Influence of Hardness and Dissolved Organic Carbon on the Acute Toxicity of Copper to Zebrafish (Danio rerio) at Different Life Stages

  • Wei Liao
  • Chenglian FengEmail author
  • Na Liu
  • Daqing Liu
  • Zhenfei Yan
  • Yingchen Bai
  • Hengwang Xie
  • Hong Shi
  • Daishe Wu


Copper (Cu) bioavailability varies under water conditions. In the present study, the whole life of zebrafish was divided into three different life stages (larvae, juvenile and adult) based on the growth curve, then the influences of water hardness and dissolved organic carbon (DOC) concentration on the acute toxicity of zebrafish were respectively investigated. The results indicated that the life stages had significant effects on Cu toxicity. The larvae stage was less sensitive to Cu than both the juvenile and adult stages. With the increase of water hardness, the toxicity of Cu on zebrafish was decreased, a linear relationship was observed between water hardness and Cu toxicity, and the same was true for DOC concentration. The results showed that taking the 24 days juvenile zebrafish to study the water quality criteria of Cu was stable, sensitive and economical.


Zebrafish Life stage DOC Hardness Acute toxicity Water quality criteria 



This study is supported by the National Natural Scientific Foundation of China (41773085) and the China National Project of Water Pollution Control and Treatment (Project No. 2017ZX07301005-001).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

128_2019_2721_MOESM1_ESM.docx (26 kb)
Electronic supplementary material 1 (DOCX 26 kb)


  1. Chen WY, Lin CJ, Ju YR, Tsai JW, Liao CM (2012) Assessing the effects of pulsed waterborne copper toxicity on life-stage tilapia populations. Sci Total Environ 417–418(4):129–137CrossRefGoogle Scholar
  2. Del Signore A, Hendriks AJ, Lenders HJR, Leuven RSEW, Breure AM (2016) Development and application of the SSD approach in scientific case studies for ecological risk assessment. Environ Toxicol Chem 35(9):2149–2161CrossRefGoogle Scholar
  3. De Schamphelaere KAC, Janssen CR (2002) A biotic ligand model predicting acute copper toxicity for Daphnia magna: the effects of calcium, magnesium, sodium, potassium, and pH. Environ Sci Technol 36(1):48–54CrossRefGoogle Scholar
  4. De Schamphelaere KAC, Janssen CR (2006) Bioavailability models for predicting copper toxicity to freshwater green microalgae as a function of water chemistry. Environ Sci Technol 40(14):4514–4522CrossRefGoogle Scholar
  5. Donnachie RL, Johnson AC, Moeckel C, Pereira MG, Sumpter JP (2014) Using risk-ranking of metals to identify which poses the greatest threat to freshwater organisms in the UK. Environ Pollut 194(7):17–23CrossRefGoogle Scholar
  6. Erickson RJ (2013) The biotic ligand model approach for addressing effects of exposure water chemistry on aquatic toxicity of metals: genesis and challenges. Environ Toxicol Chem 32(6):1212–1214CrossRefGoogle Scholar
  7. Erickson RJ, Benoit DA, Mattson VR, Leonard EN Jr HPN (2010) The effects of water chemistry on the toxicity of copper to fathead minnows. Environ Toxicol Chem 15(2):181–193CrossRefGoogle Scholar
  8. Feng CL, Wang H, Wang Y, Wu FC (2015) Predicted no effect concentration of Bisphenol A (BPA) based on different toxicological endpoints. Asian J Ecotoxicol 10(1):119–129 (in Chinese) Google Scholar
  9. Festa RA, Thiele DJ (2011) Copper: an essential metal in biology. Curr Biol 21(21):R877–R883CrossRefGoogle Scholar
  10. Fu ZY, Wu FC, Chen LL, Xu BS, Feng CL, Bai YC, Liao HQ, Sun SY, Giesy JP, Guo WJ (2016) Copper and zinc, but not other priority toxic metals, pose risks to native aquatic species in a large urban lake in Eastern China. Environ Pollut 219:1069–1076CrossRefGoogle Scholar
  11. Grosell M, Gerdes R, Brix KV (2006) Influence of Ca, humic acid and pH on lead accumulation and toxicity in the fathead minnow during prolonged water-borne lead exposure. Comp Biochem Physiol C 143(4):473–483CrossRefGoogle Scholar
  12. Jiang JH, Wu SG, Chen JB, Wu CX, Cai LM, Zhao XP (2015) Acute toxicity effects of triadimefon on different life stages of zebrafish (Danio rerio) and Chinese rare minnow (Gobiocypris rarus). Asian J Ecotoxicol 10(05):150–156 (in Chinese) Google Scholar
  13. Jin XW, Wang YY, Jin W, Rao KF, Giesy JP, Hollert H, Richardson KL, Wang ZJ (2014) Ecological risk of nonylphenol in China surface waters based on reproductive fitness. Environ Sci Technol 48(2):1256–1262CrossRefGoogle Scholar
  14. Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embrypnic-development of the zebrafish. Dev Dynam 203(3):253–310CrossRefGoogle Scholar
  15. Li XF, Wang PF, Feng CL, Liu DQ, Chen JK, Wu FC (2018) Acute toxicity and hazardous concentrations of Zinc to native freshwater organisms under different pH values in China. Bull Environ Contam Tox 103(1):120–126CrossRefGoogle Scholar
  16. Madhav MR, David SEM, Kumar RSS, Swathy JS, Bhuvaneshwari M, Mukherjee A, Chandrasekaran N (2017) Toxicity and accumulation of copper oxide (CuO) nanoparticles in different life stages of Artemia salina. Environ Toxicol Pharm 52:227–238CrossRefGoogle Scholar
  17. Mckim JM (1977) Evaluation of tests with early life stages of fish for predicting long-term toxicity. J Fish Res Board Can 34(8):1148–1154CrossRefGoogle Scholar
  18. McNulty HR, Anderson BS, Hunt JW, Turpen SL, Singer MM (1994) Age-specific toxicity of copper to larval topsmelt atherinops-affinis. Environ Toxicol Chem 13(3):487–492Google Scholar
  19. OECD (1992) Guidelines for the testing of chemicals, sect. 2: effects on biotic systems, test no. 203: fish, acute toxicity test. Organisation for Economic Co-operation and Development, ParisGoogle Scholar
  20. Paquin PR, Gorsuch JW, Apte SC, Batley GE, Bowles KC, Campbell PGC, Delos CG, Toro DMD, Dwyer RL, Galvez F (2002) The biotic ligand model: a historical overview. Comp Biochem Physiol C 133(1):3–35Google Scholar
  21. Peters A, Wilson I, Merrington G, Heijerick D, Baken S (2019) Assessing compliance of european fresh waters for copper: accounting for bioavailability. Bull Environ Contam Tox 102(2):153–159CrossRefGoogle Scholar
  22. Playle RC, Gensemer RW, Dixon DG (2010) Copper accumulation on gills of fathead minnows: influence of water hardness, complexation and pH of the gill micro-environment. Environ Toxicol Chem 11(3):381–391CrossRefGoogle Scholar
  23. Rüdel H, Díaz MC, Garelick H, Kandile NG, Miller BW, Pantoja ML, Purchase D, Shevah Y, Van SP (2015) Consideration of the bioavailability of metal/metalloid species in freshwaters: experiences regarding the implementation of biotic ligand model-based approaches in risk assessment frameworks. Environ Sci Pollut Res 22(10):7405–7421CrossRefGoogle Scholar
  24. Sciera KL, Isely JJ, Tomasso JR, Klaine SJ (2004) Influence of multiple water-quality characteristics on copper toxicity to fathead minnows (Pimephales promelas). Environ Toxicol Chem 23(12):2900–2905CrossRefGoogle Scholar
  25. Shi H, Feng CL, Huang H, Wu FC (2016) The correlation discussion between aluminum toxicity to aquatic organisms and water hardness. Asian J Ecotoxicol 11(1):141–152 (in Chinese) Google Scholar
  26. Shi QP, Wang M, Shi FQ, Yang LH, Guo YY, Feng CL, Liu JF, Zhou BS (2018) Developmental neurotoxicity of triphenyl phosphate in zebrafish larvae. Aquat Toxicol 203:80–87CrossRefGoogle Scholar
  27. Tan QG, Wang WX (2011) Acute toxicity of cadmium in Daphnia magna under different calcium and pH conditions: importance of influx rate. Environ Sci Technol 45(5):1970–1976CrossRefGoogle Scholar
  28. Tang S, Doering JA, Sun JX, Beitel SC, Shekh K, Patterson S, Crawford S, Giesy JP, Wiseman SB, Hecker M (2016) Linking oidative sress and mgnitude of cmpensatory rsponses with life-stage specific differences in sensitivity of white sturgeon (Acipenser transmontanus) to copper or cadmium. Environ Sci Technol 50(17):9717–9726CrossRefGoogle Scholar
  29. Toro DMD, Allen HE, Bergman HL, Meyer JS, Paquin PR, Santore RC (2001) Biotic ligand model of the acute toxicity of metals. 1. Technical basis. Environ Toxicol Chem 20(10):2383–2396CrossRefGoogle Scholar
  30. USEPA (2007a) Aquatic life ambient freshwater quality criteria-copper. US EPA Office of Water, Washington DC, pp 2–4Google Scholar
  31. Van Genderen EJ, Ryan AC, Tomasso JR, Klaine SJ (2005) Evaluation of acute copper toxicity to larval fathead minnows (Pimephales promelas) in soft surface waters. Environ Toxicol Chem 24(2):408–414CrossRefGoogle Scholar
  32. Verriopoulos G, Moraïtou-Apostolopoulou M (1982) Differentiation of the sensitivity to copper and cadmium in different life stages of a copepod. Mar Pollut Bull 13(4):123–125CrossRefGoogle Scholar
  33. Wang CY, Chen H, Wu KB, An LH, Zheng BH (2011) Application of the biotic ligand model to predict copper acute toxicity to Medaka fish in typical Chinese rivers. Water Sci Technol 64(6):1277–1283CrossRefGoogle Scholar
  34. Wang Z, Meador JP, Leung KMY (2016) Metal toxicity to freshwater organisms as a function of pH: a meta-analysis. Chemosphere 144:1544–1552CrossRefGoogle Scholar
  35. Wu FC, Feng CL, Cao YJ, Zhang RQ (2011) Aquatic life ambient freshwater quality criteria for copper in China. Asian J Ecotoxicol 6(6):617–628 (in Chinese) Google Scholar
  36. Yang Y, Qi SZ, Wang DH, Wang K, Zhu LZ, Chai TT, Wang CJ (2016) Toxic effects of thifluzamide on zebrafish (Danio rerio). J Hazard Mater 307:127–136CrossRefGoogle Scholar
  37. Zhang YH, Zang WC, Qin LM, Zheng L, Cao Y, Yan ZZ, Yi XL, Zeng HH, Liu ZT (2017) Water quality criteria for copper based on the BLM approach in the freshwater in China. PLoS ONE 12(2):e0170105CrossRefGoogle Scholar

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

Authors and Affiliations

  1. 1.Key Laboratory of Poyang Lake Environment and Resource Utilization, Ministry of Education, School of Environmental and Chemical EngineeringNanchang UniversityNanchangChina
  2. 2.State Key Laboratory of Environmental Criteria and Risk AssessmentChinese Research Academy of Environmental SciencesBeijingChina
  3. 3.Jiangxi Irrigation Experiment Central StationNanchangChina

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