Estimating the Threat Posed by the Crayfish Plague Agent
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Estimating the threat posed by the crayfish plague agent Aphanomyces astaci to crayfish species of Europe and North America — Introduction pathways, distribution and genetic diversity by Jörn Panteleit from Aachen, 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, Germany Dr. Japo Jussila, University of Eastern Finland, Kuopio, Suomi-Finland Date of oral examination: January 17th, 2019 TABLE OF CONTENTS 1. LIST OF PUBLICATIONS ........................................................................................................................ 3 2. ABSTRACT ............................................................................................................................................ 4 2.1 Zusammenfassung ......................................................................................................................... 5 3. ABBREVIATIONS .................................................................................................................................. 6 4. INTRODUCTION ................................................................................................................................... 7 4.1 Invasive species ............................................................................................................................. 7 4.2 Freshwater crayfish in Europe ....................................................................................................... 8 4.3 The crayfish plague agent Aphanomyces astaci ........................................................................... 9 4.4 Genetic methods to detect and characterize Aphanomyces astaci ............................................ 11 5. OBJECTIVES OF THE THESIS ............................................................................................................... 15 6. DESCRIPTION OF METHODS .............................................................................................................. 17 6.1 DNA isolation ............................................................................................................................... 17 6.2 Aphanomyces astaci detection ................................................................................................... 17 6.3 Genetic characterization of Aphanomyces astaci ....................................................................... 18 7. SUMMARY OF RESULTS ..................................................................................................................... 19 8. DISCUSSION ....................................................................................................................................... 21 8.1 Crayfish pet trade in Europe ....................................................................................................... 21 8.2 Presence of different Aphanomyces astaci strains in Europe ..................................................... 22 8.3 Presence of Aphanomyces astaci in North America ................................................................... 25 9. CONCLUSION AND FUTURE PERSPECTIVE ......................................................................................... 26 10. REFERENCES .................................................................................................................................... 28 11. DANKSAGUNG/ACKNOWLEDGMENT .............................................................................................. 38 12. APPENDIX ........................................................................................................................................ 40 12.1 Appendix I .................................................................................................................................. 40 12.2 Appendix II ................................................................................................................................. 59 12.3 Appendix III ................................................................................................................................ 72 12.4 Appendix IV ............................................................................................................................... 98 12.5 Appendix V .............................................................................................................................. 122 12.6 Appendix VI ............................................................................................................................. 142 13. DECLARATION ................................................................................................................................ 166 14. CURRICULUM VITAE ...................................................................................................................... 168 1. LIST OF PUBLICATIONS I. Panteleit, J., Keller, N. S., Kokko, H., Jussila, J., Makkonen, J., Theissinger, K., and Schrimpf, A. (2017): Investigation of ornamental crayfish reveals new carrier species of the crayfish plague pathogen (Aphanomyces astaci). Aquatic Invasions 12(1): 77–83. DOI: 10.3391/ai.2017.12.1.08. II. Pârvulescu, L., Togor, A., Lele, S., Scheu, S., Şinca, D., and Panteleit, J. (2017): First established population of marbled crayfish Procambarus fallax (Hagen, 1870) f. virginalis (Decapoda, Cambaridae) in Romania. BioInvasions Records 6(4): 357–362. DOI: 10.3391/bir.2017.6.4.09. III. Makkonen, J., Jussila, J., Panteleit, J., Keller, N. S., Schrimpf, A., Theissinger, K., Kortet, R., Martín-Torrijos, L., Sandoval-Sierra, J., Diéguez-Uribeondo, J., and Kokko, H. (2018): MtDNA allows the sensitive detection and haplotyping of the crayfish plague disease agent Aphanomyces astaci showing clues about its origin and migration. Parasitology Feb 26, 1–9. DOI: 10.1017/S0031182018000227. IV. Panteleit, J., Keller, N. S., Diéguez-Uribeondo, J., Makkonen, J., Martín- Torrijos, L., Patrulea, V., Pîrvu, M., Preda, C., Schrimpf, A., and Pârvulescu, L. (2018): Hidden sites in the distribution of the crayfish plague pathogen Aphanomyces astaci in eastern Europe: relicts of genetic groups from older outbreaks? Journal of Invertebrate Pathology. DOI: 10.1016/j.jip.2018.05.006. V. Panteleit, J., Jussila, J., Keller, N. S., Kokko, H., Makkonen, J., Schulz, R., Theissinger, K., and Schrimpf, A.: Three haplotypes of the crayfish plague agent Aphanomyces astaci found to be responsible for recent crayfish plague outbreaks in Germany and Austria. Submitted to Knowledge and Management of Aquatic Ecosystems. VI. Panteleit, J., Horvath, T., Jussila, J., Makkonen, J., Perry, W., Schulz, R., Theissinger, K., and Schrimpf, A. (2019): Invasive rusty crayfish (Faxonius rusticus) populations in North America are infected with the crayfish plague disease agent (Aphanomyces astaci). Freshwater Science 38(2). DOI: DOI: 10.1086/703417. 3 2. ABSTRACT Invasive species often have a significant impact on the biodiversity of ecosystems and the species native to it. One of the worst invaders worldwide is Aphanomyces astaci, the causative agent of the crayfish plague, an often fatal disease to crayfish species not native to North America. Aphanomyces astaci originates from North America and was introduced to Europe in the midst of the 19th century. Since then, it spread throughout Europe diminishing the European crayfish populations. The overall aim of this thesis was to evaluate the threat that A. astaci still poses to European crayfish species more than 150 years after its introduction to Europe. In the first part of the thesis, crayfish specimens, which are available in the German pet trade, were tested for infections with A. astaci. Around 13% of the tested crayfish were clearly infected with A. astaci. The study demonstrated the potential danger the pet trade poses for biodiversity through the import of alien species and their potential pathogens, in general. In the second part of the thesis, the A. astaci infection prevalence of crayfish species in wild populations in Europe was tested. While the stone crayfish, Austropotamobius torrentium, showed high susceptibility to different haplogroups of A. astaci, the narrow-clawed crayfish, Astacus leptodactylus, was able to survive infections, even by haplogroup B, which is considered to be highly virulent. In the last part of the thesis, A. astaci was traced back to its original distribution area of North America. While the crayfish plague never had such a devastating effect on crayfish in North America as it had in Europe, the reasons for the success of invasive crayfish within North America are not yet fully understood. It is possible that A. astaci increases the invasion success of some crayfish species. Several populations of the rusty crayfish, Orconectes rusticus, in the Midwest of North America were confirmed to be infected with A. astaci and a new genotype was identified, possibly indicating that each crayfish host is vector of a unique A. astaci genotype, even in North America. Overall, the present thesis provides evidence that A. astaci is still a major threat to the crayfish species indigenous to Europe. Crayfish mass mortalities still occur in susceptible crayfish species like A. torrentium even 150 years after the first introduction of A. astaci. While there are some indications for increased resistances through processes of co-evolution, the continuous introduction