Natural Antifouling Defences of Tropical Sea Stars
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This file is part of the following reference: Guenther, Jana (2007) Natural antifouling defences of tropical sea stars. PhD thesis, James Cook University. Access to this file is available from: http://eprints.jcu.edu.au/3223 Natural antifouling defences of tropical sea stars Thesis submitted by Jana Guenther BSc (Hons) October 2007 for the degree of Doctor of Philosophy in the School of Marine and Tropical Biology James Cook University Statement of Access _______________________________________________ I, the undersigned, author of this work, understand that James Cook University will make this thesis available for use within the University Library and, via the Australian Digital Theses network, for use elsewhere. I understand that, as an unpublished work, a thesis has significant protection under the Copyright Act and I do not wish to place any further restriction on access to this work. _____________________________________ ______________________ Signature Date ii Statement of Sources Declaration _______________________________________________ I declare that this thesis is my own work and has not been submitted in any form for another degree or diploma at any university or other institution of tertiary education. Information derived from the published or unpublished work of others has been acknowledged in the text and a list of references is given. _____________________________________ ______________________ Signature Date iii Electronic Copy _______________________________________________ I, the undersigned, the author of this work, declare that the electronic copy of this thesis provided to the James Cook University Library is an accurate copy of the print thesis submitted, within the limits of the technology available. _____________________________________ ______________________ Signature Date iv Statement on the Contribution of Others _______________________________________________ Financial support for this study was provided by the Australian Defence Science and Technology Organisation – Biomimetic Fouling Control Program, the School of Marine and Tropical Biology and the Graduate School of James Cook University, CRC Reef, AIMS@JCU and the Institute of Marine Engineering, Science and Technology. A James Cook University Postgraduate Research Scholarship provided stipend support. Alan Dartnall (James Cook University, Townsville), Genefor Walker-Smith (Tasmanian Museum and Art Gallery, Hobart) and Anders Warén (Swedish Museum of Natural History, Stockholm), assisted with the identification of the sea stars Astropecten velitaris and A. indicus, the surface-associated copepods Stellicola illgi and Paramolgus sp., and the eulimid Thyca (Granulithyca) nardoafrianti, respectively (Chapter 2). Kirsten Heimann (James Cook University, Townsville) supplied the chrysophyte alga Chrysocystis fragilis (Chapter 3). Alex Wu (University of New South Wales, Sydney) measured the contact angles of control surfaces using a goniometer (Chapter 4). Kathy Burns (Australian Institute of Marine Sciences, Townsville) assisted with gas chromatography – mass spectrometry (Chapter 5 and 6). v Acknowledgements _______________________________________________ Many wonderful people have supported me during this study and have therefore contributed to its success. Firstly, I would like to thank my supervisor Rocky de Nys. He has been an excellent supervisor. He readily discussed my work and provided valuable guidance throughout this study. His seemingly endless enthusiasm and motivation has often helped me keep going. I am very grateful to have been one of his students. My associate supervisor Tony Wright at the Australian Institute of Marine Science, Townsville, provided valuable advice on natural products chemistry. He has been very encouraging and reminded me to ‘keep things as simple as possible’. Thank you to all the members of the de Nys lab, especially Andrew Scardino, Raymond Bannister, Steve Whalan, Piers Ettinger-Epstein, Nick Paul and Michael Horne, who have made working in this lab very enjoyable. Thank you to all the members of the chemistry lab at the Australian Institute of Marine Sciences, Townsville. Jonathon Nielson, Marnie Freckelton, Piers Ettinger-Epstein, Cherie Motti and Rick Willis have given me advice on extractions, chromatography and analysis with nuclear magnetic resonance. Kathy Burns and Catherine Pratt have helped with the analysis of samples using gas chromatography, which was much appreciated. Bruce Bowden has helped with the oxidation of a sterol. Numerous people have helped with the collection of sea stars by SCUBA diving and snorkel, thank you to Raymond Bannister, Alicia Crawley, Clare Omodei, Cameron Crothers, Dominica Loong, Daniel Louden, Nick Paul, Andrew Scardino, Steve Whalan and the crown-of-thorns starfish control team (Association of Marine Park Tourism Operators), especially Craig Zytkow. Thank you also to the staff at the Marine and Aquaculture Research Facilities Unit and Lizard Island Research Station, who provided logistical help with maintaining sea stars in tanks. Genefor Walker-Smith at the Tasmanian Museum and Art Gallery, Hobart, Anders Waren at the Swedish Museum of Natural History, Stockholm, and Alan Dartnall at James Cook University have helped me with the taxonomic identification of some sea stars and surface-associated organisms. vi Thank you to Hua Zhang, Alex Wu and Tim Charlton at the University of New South Wales, Sydney, for their logistical help with contact angle measurements. Kevin Blake at the Advanced Analytical Centre was very helpful with the use of the scanning electron microscope. Rob Gegg at the workshop was also helpful with making PVC panels. Thank you to Kirsten Heimann and Stanley Hudson for supplying the alga Chrysocystis fragilis and Erika Martinez Fernandez and Megan Stride for advice on culturing microalgae. Megan Huggett at the University of Hawaii, Honolulu, has given me valuable advice on culturing Hydroides elegans larvae. Nick Paul and David Donald have helped with the statistical analyses, which was very much appreciated. I would like to thank Rachel James, Maya Holme, Malwine Lober and Kiki Tziouveli in the Aquaculture department as well as Tuyet-Nga Hua, Caroline Hay, Jessica Gautherot and Lauren Scardino for their friendship and support. Finally, I would like to thank my family. My fiancé Raymond Bannister has given me much love, understanding and encouragement. He has shared my joy and dried my tears – life is wonderful with you by my side! My parents, Joachim and Gisela Guenther, have always encouraged me to fulfil my ambitions in life and have strongly supported me during my tertiary studies in Australia. I would not have made it this far without my family, and I dedicate this thesis to them. vii Abstract _______________________________________________ Qualitative evidence suggests that sea stars have remarkably clean epidermal surfaces. To quantify surface-associated micro- and macroorganisms, field surveys were conducted in northern Queensland, Australia, during the wet and dry seasons. Mean bacterial abundances on 7 sea star species were approximately 104 to 105 cells cm-2 during both seasons. There were no consistent trends in bacterial abundances with season, species and aboral positions on sea star arms. Low numbers of parasitic and commensal macroorganisms were found on 6 sea star species, and no common generalist macrofouling organisms, such as algae, barnacles, serpulid polychaetes, bryozoans and ascidians, were discovered on any specimens. Sea stars therefore offered an excellent model to investigate the mechanisms driving fouling-resistant surfaces. Subsequently, 5 sea star species belonging to the order Valvatida were chosen for this study to investigate mechanical, physical and chemical fouling deterrence: Acanthaster planci (Family Acanthasteridae), Linckia laevigata and Fromia indica (both Family Ophidiasteridae), Cryptasterina pentagona (Family Asterinidae) and Archaster typicus (Family Archasteridae). It has been proposed that mechanical antifouling defence mechanisms of sea stars are linked with pedicellariae, which are pincer-like appendages made of calcareous ossicles. In this study, the role of pedicellariae of the sea star A. planci in fouling control was investigated. The morphology and distribution of its pedicellariae were measured to determine if larvae or propagules of fouling organisms could settle between pedicellariae without being in their physical range. The elementary and straight pedicellariae of A. planci had a mean length of 0.7 mm and a mean distance of 2.6 mm. The total number of pedicellariae was proportional to the estimated surface area of A. planci. To determine how pedicellariae respond to tactile stimulation, pedicellariae were stimulated by touching either inner, outer or basal sites of pedicellariae with a hypodermic needle. Pedicellariae closed rapidly on touch and closed for significantly longer when touched at their inner sites (8.9 s) than outer (6.7 s) and basal (7.9 s) sites. Settling larvae were simulated by dropping silica beads (size: 50.2 μm, 181.5 μm, 255.7 μm and 510.7 μm, density: 2.5 g ml-1) and zirconium/silica beads (size: 191.2 μm and 507.6 μm, density: 3.7 g ml-1) over the pedicellariae. The percentage of responding pedicellariae increased proportionally with increasing size of the silica beads. However, the percentage also increased when zirconium/silica beads of similar size but higher density were used, demonstrating that the mass, not size, of the beads was the main driving factor for