1986) Endospore forming Gram positive rods and cocci. In: Sneath PHA, Mair N, Sharpe M, Holt J (eds) Bergey’s manual of systematic bacteriology, vol II. Williams Wilkins, Baltimore, pp 1104–1207Google Scholar
  49. Sourisseau M, Schilte C, Casartelli N (2007) Characterization of reemerging chikungunya virus. PLoS Pathog 3:89CrossRefGoogle Scholar
  50. Su T, Mulla MS (2004) Documentation of high-level Bacillus sphaericus 2362 resistance in field populations of Culex quinquefasciatus breeding in polluted water in Thailand. J Am Mosq Control Assoc 20:405–411PubMedGoogle Scholar
  51. Surendran A, Vennison SJ (2011) Occurrence and distribution of mosquitocidal Bacillus sphaericus in soil. Acad J Entomol 4(1):17–22Google Scholar
  52. Teng HJ, Lu LC, Wu YL (2005) Evaluation of various control agents against mosquito larvae in rice paddies in Taiwan. J Vector Ecol 30:126–132PubMedGoogle Scholar
  53. Tyrell DJ, Lee A, Bulla JR (1981) Characterization of spore coat proteins of Bacillus thuringiensis and Bacillus cereus. Comp Biochem Physiol Part B: Biochem Mol Biol 70(3):535–539CrossRefGoogle Scholar
  54. Wirth MC, Georghiou GP, Federici BA (1997) CytA enables CryIV endotoxins of Bacillus thuringiensis to overcome high levels of CryIV resistance in the mosquito, Culex quinquefasciatus. Proc Natl Acad Sci USA 94:10536–10540CrossRefGoogle Scholar
  55. Wirth MC, Delecluse A, Walton WE (2004) Laboratory selection for resistance to Bacillus thuringiensis subsp. Jegathesan or a component toxin, Cry 11B, in Culex quinquefasciatus (Diptera: Culicidae). J Med Entomol 41(3):435–441CrossRefGoogle Scholar
  56. Wirth MC, Zaritsky A, Ben-Dov E, Khasdan R, Boussiba S, Walton S (2007) Cross resistance spectra of Culex quinquefasciatus resistant to mosquitocidal toxins of Bacillus thuringiensis towards recombinant Escherichia coli expressing genes from B. thuringiensis spp. Israelensis. Environ Microbiol 9:1393–1401CrossRefGoogle Scholar
  57. World Health Organization (1992) Vector resistance to pesticides. Fifth report of the WHO expert committee on vector biology and control. Technical Report Series No. 818. Geneva, Switzerland, WHOGoogle Scholar
  58. World Health Organization (2009) Mosquito wars. Bull World Health Organ 87:167–168Google Scholar
  59. Yuan Z, Zhang YM, Cali Q, Liu EY (2000) High-level field resistance to Bacillus sphaericus C3-41 in Culex quinquefasciatus from southern China. Biocontrol Sci Technol 10:41–49CrossRefGoogle Scholar
  60. Zhang WJ, Jiang FB, Ou JF (2011) Global pesticide consumption and pollution: with China as a focus. Proc Int Acad Ecol Environ Sci 1(2):125–144Google Scholar

Copyright information

© Indian Society for Parasitology 2013

Authors and Affiliations

  1. 1.Centre of Advanced Study in Marine Biology, Faculty of Marine SciencesAnnamalai UniversityParangipettaiIndia

Personalised recommendations