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University of Veterinary Medicine Hannover University of Veterinary Medicine Hannover Analysis of the reproductive biology of harbour porpoises (Phocoena phocoena) from the North and Baltic Sea on a long-term basis Inaugural-Dissertation in fulfillment of the requirements of the degree of - Doctor medicinae veterinariae - (Dr. med. vet.) submitted by Tina Kesselring Rostock Hannover 2019 Academic supervision: 1st TiHo supervisor: Prof. Prof. h. c. Dr. Ursula Siebert, Institute for Terrestrial and Aquatic Wildlife Research, University of Veterinary Medicine Hannover, Foundation 2nd TiHo Supervisor: Prof. Dr. Ralph Brehm, Institute for Anatomy, University of Veterinary Medicine Hannover, Foundation 1st Referees: Prof. Prof. h. c. Dr. Ursula Siebert, Institute for Terrestrial and Aquatic Wildlife Research, University of Veterinary Medicine Hannover, Foundation Prof. Dr. Ralph Brehm, Institute for Anatomy, University of Veterinary Medicine Hannover, Foundation 2nd Referee: Prof. Dr. Andreas Beineke, Institute for Pathology, University of Veterinary Medicine Hannover, Foundation Day of the oral examination: 01.04.1019 Funding statement: This project was partly funded by the Ministry of Energy, Agriculture, the Environment, Nature and Digitalization Schleswig Holstein (MELUND) and the state centre for coastal protection, national park and ocean protection Schleswig – Holstein. To my mother and my grandparents ‘For most of history, man has had to fight nature to survive; in this century he is beginning to realize that, in order to survive, he must protect it.’ Jacques Yves Cousteau Parts of this thesis have been published in peer reviewed journals: Publication I Kesselring T, Viquerat S, Brehm R, Siebert U. Coming of age: - Do female harbour porpoises (Phocoena phocoena) from the North Sea and Baltic Sea have sufficient time to reproduce in a human influenced environment? Published in October 2017: PLoS One 2017; 12. doi:10.1371/journal.pone.0186951. Corrected version was published in June 20, 2018. Correction: Coming of age: - Do female harbour porpoises (Phocoena phocoena) from the North Sea and Baltic Sea have sufficient time to reproduce in a human influenced environment? PLoS ONE 13(6): e0199633. https://doi.org/10.1371/journal.pone.0199633 Publication II Kesselring, T, Viquerat, S, IJsseldijk, LL, Langeheine, M, Wohlsein, P, Gröne, A, Bergmann, M, Siebert, U, Brehm, R. Testicular morphology and spermatogenesis in harbour porpoises (Phocoena phocoena) Published online in November 2018 in Theriogenology, Theriogenology 126 (2019) 177-186 DOI: 10.1016/j.theriogenology.2018.11.031 Parts of this thesis have been presented in poster presentations and an oral presentation at the following conferences: Poster presentations: Kesselring, T.; Viquerat, S.; Wohlsein, P.; Langeheine, M.; Gröne, A.; Ijsseldijk, L.; Bergmann, M.; Siebert, U.; Brehm, R. : Seasonal effects on testicular development of harbour porpoises from the North and Baltic Sea. In: Reproduction in Domestic Animals, special Issue: 50th Annual Conference of Physiology and Pathology of Reproduction and 42nd Mutual Conference on Veterinary and Human Reproductive Medicine, Munich, Germany, 15th - 17th February 2017. Volume 52, Issue Supplement S1, Pp 28-29 31st Annual Conference of the European Cetacean Society, Middelfart, Denmark, 29th April - 03rd Mai, 2017 Kesselring, T.1, Viquerat, S. 1, Brehm, R. 2, Siebert, U. 1: Identifying sexual maturity in female harbour porpoises (Phocoena phocoena) from the North and Baltic Seas using ovarian characteristics 22nd Biennial Society for Marine Mammalogy Conference on the Biology of Marine Mammals, Halifax, Canada, 22nd - 27th October 2017 Kesselring, T.; Viquerat, S.; Bergmann, M.; Isseldijk, L.; Gröne, A.; Langeheine, M.; Wohlsein, P.; Brehm, R.; Siebert, U.: New insights into the reproductive biology of harbour porpoises (Phocoena phocoena) from the North and Baltic Sea. DOI: 10.13140/RG.2.2.32302.20806 Oral presentation: Kesselring, T.; Viquerat, S.; Wohlsein, P.; Langeheine, M.; Gröne, A.; Ijsseldijk, L.; Bergmann, M.; Siebert, U.; Brehm, R. Assessment of spermatogenesis in harbour porpoises (Phocoena phocoena) from the North and Baltic Seas. In: Reproduction in Domestic Animals, special Issue: 51th Annual Conference of Physiology and Pathology of Reproduction and 43rd Mutual Conference of Veterinary and Human Reproductive Medicine, Hanover, Germany, 21st – 23rd February 2018. Volume 53, Issue Supplement S1, P 21, DOI: 10.1111/rda.13127 . List of contents Chapter 1: Introduction ...................................................................................... - 1 - 1.1. General background ............................................................................... - 1 - 1.2. First part of the thesis: study of age at sexual maturity and demography in female harbour porpoises ................................................................................ - 4 - 1.3. Second part of the thesis: study of testicular morphology and spermatogenesis in male harbour porpoises ................................................... - 6 - Chapter 2: Publication I: Correction: Coming of Age: - Do female harbour porpoises (Phocoena phocoena) from the North Sea and Baltic Sea have sufficient time to reproduce in a human influenced environment? ......................................... - 9 - 2.1. Abstract ................................................................................................ - 10 - 2.2. Introduction .......................................................................................... - 11 - 2.3. Materials and Methods ......................................................................... - 13 - 2.3.1. Sample collection........................................................................... - 13 - 2.3.2. Age assessment ............................................................................ - 13 - 2.3.3. Ovarian examination ...................................................................... - 14 - 2.3.4. Data analysis ................................................................................. - 14 - 2.4. Results ................................................................................................. - 16 - 2.5. Discussion ............................................................................................ - 24 - 2.6. Acknowledgements .............................................................................. - 29 - 2.7. References ........................................................................................... - 30 - Chapter 3: Publication II: Testicular development and spermatogenesis in harbour porpoises (Phocoena phocoena) .......................................................................... - 37 - 3.1. Abstract ................................................................................................ - 38 - 3.2. Introduction .......................................................................................... - 39 - 3.3. Materials and Methods ......................................................................... - 41 - 3.3.1. Sample collection........................................................................... - 41 - 3.3.2. Histological preparation ................................................................. - 44 - 3.3.3. Age determination.......................................................................... - 44 - 3.3.4. Histological examination ................................................................ - 44 - 3.3.5. Modelling the diameter of the convoluted seminiferous tubules over the Julian day of the year ................................................................................. - 45 - 3.4. Results ................................................................................................. - 47 - 3.4.1. Macroscopic findings of the testes during the course of the year .. - 47 - 3.4.2. Testicular maturation and staging of the seminiferous epithelium . - 48 - 3.4.3. Morphometrical analysis and maturity classes .............................. - 56 - 3.5. Discussion ............................................................................................ - 60 - 3.5.1. Testicular maturation and staging of the seminiferous epithelium . - 60 - 3.5.2. Morphometrical analysis and age groups ...................................... - 61 - 3.6. Conclusions .......................................................................................... - 63 - 3.7. Acknowledgements .............................................................................. - 64 - 3.8. References ........................................................................................... - 65 - Chapter 4: Discussion ..................................................................................... - 73 - 4.1. General discussion ............................................................................... - 73 - 4.2. First part of the thesis: study of age at sexual maturity and demography in female harbour porpoises .............................................................................. - 75 - 4.3. Second part of the thesis: study of testicular morphology and spermatogenesis in male harbour porpoises ................................................. - 80 - Chapter 5: Summary ....................................................................................... - 85 - Chapter 6: Zusammenfassung ........................................................................ - 87 - References ..........................................................................................................
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