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Study of the Reproductive Biology and Chemical Communication of Sea Cucumbers (Holothuria Arguinensis and H

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Study of the Reproductive Biology and Chemical Communication of Sea Cucumbers (Holothuria Arguinensis and H NATHALIE MARQUET Study of the reproductive biology and chemical communication of sea cucumbers (Holothuria arguinensis and H. mammata) UNIVERSIDADE DO ALGARVE Faculdade de Ciências e Tecnologia Faro, 2017 NATHALIE MARQUET Study of the reproductive biology and chemical communication of sea cucumbers (Holothuria arguinensis and H. mammata) Doutoramento em Ciências do Mar, da Terra e do Ambiente Ramo Ciências do Mar Especialidade em Ecologia Marinha Orientadores: Prof. Adelino V.M. Canário Dr. Peter C. Hubbard Dr. Mercedes González-Wangüemert UNIVERSIDADE DO ALGARVE Faculdade de Ciências e Tecnologia Faro, 2017 NATHALIE MARQUET Study of the reproductive biology and chemical communication of sea cucumbers (Holothuria arguinensis and H. mammata) Doctoral program in Marine, Earth and Environmental Sciences Marine Sciences Branch Specialty in Marine Ecology Supervisors: Prof. Adelino V.M. Canário Dr. Peter C. Hubbard Dr. Mercedes González-Wangüemert UNIVERSIDADE DO ALGARVE Faculdade de Ciências e Tecnologia Faro, 2017 Título | Thesis title Study of the reproductive biology and chemical communication of sea cucumbers (Holothuria arguinensis and H. mammata) Declaração de autoria de trabalho: Declaro ser a autora deste trabalho que é original e inédito. Autores e trabalhos consultados estão devidamente citados no texto e constam da listagem de referências incluída. Declaration of authorship: This work has not previously been submitted for a degree in any university. To the best of my knowledge and belief, the thesis contains no material previously published or written by another person except where due reference is made in the thesis itself. Nathalie Marquet i Copyright : Nathalie Marquet A Universidade do Algarve reserva para si o direito, em conformidade com o disposto no Código do Direito de Autor e dos Direitos Conexos, de arquivar, reproduzir e publicar a obra, independentemente do meio utilizado, bem como de a divulgar através de repositórios científicos e de admitir a sua cópia e distribuição para fins meramente educacionais ou de investigação e não comerciais, conquanto seja dado o devido crédito ao autor e editor respetivos”. According to the code of authors copyright and related rights, the University of Algarve reserves the legal right, to file, reproduce and publish this work, regardless the means of disclosure, as well to divulge through scientific repositories and to allow its copy and distribution, for educational or research purposes, if due credit is given to the respective author and publisher. ii Support This study was funded by the Portuguese Foundation for Science and Technology (FCT) through a PhD grant (SFRH/BD/90761/2012). Apoio Este estudo foi suportado através de uma bolsa de Doutoramento concedida pela Fundação para a Ciência e Tecnologia (FCT): Bolsa de Doutoramento (Ref: SFRH/BD/90761/2012). iii iv Acknowledgements Many people helped me to overcome and complete the doctoral adventure during these last four years. I hope they will find in the following lines all the gratitude I have toward them for their patience, guidance, advice, mental support and friendship. First of all, I would like to thank my supervisors; Prof. Adelino Canário who offered me the opportunity to work on the reproductive biology in sea cucumbers, and who was always there, although he had a busy schedule, to help me in solving administrative and thesis related problems; Dr. Peter Hubbard for his guidance, listening, advice, and his sense of humor (British humor is not that bad after all). Thank you very much to both of them for their confidence, support, and patience, particularly during these last months, and for their revisions of my thesis that improved considerably its quality. Thank you also to Dr. Mercedes González-Wangüemert who welcomed me at the beginning of the PhD. Thank you very much for the collection of the samples of H. mammata in Spain and for the revision of some chapters of my thesis. I would like to thank Prof. Deborah Power who guided me in my histology and immunohistochemistry work. A special thank you for her listening, her constant positive attitude and, her corrections and suggestions for some chapters of my thesis. Thank you very much to Dr. Rita Costa, Dr. Ricardo Alves, Dr. Marco Campinho, Nadia Silvia and Marlene Trindade who helped and advised me when I had problems related to slides preparation, techniques and microscopes. A particular thank you to Prof. José Paulo Da Silva who helped me in the chemical analysis of the spawning water. I am very grateful for everything he taught me. Thank you very much for his patience (I realized how hard it is difficult for a chemist to speak to a biologist), his time and, his revisions of the bioassays chapter. I really appreciated to collaborate with him. Thank you very much to Dr. João Cardoso who guided me in the GPCR world; thank you for his patience, his great help and the corrections of that “crazy” chapter. Thank you to Dr. Bruno Louro who performed the assembly in the transcriptome analysis. Thank you also to Dr. Rute Martins who guided me in the RNA extraction. An enormous thank you to João Reis and Cristóvão Nunes for their help in the preparation of the tanks and experiments, and in the maintenance of the sea cucumbers in Ramalhete. Thank you very much for their advice and time. I would like to thank Prof. Ester Serrão who helped me in the beginning of the thesis, and all the nice people and friends I met in her lab: Cristina, Licinia, Carla Lourenço, Sara Valente, Neusa, Ana Balau, Nelson, Xana, Rita Jacinto, João Neiva, Buga, Tania Aires, Maria Klein. Thank you very much for their help, support and friendship. A special thank you to Dr Katy Nicastro and Dr Gerardo Zardi who helped me to find solutions to go on my sampling sites in the beginning of thesis and who were all time opened for discussion. Thank you very much for your help! Thank you very much to the nice colleagues I had in the office: Flavia, Alex, Soraia, Carmen, Lijie, Victoria. It was very nice to talk and relax with you all, especially during our lunch break. v Then, I would like to thank all the people I met in CCMAR during these four years: Barry, Ross, Adrian, Iarre, Inne, Gangadhar, Nadege, Patricia Diogo, Patricia Mateus, Zélia, Ricardo, Nour, Mona, Xushuai, Tom, Amrit, Roberto…and I am sure I forgot a lot of people…but be sure you are in my heart. A special thank you to the CCMAR administrative team: Liliane Leman, Cristina Inacio, Marco Granja, Andrea Pinto who were of a great help to solve any administrative problems. I would like to thank my “old” friends who were all the time present to listen to me and to motivate me: Virginie, Delphine, Nathalie and Belgin. It is being a long time now since we finished our study at the ULB, but we can still count on each other. Thanks for everything girls! A very particular thank you to my friend Farah. It was really a pleasure to meet you here and I am very happy we shared all these moments together. I did not get only a best friend during the PhD adventure but I got a best friend for life. Thank you very much for your support, your joke, your help, and your friendship. I would like to thank my family, and specially my parents who are always supporting me in everything I undertake, and who are being present for me all the time. Thank you for everything and specially for your encouragement during all these years, not only those from the PhD, but also those from before. And, as you said so well Papa, I would probably end up as a centenarian student ☺ Merci du fond du cœur, je ne serais jamais arrivée là où je suis aujourd’hui sans vous, vous êtes des parents en or, et je vous aime très fort. Finally, the last but not least, I would like to address my last acknowledgements to João Afonso. I am very glad I met you here and I am very thankful for everything you did for me during these last years. Thank you very much for your mental support and your positive attitude in all circumstances, your amazing help, your patience and your love. You were definitively my pillar in this PhD adventure, and for sure it would not have been the same without you. Amo-te muito. vi To myself I am only a child playing on the beach, while vast oceans of truth lie undiscovered before me. Isaac Newton vii viii Summary New sea cucumber fisheries are emerging in the Mediterranean Sea and Atlantic Ocean in response to a strong Chinese market demand. However, little is known about the biology of the new target species, hindering decisions on their management. The main objective of the present thesis was to study the reproductive biology and the role played by chemical communication and chemosensory systems in Holothuria arguinensis and Holothuria mammata. The different populations sampled in a narrow range along the Iberian Peninsula varied in size/weight, gonadal production, and maturity profile within each species, suggesting the influence of singular features of each location. However, they had all the same general reproductive pattern with a summer-autumn spawning. These results, essential to manage populations, were also useful to determine when to develop bioassays to test whether and how these species communicate during reproduction. Male sea cucumbers, but not females, release chemicals that attract and induce spawning in both sexes. A preliminary analysis of the male spawning water suggests a pheromone with multiple components, among them possibly phosphatidylcholine derivatives. Histology, histochemistry and immunohistochemistry of the potential chemosensory structures involved in the detection of these cues – tentacles, papillae and tube feet – show no obvious differences between them.
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