Are in vitro methods for the detection of endocrine potentials in the aquatic environment predictive for in vivo effects? Outcomes of the Projects SchussenAktiv and SchussenAktivplus in the Lake Constance Area, Germany

PLoS One. 2014 Jun 5;9(6):e98307. doi: 10.1371/journal.pone.0098307. eCollection 2014.

Abstract

Many studies about endocrine pollution in the aquatic environment reveal changes in the reproduction system of biota. We analysed endocrine activities in two rivers in Southern Germany using three approaches: (1) chemical analyses, (2) in vitro bioassays, and (3) in vivo investigations in fish and snails. Chemical analyses were based on gas chromatography coupled with mass spectrometry. For in vitro analyses of endocrine potentials in water, sediment, and waste water samples, we used the E-screen assay (human breast cancer cells MCF-7) and reporter gene assays (human cell line HeLa-9903 and MDA-kb2). In addition, we performed reproduction tests with the freshwater mudsnail Potamopyrgus antipodarum to analyse water and sediment samples. We exposed juvenile brown trout (Salmo trutta f. fario) to water downstream of a wastewater outfall (Schussen River) or to water from a reference site (Argen River) to investigate the vitellogenin production. Furthermore, two feral fish species, chub (Leuciscus cephalus) and spirlin (Alburnoides bipunctatus), were caught in both rivers to determine their gonadal maturity and the gonadosomatic index. Chemical analyses provided only little information about endocrine active substances, whereas the in vitro assays revealed endocrine potentials in most of the samples. In addition to endocrine potentials, we also observed toxic potentials (E-screen/reproduction test) in waste water samples, which could interfere with and camouflage endocrine effects. The results of our in vivo tests were mostly in line with the results of the in vitro assays and revealed a consistent reproduction-disrupting (reproduction tests) and an occasional endocrine action (vitellogenin levels) in both investigated rivers, with more pronounced effects for the Schussen river (e.g. a lower gonadosomatic index). We were able to show that biological in vitro assays for endocrine potentials in natural stream water reasonably reflect reproduction and endocrine disruption observed in snails and field-exposed fish, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Ecotoxicology / methods*
  • Endocrine Disruptors / analysis
  • Endocrine Disruptors / chemistry*
  • Environmental Monitoring / methods*
  • Fishes
  • Germany
  • Humans
  • In Vitro Techniques
  • Rivers / chemistry*
  • Snails
  • Vitellogenins / chemistry
  • Water Pollutants, Chemical / analysis*

Substances

  • Endocrine Disruptors
  • Vitellogenins
  • Water Pollutants, Chemical

Grants and funding

The project SchussenAktiv was funded by the Ministerium für Umwelt, Naturschutz und Verkehr Baden-Württemberg (UVM), and the foundation “Natur und Umwelt“ of the Landesbank Baden-Württemberg (LBBW). We acknowledge support by Deutsche Forschungsgemeinschaft and Open Access Publishing Fund of Tubingen University. The project SchussenAktivplus is funded by the Federal Ministry for Education and Research (BMBF) and cofounded by the Ministry of Environment Baden-Württemberg. In addition, the city of Ravensburg, the AZV Mariatal and the AV Unteres Schussental financially contribute to the project. The commercial companies (Jedele & Partner GmbH and Ökonsult GbR) provided funding to this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials. SchussenAktivplus is connected to the BMBF action plan “Sustainable water management (NaWaM)” and is integrated in the BMBF frame programme “Research for sustainable development FONA”. Contract period: 1/2012 to 12/2014, Funding number: 02WRS1281A. All funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.