Acute toxicity of copper to the larval stage of three species of ambystomatid salamanders

  • Scott M. WeirEmail author
  • Shuangying Yu
  • David E. Scott
  • Stacey L. Lance


Copper (Cu) appears to be consistently more toxic to anuran species relative to other vertebrate taxa. There are limited Cu toxicity data for salamanders; of the few studies conducted on salamanders, most examined Cu effects on the embryonic, but not the larval, stage. We performed acute toxicity experiments, to quantify LC50s, on Harrison stage 46 larvae (free swimming hatchlings with egg yolk completely absorbed) of three ambystomatid salamander species. Each LC50 experiment used exposure concentrations of 10, 20, 30, 40, 50, and 60 µg/L with 10 replicates per concentration each containing one larva. We found very high toxicity for all species compared to previously published research on the embryo stage. Specifically, the 4-d LC50s for Ambystoma tigrinum and A. opacum were 35.3 and 18.73 µg/L, respectively. The same Cu concentrations caused similar toxicity to A. talpoideum (LC50 = 47.88 µg/L), but exposures required up to 48 d to elicit the same level of mortality. A time-to-event analysis indicated that time to mortality was significantly affected by Cu concentration. Additionally, for A. talpoideum, we observed that elevated levels of Cu decreased growth rate. Comparisons with previously reported Cu toxicity for embryos suggest that, as with fish, Cu may be more toxic to larval salamander stages than for embryos. Further, our data suggest that Cu is an important environmental contaminant that deserves increased scrutiny on the potential for population-level effects where contamination has occurred in wetlands and streams inhabited by salamanders.


Amphibians Metals Mortality LC50 Caudata Sublethal effect 



We thank A. Coleman for assistance in the lab and field collections. Animals were collected under SCDNR permit #G-09-03 following IACUC procedures (A2012-12-003-Y3-A3) from the University of Georgia. We thank three anonymous reviewers for comments that improved the manuscript.


This research was partially supported by United States Department of Energy under Award Numbers DE­FC09­07SR22506 and DE-EM0004391 to the University of Georgia Research Foundation, and was also made possible by the status of the Savannah River Site as a National Environmental Research Park, as well as the protection of research wetlands in the SRS Set-Aside Program. Project funding was provided by the DOE National Nuclear Security Administration.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.


  1. Anderson BS, Middaugh DP, Hunt JW, Turpen SL (1991) Copper toxicity to sperm, embryos and larvae of topsmelt Atherinops affinis, with notes on induced spawning. Mar Environ Res 31:17–35CrossRefGoogle Scholar
  2. Biek R, Funk WC, Maxwell BA, Mills LS (2002) What is missing in amphibian decline research: insights from ecological sensitivity analysis. Conserv Biol 16:728–734CrossRefGoogle Scholar
  3. Birge WJ, Black JA (1979) Effects of copper on embryonic and juvenile stages of aquatic animals. In: Nriagu JO (ed) Copper in the environment: Part II Health effects. John Wiley and Sons, New York, NY, p 373–399Google Scholar
  4. Birge WJ, Westerman AG, Spromberg JA (2000) Comparative toxicology and risk assessment of amphibians. In: Sparling DW, et al., (eds) Ecotoxicology of amphibians and reptiles, 1st edn. SETAC Press, Pensacola, p 727–791Google Scholar
  5. Bridges CM, Dwyer FJ, Hardesty DK, Whites DW (2002) Comparative contaminant toxicity: are amphibian larvae more sensitive than fish? Bull Environ Contam Toxicol 69:562–569CrossRefGoogle Scholar
  6. Chapman GA (1985) Acclimation as a factor influencing metal criteria. In: Bahner RC, Hansen DJ (eds) Aquatic toxicology and hazard assessment: eighth symposium. American Society for Testing of Materials, Philadelphia, p 119–136CrossRefGoogle Scholar
  7. Collins JP, Storfer A (2003) Global amphibian declines: sorting the hypotheses. Divers Distrib 9:89–98CrossRefGoogle Scholar
  8. Davidson C (2004) Declining downwind: amphibian population declines in California and historical pesticide use. Ecol Appl 14:1892–1902CrossRefGoogle Scholar
  9. Duellman WE, Trueb L (1986) Biology of amphibians. Johns Hopkins University Press, BaltimoreGoogle Scholar
  10. Duncan DA, Klaverkamp JF (1983) Tolerance and resistance to cadmium in white suckers (Catostomus commersoni) previously exposed to cadmium, mercury, zinc, or selenium. Can J Fish Aquat Sci 40:128–138CrossRefGoogle Scholar
  11. Eisler R (1998) Copper hazards to fish, wildlife, and invertebrates: a synoptic review. US Geological Survey, Washington, DC, USGS/BRD/BSR 1997–2002Google Scholar
  12. Fedorenkova A, Vonk JA, Lenders HJR, Creemers RCM, Breure AM, Hendriks AJ (2012) Ranking ecological risks of multiple chemical stressors on amphibians. Environ Toxicol Chem 31:1416–1421CrossRefGoogle Scholar
  13. Flynn RW, Scott DE, Kuhne W, Soteropoulos D, Lance SL (2015) Lethal and sublethal measures of chronic copper toxicity in the eastern narrowmouth toad, Gastrophryne carolinensis, reveal within and among population variation. Environ Toxicol Chem 34:575–582CrossRefGoogle Scholar
  14. García-Muñoz E, Guerrero F, Parra G (2010) Intraspecific and interspecific tolerance to copper sulphate in five iberian amphibian species at two developmental stages. Archives of Environmental Contamination and Toxicology 59:312–321CrossRefGoogle Scholar
  15. Glass GV, Peckham PD, Sander JR (1972) Consequences of failure to meet assumptions underlying the fixed effects analysis of variance and covariance. Rev Educ Res 42:237–288CrossRefGoogle Scholar
  16. Horne MT, Dunson WA (1994) Exclusion of the Jefferson salamander, Ambystoma jeffersonianum, from some potential breeding ponds in Pennsylvania: effects of pH, temperature, and metals on embryonic development. Arch Environ Contam Toxicol 27:323–330Google Scholar
  17. Horne MT, Dunson WA (1995) Effects of low pH, metals, and water hardness on larval amphibians. Arch Environ Contam Toxicol 29:500–505Google Scholar
  18. Lance SL, Erickson MR, Flynn RW, Mills GL, Tuberville TD, Scott DE (2012) Effects of chronic copper exposure on development and survival of in the southern leopard frog (Lithobates [Rana] sphenocephalus). Environ Toxicol Chem 31:1587–1594CrossRefGoogle Scholar
  19. Lance SL, Flynn RW, Erickson MR, Scott DE (2013) Within- and among-population differences in response to chronic copper exposure in southern toads, Anaxyrus terrestris. Environ Pollut 177:135–142CrossRefGoogle Scholar
  20. McKim JM, Eaton JG, Holcombe GW (1978) Metal toxicity to embryos and larvae of eight species of freshwater fish–II: copper. Bull Environ Contam Toxicol 19:608–616CrossRefGoogle Scholar
  21. Morey S, Reznick D (2001) Effects of larval density on postmetamorphic spadefoot toads (Spea hammondii). Ecol 82:510–522CrossRefGoogle Scholar
  22. Pechenik JA (2006) Larval experience and latent effects: metamorphosis is not a new beginning. Integr Comp Biol 46:323–333CrossRefGoogle Scholar
  23. Perschbacher PW, Wurts WA (1999) Effects of calcium and magnesium hardness on acute copper toxicity to juvenile channel catfish Ictalurus punctatus. Aquaculture 172:275–280CrossRefGoogle Scholar
  24. Plautz SC, Salice CJ (2013) Plasticity in offspring contaminant tolerance traits: developmental cadmium exposure trumps parental effects. Ecotoxicology 22:847–853CrossRefGoogle Scholar
  25. R Core Team (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria, Accessed 10 Feb 2014
  26. Scott DE (1994) The effect of larval density on adult demographic traits in Ambystoma opacum. Ecology 75:1383–1396CrossRefGoogle Scholar
  27. Scott DE, Casey ED, Donovan MF, Lynch TK (2007) Amphibian lipid levels at metamorphosis correlate to post-metamorphic terrestrial survival. Oecologia 153:521–532CrossRefGoogle Scholar
  28. Semlitsch RD, Scott DE, Pechmann JHK (1988) Time and size at metamorphosis related to adult fitness in Ambystoma talpoideum. Ecology 69:184–192CrossRefGoogle Scholar
  29. Smith DC (1987) Adult recruitment in chorus frogs: effect of size and date at metamorphosis. Ecology 68:344–350CrossRefGoogle Scholar
  30. Soteropoulos DL, Lance SL, Flynn RW, Scott DE (2014) Effects of copper exposure on hatching success and early larval survival in marbled salamanders, Ambystoma opacum. Environ Toxicol Chem 33:1631–1637CrossRefGoogle Scholar
  31. Sparling DW, Linder G, Bishop CA, Krest S (2010) Recent advances in amphibian and reptile ecotoxicology. In: Sparling DW, Linder G, Bishop CA, Krest S (eds) Ecotoxicology of amphibians and reptiles, 2nd edn. CRC Press, Pensacola, FL, USA, p 1–13CrossRefGoogle Scholar
  32. Therneau T, Lumley T (2009) Survival: survival analysis, including penalized likelihood. R package version 2.35–2.4. Accessed 24 Mar 2017
  33. Venables WN, Ripley BD (2002) Modern applied statistics with S, 4th edn. Springer, New York, NYCrossRefGoogle Scholar
  34. von Westernhagen H (1988) Sublethal effects of pollutants on fish eggs and larvae. In: Hoar WS, Randall DJ (eds) Fish physiology, vol. 11: The physiology of developing fish. Academic Press, New York, p 253–346Google Scholar
  35. Watson S, Russell AP (2000) A posthatching developmental staging table for the Long-toed Salamander, Ambystoma macrodactylum krausei. Amphibia-Reptilia 21:143–154CrossRefGoogle Scholar
  36. Weir SM, Scott DE, Salice CJ, Lance SL (2016) Integrating copper toxicity and climate change to understand extinction risk to two species of pond-breeding anurans. Ecol Appl 26:1721–1732CrossRefGoogle Scholar
  37. Weltje L, Simpson P, Gross M, Crane M, Wheeler JR (2013) Comparative acute and chronic sensitivity of fish and amphibians: a critical review of data. Environ Toxicol Chem 32:984–994CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Scott M. Weir
    • 1
    • 2
    Email author
  • Shuangying Yu
    • 1
    • 3
  • David E. Scott
    • 1
  • Stacey L. Lance
    • 1
  1. 1.Savannah River Ecology LaboratoryUniversity of GeorgiaAikenUSA
  2. 2.Department of BiologyQueens University of CharlotteCharlotteUSA
  3. 3.Sciences DivisionCentral Piedmont Community CollegeCharlotteUSA

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