Skip to main content
SpringerLink
Log in

Using earthworm avoidance behaviour to assess the toxicity of formulated herbicides and their active ingredients on natural soils

  • SOILS, SEC 4 • ECOTOXICOLOGY • RESEARCH ARTICLE
  • Published:
Journal of Soils and Sediments Aims and scope Submit manuscript

Abstract

Background, aim, and scope

Soil quality has been threatened by intensive agricultural practises, namely those relying on the application of pesticides, such as herbicides. Among the non-target terrestrial organisms exposed to such scenarios, earthworms are key ecological receptors widely used in ecotoxicological studies. As such, this work aims to assess the effects of two herbicide active ingredients (a.i.)—sulcotrione and penoxsulam—and their respective commercial formulations—MIKADO® and VIPER® (referred as Mikado and Viper)—on the avoidance behaviour of Eisenia andrei. In an attempt to enhance the ecological relevance of the generated toxicity data, the avoidance tests were run with standard (LUFA 2.2; L) and natural soils (from corn and rice fields), as long as their habitat function did not constrain the earthworm behaviour.

Methodology

Earthworms were bred in the lab before test conductance. The natural soils used as substrates were collected before the cropping season on corn (C) and rice (R) fields, which are integrated in a wide area exploited for agriculture. Their physico-chemical characterization evolved the determination of pH (H2O, KCl), conductivity, organic matter (OM) and clay/silt contents, and water-holding capacity (WHC). The avoidance tests intended to ascertain (1) the random distribution of earthworms in the natural soils C and R (dual-control tests), (2) the habitat function of natural soils against each other and against L soil, (3) the effect of active ingredients and formulated herbicides on E. andrei behaviour. Avoidance tests with the a.i.s were only performed in L soil. Data evaluation followed ISO (2005) guidelines.

Results

C and R soils presented higher OM (5.1% and 4.5%, respectively) and clay/silt (53.3 and 43.1, respectively) contents and WHC (107.2 and 109.9%, respectively) than L soil (4.1, 21.4 and 48.0%, correspondingly). Earthworms distributed randomly in dual-control tests, but preferred R soil significantly, relative to L or C soils. The LOEC and EC50 values calculated for sulcotrione (>1,000.0 and 1,263.3 mg a.i. kg-1, respectively) and Mikado (1,012.8 and 1,301.3 mg a.i. kg-1, respectively) were much higher than those calculated for penoxsulam (100 and 80.6 mg a.i. kg-1, respectively) or Viper (52.7 and 51.5 mg a.i. kg-1, respectively), when L soil was used as substrate. Moreover, the habitat function of L soil contaminated with the formulated herbicide Viper was more constrained relative to that of the a.i. penoxsulam. Viper induced higher % avoidance on E. andrei exposed to the contaminated L soil compared to that under the R soil.

Discussion

The response of earthworms to R (attraction) and C (avoidance) soils could be related, not only to the quantity of OM content, but also to the quality of organic and inorganic fractions of soil, beyond other intrinsic properties of soils. Both Mikado and sulcotrione impacted the behaviour of E. andrei only slightly. This endpoint was more affected under penoxsulam or Viper exposures on L soil, being the latter-formulated product even more repellent for E. andrei than the a.i. The effect of adjuvants added to the commercial formulation of Viper, may have increased the toxicity of the a.i. Thereby, our results reinforce the need for a careful assessment of the impacts of formulated products. Furthermore, since there was a reduction in earthworm % avoidance under Viper exposures on the natural soil R, it was possible that pesticide bioavailability had been reduced by its sorption to OM and clay mineral sorption sites.

Conclusions

Though the standard L soil should be used for reproducibility and comparison means, other natural soils should be added to the assessment of chemicals, for sake of ecological relevance. Both herbicides induced avoidance behaviour on E. andrei, albeit stronger effects were denoted by penoxsulam and its respective formulated product, Viper. Overall, avoidance tests provided a sensitive, valuable and feasible response either to compare the habitat function of different standard and agricultural natural soils or to test the effect of herbicides.

Recommendations and perspectives

An effort should be made to enlarge the terrestrial ecotoxicological database as a way to fulfil the huge lack of information available for this ecosystem. In this context, additional research congregating a potential linkage between physiological activities sustaining the regular metabolism of earthworms and their avoidance behaviour or even their reproductive effects would be welcomed, especially in what regards formulated pesticides. Such approach would provide a robust and comprehensive understanding of chemical effects. Furthermore, it is encouraged that natural soils should be used to improve the reliability of chemical testing.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Achazi RK (2002) Invertebrates in risk assessment: development of a test battery and of short term biotests for ecological risk assessment of soil. J Soils Sediments 2(4):174–178

    Article  CAS  Google Scholar 

  • Amorim MJB, Römbke J, Soares AMVM (2005) Avoidance behaviour of Enchytraeus albidus: effects of benomyl, carbendazim, phenmediphan and different soil types. Chemosphere 59:501–510

    Article  CAS  Google Scholar 

  • Antunes SC, Castro BB, Pereira R, Gonçalves F (2008) Contribution for tier 1 of the ecological risk assessment of Cunha Baixa uranium mine (Central Portugal): II. Soil ecotoxicological screening. Sci Total Environ 390(2–3):387–395

    CAS  Google Scholar 

  • Bayer CropScience (2004) MIKADO Safety Data Sheet according to Directive EEC 2001/58. Online at http://www.bayercropscience.pt

  • Bird J, Beer A, Guthrie L (2006) Products: new product pipelines slim down. In: Jarvis P (ed) Agrow Magazine, Issue 1. Leverkusen, Germany, p 44

    Google Scholar 

  • CEC (Commission of the European Communities) (2006) Proposal for a Directive of the European Parliament and of the Council Establishing a Framework for the Protection of Soil and Amending Directive 2004/35/EC. COM(2006) 232 final, 2006/0086 (COD), Brussels

  • Chaabane H, Vulliet E, Joux F, Lantoine F, Conan P, Cooper JF, Coste CM (2007) Photodegradation of sulcotrione in various aquatic environments and toxicity of its photoproducts for some marine micro-organisms. Water Res 41(8):1781–1789

    Article  CAS  Google Scholar 

  • Cox C, Surgan M (2006) Unidentified inert ingredients in pesticides: implications for human and environmental health. Environ Health Persp 114(12):1803–1806

    CAS  Google Scholar 

  • EC (European Commission) (2002) Guidance Document on Terrestrial Ecotoxicology Under Council Directive 91/414/EEC. Draft Working Document. European Commission, Health & Consumer Protection Directorate-General. SANCO/10329/2002, rev. 2 final, 17 October 2002, 39 pp

  • EC (European Commission) (2003) Technical Guidance Document on Risk Assessment in support of Commission Directive 93/67/EEC on Risk Assessment for new notified substances, Commission Regulation (EC) No 1488/94 on Risk Assessment for existing substances, Directive 98/8/EC of the European Parliament and of the Council concerning the placing of biocidal products on the market. Part II., Joint research Centre, Ispra

  • ECPA (European Crop Protection Association) (2003) ECPA Statistical Review 2002. D/03/EJ/12560, Brussels, 31 pp

  • Edwards CA, Bohlen PJ (1996) Biology and ecology of earthworms. Chapman and Hall, London

    Google Scholar 

  • EEC (European Environment Commission) (1991) Council directive no. 91/414/EEC concerning the placing of plant protection products on the market. Official Journal of the European Union L230 of 19.08.1991:1–32

  • FAOUN (Food and Agriculture Organization of the United Nations) (1984) Physical and chemical methods of soil and water analysis. Soils Bull 10:1–275

    Google Scholar 

  • Farenhorst A (2006) Importance of soil organic matter fractions in soil-landscape and regional assessments of pesticide sorption and leaching in soil. Soil Sci Soc Am J 70:1005–1012

    Article  CAS  Google Scholar 

  • Finney DJ (1971) Probit analysis. Cambridge University Press, pp 333

  • FOOTPRINT PPDB (The FOOTPRINT Pesticide Properties Database) (2008) FOOTPRINT: creating tools for pesticide risk assessment and management in Europe. Research Project in the 6th Framework Program for Research and Technological Development. [Online]. Retrieved on August from: http://sitem.herts.ac.uk/aeru/footprint/en/index.htm

  • Garcia M, Römbke J, de Brito MT, Scheffczyk A (2008) Effects of three pesticides on the avoidance behavior of earthworms in laboratory tests performed under temperate and tropical conditions. Environ Pollut 153(2):450–456

    Article  CAS  Google Scholar 

  • Gerrard J (2000) Fundamentals of soils. Routledge Fundamentals of Physical Geography, Routledge, Taylor & Francis Group, London, Great Britain, 230 pp

  • Hund-Rinke K, Wiechering H (2001) Earthworm avoidance test for soil assessments—an alternative for acute and reproduction tests. J Soil Sediments 1(1):15–20

    Article  CAS  Google Scholar 

  • Hund-Rinke K, Lindemann M, Simon M (2005) Experiences with novel approaches in earthworm testing alternatives. J Soils Sediments 5(4):233–239

    Article  CAS  Google Scholar 

  • ISO (International Organization for Standardization) (2005) Draft ISO-17512: Soil Quality and Avoidance test for evaluating the quality of soils and the toxicity of chemicals. Test with earthworms (Eisenia fetida/andrei). Geneve, Switzerland

  • Jabusch TW, Tjeerdema RS (2005) Partitioning of penoxsulam, a new sulfonamide herbicide. J Agric Food Chem 53:7179–7183

    Article  CAS  Google Scholar 

  • Jänsch S, Amorim MJ, Römbke J (2005) Identification of the ecological requirements of important terrestrial ecotoxicological test species. Environ Rev 13:51–83

    Article  Google Scholar 

  • Lavelle P, Decaëns T, Aubert M, Barot S, Blouin M, Bureau F, Margerie P, Mora P, Rossi J-P (2006) Soil invertebrates and ecosystem services. Eur J Soil Biol 42:S3–S15

    Article  Google Scholar 

  • Løkke H, van Gestel CAM (1998) Handbook of soil invertebrate toxicity tests. Wiley, Chichester, UK

    Google Scholar 

  • Loureiro S, Soares AMVM, Nogueira AJA (2005) Terrestrial avoidance behaviour tests as screening tool to assess soil contamination. Environ Pollut 138(1):121–131

    Article  CAS  Google Scholar 

  • Matringe M, Sailland A, Pelissier B, Rolland A, Zink O (2005) p-Hydroxyphenylpyruvate dioxygenase inhibitor-resistant plants. Pest Manag Sci 61:269–276

    Article  CAS  Google Scholar 

  • Meazza G, Scheffler BE, Tellez MR, Rimando AM, Romagni JG, Duke SO, Nanayakkara D, Khan IA, Abourashed EA, Dayan FE (2002) The inhibitory activity of natural products on plant p-hydroxyphenylpyruvate dioxygenase. Phytochemistry 59:281–288

    Article  Google Scholar 

  • Muthukaruppan G, Janardhanan S, Vijayalakshmi GS (2005) Sublethal toxicity of the Herbicide Butachlor on the earthworm Perionyx sansibaricus and its histological changes. J Soils Sediments 5(2):82–86

    Article  CAS  Google Scholar 

  • Natal-da-Luz T, Ribeiro R, Sousa JP (2004) Avoidance tests with Collembola and earthworms as early screening tools for site-specific assessment of polluted soils. Environ Toxicol Chem 23(9):2188–2193

    Article  Google Scholar 

  • Natal-da-Luz T, Amorim M, Römbke J, Sousa JP (2008) Avoidance tests with earthworms and springtails: Defining the minimum exposure time to observe a significant response. Ecotox Environ Saf, doi:10.1016/j.ecoenv.2007.09.005

  • O’Halloran K (2006) Toxicological considerations of contaminants in the terrestrial environment for ecological risk assessment. Hum Ecol Risk Assess 12:74–83

    Article  CAS  Google Scholar 

  • Pereira R, Antunes SC, Marques S, Gonçalves F (2008) Contribution for tier 1 of the ecological risk assessment of Cunha Baixa uranium mine (Central Portugal): I Soil chemical characterization. Sci Total Environ 390:377–386

    Article  CAS  Google Scholar 

  • Reinecke SA, Reinecke AJ (2007) The impact of organophosphate pesticides in orchards on earthworms in the Western Cape, South Africa. Ecotox Environ Saf 66:244–251

    Article  CAS  Google Scholar 

  • Reinecke AJ, Maboeta MS, Vermeulen LA, Reinecke SA (2002) Assessment of lead nitrate and mancozeb toxicity in earthworms using the avoidance response. Bull Environ Contam Toxicol 68:779–786

    Article  CAS  Google Scholar 

  • Roberts DW, Knuteson JA, Jackson R (2003) The dissipation of penoxsulam in flooded rice fields. In: Pesticides in Air, Plant, Soil & Water Systems: XII Symposium Pesticide Chemistry, Institute of Agricultural and Environmental Chemistry of the Catholic University, Piacenza, Italy, pp 349–357

  • Römbke J (2006) Tools and techniques for the assessment of ecotoxicological impacts of contaminants in the terrestrial environment. Hum Ecol Risk Assess 12:84–101

    Article  Google Scholar 

  • Römbke J, Jänsch S, Didden W (2005) The use of earthworms in ecological soil classification and assessment concepts. Ecotox Environ Saf 62:249–265

    Article  Google Scholar 

  • Smith R, Pollard SJT, Weeks JM, Nathanail CP (2006) Assessing significant harm to terrestrial ecosystems from contaminated land. Soil Use Manage 21:527–540

    Article  Google Scholar 

  • Sousa A, Pereira R, Antunes SC, Cachada A, Pereira E, Duarte AC, Gonçalves F (2008) Validation of avoidance assays for the screening assessment of soils under different anthropogenic disturbances. Ecotox Environ Saf 71:661–670

    Article  CAS  Google Scholar 

  • SPAC (Soil and Plant Analysis Council) (2000) Handbook of reference methods. CRC, Boca Raton, Florida

    Google Scholar 

  • ter Halle A, Drncova D, Richard C (2006) Phototransformation of the herbicide sulcotrione on maize cuticular wax. Environ Sci Technol 40(9):2989–2995

    Article  Google Scholar 

  • Tominack RL (2000) Herbicide formulations. Clin Toxicol 38(2):129–135

    Article  CAS  Google Scholar 

  • Tomlin CDS (2000) The pesticide manual, 12th edn. British Crop Protection Council, Surrey

    Google Scholar 

  • Tsui MTK, Chu LM (2003) Aquatic toxicity of glyphosate-based formulations: comparison between different organisms and the effects of environmental factors. Chemosphere 52(7):1189–1197

    Article  CAS  Google Scholar 

  • U.S. EPA (U.S. Environmental Protection Agency) (2007) Penoxsulam. Human Health Risk Assessment for Proposed Uses on Fish and Shellfish. PC Code: 119031, Petition no. 5F7012, DP Num: 325461. U.S.EPA Office of Prevention, Pesticides and Toxic Substances, Washington, DC 20460

  • Vermeulen LA, Reinecke AJ, Reinecke SA (2001) Evaluation of the fungicide manganese-zinc ethylene bis(dithiocarbamate) (Mancozeb) for sublethal and acute toxicity to Eisenia fetida (Oligochaeta). Ecotox Environ Saf 48:183–189

    Article  CAS  Google Scholar 

  • WHO (World Health Organization) (2005) The WHO Recommended Classification of Pesticides by Hazard and Guidelines to Classification. International Program on Chemical Safety, NLM Classification: WA 240, WHO Library Cataloguing-in-Publication Data, Geneva, Switzerland

  • Yeardley RB Jr, Lazorchak JM, Gast LC (1996) The potential of an earthworm avoidance test for evaluation of hazardous waste sites. Environ Toxicol Chem 15(9):1532–1537

    Article  CAS  Google Scholar 

  • Ying G-G, Williams B (2000) Laboratory study on the interaction between herbicides and sediments in water systems. Environ Pollut 107:399–405

    Article  CAS  Google Scholar 

  • Zar JH (1996) Biostatistical Analysis, 3rd edn. Prentice-Hall, Inc., USA, p 662

    Google Scholar 

  • Zhou S-P, C-q Duan, Fu H, Chen Y-H, Wang X-H, Yu Z-F (2007) Toxicity assessment for chlorpyrifos-contaminated soil with three different earthworm test methods. Journal of Environmental Sciences 19:854–858

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Bayer CropScience and Dow AgroSciences LLC for the free supply of the technical ingredients sulcotrione and penoxsulam, respectively. Furthermore, we still would like to thank Dow AgroSciences LLC for the provided formulated product Viper. CR Marques was supported by a PhD grant from FCT (Fundação para a Ciência e Tecnologia).

Author information

Authors and Affiliations

  1. Department of Biology, Centre for Environmental and Marine Studies (CESAM), Campus Universitário de Santiago, Universidade de Aveiro, P-3810-193, Aveiro, Portugal

    Catarina Marques, Ruth Pereira & Fernando Gonçalves

Authors
  1. Catarina Marques
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ruth Pereira
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fernando Gonçalves
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Catarina Marques.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marques, C., Pereira, R. & Gonçalves, F. Using earthworm avoidance behaviour to assess the toxicity of formulated herbicides and their active ingredients on natural soils. J Soils Sediments 9, 137–147 (2009). https://doi.org/10.1007/s11368-009-0058-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11368-009-0058-0

Keywords

Search

Navigation