Environmental Science and Pollution Research

, Volume 25, Issue 14, pp 13868–13880 | Cite as

Ecotoxicological impacts of surface water and wastewater from conventional and advanced treatment technologies on brood size, larval length, and cytochrome P450 (35A3) expression in Caenorhabditis elegans

  • Aennes AbbasEmail author
  • Lucie Valek
  • Ilona Schneider
  • Anna Bollmann
  • Gregor Knopp
  • Wolfram Seitz
  • Ulrike Schulte-Oehlmann
  • Jörg Oehlmann
  • Martin Wagner
Research Article


Anthropogenic micropollutants and transformation products (TPs) negatively affect aquatic ecosystems and water resources. Wastewater treatment plants (WWTP) represent major point sources for (micro)pollutants and TPs in urban water cycles. The aim of the current study was to assess the removal of micropollutants and toxicity during conventional and advanced wastewater treatment. Using wild-type and transgenic Caenorhabditis elegans, the endpoint reproduction, growth, and cytochrome P450 (CYP) 35A3 induction (via cyp-35A3::GFP) were assessed. Samples were collected at four WWTPs and a receiving surface water. One WWTP included the advanced treatments: ozonation followed by granular activated carbon (GAC) or biological filtration (BF), respectively. Relevant micropollutants and WWTP parameters (n = 111) were included. Significant reproductive toxicity was detected for one WWTP effluent (31–83% reduced brood size). Three of four effluents significantly promoted the growth of C. elegans larvae (49–55% increased lengths). This effect was also observed for the GAC (34–41%) and BF (30%) post-treatments. Markedly, significant cyp-35A3::GFP induction was detected for one effluent before and after ozonation, being more pronounced for the ozonated samples (5- and 7.4-fold above controls). While the advanced treatments decreased the concentrations of most micropollutants, the observed effects may be attributed to effects of residual target compounds and/or compounds not included in the target chemical analysis. This highlights the need for an integrated assessment of (advanced) wastewater treatment covering both biological and chemical parameters.


Municipal effluents Contaminants of emerging concern CYP biomarker Persistent organic pollutants (POPs) Toxic effects Transformation products In vivo bioassay Ozonation 



This work was partly supported by the German Federal Ministry of Education and Research (BMBF) within the project TransRisk [02WRS1275A] which is gratefully appreciated. The authors further thank Ralph Menzel (Humboldt University Berlin), Wolfgang Ahlf (Technical University Hamburg-Harburg), and all TransRisk project partners for the fruitful discussions and collaboration that greatly helped to improve this manuscript. We also thank the Caenorhabditis Genetics Center, funded by the National Institutes of Health Office of Research Infrastructure Programs [P40 OD010440] (USA), for supplying the Caenorhabditis elegans N2 and Escherichia coli OP50 strain.

Supplementary material

11356_2018_1605_MOESM1_ESM.pdf (348 kb)
ESM 1 (PDF 347 kb)
11356_2018_1605_MOESM2_ESM.pdf (524 kb)
ESM 2 (PDF 523 kb)
11356_2018_1605_MOESM3_ESM.pdf (687 kb)
ESM 3 (PDF 686 kb)
11356_2018_1605_MOESM4_ESM.pdf (392 kb)
ESM 4 (PDF 392 kb)
11356_2018_1605_MOESM5_ESM.pdf (351 kb)
ESM 5 (PDF 351 kb)
11356_2018_1605_MOESM6_ESM.pdf (386 kb)
ESM 6 (PDF 386 kb)
11356_2018_1605_MOESM7_ESM.pdf (391 kb)
ESM 7 (PDF 391 kb)
11356_2018_1605_MOESM8_ESM.pdf (368 kb)
ESM 8 (PDF 368 kb)
11356_2018_1605_MOESM9_ESM.pdf (166 kb)
ESM 9 (PDF 165 kb)


  1. Allard P, Kleinstreuer NC, Knudsen TB, Colaiácovo MP (2013) A C. elegans screening platform for the rapid assessment of chemical disruption of germline function. Environ Health Perspect 121(6):717–724CrossRefGoogle Scholar
  2. Anbalagan C, Lafayette I, Antoniou-Kourounioti M, Gutierrez C, Martin JR, Chowdhuri DK, De Pomerai DI (2013) Use of transgenic GFP reporter strains of the nematode Caenorhabditis elegans to investigate the patterns of stress responses induced by pesticides and by organic extracts from agricultural soils. Ecotoxicology 22:72–85CrossRefGoogle Scholar
  3. Boyd WA, McBride SJ, Rice JR, Snyder DW, Freedman JH (2010) A high throughput method for assessing chemical toxicity using a Caenorhabditis elegans reproduction assay. Toxicol Appl Pharmacol 245(2):153–159CrossRefGoogle Scholar
  4. Burton GA, Pitt R, Clark S (2000) The role of traditional and novel toxicity test methods in assessing stormwater and sediment contamination. Crit Rev Environ Sci Technol 30(4):413–447CrossRefGoogle Scholar
  5. Caylor RC, Jin Y, Ackley BD (2013) The Caenorhabditis elegans voltage-gated calcium channel subunits UNC-2 and UNC-36 and the calcium-dependent kinase UNC-43/CaMKII regulate neuromuscular junction morphology. Neural Dev 8:10CrossRefGoogle Scholar
  6. Chalew TE, Halden RU (2009) Environmental exposure of aquatic and terrestrial biota to triclosan and triclocarban. J Am Water Works Assoc 45(1):4–13CrossRefGoogle Scholar
  7. Eom HJ, Kim H, Kim BM, Chon TS, Choi J (2014) Integrative assessment of benzene exposure to Caenorhabditis elegans using computational behavior and toxicogenomic analyses. Environ Sci Technol 48(14):8143–8151CrossRefGoogle Scholar
  8. European Commission (EC) (2000) Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 establishing a framework for community action in the field of water policy. EU Water Framework Directive Off J Eur Union (OJ) L 327:1–73Google Scholar
  9. Félix MA, Braendle C (2010) The natural history of Caenorhabditis elegans. Curr Biol 20(22):965–969CrossRefGoogle Scholar
  10. Forsgren AJ (2015) Wastewater treatment: occurrence and fate of polycyclic aromatic hydrocarbons (PAHs). CRC Press, Boca RatonGoogle Scholar
  11. Giebner S, Ostermann S, Straskraba S, Oetken M, Oehlmann J, Wagner M (2016) Effectivity of advanced wastewater treatment: reduction of in vitro endocrine activity and mutagenicity but not of in vivo reproductive toxicity. Environ Sci Pollut Res Int 25:3965–3976. CrossRefGoogle Scholar
  12. Haegerbaeumer A, Höss S, Heininger P, Traunspurger W (2018) Is Caenorhabditis elegans representative of freshwater nematode species in toxicity testing? Environ Sci Pollut Res 25(3):2879–2888CrossRefGoogle Scholar
  13. Hägerbäumer A, Höss S, Heininger P, Traunspurger W (2015) Experimental studies with nematodes in ecotoxicology: an overview. J Nematol 47(1):11–27Google Scholar
  14. Hartman PS, Freedman M (2005) Getting Off on DEET: C. elegans as a model system to study an insect repellent. Conference-Abstract. International Worm Meeting, 2005Google Scholar
  15. Hitchcock DR, Black MC, Williams PL (1997) Investigations into using the nematode Caenorhabditis elegans for municipal and industrial wastewater toxicity testing. Arch Environ Contam Toxicol 33:252–260CrossRefGoogle Scholar
  16. Hitchcock DR, Law SE, Wu J, Williams PL (1998) Determining toxicity trends in the ozonation of synthetic dye wastewaters using the nematode Caenorhabditis elegans. Arch Environ Contam Toxicol 34:259–264CrossRefGoogle Scholar
  17. Höss S, Weltje L (2007) Endocrine disruption in nematodes: effects and mechanisms. Ecotoxicology 16(1):15–28CrossRefGoogle Scholar
  18. Höss S, Bergtold M, Haitzer M, Traunspurger W, Steinberg CE (2001) Refractory dissolved organic matter can influence the reproduction of Caenorhabditis elegans (Nematoda). Freshw Biol 46:1–10CrossRefGoogle Scholar
  19. Höss S, Ahlf W, Bergtold M, Bluebaum-Gronau E, Brinke M, Donnevert G, Menzel R, Möhlenkamp C, Ratte HT, Traunspurger W, von Danwitz B, Pluta HJ (2012) Interlaboratory comparison of a standardized toxicity test using the nematode Caenorhabditis elegans (ISO 10872). Environ Toxicol Chem 31(7):1525–1535CrossRefGoogle Scholar
  20. Inokuchi A, Nihira M, Minakoshi M, Yamamoto R, Ishibashi H, Tominaga N, Arizono K (2014) Comparative study of the biological effects of antimicrobials, triclosan and trichlocarban, for C. elegans. J Environ Safety 5(2):95–98Google Scholar
  21. Jones LM, Rayson SJ, Flemming AJ, Urwin PE (2013) Adaptive and specialised transcriptional responses to xenobiotic stress in Caenorhabditis elegans are regulated by nuclear hormone receptors. PLoS One 8(7):e69956CrossRefGoogle Scholar
  22. Kao A, Liu R, Gupta D, Feng Z (2016) Amantadine alleviates the motor and dopamine receptor changes in a levodopa-induced dyskinesia (LID) model of C. elegans (P5.373). Neurology 86(16 Supplement):P5.373Google Scholar
  23. Knopp G, Prasse C, Ternes TA, Cornel P (2016) Elimination of micropollutants and transformation products from a wastewater treatment plant effluent through pilot scale ozonation followed by various activated carbon and biological filters. Water Res 100:580–592CrossRefGoogle Scholar
  24. Leung MCK, Williams PL, Benedetto A, Au C, Helmcke KJ, Aschner M, Meyer JN (2008) Caenorhabditis elegans: an emerging model in biomedical and environmental toxicology. Toxicol Sci 106(1):5–28CrossRefGoogle Scholar
  25. Leung MCK, Goldstone J, Boyd W, Freedman J, Meyer J (2010) Caenorhabditis elegans generates biologically relevant levels of genotoxic metabolites from aflatoxin B1 but not benzo[a]pyrene in vivo. Toxicol Sci 118(2):444–453CrossRefGoogle Scholar
  26. Lindblom TH, Dodd AK (2006) Xenobiotic detoxification in the nematode Caenorhabditis elegans. J Exp Zool A Comp Exp Biol 305((9):720–730. CrossRefGoogle Scholar
  27. Liu S, Saul N, Pan B, Menzel R, Steinberg CE (2013) The non-target organism Caenorhabditis elegans withstands the impact of sulfamethoxazole. Chemosphere 93(10):2373–2380CrossRefGoogle Scholar
  28. Loos R, Carvalho R, António DC, Comero S, Locoro G, Tavazzi S, Paracchini B, Ghiani M, Lettieri T, Blaha L, Jarosova B, Voorspoels S, Servaes K, Haglund P, Fick J, Lindberg RH, Schwesig D, Gawlik BM (2013) EU wide monitoring survey on waste water treatment plant effluents. Water Res 47(17):6475–6487CrossRefGoogle Scholar
  29. Magdeburg A, Stalter D, Oehlmann J (2012) Whole effluent toxicity assessment at a wastewater treatment plant upgraded with a full-scale post-ozonation using aquatic key species. Chemosphere 88:1008–1014CrossRefGoogle Scholar
  30. McLaggan D, Amezaga MR, Petra E, Frost A, Duff EI, Rhind SM, Fowler PA, Glover LA, Lagido C (2012) Impact of sublethal levels of environmental pollutants found in sewage sludge on a novel Caenorhabditis elegans model biosensor. PLoS One 7(10):e46503CrossRefGoogle Scholar
  31. Menzel R, Bogaert T, Achazi R (2001) A systematic gene expression screen of Caenorhabditis elegans cytochrome P450 genes reveals CYP35 as strongly xenobiotic inducible. Arch Biochem Biophys 395:158–168CrossRefGoogle Scholar
  32. Menzel R, Rodel M, Kulas J, Steinberg CE (2005) CYP35: xenobiotically induced gene expression in the nematode Caenorhabditis elegans. Arch Biochem Biophys 438:93–102CrossRefGoogle Scholar
  33. Menzel R, Yeo HL, Rienau S, Li S, Steinberg CE, Sturzenbaum SR (2007) Cytochrome P450s and short-chain dehydrogenases mediate the toxicogenomic response of PCB52 in the nematode Caenorhabditis elegans. J Mol Biol 370:1–13CrossRefGoogle Scholar
  34. Menzel R, Swain SC, Höss S, Claus E, Menzel S, Steinberg CE, Reifferscheid G, Stürzenbaum SR (2009) Gene expression profiling to characterize sediment toxicity—a pilot study using Caenorhabditis elegans whole genome microarrays. BMC Genomics 10:160–174CrossRefGoogle Scholar
  35. Min H, Kawasaki I, Gong J, Shim YH (2015) Caffeine induces high expression of cyp-35A family genes and inhibits the early larval development in Caenorhabditis elegans. Mol Cells 38(3):236–242CrossRefGoogle Scholar
  36. Offermann K, Matthäi A, Ahlf W (2009) Assessing the importance of dietborne cadmium and particle characteristics on bioavailability and bioaccumulation in the nematode Caenorhabditis elegans. Environ Toxicol Chem 28(6):1149–1158CrossRefGoogle Scholar
  37. Peris-Vicente J, Roca-Genovés P, Tayeb-Cherif K, Esteve-Romero J (2016) Development and validation of a method to determine thiabendazole and o-phenylphenol in wastewater using micellar liquid chromatography-fluorescence detection. Electrophoresis 37(19):2517–2521CrossRefGoogle Scholar
  38. Peter E, Candido M, Jones D (1996) Transgenic Caenorhabditis elegans strains as biosensors. Trends Biotechnol 14(4):125–129CrossRefGoogle Scholar
  39. Petersen CI, McFarland TR, Stepanovic SZ, Yang P, Reiner DJ, Hayashi K, George AL, Roden DM, Thomas JH, Balser JR (2004) In vivo identification of genes that modify ether-a-go-go-related gene activity in Caenorhabditis elegans may also affect human cardiac arrhythmia. Proc Natl Acad Sci 101(32):11773–11778CrossRefGoogle Scholar
  40. Petrie B, Barden R, Kasprzyk-Hordern B (2015) A review on emerging contaminants in wastewaters and the environment: current knowledge, understudied areas and recommendations for future monitoring. Water Res 72:3–27CrossRefGoogle Scholar
  41. Prasse C, Stalter D, Schulte-Oehlmann U, Oehlmann J, Ternes TA (2015) Spoilt for choice: a critical review on the chemical and biological assessment of current wastewater treatment technologies. Water Res 87:237–270CrossRefGoogle Scholar
  42. Quevauviller P, Thomas O, Van Der Beken A (2007) Wastewater quality monitoring and treatment, vol 13. Wiley, HobokenGoogle Scholar
  43. Reichert K, Menzel R (2005) Expression profiling of five different xenobiotics using a Caenorhabditis elegans whole genome microarray. Chemosphere 61:229–237CrossRefGoogle Scholar
  44. Ristau K, Akgül Y, Bartel AS, Fremming J, Müller MT, Reiher L, Stapela F, Splett JP, Spann N (2015) Toxicity in relation to mode of action for the nematode Caenorhabditis elegans: acute-to-chronic ratios and quantitative structure–activity relationships. Environ Toxicol Chem 34(10):2347–2353CrossRefGoogle Scholar
  45. Roh J, Lee H, Kwon J (2014) Changes in the expression of cyp35a family genes in the soil nematode Caenorhabditis elegans under controlled exposure to chlorpyrifos using passive dosing. Environ Sci Technol 48(17):10475–10481CrossRefGoogle Scholar
  46. Seitz W, Winzenbacher R (2017) A survey on trace organic chemicals in a German water protection area and the proposal of relevant indicators for anthropogenic influences. Environ Monit Assess 189(6):244CrossRefGoogle Scholar
  47. Sinclair CJ, Boxall AB (2003) Assessing the ecotoxicity of pesticide transformation products. Environ Sci Technol 37(20):4617–4625CrossRefGoogle Scholar
  48. Stalter D, Peters LI, O'Malley E, Tang JY, Revalor M, Farré MJ, Watson K, von Gunten U, Escher BI (2016) Sample enrichment for bioanalytical assessment of disinfected drinking water: concentrating the polar, the volatiles, and the unknowns. Environ Sci Technol 50(12):6495–6505CrossRefGoogle Scholar
  49. Tang JY, Busetti F, Charrois JW, Escher BI (2014) Which chemicals drive biological effects in wastewater and recycled water? Water Res 60:289–299CrossRefGoogle Scholar
  50. Völker J, Vogt T, Castronovo S, Wick A, Ternes TA, Joss A, Oehlmann J, Wagner M (2017) Extended anaerobic conditions in the biological wastewater treatment: higher reduction of toxicity compared to target organic micropollutants. Water Res 116:220–230CrossRefGoogle Scholar
  51. Wernersson AS, Carere M, Maggi C, Tusil P, Soldan P, James A, Sanchez W, Dulio V, Broeg K, Reifferscheid G, Buchinger S, Maas H, Van Der Grinten E, O’Toole S, Ausili A, Manfra L, Marziali L, Polesello S, Lacchetti I, Mancini L, Lilja K, Linderoth M, Lundeberg T, Fjällborg B, Porsbring T, Larsson J, Bengtsson-Palme J, Förlin L, Kienle C, Kunz P, Vermeirssen E, Werner I, Robinson CD, Lyons B, Katsiadaki I, Whalley C, den Haan K, Messiaen M, Clayton H, Lettieri T, Negrão Carvalho R, Gawlik BM, Hollert H, Di Paolo C, Brack W, Kammann U, Kase R (2015) The European technical report on aquatic effect-based monitoring tools under the water framework directive. Environ Sci Eur 27(1):1–11CrossRefGoogle Scholar
  52. Wilson MJ, Khakouli-Duarte T (2009) Nematodes as environmental indicators. CABI, WallingfordCrossRefGoogle Scholar
  53. Xiong H, Pears C, Woollard A (2017) An enhanced C. elegans based platform for toxicity assessment. Sci Rep 7(1):9839CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Ecology, Diversity and EvolutionGoethe Universität FrankfurtFrankfurtGermany
  2. 2.Department of Clinical PharmacologyGoethe-University HospitalFrankfurtGermany
  3. 3.Zweckverband LandeswasserversorgungLangenauGermany
  4. 4.Department of Wastewater Technology and Water ReuseTechnische Universität DarmstadtDarmstadtGermany
  5. 5.Department of BiologyNorwegian University of Science and TechnologyTrondheimNorway

Personalised recommendations