Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Toxicity of phenmedipham and carbendazim to Enchytraeus crypticus and Eisenia andrei (Oligochaeta) in Mediterranean soils

  • 390 Accesses

  • 13 Citations



The main objective of the present study was to evaluate the toxicity of two reference chemicals, Carbendazim and Phenmedipham, for the compostworm Eisenia andrei (effects of Carbendazim) and the potworm Enchytraeus crypticus (effects of Phenmedipham) in 12 Mediterranean soils with contrasting soil properties. The observed toxicity was also compared to that obtained for OECD standard soil, used as a control.

Materials and methods

The soils were selected to be representative for the Mediterranean region and to cover a broad range of soil properties. The evaluated endpoints were avoidance behavior and reproduction. Soils were also assembled in two groups according to their pedological properties.

Results and discussion

Toxicity benchmarks (AC50s) obtained for E. andrei avoidance behavior in carbendazim-contaminated soils were generally higher for sandy soils with low pH. The toxic effects on the reproduction of the compostworms were similar in the six tested soils, indicating a low influence of soil properties. The avoidance response of E. crypticus towards Phenmedipham was generally highly variable in all tested soils. Even though, a higher toxicity was observed for more acidic soils. The EC50s for reproduction of the latter species varied by a factor of 9 and Phenmedipham toxicity also tended to be increasing in soils with lower pH, except for the soils with extreme organic matter content (0.6 and 5.8%).


A soil effect on chemical toxicity was clearly confirmed, highlighting the influence that test soils can have in site-specific ecological risk assessment. Despite some relationships between soil properties and toxicity were outlined, a clear and statistically significant prediction of chemical toxicity could not be established. The range of soil properties was probably narrow to give clearer and more consistent insights on their influence. For the four groups of tests, the toxicity observed for OECD soil was either similar, lower, or generally higher if compared with Mediterranean soils. Moreover, it did represent neither the organic matter content found in Mediterranean soils nor their textural classes.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Achazi RK, Chroszcz G, Pilz B, Rothe B, Steudel I, Throl C (1996) Der Einfluss des pH-Werts und von PCB52 auf Reproduktion und Besiedlungsaktivität von terrestrischen Enchytraeen in PAK-, PCB- und schwermetallbelasteten Rieselfeldböden. Verh Ges Ö kol 26:37–42

  2. Achazi RK, Fröhlich E, Henneken M, Pilz C (1999) The effect of soil from former irrigation fields and of sewage sludge on dispersal activity and colonizing success of the annelid Enchytraeus crypticus Westheide & Graefe, 1992 (Enchytraeidae, Oligochaeta). Newsl Enchytraeidae 6:117–126

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

  4. Amorim MJB, Römbke J, Scheffczyk A, Soares AMVM (2005b) Effect of different soil types on the enchytraeids Enchytraeus albidus and Enchytraeus luxuriosus using the herbicide Phenmedipham. Chemosphere 61:1102–1114

  5. Amorim MJB, Römbke J, Schallnaβ H-J, Soares AMVM (2005c) Effect of soil properties and aging on the toxicity of copper for Enchytraeus albidus, Enchytraeus luxuriosus and Folsomia candida. Environ Toxicol Chem 24:1875–1885

  6. Amorim MJB, Rombke J, Scheffczyk A, Nogueira AJA, Soares AMVM (2005d) Effects of different soil types on the collembolans Folsomia candida and Hypogastrura assimilis using the herbicide phenmedipham. Arch Environ Contam Toxicol 49:343–352

  7. Amorim MJB, Novais S, Rombke J, Soares AMVM (2008a) Avoidance test with Enchytraeus albidus (Enchytraeidae): effects of different exposure time and soil properties. Environ Pollut 155:112–116

  8. Amorim MJB, Novais S, Römbke J, Soares AMVM (2008b) Enchytraeus albidus (Enchytraeidae): a test organism in a standardised avoidance test? Effects of different chemical substances. Environ Int 34:363–371

  9. Arias-Estevez M, Lopez-Periago E, Martinez-Carballo E et al (2008) The mobility and degradation of pesticides in soils and the pollution of groundwater resources. Agric Ecosyst Environ 123:247–260

  10. Autio S, Siimes K, Laitinen P et al (2004) Adsorption of sugar beet herbicides to Finnish soils. Chemosphere 55:215–226

  11. Berglöf T, Van Dung T, Kylin H, Nilsson I (2002) Carbendazim sorption–desorption in Vietnamese soils. Chemosphere 48:267–273

  12. Breitholtz M, Rudén C, Hansson SO, Bengtsson B-E (2006) Ten challenges for improved ecotoxicological testing in environmental risk assessment. Ecotoxicol Environ Saf 63:324–335

  13. Chelinho S, Domene X, Campana P et al (2011) Improving ecological risk assessment in the Mediterranean area: selection of reference soils and evaluating the influence of soil properties on avoidance and reproduction of two oligochaete species. Environ Toxicol Chem 30:1050–1058

  14. Daam MA, Silva E, Leitão S, Cerejeira MJ (2011) Environmental risk assessment of pesticides in mediterranean portugal: status and research needs. In: Daniels JA (ed) Advances in environmental research, vol 9. Nova Science Publishers, Inc., New York, pp 213–238

  15. De Silva PMCS, Van Gestel CAM (2009) Comparative sensitivity of Eisenia andrei and Perionyx excavatus in earthworm avoidance tests using two soil types in the tropics. Chemosphere 77:1609–1613

  16. De Silva PMCS, Pathiratne A, Van Gestel CAM (2009) Influence of temperature and soil type on the toxicity of three pesticides to Eisenia andrei. Chemosphere 76:1410–1415

  17. Didden W, Römbke J (2001) Enchytraeids as indicator organisms for chemical stress in terrestrial ecosystems. Ecotoxicol Environ Saf 50:25–43

  18. Dios Cancela G, Romero Taboada E, Sanchez-Rasero F (1992) Carbendazim adsorption on montmorillonite, peat and soils. J Soil Sci 43:99–111

  19. Domene X, Colón J, Uras MV et al (2010) Role of soil properties in sewage sludge toxicity to soil collembolans. Soil Biol Biochem 42:1982–1990

  20. Domene X, Chelinho S, Campana P et al (2012) Applying a GLM-based approach to model the influence of soil properties on the toxicity of phenmedipham to Folsomia candida. J Soils Sediments 12:888–899

  21. Ellis SR, Hodson ME, Wege P (2007) The influence of different artificial soil types on the acute toxicity of carbendazim to the earthworm Eisenia fetida in laboratory toxicity tests. Eur J Soil Biol 43:S239–S245

  22. Environment Canada (2007) Biological test method: test for measuring survival and reproduction of springtails exposed to contaminants in soil. Environmental Technology Centre, Report EPS 1/RM/47, Environment Canada, Ottawa, Ontario. http://publications.gc.ca/collections/collection_2012/ec/En49-7-1-47-eng.pdf. Accessed 7 Nov 2013

  23. EPA (2005) Reregistration eligibility decision for phenmedipham, EPA 738-R-05-007 United States Environmental Protection Agency. http://www.epa.gov/oppsrrd1/REDs/phenmedipham_red.pdf. Accessed 13 Sept 2013

  24. European Commission (2004) Review report for the active substance phenmedipham. Health & Consumer Directorate—General. Report SANCO/4060/2001-final. http://ec.europa.eu/comm/food/plant/protection/evaluation/existactive/list_phenmedipham.pdf. Accessed 23 May 2013

  25. European Commission (2005) Soil atlas of Europe, European Soil Bureau Network, Office for Official Publications of the European Communities, L-2995, Luxembourg. http://eusoils.jrc.ec.europa.eu/projects/Soil_Atlas/Download.cfm. Accessed 7 Nov 2013

  26. 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 153:450–456

  27. Hofman J, Hovorkova I, Machat J (2009) Comparison and characterization of OECD artificial soils. In: Moser H, Roembke J (eds) Ecotoxicological characterisation of waste. Results and experiences from an European ring test. Springer, New York, pp 223–229

  28. Hund-Rinke K, Wiechering H (2001) Earthworm avoidance test for soil assessment. J Soils Sediments 1:15–20

  29. International Organization for Standardization (2004) Soil quality—effects of pollutants on Enchytraeidae (Enchytraeus sp.). Determinations of effects on reproduction and survival. ISO 16387. ISO, Geneva

  30. International Organization for Standardization (2008) Avoidance test for determining the quality of soils and effects of chemicals on behaviour. Part 1: Test with earthworms (Eisenia fetida and Eisenia andrei). ISO 17512. ISO, Geneva

  31. International Organization for Standardization (2012) Soil quality—effects of pollutants on earthworms. Part 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei. ISO 11268-2. ISO, Geneva

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

  33. Kördel W, Römbke J (2001) Requirements on physical, chemical and biological testing methods for estimating the quality of soils and soil substrates. J Soils Sediments 1:98–104

  34. Kördel W, Peijnenburg W, Klein CL et al (2009) The reference-matrix concept applied to chemical testing of soils. Trends Anal Chem 28:51–63

  35. Kuhnt G, Vetter L (1999) Field work of EUROSOILS: sampling sites, profile properties and collection procedures. In: Gawlik BM, Muntau H (eds) EUROSOILS II—laboratory reference materials for soil-related studies. European Commission, Luxembourg, pp 41–56 http://bookshop.europa.eu/en/eurosoils-ii-pbCLNA18983/. Accessed 7 Nov 2013

  36. Kuperman RG, Amorim MJB, Römbke J et al (2006) Adaptation of the Enchytraeid toxicity test for use with natural soil types. Eur J Soil Biol 42:S234–S243

  37. Kuperman RG, Checkai RT, Garcia MVB et al (2009) State of the science and the way forward for the ecotoxicological assessment of contaminated land. Pesq Agropec Bras 44:811–824

  38. Liu K, Pan X, Han Y et al (2012) Estimating the toxicity of the weak base carbendazim to the earthworm (Eisenia fetida) using in situ pore water concentrations in different soils. Sci Total Environ 438:26–32

  39. Maliszewska-Kordybach B, Klimkowicz-Pawlas A, Smreczak B (2008) Soil reference materials in ecotoxicity testing—application of the concept of EURO-Soils to soils from Poland. Pol J Environ Stud 17:257–266, http://www.pjoes.com/pdf/17.2/257-266.pdf

  40. Natal-da-Luz T, Amorim MJB, Römbke J, Sousa JP (2008) Avoidance tests with earthworms and springtails: defining the minimum exposure time to observe a significant response. Ecotoxicol Environ Saf 71:545–551

  41. Novais SC, Soares AMVM, Amorim MJB (2010) Can avoidance in Enchytraeus albidus be used as a screening parameter for pesticides testing? Chemosphere 79:233–237

  42. Organisation for Economic Co-Operation and Development (1984) Guideline for testing of chemicals—earthworm acute toxicity tests. OECD Guideline for the testing of chemicals 207, Paris, France http://www.oecd.org/chemicalsafety/risk-assessment/1948293.pdf. Accessed 7 Nov 2013

  43. Organisation for Economic Co-Operation and Development (2008) Predatory mite (Hypoaspis (Geolaelaps) aculeifer) reproduction test in soil. OECD guideline for the testing of chemicals 226, Paris, France http://www.oecd.org/chemicalsafety/testing/38972970.pdf. Accessed 7 Nov 2013

  44. Paszko T (2012) Effect of pH on the adsorption of carbendazim in Polish mineral soils. Sci Total Environ 435–436:222–229

  45. Peijnenburg WJGM, Baerselman R, Groot AC et al (1999) Relating environmental availability to bioavailability: soil-type-dependent metal accumulation in the Oligochaete Eisenia andrei. Ecotoxicol Environ Saf 44:294–310

  46. Ramos C, Carbonell G, Baudýn JMG, Tarazona JV (2000) Ecological risk assessment of pesticides in the Mediterranean region. The need for crop-specific scenarios. Sci Total Environ 247:269–278

  47. Roberts TR, Hutson DH, Philip WL, Nicholls PH, Plimmer JR (eds) (1998) Metabolic pathways of agrochemicals: herbicides and plant-growth regulators. The Royal Society of Chemistry. Bookcraft, Stroud

  48. Rocheleau S, Kuperman RG, Simini M et al (2010) Toxicity of 2,4-dinitrotoluene to terrestrial plants in natural soils. Sci Total Environ 408:3193–3199

  49. Römbke J, Amorim M (2004) Tackling the heterogeneity of soils in ecotoxicological testing: an EURO-soil based approach. J Soils Sediments 4:276–281

  50. Römbke J, Moser T (2002) Validating the enchytraeid reproduction test: organisation and results of an international ringtest. Chemosphere 46:1117–1140

  51. Römbke J, Jänsch S, Junker T et al (2006) Improvement of the applicability of ecotoxicological tests with earthworms, springtails and plants for the assessment of metals in natural soils. Environ Toxicol Chem 25:776–787

  52. Römbke J, Jänsch S, Junker T et al (2007) The effect of tributyltin-oxide on earthworms, springtails, and plants in artificial and natural soils. Arch Environ Contam Toxicol 52:525–534

  53. Sakuma M (1998) PriProbitNM 1.63. http://www.ars.usda.gov/Services/docs.htm?docid=11284. Accessed 23 May 2013

  54. Schäfer RK (2001) Evaluation of the ecotoxicological threat of ammunition derived compounds to the habitat function of soil. Ph.D. Dissertation, Freien Universität Berlin, Germany. http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000000563. Accessed 7 Nov 2013

  55. Sijm D, Kraaij R, Belfroid A (2000) Bioavailability in soil or sediment: exposure of different organisms and approaches to study it. Environ Pollut 108:113–119

  56. Smit CE, Van Gestel CAM (1998) Effects of soil type, prepercolation, and ageing on bioaccumulation and toxicity of zinc for the springtail Folsomia candida. Environ Toxicol Chem 17:1132–1141

  57. StatSoft Inc (2004) STATISTICA (data analysis software system), version 7.0. www.statsoft.com. Accessed 7 Nov 2013

  58. Stephenson GL, Koper N, Atkinson GF et al (2000) Use of nonlinear regression techniques for describing concentration–response relationships of plant species exposed to contaminated site soils. Environ Toxicol Chem 19:2968–2981

  59. Ter Braak CJF, Smilauer P (2002) CANOCO reference manual and CanoDraw for Windows users guide: software for canonical community ordination (version 4.5). Microcomputer Power, Ithaca

  60. Tibberg E (1998) Nordic reference soils: 1. Characterisation and classification of 13 typical nordic soils; 2. Sorption of 2,4-d, atrazine and glyphosate. TemaNord 537, Nordic Council of Ministers, Copenhagen

  61. Van Gestel CAM (2012) Soil ecotoxicology: state of the art and future directions. ZooKeys 296(176):275–296

  62. Van Gestel CAM, Weeks JM (2004) Recommendations of the 3rd International Workshop on Earthworm Ecotoxicology, Aarhus, Denmark, August 2001. Ecotoxicol Environ Saf 57:100–105

  63. Van Gestel CAM, Breemen EMD-V, Baerselman R et al (1992) Comparison of sublethal and lethal criteria for nine different chemicals in standardized toxicity tests using the earthworm Eisenia andrei. Ecotoxicol Environ Saf 23:206–220

  64. Westheide W, Graefe U (1992) Two new terrestrial Enchytraeus species (Oligochaeta, Annelida). J Nat Hist 26:479–488

  65. Zar JH (1999) Biostatistical analysis. Prentice Hall International, London

Download references


This study has been funded by Fundação para a Ciência e Tecnologia–Portugal (grant to Sónia Chelinho—SFRH/BM/18844/2004) and by CRUP—Acções Integradas Luso-Espanholas (Action nr. E-5/2005), the LODOTOX project of the Spanish Ministry of Science and Technology (AGL2002- 03297). The authors would like to thank the following institutions for the facilities in the Portuguese soils selection/collection: ADPM—Mértola, Companhia das Lezírias, Escola Superior Agrária de Beja, LPN—Liga para a Protecção da Natureza and Universidade de Évora.

Author information

Correspondence to Sonia Chelinho.

Additional information

Responsible editor: Winfried Schroeder

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Chelinho, S., Domene, X., Campana, P. et al. Toxicity of phenmedipham and carbendazim to Enchytraeus crypticus and Eisenia andrei (Oligochaeta) in Mediterranean soils. J Soils Sediments 14, 584–599 (2014). https://doi.org/10.1007/s11368-013-0818-8

Download citation


  • Carbendazim
  • Compost worms
  • Phenmedipham
  • Potworms avoidance test
  • Reproduction test
  • Soil properties