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Validation of the OECD reproduction test guideline with the New Zealand mudsnail Potamopyrgus antipodarum using trenbolone and prochloraz

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Abstract

The Organisation for Economic Cooperation and Development (OECD) provides several standard test methods for the environmental hazard assessment of chemicals, mainly based on primary producers, arthropods, and fish. In April 2016, two new test guidelines with two mollusc species representing different reproductive strategies were approved by OECD member countries. One test guideline describes a 28-day reproduction test with the parthenogenetic New Zealand mudsnail Potamopyrgus antipodarum. The main endpoint of the test is reproduction, reflected by the embryo number in the brood pouch per female. The development of a new OECD test guideline involves several phases including inter-laboratory validation studies to demonstrate the robustness of the proposed test design and the reproducibility of the test results. Therefore, a ring test of the reproduction test with P. antipodarum was conducted including eight laboratories with the test substances trenbolone and prochloraz and results are presented here. Most laboratories could meet test validity criteria, thus demonstrating the robustness of the proposed test protocol. Trenbolone did not have an effect on the reproduction of the snails at the tested concentration range (nominal: 10–1000 ng/L). For prochloraz, laboratories produced similar EC10 and NOEC values, showing the inter-laboratory reproducibility of results. The average EC10 and NOEC values for reproduction (with coefficient of variation) were 26.2 µg/L (61.7%) and 29.7 µg/L (32.9%), respectively. This ring test shows that the mudsnail reproduction test is a well-suited tool for use in the chronic aquatic hazard and risk assessment of chemicals.

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References

  • Ankley GT, Jensen KM, Makynen EA, Kahl MD, Korte JJ, Hornung MW, Henry TR, Denny JS, Leino RL, Wilson VS, Cardon MC, Hartig PC, Gray LE (2003) Effects of the androgenic growth promoter 17-β-trenbolone on fecundity and reproductive endocrinology of the fathead minnow. Environ Toxicol Chem 22:1350–1360

    Article  CAS  Google Scholar 

  • Baumann L, Knorr S, Keiter S, Nagel T, Rehberger K, Volz S, Oberrauch S, Schiller V, Fenske M, Holbech H, Segner H, Braunbeck T (2014) Persistence of endocrine disruption in zebrafish (Danio rerio) after discontinued exposure to the androgen 17β-trenbolone. Environ Toxicol Chem 33:2488–2496

    Article  CAS  Google Scholar 

  • Bettin C, Oehlmann J, Stroben E (1996) TBT-induced imposex in marine neogastropods is mediated by an increasing androgen level. Helgolander Meeresun 50:299–317

    Article  Google Scholar 

  • Charles S, Ducrot V, Azam D, Benstead R, Brettschneider D, De Schamphelaere K, Filipe Goncalves S, Green JW, Holbech H, Hutchinson TH, Faber D, Laranjeiro F, Matthiessen P, Norrgren L, Oehlmann J, Reategui-Zirena E, Seeland-Fremer A, Teigeler M, Thome J-P, Tobor Kaplon M, Weltje L, Lagadic L (2016) Optimaizing the design of a reproduction toxicity test with the pond snail Lymnaea stagnalis. Regul Toxicol Pharm 81:47–56

    Article  CAS  Google Scholar 

  • Christensen ER, Kusk KO, Nyholm N (2009) Dose–response regressions for algal growth and similar continuous endpoints: calculation of effective concentrations. Environ Toxicol Chem 28:826–835

    Article  CAS  Google Scholar 

  • Ducrot V, Askem C, Azam D, Brettschneider D, Brown R, Charles S, Coke M, Collinet M, Delignette-Muller M-L, Forfait-Dubuc C, Holbech H, Hutchinson T, Jach A, Kinnberg KL, Lacoste C, Le Page G, Matthiessen P, Oehlmann J, Rice L, Roberts E, Ruppert K, Davis JE, Veauvy C, Weltje L, Wortham R, Lagadic L (2014) Development and validation of an OECD reproductive toxicity test guideline with the pond snail Lymnaea stagnalis (Mollusca, Gastropoda). Regul Toxicol Pharm 70:605–614

    Article  CAS  Google Scholar 

  • Duft M, Schmitt C, Bachmann J, Brandelik C, Schulte-Oehlmann U, Oehlmann J (2007) Prosobranch snails as test organisms for the assessment of endocrine active chemicals - an overview and a guideline proposal for a reproduction test with the freshwater mudsnail Potamopyrgus antipodarum. Ecotoxicology 16:169–182

    Article  CAS  Google Scholar 

  • EFSA (2011) Conclusion on the peer review of the pesticide risk assessment of the active substance prochloraz. European food saftey authority. EFSA J 2011 9:2323

    Google Scholar 

  • Fretter V, Graham A (1994) British prosobranch mollusc. Their functional anatomy and ecology. The Ray Society, London, England

    Google Scholar 

  • Geiß C, Ruppert K, Heidelbach T, Oehlmann J (2016) The antimicrobial agents triclocarban and triclosan as potent modulators of reproduction in Potamopyrgus antipodarum (Mollusca: Hydrobiidae). J Environ Sci Heal A 51:1173–1179

    Article  Google Scholar 

  • Gourmelon A, Ahtiainen J (2007) Developing test guidelines on invertebrate development and reproduction for the assessment of chemicals, including potential endocrine active substances - the OECD perspective. Ecotoxicology 16:161–167

    Article  CAS  Google Scholar 

  • Gray LE, Wilson VS, Stoker T, Lambright C, Furr J, Noriega N, Howdeshell K, Ankley GT, Guillette L (2006) Adverse effects of environmental antiandrogens and androgens on reproductive development in mammals. Int J Androl 29:96–104

    Article  CAS  Google Scholar 

  • Green J, Wheeler JR (2013) The use of carrier solvents in regulatory aquatic toxicology testing: practical, statistical and regulatory considerations. Aquat Toxicol 144-145:242–249

    Article  CAS  Google Scholar 

  • Gruner H-E (1993) Lehrbuch der speziellen Zoologie, Band 1: Wirbellose Tiere, 3. Teil: Mollusca, Echiurida, Annelida, Onychophora, Tardigrada, Pentastomida. Gustav Fischer, Jena, Stuttgart, New York

    Google Scholar 

  • Gust M, Garric J, Giamberini L, Mons R, Abbaci K, Garnier F, Buronfosse T (2010) Sensitivity of New Zealand mudsnail Potamopyrgus antipodarum (Gray) to a specific aromatase inhibitor. Chemosphere 79:47–53

    Article  CAS  Google Scholar 

  • Hauser L, Carvalho GR, Hughes RN, Carter RE (1992) Clonal structure of the introduced freshwater snail Potamopyrgus antipodarum (Prosobranchia, Hydrobiidae), as revealed by DNA fingerprinting. P Roy Soc B-Biol Sci 249:19–25

    Article  CAS  Google Scholar 

  • Henry MJ, Sisler HD (1984) Effects of sterol biosynthesis-inhibiting (SBI) fungicides on cytochrome P-450 oxygenations in fungi. Pestic Biochem Phys 22:262–275

    Article  CAS  Google Scholar 

  • Holbech H, Kinnberg K, Petersen GI, Jackson P, Hylland K, Norrgren L, Bjerregaard P (2006) Detection of endocrine disrupters: evaluation of a fish sexual development test (FSDT). Comp Biochem Physiol C-Toxicol Pharmacol 144:57–66

    Article  Google Scholar 

  • Hornung MW, Jensen KM, Korte JJ, Kahl MD, Durhan EJ, Denny JS, Henry TR, Ankley GT (2004) Mechanistic basis for estrogenic effects in fathead minnow (Pimephales promelas) following exposure to the androgen 17α-methyltestosterone: conversion of 17α-methyltestosterone to 17α-methylestradiol. Aquat Toxicol 66:15–23

    Article  CAS  Google Scholar 

  • Jacobsen R, Forbes VE (1997) Clonal variation in life-history traits and feeding rates in the gastropod, Potamopyrgus antipodarum: performance across a salinity gradient. Funct Ecol 11:260–267

    Article  Google Scholar 

  • Janer G, Bachmann J, Oehlmann J, Schulte-Oehlmann U, Porte C (2006a) The effect of organotin compounds on gender specific androstenedione metabolism in the freshwater ramshorn snail Marisa cornuarietis. J Steroid Biochem Mol Biol 99:147–156

    Article  CAS  Google Scholar 

  • Janer G, Lyssimachou A, Bachmann J, Oehlmann J, Schulte-Oehlmann U, Porte C (2006b) Sexual dimorphism in esterified steroid levels in the gastropod Marisa cornuarietis: the effect of xenoandrogenic compounds. Steroids 71:435–444

    Article  CAS  Google Scholar 

  • Jobling S, Casey D, Rodgers-Gray T, Oehlmann J, Schulte-Oehlmann U, Pawlowski S, Baunbeck T, Turner AP, Tyler CR (2003) Comparative responses of molluscs and fish to environmental estrogens and an estrogenic effluent. Aquat Toxicol 65:205–220

    Article  CAS  Google Scholar 

  • Kinnberg K, Holbech H, Petersen GI, Bjerregaard P (2007) Effects of the fungicide prochloraz on the sexual development of zebrafish (Danio rerio). Comp Biochem Phys C-Toxicol Pharmacol 145:165–170

    Article  Google Scholar 

  • Lively CM (1987) Evidence from a New-Zealand snail for the maintenance of sex by parasitism. Nature 328:519–521

    Article  Google Scholar 

  • Matthiessen P (2008) An assessment of endocrine disruption in mollusks and the potential for developing internationally standardized mollusk life cycle test guidelines. Integr Environ Assess Manag 4:274–284

    Article  CAS  Google Scholar 

  • Matthiessen P, Gibbs PE (1998) Critical appraisal of the evidence for tributyltin-mediated endocrine disruption in mollusks. Environ Toxicol Chem 17:37–43

    Article  CAS  Google Scholar 

  • Matthiessen P, Weltje L (2015) A review of the effects of azole compounds in fish and their possible involvement in masculinization of wild fish populations. Crit Rev Toxicol 45:453–467

    Article  CAS  Google Scholar 

  • McClellan-Green PD (2013) Chapter 6: Endocrine disruption in molluscs: process and testing. In: Matthiessen P (ed) Endocrine disrupters - hazard testing and assessment methods. Wiley, Hoboken, New Jersey, pp 143–184

    Chapter  Google Scholar 

  • Nentwig G, Oetken M, Oehlmann J (2004) Effects of pharmaceuticals on aquatic invertebrates - the example of carbamazepine and clofibric acid. In: Kümmerer K (ed) Pharmaceuticals in the environment. Sources, fate, effects and risks, 2nd edn. Springer, Berlin, Heidelberg, pp 195–208, Chapter 16.

    Google Scholar 

  • OECD (2004) OECD guideline for the testing of chemicals. Sediment-water Chironomid toxicity test using spiked water. Organisation for Economic Co-operation and Development No. 219: Paris, France

  • OECD (2005) Guidance document on the validation and international acceptance of new or updated test methods for harzard assessment. Organisation for Economic Co-operation and Development No. 34: Paris, France

  • OECD (2006a) Report of the initial work towards the validation of the 21-day fish screening assay for the detection of endocrine active substances (phase 1A). Organisation for Economic Co-operation and Development No. 60: Paris, France

  • OECD (2006b) Report of the initial work towards the validation of the 21-day fish screening assay for the detection of endocrine active substances (phase 1B). Organisation for Economic Co-operation and Development No. 61: Paris, France

  • OECD (2007) OECD guideline for the testing of chemicals. Sediment-water Lumbriculus toxicity test using spiked sediment. No. 225. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2010a) Detailed review paper on molluscs life-cycle toxicity testing. No. 121. ENV/JM/MONO(2010)9. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2010b) OECD guideline for the testing of chemicals. Sediment-water Chironomid life-cycle toxicity test using spiked water or sediment. No. 233. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2010c) Validation report of the chironomid full life-cycle toxicity test. No. 136. ENV/JM/MONO(2010)35. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2011) Validation report (phase 1) for the fish sexual development test for the detection of endocrine active substances. Organisation for Economic Co-operation and Development No. 141: Paris, France

  • OECD (2012a) Guidance document on standardised test guidelines for evaluating chemicals for endocrine disruption. Organisation for Economic Co-operation and Development No. 150: Paris, France

  • OECD (2012b) OECD guideline for the testing of chemicals. Daphnia magna reproduction test. No. 211. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2016a) OECD guideline for the testing of chemicals. Potamopyrgus antipodarum reproduction test. No. 242. Organisation for Economic Co-operation and Development: Paris, France

  • OECD (2016b) OECD guideline for the testing of chemicals. Lymnaea stagnalis reproduction test. No. 243. Organisation for Economic Co-operation and Development: Paris, France

  • Oehlmann J, Di Benedetto P, Tillmann M, Duft M, Oetken M, Schulte-Oehlmann U (2007) Endocrine disruption in prosobranch molluscs: evidence and ecological relevance. Ecotoxicology 16:29–43

    Article  CAS  Google Scholar 

  • Olmstead AW, Kosian PA, Johnson R, Blackshear PE, Haselman J, Blanksma C, Korte JJ, Holcombe GW, Burgess E, Lindberg-Livingston A, Bennett BA, Woodis KK, Degitz SJ (2012) Trenbolone causes mortality and altered sexual differentiation in Xenopus tropicalis during larval development. Environ Toxicol Chem 31:2391–2398

    Article  CAS  Google Scholar 

  • Ponder WF (1988) Potamopyrgus antipodarum - a molluscan colonizer of Europe and Australia. J Mollus Stud 54:271–285

    Article  Google Scholar 

  • Robson GC (1923) Parthenogenesis in the mollusc Paludestrina jenkinsi. Br J Exp Biol 1:65–78

    Google Scholar 

  • Ruppert K, Geiß C, Askem C, Benstead R, Brown R, Coke M, Ducrot V, Egeler P, Holbech H, Hutchinson TH, Kinnberg KL, Lagadic L, Le Page G, Lorenz P, Macken A, Matthiessen P, Ostermann S, Planojevic I, Schimera A, Schmitt C, Seeland-Fremer A, Smith A, Weltje L, Oehlmann J (submitted) Development and validation of an OECD reproductive toxicity test guideline with the mudsnail Potamopyrgus antipodarum (Mollusca, Gastropoda).

  • Ruppert K, Geiß C, Ostermann S, Theis C, Oehlmann J (2016) Comparative sensitivity of juvenile and adult Potamopyrgus antipodarum (Mollusca: Hydrobiidae) under chronic exposure to cadmium and tributyltin. J Environ Sci Heal A 51:736–743

    Article  CAS  Google Scholar 

  • Scott AP (2013) Do mollusks use vertebrate sex steroids as reproductive hormones? II. Critical review of the evidence that steroids have biological effects. Steroids 78:268–281

    Article  CAS  Google Scholar 

  • Sieratowicz A, Stange D, Schulte-Oehlmann U, Oehlmann J (2011) Reproductive toxicity of bisphenol A and cadmium in Potamopyrgus antipodarum and modulation of bisphenol A effects by different test temperature. Environ Pollut 159:2766–2774

    Article  CAS  Google Scholar 

  • Städler T, Frye M, Neiman M, Lively CM (2005) Mitochondrial haplotypes and the New Zealand origin of clonal European Potamopyrgus, an invasive aquatic snail. Mol Ecol 14:2465–2473

    Article  Google Scholar 

  • Taenzler V, Bruns E, Dorgerloh M, Pfeifle V, Weltje L (2007) Chironomids: suitable test organisms for risk assessment investigations on the potential endocrine disrupting properties of pesticides. Ecotoxicology 16:221–230

    Article  CAS  Google Scholar 

  • Tassou KT, Schulz R (2009) Effects of the insect growth regulator pyriproxyfen in a two-generation test with Chironomus riparius. Ecotox Environ Safe 72:1058–1062

    Article  CAS  Google Scholar 

  • Thorpe KL, a Marca Pereira ML, Schiffer H, Burkhardt-Holm P, Weber K, Wheeler JR (2011) Mode of sexual differentiation and its influence on the relative sensitivity of the fathead minnow and zebrafish in the fish sexual development test. Aquat Toxicol 105:412–420

    Article  CAS  Google Scholar 

  • Vinggaard AM, Hass U, Dalgaard M, Andersen HR, Bonefeld-JØRgensen EVA, Christiansen S, Laier P, Poulsen ME (2006) Prochloraz: an imidazole fungicide with multiple mechanisms of action. Int J Androl 29:186–192

    Article  CAS  Google Scholar 

  • Wallace C (1979) Notes on the occurrence of males in populations of Potamopyrgus jenkinsi. J Mollus Stud 45:61–67

    Google Scholar 

  • Wilson VS, Lambright C, Ostby J, Gray LE (2002) In vitro and in vivo effects of 17β-trenbolone: a feedlot effluent contaminant. Toxicol Sci 70:202–211

    Article  CAS  Google Scholar 

  • Winterbourn M (1970) The New Zealand species of Potamopyrgus (Gastropoda: Hydrobiidae). Malacologia 10:283–321

    Google Scholar 

  • Yarrow JF, McCoy SC, Borst SE (2010) Tissue selectivity and potential clinical applications of trenbolone (17β-hydroxyestra-4,9,11-trien-3-one): a potent anabolic steroid with reduced androgenic and estrogenic activity. Steroids 75:377–389

    Article  CAS  Google Scholar 

  • Zhang XW, Hecker M, Jones PD, Newsted J, Au D, Kong R, Wu RSS, Giesy JP (2008) Responses of the medaka HPG axis PCR array and reproduction to prochloraz and ketoconazole. Environ Sci Technol 42:6762–6769

    Article  CAS  Google Scholar 

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Acknowledgements

We are grateful to the German Environment Agency (project code 371165417), the United Kingdom’s Department for Environment, Food and Rural Affairs, the Danish Ministry of the Environment and the Spanish Government (project code CTM2013-48194-C3-3-R) for the financial support and to all the laboratories that used their own funds. We would like to thank all participating laboratories for their dedicated work so that the project could be successfully carried out. Furthermore, we thank Bente Frost Holbech (University of Southern Denmark) for performing the chemical analysis within this project. This study was funded by the German Environment Agency (project code 371165417), the United Kingdom’s Department for Environment, Food and Rural Affairs, the Danish Ministry of the Environment and the Spanish Government (project code CTM2013-48194-C3-3-R).

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Authors and Affiliations

  1. Department Aquatic Ecotoxicology, Goethe University Frankfurt am Main, Max-von-Laue-Str. 13, Frankfurt am Main, 60438, Germany

    Cornelia Geiß, Katharina Ruppert & Jörg Oehlmann

  2. Centre for Environment Fisheries and Aquaculture Science Lowestoft Laboratory, Pakefield Road, Lowestoft, NR33 OHT, UK

    Clare Askem & Steve Morris

  3. Department of Biology and CESAM, University of Aveiro, Aveiro, 3810-193, Portugal

    Carlos Barroso & Paula Sanchez-Marin

  4. Bayer CropScience AG, Environmental Safety/Ecotoxicology, Alfred-Nobel-Str. 50, Monheim am Rhein, 40789, Germany

    Daniel Faber

  5. INRA, UMR Ecologie et Santé des Ecosystèmes, Agrocampus Ouest, 65 rue de Saint-Brieuc, CS 84215, Rennes Cedex, F-35042, France

    Virginie Ducrot & Laurent Lagadic

  6. Department of Biology, University of Southern Denmark, Campusvej 55, 5230, Odense M, Denmark

    Henrik Holbech & Karin Lund Kinnberg

  7. University of Plymouth, Drake Circus, Plymouth, PL4 8AA, UK

    Thomas H. Hutchinson

  8. Department of Environmental Sciences, University of Helsinki, Niemenkatu 73, Lahti, 15140, Finland

    Paula Kajankari & Olli-Pekka Penttinen

  9. Old School House, Brow Edge, Backbarrow, Ulverston, Cumbria, LA128QX, UK

    Peter Matthiessen

  10. Department of Biology, University of Iowa, 143 Biology Building, Iowa City, 52242, IA, USA

    Maurine Neiman

  11. University of Vigo, ECIMAT, Illa de Toralla s/n, 36331 Coruxo-Vigo, Galicia, Spain

    Paula Sanchez-Marin

  12. Fraunhofer Institute for Molecular Biology and Applied Ecology, Auf dem Aberg 1, Schmallenberg, 57392, Germany

    Matthias Teigeler

  13. BASF SE, Crop Protection - Ecotoxicology, Speyerer Straße 2, Limburgerhof, 67117, Germany

    Lennart Weltje

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  1. Cornelia Geiß
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  2. Katharina Ruppert
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  3. Clare Askem
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  4. Carlos Barroso
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  19. Jörg Oehlmann
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Correspondence to Cornelia Geiß.

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Conflict of interest

Some of the authors are employed by industry as disclosed in the affiliations of the authors. The test method described here could potentially be used in the future for the environmental risk assessment of chemicals that are produced by industries.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Present address: Virginie Ducrot and Laurent Lagadic, Bayer CropScience AG, Environmental Safety/Ecotoxicology, Alfred-Nobel-Str. 50, Monheim am Rhein 40789, Germany

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Geiß, C., Ruppert, K., Askem, C. et al. Validation of the OECD reproduction test guideline with the New Zealand mudsnail Potamopyrgus antipodarum using trenbolone and prochloraz. Ecotoxicology 26, 370–382 (2017). https://doi.org/10.1007/s10646-017-1770-y

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