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Survey of Estrogenic Activity in the Danish Aquatic Environment

Contents

Preface

Summary and conclusions

Sammenfatning og konklusioner

1 Background

• 1.1 Estrogens as an Environmental problem
• 1.2 Previous Danish studies on estrogens
  • 1.2.1 Field studies of intersex
  • 1.2.2 Preliminary survey of WWTP effluents
  • 1.2.3 Characterisation of WWTP influents and effluents
  • 1.2.4 Estrogen elimination processes at WWTPs
• 1.3 Project objectives

2 Strategy and design of the study

• 2.1 Overall strategy
  • 2.1.1 Selection of scenarios and locations
  • 2.1.2 Sampling
  • 2.1.3 Biological testing and chemical analysis
• 2.2 Selection of sampling sites
  • 2.2.1 Wastewater treatment plants
  • 2.2.2 Other potential sources of pollution (open land sources)
  • 2.2.3 The freshwater environment
  • 2.2.4 Special investigations

3 Overview of activities

• 3.1 Sampling methodology
• 3.2 Analysis of steroid estrogens and estrogenic activity
  • 3.2.1 Sample pre-treatment
  • 3.2.2 Biological testing
  • 3.2.3 Chemical analysis
  • 3.2.4 Data treatment and E2 response factors
• 3.3 Quality Assurance
  • 3.3.1 Quality Control samples
  • 3.3.2 Inter-laboratory comparison

4 Pre-monitoring tests and quality assurance

• 4.1 Sampling procedure
• 4.2 Enzymatic treatment of conjugated estrogens
  • 4.2.1 Background
  • 4.2.2 Investigation of loss of conjugated estrogens
  • 4.2.3 Losses during transport and storage
  • 4.2.4 Losses during sample pre-treatment
• 4.3 Quality control
  • 4.3.1 Internal quality control scheme
  • 4.3.2 Precision and Limit of Detection (LOD)
  • 4.3.3 Inter-laboratory comparison
  • 4.3.4 Comparison between control sample results from biological and chemical measurements

5 Presentation of field data

6 Sources of estrogens

• 6.1 Wastewater treatment plants (W WTPs)
  • 6.1.1 Influents
  • 6.1.2 Effluents
  • 6.1.3 Species of estrogens and their distribution
• 6.2 Conditions in water courses receiving WWTP effluents
  • 6.2.1 Upstream and downstream estrogenic activity
  • 6.2.2 Seasonal variation in receiving waters
• 6.3 Open land sources

7 State of the environment

• 7.1 Overall results
• 7.2 Geographical distribution
• 7.3 Seasonal/temporal variation

8 Discussion of the results

• 8.1 Sources of estrogens
  • 8.1.1 Wastewater treatment plants
  • 8.1.2 Drains from manure treated fields
  • 8.1.3 Other open land sources
  • 8.1.4 Geographical variation
  • 8.1.5 Seasonal variation
• 8.2 State of the aquatic environment
  • 8.2.1 General state of the aquatic environment
  • 8.2.2 Streams receiving WWTP effluents
  • 8.2.3 Reference streams and lakes
  • 8.2.4 Geographical variation
  • 8.2.5 Seasonal variation
• 8.3 Effect levels of estrogens in freshwater environments
• 8.4 Occurrence of the synthetic hormone Ethynylestradiol
  • 8.4.1 Samples with non-steroid estrogenic activity

9 Conclusions

• 9.1 Sources of estrogens
  • 9.1.1 Wastewater treatment plants
  • 9.1.2 Sources in the open land
• 9.2 Estrogenic activity in the aquatic environment
  • 9.2.1 General state
  • 9.2.2 Streams receiving wastewater effluents
  • 9.2.3 Background locations
• 9.3 Types and forms of estrogens
  • 9.3.1 Contribution from the steroid estrogens
  • 9.3.2 Conjugated versus free estrogens
  • 9.3.3 Other chemicals may contribute to estrogenic activity
• 9.4 Geographical and seasonal variations
• 9.5 Environmental significance

10 References

Appendices

1. Pre-survey testing of procedures and equipment

2. Chemical analysis

3. In-vitro test - yes assay standard curves

4. Quality assurance data for yes assay and chemical analysis

5. Steroid estrogens versus estrogenic activity

6. Results in novana format

7. Locations and sampling scheme

8. Survey data

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Version 1.0 Januar 2005, © Danish Environmental Protection Agency