Environmental Science and Pollution Research

, Volume 18, Issue 3, pp 446–460 | Cite as

The endocrine disrupting potential of sediments from the Upper Danube River (Germany) as revealed by in vitro bioassays and chemical analysis

  • Stefanie GrundEmail author
  • Eric Higley
  • René Schönenberger
  • Marc J-F. Suter
  • John P. Giesy
  • Thomas Braunbeck
  • Markus Hecker
  • Henner Hollert
Research Article



The present study was part of a comprehensive weight-of-evidence approach with the goal of identifying potential causes for the declines in fish populations, which have been observed during the past decades in the Upper Danube River.


The specific goal was the investigation of the endocrine disrupting potential of sediment extracts from different sites along the Danube River. Parallel to the identification and quantification of target estrogens, two in vitro bioassays were employed to assess the estrogenic potential (yeast estrogen screen, YES) of the sediment samples and to evaluate their effects on the production of testosterone (T) and E2 (H295R Steroidogenesis Assay). Using a potency balance approach, the contribution of the measured compounds (Chem-EEQs) to the total endocrine activity measured by the YES (YES-EEQs) was calculated.

Results and discussion

Of the nine sediment extracts tested five extracts exhibited significant estrogenic activities in the YES, which suggested the presence of ER agonists in these samples. The xenoestrogens nonylphenol (NP) and bisphenol A (BPA) and the natural estrogen estrone (E1) were detected while concentrations of 17β-estradiol (E2) and ethinylestradiol (EE2) were less than their respective limits of quantification in all sediment extracts. A comparison of the measured YES-EEQs and the calculated Chem-EEQs revealed that as much as 6% of estrogenic activity in extracts of most sediments could be explained by two xeno- and one natural estrogen. Exposure of H295R cells to sediment extracts from four different locations in the Danube River resulted in significantly increased concentrations of E2, but only slight inhibition of T synthesis. Furthermore, application of the H295R Steroidogenesis Assay provided evidence for endocrine disrupting potencies in sediment samples from the Upper Danube River, some of which were not detectable with the YES. In conclusion, differential endocrine activities were associated with several sediments from the Upper Danube River. Further investigations will have to show whether the observed activities are of biological relevance with regard to declines in fish populations in the Upper Danube River.


Endocrine disruptors Sediment Yeast estrogen screen H295R Steroidogenesis Assay Testosterone Estradiol 



This study was supported by a personal grant to S. Grund by the scholarship program of the German Federal Environmental Foundation (DBU, Deutsche Bundesstiftung Umwelt). The authors would like to thank Prof. J.P. Sumpter of Brunel University, United Kingdom, for supplying the yeast cells for the YES assay, and Martin Wagner (University of Frankfurt) and Sibylle Maletz (RWTH Aachen) for introduction into the modified YES protocol. The authors are particularly grateful for the support and proof-reading of the manuscript by Nadja Seitz. Prof. Giesy was supported by the Canada Research Chair program and an at large Chair Professorship at the Department of Biology and Chemistry and Research Centre for Coastal Pollution and Conservation, City University of Hong Kong. The research was supported by a < LOQ 6971 and 6807). The authors wish to acknowledge the support of an instrumentation grant from the Canada Foundation for Infrastructure.


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Stefanie Grund
    • 1
    • 10
    Email author
  • Eric Higley
    • 2
  • René Schönenberger
    • 3
  • Marc J-F. Suter
    • 3
  • John P. Giesy
    • 2
    • 4
    • 5
    • 6
    • 7
  • Thomas Braunbeck
    • 1
  • Markus Hecker
    • 2
    • 8
  • Henner Hollert
    • 1
    • 9
  1. 1.Aquatic Ecology and Toxicology Section, Department of ZoologyUniversity of HeidelbergHeidelbergGermany
  2. 2.Department of Veterinary Biomedical Sciences and Toxicology CentreUniversity of SaskatchewanSaskatoonCanada
  3. 3.Eawag, Swiss Federal Institute of Aquatic Science and TechnologyDuebendorfSwitzerland
  4. 4.Department of ZoologyMichigan State UniversityEast LansingUSA
  5. 5.Centre for Coastal Pollution and Conservation and Department of Biology and ChemistryCity University of Hong KongHong KongChina
  6. 6.State Key Laboratory of Pollution Control and Resource Reuse, School of the EnvironmentNanjing UniversityNanjingPeople’s Republic of China
  7. 7.State Key Laboratory of Marine Environmental Science, College of Oceanography and Environmental ScienceXiamen UniversityXiamenPeople’s Republic of China
  8. 8.ENTRIX, IncSaskatoonCanada
  9. 9.Department of Ecosystem AnalysisRWTH Aachen University, Institute for Environmental Research (Biology V)AachenGermany
  10. 10.Department of Zoology, Aquatic Ecology and Toxicology SectionUniversity of HeidelbergHeidelbergGermany

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