Chlorpyrifos-Induced Toxicity in Duttaphrynus melanostictus (Schneider 1799) Larvae

  • M. R. WijesingheEmail author
  • M. G. D. K. Bandara
  • W. D. Ratnasooriya
  • G. P. Lakraj


This study investigates the effects of continuous exposure to a widely used organophosphate pesticide, chlorpyrifos, on survival, growth, development, and activity of larvae of the Asian common toad, Duttaphrynus melanostictus Schneider 1799. Larvae were continuously exposed to six different concentrations (1–1,500 μg l−1) of commercial-grade chlorpyrifos for 14 days and monitored for 1 additional week. Chlorpyrifos at ≥1,000 μg l−1 caused significantly high and dose-dependent mortality, and the weekly LC507 day–21 day values ranged from 3,003 to 462 μg l−1. Larvae surviving exposure to ≥500 μg l−1 chlorpyrifos showed significant growth impairment, delays in metamorphosis, and decreased swimming activity. Tail abnormalities were the most common morphologic deformity at concentrations of 1,000 and 1,500 μg l−1 chlorpyrifos. The findings of the present study highlight the need to recognize the potential risk that agrochemicals pose to amphibians inhabiting agricultural landscapes in Sri Lanka and other Asian countries.


Chlorpyrifos Pesticide Concentration Dissolve Oxygen Level Mortality Level Organophosphate Pesticide 
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.



We are grateful to the Department of Zoology, University of Colombo, for providing funds and necessary facilities to carry out this project and for the technical assistance provided by A. de Silva, J. R. A. C. Jayakody, and S. Abeysinghe of the Department of Zoology, University of Colombo.


  1. Agency for Toxic Substances, Disease Registry (1997) Toxicological profile for chlorpyrifos. ASTDR, Atlanta, GAGoogle Scholar
  2. Bandara DK, Wijesinghe MR, Ratnasooriya WD, Priyani AAH (2008) Chlorpyrifos induced histopathological changes in larvae of Duttaphrynus melanostictus. 28th Annual Sessions of the Institute of Biology, Sri Lanka, held in September in Colombo. p 41 Google Scholar
  3. Bonfanti P, Colombo A, Orsi F, Nizzetto I, Andrioletti M, Bacchetta R et al (2004) Comparative teratogenicity of chlorpyrifos and malathion on Xenopus laevis development. Aquat Toxicol 70:189–200CrossRefGoogle Scholar
  4. Bridges CM (2000) Long-term effects of pesticide exposure at various life stages of the Southern leopard frog (Rana sphenocephala). Arch Environ Contam Toxicol 39:91–96CrossRefGoogle Scholar
  5. Cowman DF, Manzanti LE (2000) Ecotoxicology of “new generation” pesticides to amphibians. In: Sparling D, Linder G, Bishop C (eds) Ecotoxicology of amphibians and reptiles. SETAC Tech. Pub. Series, p 233–268Google Scholar
  6. Cremlyn R (1991) Synthetic insecticides II organophosphorous and carbamate compounds. Agrichemical: Preparation and Mode of Action. John Wiley and Sons, Chichester, UKGoogle Scholar
  7. Crossland MR (1998) Predation by tadpoles on toxic toad eggs: the effect of tadpole size on predation success and tadpole survival. J Herpetol 32:443–446CrossRefGoogle Scholar
  8. Food and Agriculture Organization/World Health Organization (1999) Pesticide residues in food. Toxicological evaluations. International Programme on Chemical Safety. Joint FAO/WHO Meeting on Pesticide Residues (JMPR) Publications, Rome, pp 1–61Google Scholar
  9. Forester DC, Czarnowsky R (1985) Sexual selection in the springpeeper, Hyla crucifer (Amphibia: Anura): role of the advertisement call. Behaviour 92:112–128CrossRefGoogle Scholar
  10. Gallo-Delgado SM, Palacio-Baena JA, Gutierrer-C PDA (2006) Effects of insecticide chlorpyrifos on growth rate and metamorphosis of Smilisca phaeota (Cope, 1862) (Anura: Hylidae). Actual Biol 28:51–58Google Scholar
  11. Global amphibian assessment (2004) IUCN, Conservation International, Nature Serve, Washington DC. Available at: Accessed 23 July 2010
  12. Gosner KL (1960) A simplified table for staging anuran embryos and larvae with notes on identification. Herpetol Sci 16:183–190Google Scholar
  13. Howard J, Julian S, Ferrigan J (2002) Golf course design and maintenance: impacts on amphibians. USGA Turfgrass Environ Res Online 6:0277–1541Google Scholar
  14. International Union for Conservation of Nature (2007) The 2007 red list of threatened fauna and flora of Sri Lanka. International Union for Conservation of Nature, Colombo, Sri LankaGoogle Scholar
  15. Kerby JL, Richards-Hrdlicka KL, Storfer A, Skelly DK (2010) An examination of amphibian sensitivity to environmental contaminants: are amphibians poor canaries? Ecol Lett 13:60–67CrossRefGoogle Scholar
  16. Kirtisinghe P (1955) The amphibians of Ceylon, 1st edn. Charles Circus, Colombo, Sri LankaGoogle Scholar
  17. Manamendra-Arachchi K, Pethiyagoda R (2006) Sri Lankawe ubhaya jeeveen (Amphibians of Sri Lanka). WHT Publications (Pvt) Ltd, Colombo, Sri LankaGoogle Scholar
  18. Marian MP, Ami IV, Pandian TJ (1983) Acute and chronic effects of carbaryl on survival, growth, and metamorphosis in the bullfrog (Rana tigrina). Arch Environ Contam Toxicol 12:271–275CrossRefGoogle Scholar
  19. Mathew M, Andrews MI (2001) Effect of hinosan on the hatching rate and larval survival of Duttaphrynus Melanostictus Schneider. Pollut Res 20:31–34Google Scholar
  20. Miles JRW, Harris CR, Tu CM (1983) Influence of temperature on the persistence of chlorpyrifos and chlorfenvinphos in sterile and natural mineral and organic soils. J Environ Sci Health B 18:705–712CrossRefGoogle Scholar
  21. National Registration Authority review of chlorpyrifos (2000) Volume 1, NRA Review Series 00.5. National Registration Authority for Agricultural and Veterinary Chemicals, Canberra, AustraliaGoogle Scholar
  22. Ortiz ME, Macro A, Fernandez MJ, Lizana M (2004) Influence of age on amphibian sensitivity to fertilizers. Environ Toxicol Chem 12:345–349Google Scholar
  23. Ozmen M, Sener S, Mete A, Kucukbay H (1999) In vitro and in vivo acetylcholinesterase-inhibiting effect of new classes of organophosphate compounds. Environ Toxicol Chem 18:241–246CrossRefGoogle Scholar
  24. Pethiyagoda R, Manamendra-Arachchi K (1998) Evaluating Sri Lanka’s amphibian diversity, Occasional papers, No. 2. The Wildlife Heritage Trust of Sri LankaGoogle Scholar
  25. Pethiyagoda R, Manamendra-Arachchi K, Bahir MM, Meegaskumbura M (2006) Sri Lankan amphibians: diversity, uniqueness and conservation. In: Bambaradeniya CNB (ed) The fauna of Sri Lanka. Status of taxonomy, research and conservation. The World Conservation Union, Colombo, Sri Lanka, pp 216–231Google Scholar
  26. Ragnarsdottir K (2000) Environmental fate and toxicology of organophosphate pesticides. J Geol Soc Lond 157:859–876CrossRefGoogle Scholar
  27. Relyea RA (2009) A cocktail of contaminants: how mixtures of pesticides at low concentrations affect aquatic communities. Oecologia 159:363–376CrossRefGoogle Scholar
  28. Relyea RA, Jones DK (2009) The toxicity of Roundup Original Max to 13 species of larval amphibians. Environ Toxicol Chem 28:2004–2008CrossRefGoogle Scholar
  29. Rouse JD, Bishop CA, Struger J (1999) Nitrogen pollution: an assessment of its threat to amphibian survival. Environ Health Perspect 107:799–803CrossRefGoogle Scholar
  30. Rowe CL, Kinney OM, Nagle RD, Congdon JC (1998) Elevated maintenance costs in an anura exposed to a mixture of trace elements during the embryonic and early larval periods. Physiol Zool 71:27–35CrossRefGoogle Scholar
  31. Sayim F (2008) Acute toxic effects of malathion on the 21 stage larvae of the marsh frog. Turk J Zool 32:99–106Google Scholar
  32. Semlitsch RD, Scott DE, Pechmann JHK (1988) Time and size at metamorphosis related to adult fitness in Ambystoma talpoideum. Ecology 69:184–192CrossRefGoogle Scholar
  33. Sheffield SR (2000) Lethal and sub lethal effects of the OP insecticide chlorpyrifos in larval amphibians. Toxicology poster session, SETAC, PWA062Google Scholar
  34. Smith DC (1987) Adult recruitment in chorus frogs: effects of size and date at metamorphosis. Ecology 68:344–350CrossRefGoogle Scholar
  35. Sparling DW, Fellers GM (2007) Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii. Environ Pollut 147:535–539CrossRefGoogle Scholar
  36. Sparling DW, Fellers GM (2009) Toxicity of two insecticides to California, USA, amphibians and its relevance to declining amphibian populations. Environ Toxicol Chem 28:1696–1703CrossRefGoogle Scholar
  37. Sparling DW, Fellers GM, McConnell LL (2001) Pesticides and amphibian population declines in California, USA. Environ Toxicol Chem 20:1591–1595CrossRefGoogle Scholar
  38. Stuart SN, Chanson JS, Cox NA, Young BE, Rodrigues ASL, Fischman DL et al (2004) Status and trends of amphibian declines and extinctions worldwide. Science 306:1783–1786CrossRefGoogle Scholar
  39. Sumanadasa DM, Wijesinghe MR, Ratnasooriya WD (2008a) Effects of diazinon on larvae of the Asian common toad (Duttaphrynus melanostictus Schneider 1799). Environ Toxicol Chem 27:157–162CrossRefGoogle Scholar
  40. Sumanadasa DM, Wijesinghe MR, Ratnasooriya WD (2008b) Effects of diazinon on survival, growth rate of two amphibian larvae. J Natl Sci Found Sri Lanka 36:165–169Google Scholar
  41. United States Environmental Protection Agency (2002) Interim reregistration eligibility decision for chlorpyrifos. Prevention, pesticides and toxic substances. (7508C). EPAR-01-00. Available at: Accessed Feb 2002
  42. Wijesinghe MR, Kariyawasama G, Ratnasooriya WD (2006) The effect of gramaxone (main ingredient paraquat) on the growth and survival of larvae of the endemic frog, Polypedates crucuger. In: Proceedings of the 62nd Annual Scientific Sessions of the Sri Lanka Association for the Advancement of Science, Sri Lanka, p 72Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • M. R. Wijesinghe
    • 1
    Email author
  • M. G. D. K. Bandara
    • 1
  • W. D. Ratnasooriya
    • 1
  • G. P. Lakraj
    • 2
  1. 1.Department of ZoologyUniversity of ColomboColombo 03Sri Lanka
  2. 2.Department of StatisticsUniversity of ColomboColombo 03Sri Lanka

Personalised recommendations