Skip to main content
Log in

Getting More Ecologically Relevant Information from Laboratory Tests: Recovery of Lemna minor After Exposure to Herbicides and Their Mixtures

  • Published:
Archives of Environmental Contamination and Toxicology Aims and scope Submit manuscript

Abstract

Recovery after exposure to herbicides—atrazine, isoproturon, and trifluralin—their binary and ternary mixtures, was studied under laboratory conditions using a slightly adapted standard protocol for Lemna minor. The objectives of the present study were (1) to compare empirical to predicted toxicity of selected herbicide mixtures; (2) to assess L. minor recovery potential after exposure to selected individual herbicides and their mixtures; and (3) to suggest an appropriate recovery potential assessment approach and endpoint in a modified laboratory growth inhibition test. The deviation of empirical from predicted toxicity was highest in binary mixtures of dissimilarly acting herbicides. The concentration addition model slightly underestimated mixture effects, indicating potential synergistic interactions between photosynthetic inhibitors (atrazine and isoproturon) and a cell mitosis inhibitor (trifluralin). Recovery after exposure to the binary mixture of atrazine and isoproturon was fast and concentration-independent: no significant differences between relative growth rates (RGRs) in any of the mixtures (IC10Mix, 25Mix, and 50Mix) versus control level were recorded in the last interval of the recovery phase. The recovery of the plants exposed to binary and ternary mixtures of dissimilarly acting herbicides was strictly concentration-dependent. Only plants exposed to IC10Mix, regardless of the herbicides, recovered RGRs close to control level in the last interval of the recovery phase. The inhibition of the RGRs in the last interval of the recovery phase compared with the control level is a proposed endpoint that could inform on reversibility of the effects and indicate possible mixture effects on plant population recovery potential.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Altenburger R, Backhaus T, Boedeker W, Faust M, Scholze M (2013) Simplifying complexity: mixture tocixity assessment in the last 20 years. Environ Toxicol Chem 32(8):1685–1687

    Article  CAS  Google Scholar 

  • Altenburger R, Ait-Aissa S, Antczak P, Backhaus T, Barcelo D, Seiler TB, Brion F, Busch W, Chipman K, Lopez de Alda M, Umbuzeiro Gde A, Escher BI, Falciani F, Faust M, Focks A, Hilscherova K, Hollender J, Hollert H, Jager F, Jahnke A, Kortenkamp A, Krauss M, Lemkine GF, Munthe J, Neumann S, Schymanski EL, Scrimshaw M, Segner H, Slobodnik J, Smedes F, Kughathas S, Teodorovic I, Tindall AJ, Tollefsen KE, Walz KH, Williams TD, Van den Brink PJ, van Gils J, Vrana B, Zhang X, Brack W (2015) Future water quality monitoring–adapting tools to deal with mixtures of pollutants in water resource management. Sci Total Environ 512–513:540–551

    Article  Google Scholar 

  • Appenroth KJ, Krech K, Keresztes A, Fischer W, Koloczek H (2010) Effects of nickel on the chloroplasts of the duckweeds Spirodela polyrhiza and Lemna minor and their possible use in biomonitoring and phytoremediation. Chemosphere 78:216–223

    Article  CAS  Google Scholar 

  • Arrhenius A, Gronvall F, Scholze M, Backhaus T, Blanck H (2004) Predictability of the mixture toxicity of 12 similarly acting congeneric inhibitors of photosystem II in marine periphyton and epipsammon communities. Aquat Toxicol 68:351–367

    Article  CAS  Google Scholar 

  • Backhaus T, Faust M (2012) Predictive environmental risk assessment of chemical mixtures: a conceptual framework. Environ Sci Technol 46:2564–2573

    Article  CAS  Google Scholar 

  • Backhaus T, Arrhenius A, Blanck H (2004a) Toxicity of a mixture of dissimilarly acting substances to natural algal communities: predictive power and limitations of independent action and concentration addition. Environ Sci Technol 38(23):6363–6370

    Article  CAS  Google Scholar 

  • Backhaus T, Faust M, Scholze M, Gramatica P, Vigni M, Horst Grimme L (2004b) Joint algal toxicity of phenylurea herbicides is equally predictable by concentration addition and independent action. Environ Toxicol Chem 23(2):258–264

    Article  CAS  Google Scholar 

  • Belden JB, Gilliom RJ, Lydy MJ (2007) How well can we predict the toxicity of pesticide mixtures to aquatic life? Integr Environ Assess Manag 3(3):364–372

    Article  CAS  Google Scholar 

  • Boxall ABA, Fogg LA, Ashauer R, Bowles T, Sinclair C, Colyer A, Brain RA (2013) Effects of repeated pulsed herbicide axposures on the growth of aquatic macrophytes. Environ Toxicol Chem 32(1):193–200

    Article  CAS  Google Scholar 

  • Brack W, Altenburger R, Schüürmann G, Krauss M, López Herráez D, van Gils J, Slobodnik J, Munthe J, Gawlik BM, van Wezel A, Schriks M, Hollender J, Tollefsen KE, Mekenyan O, Dimitrov S, Bunke D, Cousins I, Posthuma L, van den Brink PJ, López de Alda M, Barceló D, Faust M, Kortenkamp A, Scrimshaw M, Ignatova S, Engelen G, Massmann G, Lemkine G, Teodorovic I, Walz KH, Dulio V, Jonker MTO, Jäger F, Chipman K, Falciani F, Liska I, Rooke D, Zhang X, Hollert H, Vrana B, Hilscherova K, Kramer K, Neumann S, Hammerbacher R, Backhaus T, Mack J, Segner H, Escher B, de Aragão Umbuzeiro G (2015) SOLUTIONS for present and future emerging pollutants in land and water resources management. Sci Total Environ 503–504:22–31

    Article  Google Scholar 

  • Brain RA, Hosmer AJ, Desjardins D, Kendall TZ, Krueger H, Wall SB (2012) Recovery of duckweed from time-varying exposure to atrazine. Environ Toxicol Chem 31(5):1121–1128

    Article  CAS  Google Scholar 

  • Busch W, Schmidt S, Kuhne R, Schulze T, Krauss M, Altenburger R (2016) Micropollutants in European rivers: a mode of action survey to support the development of effect-based tools for water monitoring. Environ Toxicol Chem 35(8):1887–1899

    Article  CAS  Google Scholar 

  • Carvalho RN, Arukwe A, Ait-Aissa S, Nilles AB, Balzamo S, Baun A, Belkin S, Blaha L, Brion F, Conti D, Creusot N, Essig Y, Ferrero VEV, Flander-Putrle V, Furhacker M, Grillari-Voglauer R, Hogstrand C, Jona A, Kharlyngdoh JB, Loos R, Lundebye AK, Modig C, Olsson PE, Pillai S, Polak N, Potalivo M, Sanchez W, Schifferli A, Schirmer K, Sforzini S, Sturzenbaum SR, Softeland L, Turk V, Viarengo A, Werner I, Yagur-Kroll S, Zounkova R, Lettieri T (2014) Mixtures of chemical pollutants at european legislation safety concentrations: how safe are they? Toxicol Sci 141(1):218–233

    Article  CAS  Google Scholar 

  • Cedergreen N, Madsen TV (2003) Light regulation of root and leaf NO3 uptake and reduction in the floating macrophyte Lemna minor. New Phytol 161:449–457

    Article  Google Scholar 

  • Cedergreen N, Abbaspoor M, Sørensen H, Streibig JC (2007) Is mixture toxicity measured on a biomarker indicative of what happens on a population level? A study with Lemna minor. Ecotox Environ Safe 67:323–332

    Article  CAS  Google Scholar 

  • Cedergreen N, Sørensen H, Svendsen C (2012) Can the joint effect of ternary mixtures be predicted from binary mixture toxicity results? Sci Total Environ 427–428:229–237

    Article  Google Scholar 

  • Coors A, Frische T (2011) Predicting the aquatic toxicity of commercial pesticide mixtures. Environ Sci Eur 23:22

    Article  Google Scholar 

  • EC (2009) Regulation (EC) no 1107/2009 of the European Parliament and of the Council of 21 October 2009 concerning the placing of plant protection products on the market and repealing Council Directives 79/117/EEC and 91/414/EEC. Off J Eur Union L 309:1–50

    Google Scholar 

  • EC (2013) Directive 2013/39/EU of the European Parliament and of the Council of 12 August 2013 amending Directives 2000/60/EC and 2008/105/EC as regards priority substances in the field of water policy

  • EFSA PPR Panel (European Food Safety Authority Panel on Plant Protection Products and their Residues) (2013) Guidance on tiered risk assessment for plant protection products for aquatic organisms in edge-of-field surface waters. EFSA J 11(7):3290. doi:10.2903/j.efsa.2013.3290

    Article  Google Scholar 

  • EFSA Scientific Committee (European Food Safety Authority Scientific Committee) (2016) Recovery in environmental risk assessments at EFSA. EFSA J 14(2):4313. doi:10.2903/j.efsa.2016.4313

    Article  Google Scholar 

  • EU (Commission Regulation) (2013) No 283/2013 of 1 March 2013 setting out the data requirements for active substances, in accordance with the Regulation (EC) No 1107/2009 of the European Parliament and of the Council concerning the placing of plant protection products on the market. Off J Eur Union 93, 03.04.2013. p 84

  • EU (European Commission) (2011) Directorate-general for health & consumers—scientific committee on health and environmental risks (SCHER). Scientific committee on emerging and newly identified health risks (SCENIHR) Scientific committee on consumer safety (SCCS) toxicity and assessment of chemical mixtures (final approved opinion)

  • Fairchild JF, Ruessler DS, Haverland PS, Carlson AR (1997) Comparative sensitivity of Selenastrum capricornutum and Lemna minor to sixteen herbicides. Arch Environ Con Tox 32:353–357

    Article  CAS  Google Scholar 

  • Faust M, Altenburger R, Backhaus T, Blanck H, Boedeker W, Gramatica P, Hamer V, Scholze M, Vigni M, Grimme LH (2001) Predicting the joint algal toxicity of multi-component s-triazine mixtures at low-effect concentrations of individual toxicants. Aquat Toxicol 56(1):13–32

    Article  CAS  Google Scholar 

  • Faust M, Altenburger R, Backhaus T, Blanck H, Boedeker W, Gramatica P, Hamer V, Vighi M, Grimme LH (2003) Joint algal toxicity of 16 dissimilarly acting chemicals is predictable by the concenpt of independent action. Aquat Toxicol 63(1):43–63

    Article  CAS  Google Scholar 

  • Junghaus M, Backhaus T, Faust M, Scholze M, Grimme LH (2003) Predictability of combined effects of eight chloroacetanilide herbicides on algal reproduction. Pest Manag Sci 59:1101–1110

    Article  Google Scholar 

  • Kienzler A, Berggren E, Bessems J, Bopp S, van der Linden S, Worth A (2014) Assessment of mixtures—review of regulatory requirements and guidance. JRC Science and policy reports. European Commission Joint Research Centre Institute for Health and Consumer Protection Systems Toxicology Unit and European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM). ISBN 978-92-79-38479-0

  • Knauer K, Hommen U (2012) Sensitivity, variability, and recovery of functional and structural endpoints of an aquatic community exposed to herbicides. Ecotox Environ Safe 78:178–183

    Article  CAS  Google Scholar 

  • Knauert S, Escher B, Singer H, Hollender J, Knauer K (2008) Mixture toxicity of three photosystem II inhibitors (atrazine, isoproturon, and diuron) toward photosyntesis of freshwater studied in outdoor mesocosms. Environ Sci Technol 42(17):6424–6430

    Article  CAS  Google Scholar 

  • Knauert S, Dawo U, Hollender J, Hommen U, Kauer K (2009) Effects of photosystem II Inhibitors and their mixture on freshwater phytoplankton succession in outdoor mesocosms. Environ Toxicol Chem 28(4):836–845

    Article  CAS  Google Scholar 

  • Kortenkamp A, Faust M (2010) Combined exposures to anti-androgenic chemicals: steps towards cumulative risk assessment. Int J Androl 33:463–474

    Article  CAS  Google Scholar 

  • Kortenkamp A, Backhaus T, Faust M (2009) State of the art report on mixture toxicity. Final report, study Contract number 070307/2007/485103/ETU/D.1. European Commission, Brussels, p 391

  • Mohammad M, Itoh K, Suyama K (2010) Effects of herbicides on Lemna gibba and recovery from damage after prolonged exposure. Arch Environ Con Tox 58:605–612

    Article  CAS  Google Scholar 

  • OECD (Organization for Economic Cooperation and Development) (1998) Guidelines for the testing of chemicals. Test no. 408: Repeated dose 90 Day Oral Toxicity Study in Rodents. Paris

  • OECD (Organization for Economic Cooperation and Development) (2006) Guidelines for the testing of chemicals. Test no. 221: Lemna sp. growth inhibition test. Paris

  • OECD (Organization for Economic Cooperation and Development) (2008) Guidelines for the testing of chemicals. Test no. 407: Repeated dose 28-day oral toxicity study in rodents. Paris

  • Peterson DR (1994) Calculating the aquatic toxicity of hydrocarbon mixtures. Chemosphere 29(12):2493–2506

    Article  CAS  Google Scholar 

  • Ratte M, Ratte H (2014) Myriophyllum toxicity test: result of a ring test using M. aquaticum and M. spicatum grown in a water-sediment system. OECD Environment, Health and Safety Publications (EHS), Series on testing and assessment, No. 206, OECD Publishing, Paris

  • Schmitt W, Bruns E, Dollinger M, Sowig P (2013) Mechanistic TK/TD-model simulating the effect of growth inhibitors on Lemna populations. Ecol Model 255:1–10

    Article  CAS  Google Scholar 

  • Teodorović I, Knežević V, Tunić T, Čučak M, Nikolić-Lečić J, Leovac A, Ivančev-Tumbas I (2012) Myriophyllum aquaticum versus Lemna minor: sensitivity and recovery potential after exposure to atrazine. Environ Toxicol Chem 31(2):417–426

    Article  Google Scholar 

  • TesTox (2003) Version 1.0. University of Novi Sad, Novi Sad, Serbia

  • Tunić T, Knežević V, Kerkez Đ, Tubić A, Šunjka D, Lazić S, Brkić D, Teodorović I (2015) Some arguments in favour of a Myriophyllum aquaticum growth inhibition test in a water-sediment system as an additional test in risk assessment of herbicides. Environ Toxicol Chem 34(9):2104–2115

    Article  Google Scholar 

  • US EPA (United States Environmental Protection Agency) (1995) Determination of chlorinated pesticides, herbicides, and organohalides by liquid-solid extraction and electron capture gas chromatography, revision 2.0. Method 508.1. Office of Research and Development U.S. Environmental Protection Agency, Cincinnati, OH

  • US EPA (United States Environmental Protection Agency) (2000) Determination of phenylurea compounds in drinking water by solid phase extraction and high performance liquid chromatography with UV detection. Revision 1.0. Method 532. Office of Research and Development U.S. Environmental Protection Agency, Cincinnati, OH

  • Von der Ohe PC, Liess M (2004) Relative sensitivity distribution of aquatic invertebrates to organic and metal compounds. Environ Toxicol Chem 23:150–156

    Article  Google Scholar 

  • Von der Ohe PC, de Deckere E, Prus A, Munoz I, Wolfram G, Villagrasa M, Ginebreda A, Hein M, Brack W (2009) Toward an integrated assessment of the ecological and chemical status of European river basins. Integr Environ Assess Manag 5(1):50–61

    Article  Google Scholar 

  • Wilson PC, Koch R (2013) Influence of exposure concentration and duration on effects and recovery of Lemna minor exposed to the herbicide Norflurazon. Arch Environ Con Tox 64:228–234

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The study was financially supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia via Grant No. 172028. The authors thank Mirjana Kezunović for her assistance with the laboratory work. We also are grateful to two anonymous reviewers for constructive criticism and valuable comments on the earlier version of the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Varja Knežević.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 165 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Knežević, V., Tunić, T., Gajić, P. et al. Getting More Ecologically Relevant Information from Laboratory Tests: Recovery of Lemna minor After Exposure to Herbicides and Their Mixtures. Arch Environ Contam Toxicol 71, 572–588 (2016). https://doi.org/10.1007/s00244-016-0321-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00244-016-0321-5

Keywords

Navigation