Impact of Atrazine on Chlorpyrifos Toxicity in Four Aquatic Vertebrates



Atrazine has been shown previously to potentiate chlorpyrifos toxicity in selected invertebrates. This study examined interactions of atrazine and chlorpyrifos in four aquatic vertebrates. Organisms were exposed to binary mixtures of atrazine and chlorpyrifos during toxicity bioassays. Inhibition of cholinesterase (ChE) enzyme activity and chlorpyrifos uptake kinetics were also examined with and without atrazine exposure. Atrazine alone did not affect organisms at concentrations up to 5000 μg/L; however, the presence of atrazine at 1000 μg/L did result in a significant increase in the acute toxicity of chlorpyrifos in Xenopus laevis. Mixed results were encountered with Pimephales promelas; some bioassays showed greater than additive toxicity, while others showed an additive response. No effect of atrazine on chlorpyrifos toxicity was observed for Lepomis macrochirus and Rana clamitans. Atrazine did not affect ChE activity or chlorpyrifos uptake rates, indicating that these toxicodynamic and toxicokinetic parameters may not be related to the mechanism of atrazine potentiation of chlorpyrifos toxicity. Based on the results of this study, it does not appear that a mixture toxicity of atrazine and chlorpyrifos at environmentally relevant concentrations presents a risk to the vertebrate organisms examined in this study.


  1. Andersson T, Förlin L (1992) Regulation of the cytochrome P450 enzyme system in fish. Aquat Toxicol 24:1–20CrossRefGoogle Scholar
  2. Anderson T, Lydy M (2002) Increased toxicity to invertebrates associated with a mixture of atrazine and organophosphate insecticides. Environ Toxicol Chem 21:1507–1514CrossRefGoogle Scholar
  3. Belden J, Lydy M (2000) Impact of atrazine on organophosphate insecticide toxicity. Environ Toxicol Chem 19:2266–2274CrossRefGoogle Scholar
  4. Belden J, Lydy M (2001) Effects of atrazine on acetylcholinesterase activity in midges (Chironomus tentans) exposed to organophosphorus insecticides. Chemosphere 44:1685–1689CrossRefGoogle Scholar
  5. Belden J, Hofelt C, Lydy M (2000) Analysis of multiple pesticides in urban storm water using solid-phase extraction. Arch Environ Contam Toxicol 38:7–10CrossRefGoogle Scholar
  6. Beste C (1983) Herbicide handbook of the weed society of America. Weed Society of America, Champaign, IllinoisGoogle Scholar
  7. Biagianti-Risbourg S, Bastide J (1995) Hepatic perturbations induced by an herbicide (atrazine) in juvenile grey mullet Liza ramada (Mugilidae, Teleostei): An ultrastructural study. Aquat Toxicol 31:217–229CrossRefGoogle Scholar
  8. Blaustein A, Wake D, Sousa W (1994) Amphibian declines: judging stability, persistence, and susceptibility of populations to local and global extinctions. Conserv Biol 8:60–71CrossRefGoogle Scholar
  9. Denton D, Wheelock C, Murray S, Deanovic L, Hammock B, Hinton D (2003) Joint acute toxicity of esfenvalerate and diazinon to larval fathead minnows (Pimephales promelas). Environ Toxicol Chem 22:336CrossRefGoogle Scholar
  10. Doherty M, Khan M (1981) Hepatic microsomal mixed-function oxidase in the frog, Xenopus laevis. Comp Biochem Physiol 68: 221–228Google Scholar
  11. Ellman G, Courtney K, Andres V, Featherstone R (1961) A new and rapid coloremetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefGoogle Scholar
  12. Etheridge A, Richter S (1978) Xenopus laevis: Rearing and breeding the African clawed frog. Nasco, Fort Atkinson, WisconsinGoogle Scholar
  13. Gindi T, Knowland J (1979) The activity of cholinesterase during the development of Xenopus laevis. J Embryol Exp Morph 51:209–215Google Scholar
  14. Goksøyr A, Förlin L (1992) The cytochrome P450 system in fish, aquatic toxicology and environmental monitoring. Aquat Toxicol 22:287–312CrossRefGoogle Scholar
  15. Harris M, Bishop C, Struger J, Ripley B, Bogart J (1998) The functional integrity of northern leopard frog (Rana pipiens) and green frog (Rana clamitans) populations in orchard wetlands II. Effects of pesticides and eutrophic conditions on early life stage development. Environ Toxicol Chem 17:1351–1363CrossRefGoogle Scholar
  16. Harvey PW, DJ Everett (2003) The adrenal cortex and steroidogenesis as cellular and molecular targets for toxicity: Critical omissions from regulatory endocrine disruptor screening strategies for human health? J Appl Toxicol 23:81–87CrossRefGoogle Scholar
  17. Hayes T, Collins A, Lee M, Mendozza M, Noriega N, Stuart A, Vonk A (2002) Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses. Proc Natl Acad Sci USA 99:5476–5480CrossRefGoogle Scholar
  18. Jin-Clark Y, Lydy M, Zhu K (2002) Effects of atrazine and cyanazine on chlorpyrifos toxicity in Chironomus tentans (Diptera: Chironomidae). Environ Toxicol Chem 21:598–603CrossRefGoogle Scholar
  19. Kiely T, Donaldson D, Grube A (2004) Pesticide industry sales and usage: 2000 and 2001 market estimates. US Environmental Protection Agency, Washington, DCGoogle Scholar
  20. Kiesecker J (2002) Synergism between trematode infection and pesticide exposure: A link to amphibian limb deformities in nature? Proc Natl Acad Sci USA 99:9900–9904CrossRefGoogle Scholar
  21. Klaassen C (2001) Casarett and Doull’s Toxicology. McGraw-Hill, New York, New YorkGoogle Scholar
  22. Linder G, Krest S, Sparling D (2003) Amphibian Decline: An integrated analysis of multiple stressor effects. SETAC Press, Pensacola, FloridaGoogle Scholar
  23. Londoñdo D, Siegfried B, Lydy M (2004) Molecular characterization of atrazine induction of Cytochrome P450 in Chironomus tentans (Fabricius) (Diptera: Chironomidae). Chemosphere 56:701–706CrossRefGoogle Scholar
  24. Lower N, Moore A, Scott A, Ellis T, James K, Russell I (2005) A non-invasive method to assess the impacts of electronic tag insertion on stress levels in fish. J Fish Biol 67:1202–1212CrossRefGoogle Scholar
  25. Lydy M, Linck S (2003) Assessing the impact of triazine herbicides on organophosphate insecticide toxicity to the earthworm Eisenia fetida. Arch Environ Contam Toxicol 45:343–349CrossRefGoogle Scholar
  26. Lydy M, Lasater L, Landrum P (2000) Toxicokinetic of DDE and 2-Chlorobipphenyl in Chironomus tentans. Arch Environ Contam Toxicol 38:163–168CrossRefGoogle Scholar
  27. Miota F, Siegfried B, Scharf M, Lydy M (2000) Atrazine induction of cytochrome P450 in Chironomus tentans larvae. Chemosphere 40:285–291CrossRefGoogle Scholar
  28. Mukhopadhyay I, Nazir A, Saxena D, Kar Cowdhuri D (2003) Heat shock response: hsp70 in environmental monitoring. J Biochem Mol Toxicol 17:249–254CrossRefGoogle Scholar
  29. Nieuwkoop P, Faber J (1994) The stages of Xenopus embryonic development. Garland Publishing Inc, New York, New YorkGoogle Scholar
  30. Oleksiak M, Wu S, Parkers C, Karchner S, Stegemen J, Zeldin D (2000) Identification, functional characterization, and regulation of a new cytochrome P450 subfamily, the CYP2Ns. J Biol Chem 275:2312–2321CrossRefGoogle Scholar
  31. Orme S, Kegley S (2004) PAN Pesticide database. Pesticide Action Network, San Francisco, CaliforniaGoogle Scholar
  32. Oruç E, Üner N (2002) Marker enzyme assessment in the liver of Cyprinus carpio (L.) exposed to 2,4-D and azinphosmethyl. J Biochem Mol Toxicol 16:182–188CrossRefGoogle Scholar
  33. Pape-Lindstrom P, Lydy M (1997) Synergistic toxicity of atrazine and organophosphate insecticides contravenes the response addition mixture model. Environ Toxicol Chem 16:2415–2420CrossRefGoogle Scholar
  34. Perry C (1990) Source, extent, and degradation of herbicides in a shallow water aquifer near Hesston, Kansas. Water-Resources Investigations Report 91-4019. US Geological Survey, Lawrence, KansasGoogle Scholar
  35. Pruett SB, Fan R, Zheng Q, Myers L, Hèbert P (2003) Modeling and predicting immunological effects of chemical stressors: Characterization of a quantitative biomarker for immunological changes caused by atrazine and ethanol. Toxicol Sci 75:323–354CrossRefGoogle Scholar
  36. Richards S, Kendall R (2002) Biochemical effects of chlorpyrifos on two developmental stages of Xenopus laevis. Environ Toxicol Chem 21:1826–1835CrossRefGoogle Scholar
  37. Rocha-e-Silva T, Rossa M, Rantin F, Matsumura-Tundisi T, Tundisi J, Degterev I (2004) Comparison of liver mixed-function oxygenase and antioxidant enzymes in vertebrates. Comp Biochem Phys C 137:155–165Google Scholar
  38. Rosès N, M Poquet I Munoz. 1999. Behavioral and histological effects of atrazine on freshwater mollusks (Physa acuta Drap. and Ancylus fluviatilis Mull. Gastropoda). J Appl Toxicol 19:351–356CrossRefGoogle Scholar
  39. Saito H, Ohi H, Sugata E, Murayama N, Fujita Y, Higuchi S (1997) Purification and characterization of a cytochrome p450 from liver microsomes of Xenopus laevis. Arch Biochem Biophys 345:56–64CrossRefGoogle Scholar
  40. Schuler L, Trimble A, Belden J, Lydy J (2005) Joint toxicity of triazine herbicides and organophosphate insecticides to the midge Chironomus tentans. Arch Environ Contam Toxicol 49:173–177CrossRefGoogle Scholar
  41. Schuler L, Wheeler M, Bailer A, Lydy M (2003) Toxicokinetics of sediment-sorbed benzo[a]pyrene and hexachlorobiphenyl using the freshwater invertebrates Hyalella azteca, Chironomus tentans, and Lumbriculus variegatus. Environ Toxicol Chem 22:439–449CrossRefGoogle Scholar
  42. Selim H (2003) Retention and runoff losses of atrazine and metribuzin in soil. J Environ Qual 32:1058–1071CrossRefGoogle Scholar
  43. Solomon K, Baker D, Richards R, Dixon K, Klaine S, La Point T, Kendall R, Weisskopf C, Giddings J, Giesy J, Hall L, Williams W (1996) Ecological Risk assessment of atrazine in North American surface waters. Environ Toxicol Chem 15:31–76CrossRefGoogle Scholar
  44. Tanguy A, Boutet I, Laroche J, Moraga D (2005) Molecular identification and expression study of differentially regulated genes in the Pacific oyster Crassostrea gigas in response to pesticide exposure. FEBS 272:390–403CrossRefGoogle Scholar
  45. Thurman E, Goolsby D, Meyer M, Mills M, Pomes M, Kolpin D (1992) A reconnaissance study of herbicides and their metabolites in surface water in the Midwestern United States using immunoassay and gas chromatography / mass spectrometry. Environ Sci Technol 26:2440–2447CrossRefGoogle Scholar
  46. Trimble J, Lydy M (2006) Effects of triazine herbicides on organophosphate insecticide toxicity in Hyalella azteca. Arch Environ Contam Toxicol 51:29–34CrossRefGoogle Scholar
  47. US Environmental Protection Agency (1994) Short-term methods for estimating the chronic toxicity of effluents and receiving water to freshwater organisms. EPA-600-4-91-002. US Environmental Protection Agency, Washington, DCGoogle Scholar
  48. US Geological Survey (1998) Pesticides in surface and ground water in the United States: summary of results of the National Water Quality Assessment Program (NAWQA). US Geological Survey Pesticide National Synthesis Project, Sacramento, CaliforniaGoogle Scholar
  49. Wake D (1991) Declining amphibian populations. Science 253:860CrossRefGoogle Scholar
  50. Woolhouse H (1981) Aspects of the carbon and energy requirements of photosynthesis considered in relation to environmental constraints. In: Townsend CR, Calow P (eds) Physiological ecology. Sinauer Associates, Sunderland, Massachusetts, p 51–85Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  1. 1.Fisheries and Illinois Aquaculture Center and Department of ZoologySouthern Illinois UniversityCarbondaleUSA

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