Larval Responses of Three Midwestern Anurans to Chronic, Low-Dose Exposures of Four Herbicides

  • Bethany K. WilliamsEmail author
  • Raymond D. Semlitsch


Low levels of agricultural herbicides often contaminate surface water and might persist throughout the growing season, potentially acting as stressors on aquatic organisms. Although low-dose, chronic exposures to agrochemicals are likely common for many nontarget organisms, studies addressing these effects using end-use herbicide formulations are rare. We exposed three common species of tadpoles to conservative levels of atrazine, S-metolachlor, and glyphosate end-use herbicide formulations throughout the larval period to test for survival differences or life-history trait alterations. Exposure to the glyphosate product Roundup WeatherMax® at 572 ppb glyphosate acid equivalents (a.e.) resulted in 80% mortality of western chorus frog tadpoles, likely as a result of a unique surfactant formulation. Exposure to WeatherMax® or Roundup Original Max® at 572 ppb a.e. also lengthened the larval period for American toads. Chronic atrazine and S-metolachlor exposures induced no significant negative effects on survival, mass at metamorphosis, or larval period length at the levels tested. These results highlight the importance of explicitly tying chronic tests to the natural environment and considering contributions of surfactant/adjuvant components to end-use formulation toxicities, even between very similar products.


Atrazine Glyphosate Larval Period Metolachlor American Toad 
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.


  1. Battaglin WA, Rice KC, Focazio MJ, Salmons S, Barry RX (2009) The occurrence of glyphosate, atrazine, and other pesticides in vernal pools and adjacent streams in Washington, DC, Maryland, Iowa, and Wyoming, 2005–2006. Environ Monit Assess 155:281–307CrossRefGoogle Scholar
  2. Boone MD, James SM (2003) Interactions of an insecticide, herbicide, and natural stressors in amphibian community mesocosms. Ecol Appl 13:829–841CrossRefGoogle Scholar
  3. Boone MD, James SM (2005) Aquatic and terrestrial mesocosms in amphibian ecotoxicology. Appl Herpetol 2:231–257CrossRefGoogle 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. Carlisle SM, Trevors JT (1988) Glyphosate in the environment. Water Air Soil Pollut 39:409–420Google Scholar
  6. Cauble K, Wagner RS (2005) Sublethal effects of the herbicide glyphosate on amphibian metamorphosis and development. Bull Environ Contam Toxicol 75:429–435CrossRefGoogle Scholar
  7. Chen CY, Hathaway KM, Folt CL (2004) Multiple stress effects of Vision® herbicide, pH, and food on zooplankton and larval amphibian species from forest wetlands. Environ Toxicol Chem 23:823–831CrossRefGoogle Scholar
  8. Cook ME, Moore PA (2008) The effects of the herbicide metolachlor on agonistic behavior in the crayfish, Orconectes rusticus. Arch Environ Contam Toxicol 55:94–102CrossRefGoogle Scholar
  9. DeGarady CJ, Halbrook RS (2003) Impacts from PCB accumulation on amphibians inhabiting streams flowing from the Paducah Gaseous Diffusion Plant. Arch Environ Contam Toxicol 45:525–532CrossRefGoogle Scholar
  10. Donald WW, Hjelmfelt AT Jr, Alberts EE (1998) Herbicide distribution and variability across Goodwater Creek watershed in north central Missouri. J Environ Qual 27:999–1009Google Scholar
  11. Edginton AN, Sheridan PM, Stephenson GR, Thompson DG, Boermans HJ (2004) Comparative effects of pH and Vision herbicide on two life stages of four anuran amphibian species. Environ Toxicol Chem 23:815–822CrossRefGoogle Scholar
  12. Eggen R, Behra R, Burkhardt-Holm P, Escher BI, Schweigert N (2004) Challenges in ecotoxicology. Environ Sci Technol A 38:58A–64ACrossRefGoogle Scholar
  13. Fioramonti E, Semlitsch RD, Reyer HU, Fent K (1997) Effects of triphenyltin and pH on the growth and development of Rana lessonae and Rana esculenta tadpoles. Environ Toxicol Chem 16:1940–1947Google Scholar
  14. Gianessi LP, Reigner N (2006) Pesticide use in U.S. crop protection: 2002. Crop Life Foundation, Washington, DCGoogle Scholar
  15. Giesy JP, Dobson S, Solomon KR (2000) Ecotoxicological risk assessment for Roundup herbicide. Rev Environ Contam Toxicol 167:35–120Google Scholar
  16. Gilliom RJ, Barbash JE, Crawford CG, Hamilton PA, Martin JD, Nakagaki N, Nowell LH, Scott JC, Stackelberg PE, Thelin GP, Wolock DM (2006) The quality of our nation’s waters—pesticides in the nation’s streams and ground water, 1992–2001: US Geological Survey Circular 1291Google Scholar
  17. Gosner N (1960) A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 16:183–190Google Scholar
  18. Hayes TB, Case P, Chui S, Chung D, Haeffele C, Haston K, Lee M, Mai VP, Marjuoa Y, Parker J, Tsui M (2006) Pesticide mixtures, endocrine disruption, and amphibian declines: are we underestimating the impact? Environ Health Perspect 114(S-1):40–50CrossRefGoogle Scholar
  19. Hoffmann RS, Capel PD, Larson SJ (2000) Comparison of pesticides in eight U.S. urban streams. Environ Toxicol Chem 19:2249–2258CrossRefGoogle Scholar
  20. Howe CM, Berril M, Pauli BD, Helbing CC, Werry K, Veldhoen N (2004) Toxicity of glyphosate-based pesticides to four North American frog species. Environ Toxicol Chem 23:1928–1938CrossRefGoogle Scholar
  21. Howe GE, Gillis R, Mowbray RC (1998) Effect of chemical synergy and larval stage on the toxicity of atrazine and alachlor to amphibian larvae. Environ Toxicol Chem 17:519–525CrossRefGoogle Scholar
  22. Huang YW, Niyogi DK, Nam PK, Bandeff JM (2009) Livestock hormones in aquatic ecosystems. In: Coats J, Henderson K (eds) Veterinary pharmaceuticals in the environment. American Chemical Society (in press)Google Scholar
  23. Kolpin DW, Furlong ET, Meyer MT, Thurman EM, Zaugg SD, Barber LB, Buxton HT (2002) Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000: a national reconnaissance. Environ Sci Technol 36:1202–1211CrossRefGoogle Scholar
  24. Kolpin DW, Thurman EM, Lee EA, Meyer MT, Furlong ET, Glassmeyer ST (2006) Urban contributions of glyphosate and its degradate AMPA to streams in the United States. Sci Total Environ 354:191–197CrossRefGoogle Scholar
  25. Lerch RN, Blanchard P (2003) Watershed vulnerability to herbicide transport in northern Missouri and southern Iowa streams. Environ Sci Technol 37:5518–5527CrossRefGoogle Scholar
  26. Mann RM, Bidwell JR (1999) The toxicity of glyphosate and several glyphosate formulations to four species of southwestern Australian frogs. Arch Environ Contam Toxicol 36:193–199CrossRefGoogle Scholar
  27. Mazanti L, Sparling DW, Rice C, Bialek K, Stevenson C, Teels B (2003) Synergistic effects of a combined exposure to herbicides and an insecticide in Hyla versicolor. In: Linder G, Krest S, Sparling D, Little E (eds) Multiple stressor effects in relation to declining amphibian populations. ASTM STP 1443. American Society for Testing and Materials, West Conshohocken, PA, pp 111–129CrossRefGoogle Scholar
  28. Partearroyo MA, Pilling SJ, Jones MN (1991) The lysis of isolated fish (Oncorhynchus mykiss) gill epithelial cells by surfactants. Comp Biochem Physiol 100C:381–388Google Scholar
  29. Perkins PJ, Boermans HJ, Stephenson GR (2000) Toxicity of glyphosate and triclopyr using the frog embryo teratogenesis assay—Xenopus. Environ Toxicol Chem 19:940–945Google Scholar
  30. Relyea RA (2005a) The impact of insecticides and herbicides on the biodiversity and productivity of aquatic communities. Ecol Appl 15:618–627CrossRefGoogle Scholar
  31. Relyea RA (2005b) The lethal impact of Roundup on aquatic and terrestrial amphibians. Ecol Appl 15:1118–1124CrossRefGoogle Scholar
  32. Relyea RA (2005c) The lethal impacts of Roundup and predatory stress on six species of North American tadpoles. Arch Environ Contam Toxicol 48:351–357CrossRefGoogle Scholar
  33. Relyea RA (2006) The impact of insecticides and herbicides on the biodiversity and productivity of aquatic communities: response to Thompson et al. Ecol Appl 16:2027–2034CrossRefGoogle Scholar
  34. Relyea RA (2009) A cocktail of contaminants: how mixtures of pesticides at low concentrations affect aquatic communities. Oecologia 159:363–376CrossRefGoogle Scholar
  35. Relyea RA, Mills N (2001) Predator-induced stress makes the pesticide carbaryl more deadly to gray treefrog tadpoles (Hyla versicolor). Proc Natl Acad Sci USA 98:2491–2496CrossRefGoogle Scholar
  36. Relyea RA, Schoeppner NM, Hoverman JT (2005) Pesticides and amphibians: the importance of community context. Ecol Appl 15:1125–1134CrossRefGoogle Scholar
  37. Rohr JR, Sager T, Sesterhenn TM, Palmer BD (2006) Exposure, postexposure, and density-mediated effects of atrazine on amphibians: breaking down net effects into their parts. Environ Health Perspect 114:46–50CrossRefGoogle Scholar
  38. Scribner EA, Battaglin WA, Dietze JE, Thurman EM (2003) Reconnaissance data for glyphosate, other selected herbicides, their degradation products, and antibiotics in 51 streams in nine Midwestern states, 2002. US Geological Survey Open-File Report 03–217Google Scholar
  39. Semlitsch RD, Scott DE, Pechmann JHK (1988) Time and size at metamorphosis related to adult fitness in Ambystoma talpoideum. Ecology 69:184–192CrossRefGoogle Scholar
  40. Senseman SA, Lavy TL, Mattice JD, Gbur EE, Skulman BW (1997) Trace level pesticide detections in Arkansas surface waters. Environ Sci Technol 31:395–401CrossRefGoogle Scholar
  41. Smith GR (2001) Effects of acute exposure to a commercial formulation of glyphosate on the tadpoles of two species of anurans. Bull Environ Contam Toxicol 67:483–488CrossRefGoogle Scholar
  42. Stark JD, Banks JE, Vargas R (2004) How risky is risk assessment: the role that life history strategies play in susceptibility of species to stress. Proc Natl Acad Sci USA 101:732–736CrossRefGoogle Scholar
  43. Storrs SI, Kiesecker JM (2004) Survivorship patterns of larval amphibians exposed to low concentrations of atrazine. Environ Health Perspect 112:1054–1057Google Scholar
  44. Struger J, Thompson D, Staznik B, Martin P, McDaniel T, Marvin C (2008) Occurrence of glyphosate in surface waters of southern Ontario. Bull Environ Contam Toxicol 80:378–384CrossRefGoogle Scholar
  45. Thompson DG, Wojtaszek BF, Staznik B, Chartrand DT, Stephenson GR (2004) Chemical and biomonitoring to assess potential acute effects of Vision® herbicide on native amphibian larvae in forest wetlands. Environ Toxicol Chem 23:843–849CrossRefGoogle Scholar
  46. Thompson DG, Solomon KR, Wojtaszek BF, Edginton AN, Stephenson GR (2006) The impact of insecticides and herbicides on the biodiversity and productivity of aquatic communities. Ecol Appl 16:2022–2027CrossRefGoogle Scholar
  47. Tsui MT, Chu LM (2003) Aquatic toxicity of glyphosate-based formulations: comparison between different organisms and the effects of environmental factors. Chemosphere 52:1189–1197CrossRefGoogle Scholar
  48. USDA (2006) Agricultural chemical use database. National Agricultural Statistics Service (NASS). Available from (accessed on November 19, 2007)
  49. USEPA (2002) National water quality inventory-2000. Report to Congress. EPA 841-F-02-003. Office of Water (4503-T). US Environmental Protection Agency, Washington, DCGoogle Scholar
  50. USEPA (2006) 2006 Drinking water standards and health advisories. EPA 822-R-06-013. Office of Water. US Environmental Protection Agency, Washington, DCGoogle Scholar
  51. Williams BK (2008) A multiscale investigation of ecologically relevant effects of agricultural runoff on amphibians. PhD dissertation, University of Missouri, ColumbiaGoogle Scholar
  52. Winfield TW, Bashe WJ, Baker TV (1990) Determination of glyphosate in drinking water by direct-aqueous-injection HPLC, post-column derivatization, and fluorescence detection. US Environmental Protection Agency Method 547Google Scholar
  53. WHO (2006) Guidelines for drinking water quality: 1st addendum to the 3rd edition, vol 1. World Health Organization Press, GenevaGoogle Scholar
  54. Wojtaszek BF, Staznik B, Chartrand DT, Stephenson GR, Thompson DG (2004) Effects of Vision® herbicide on mortality, avoidance response, and growth of amphibian larvae in two forest wetlands. Environ Toxicol Chem 23:832–842CrossRefGoogle Scholar
  55. Wolf MC, Moore PA (2002) The effects of the herbicide metolachlor on the perception of chemical stimuli by Orconectes rusticus. J North Am Benthol Soc 21:457–467CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Division of Biological SciencesUniversity of MissouriColumbiaUSA
  2. 2.U.S. Geological SurveyColumbia Environmental Research CenterColumbiaUSA
  3. 3.Division of Biological SciencesUniversity of MissouriColumbiaUSA

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