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WEATHERFISH Misgurnus Fossilis CONSERVATION OF THE EUROPEAN WEATHERFISH Misgurnus fossilis STOCKING MEASURES, AUTECOLOGY AND POTENTIAL THREATS by Benjamin Schreiber from Senden, Germany Accepted Dissertation thesis for the partial fulfillment of the requirements for a Doctor of Natural Sciences Fachbereich 7: Natur- und Umweltwissenschaften Universität Koblenz-Landau Thesis examiners: Prof. Dr. Ralf Schulz, University of Koblenz-Landau Prof. Dr. Henner Hollert, RWTH Aachen University Date of the oral examination: 31.07.2017 i OVERVIEW OF PUBLICATIONS As a cumulative dissertation, this thesis is based on the following four scientific articles that are either published or submitted for publication: 1. Schreiber, B., Korte, E., Schmidt, T., Schulz, R. Reintroduction and stock enhancement of European weatherfish (Misgurnus fossilis L.) in Rhineland-Palatinate and Hesse, Germany. (Submitted to Fisheries Management and Ecology) [Appendix I] 2. Schreiber, B., Monka, J., Drozd, B., Hundt, M., Weiss, M., Oswald, T., Gergs, R., Schulz, R., 2017. Thermal requirements for growth, survival and aerobic performance of weatherfish larvae Misgurnus fossilis. J. Fish Biol. 90, 1597–1608. doi:10.1111/jfb.13261 [Appendix II] 3. Schreiber, B., Petrenz, M., Monka, J., Drozd, B., Hollert, H., Schulz, R., 2017. Weatherfish (Misgurnus fossilis) as a new species for toxicity testing? Aquat. Toxicol. 183, 46–53. doi:10.1016/j.aquatox.2016.12.006 [Appendix III] 4. Schreiber, B., Fischer, J., Schiwy, S., Hollert, H., Schulz, R, 2017. Towards more ecological relevance in sediment toxicity testing with fish: evaluation of multiple bioassays with embryos of the benthic weatherfish (Misgurnus fossilis). Sci. Total Environ., Accepted. [Appendix IV] ii CONTENTS ABBREVIATION LIST ................................................................................................................... iv ABSTRACT ................................................................................................................................. vi INTRODUCTION, OBJECTIVES AND THESIS OUTLINE ..................................................................... 1 1.1 INTRODUCTION .................................................................................................................. 1 1.2 OBJECTIVES AND THESIS OUTLINE ..................................................................................... 4 METHODICAL BACKGROUND ....................................................................................................... 7 STOCKING MEASURES AND AUTECOLOGY ................................................................................... 8 3.1 REGIONAL REINTRODUCTION AND STOCK ENHANCEMENT OF WEATHERFISH ..................... 8 3.2 THERMAL REQUIREMENTS OF WEATHERFISH LARVAE ...................................................... 11 CHEMICAL STRESS AS A POTENTIAL THREAT ............................................................................. 15 4.1 FEASIBILITY OF WEATHERFISH FOR TOXICITY TESTING .................................................... 15 4.2 EVALUATION OF MULTIPLE SEDIMENT BIOASSAYS WITH WEATHERFISH ........................... 19 CONCLUSIONS AND OUTLOOK ................................................................................................... 24 REFERENCES ............................................................................................................................. 26 DANKSAGUNG ........................................................................................................................... 35 DECLARATION .......................................................................................................................... 36 APPENDICES .............................................................................................................................. 38 APPENDIX I: SCIENTIFIC PUBLICATION 1 ................................................................................ 39 APPENDIX II: SCIENTIFIC PUBLICATION 2 ............................................................................... 48 APPENDIX III: SCIENTIFIC PUBLICATION 3 .............................................................................. 65 APPENDIX IV: SCIENTIFIC PUBLICATION 4 .............................................................................. 89 APPENDIX V: CURRICULUM VITAE ....................................................................................... 122 iii ABBREVIATION LIST 3Rs Replacement, reduction and refinement of animal experiments AL Altrip ASE Accelerated solvent extraction CI Confidence interval DCA 3,4-dichloroaniline DMSO Dimethyl sulfoxide DO Dissolved oxygen dph Days post-hatch ECX Effect concentration, X% EEA European Environment Agency EEC European Economic Community ERA Ecological risk assessment ES Ehrenbreitstein EU European Union ex-ACE Acetone extract ex-PHWE Pressurized hot water extract FET Acute fish embryo toxicity test G Mean mass-specific growth rate GADM Global administrative areas GnRH Gonadotropin-releasing hormone HH Hamburg ISO International Organization for Standardization IUCN International Union for Conservation of Nature LCX Lethal concentration, X% LOEC Lowest observed effect concentration MD Dry mass ṀO2 Routine metabolic rate OCR Oxygen consumption rate OECD Organization for Economic Co-Operation and Development OECD-236 Test guideline No. 236 of the OECD PAHs Polycyclic aromatic hydrocarbons PCBs Polychlorinated biphenyls iv PCDD/Fs Polychlorinated dibenzodioxins and dibenzofurans PHWE Pressurized hot water extraction RAS Recirculating aquaculture system SCA Sediment contact assay SD Standard deviation SE Standard error ST Streitgraben TC Critical temperature TOpt Temperature optimum TOptG Optimum temperature for growth v ABSTRACT The European weatherfish (Misgurnus fossilis) is a benthic freshwater fish species belonging to the family Cobitidae, that is subjected to a considerable decline in many regions across its original distribution range. Due to its cryptic behavior and low economic value, the causes of threat to weatherfish remained partly unknown and the species is rarely at the center of conservation efforts. In order to address these concerns, the overall aim of the present thesis was to provide a comprehensive approach for weatherfish conservation, including the development of stocking measures, investigations on the species autecology and the evaluation of potential threats. The first objective was to devise and implement a regional reintroduction and stock enhancement program with hatchery-reared weatherfish in Germany. Within this program (2014-2016), a total number of 168,500 juvenile weatherfish were stocked to seven water systems. Recaptures of 45 individuals at two reintroduction sites supported the conclusion that the developed stocking strategy was appropriate. In order to broaden the knowledge about weatherfish autecology and thereby refining the rearing conditions and the selection of appropriate stocking waters, the second objective was to investigate the thermal requirements of weatherfish larvae. Here, the obtained results revealed that temperatures higher than previously suggested were tolerated by larvae, whereas low temperatures within the range of likely habitat conditions increased mortality rates. As weatherfish can be frequently found in agriculturally impacted waters (e.g. ditch systems), they are assumed to have an increased probability to be exposed to chemical stress. Since the resulting risk has not yet been investigated with a focus on weatherfish, the third objective was to provide a methodical foundation for toxicity testing that additionally complies with the requirements of alternative test methods. For this purpose, the acute fish embryo toxicity test was successfully transferred to weatherfish and first results exhibited that sensitivity of weatherfish towards a tested reference substance (3,4-dichloroaniline) was highest compared to other species. On the basis of these findings, the fourth objective was to apply weatherfish embryos for multiple sediment bioassays in order to investigate teratogenic effects derived from sediment-associated contaminants. In this context, weatherfish revealed particular sensitivity to water extractable substances, indicating that sediment contamination might pose a considerable risk. Moreover, as an endangered benthic fish species with high ecological relevance for European waters that are specifically exposed to hazardous contaminants, the weatherfish might be a prospective species for an ecological risk assessment of sediment toxicity. Overall, the present thesis contributed to the conservation of weatherfish by considering a variety of aspects that interact and reinforce one another in order to achieve improvements for the species situation. vi 1 CHAPTER INTRODUCTION, OBJECTIVES AND THESIS OUTLINE 1 1.1 INTRODUCTION Fishes are the most abundant and diverse of the vertebrate groups, including more than 30,000 described species that inhabit an enormous variety of aquatic habitats (Lévêque et al., 2008; FishBase, 2017a). The fundamental (e.g. nutrient cycling) and demand-derived ecosystem services (e.g. economic and recreational values) provided by fish populations are undoubtedly of significant value for human societies (Holmlund & Hammer, 1999; Ormerod, 2003). However, the worlds ichthyofauna is increasingly threatened by consequences of the expansion of human populations, including (i) competition for water, (ii) habitat alteration, (iii) pollution, (iv) introduction of exotic species and (v) commercial exploitation
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