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Physiological Responses to Different Environmental and Cultural Berried females Eggs auplisoma Adult Spiny lobster Lifecycle Instar I phyllosoma Juvenile Puerulus Final instar phyllosoma PHYSIOLOGICAL RESPOSES TO DIFFERET EVIROMETAL AD CULTURE CODITIOS DURIG OTOGEY OF THE SPIY LOBSTER, Sagmariasus verreauxi By Mark Andrew Jensen B.Aqua. (Hons) Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy UNIVERSITY OF TASMANIA June 2012 Mark Jensen Declarations Declarations Statement of originality I hereby declare that this thesis contains no material which has been accepted for a degree or diploma by the University or any other institution, except by way of background information and duly acknowledged in the thesis, and to the best of my knowledge and belief no material previously published or written by another person except where due acknowledgement is made in the text of the thesis, nor does the thesis contain any material that infringes copyright. Mark Jensen Date University of Tasmania 2012 Authority of access This thesis may be made available for loan and limited copying and communication in accordance with the Copyright Act 1968 . Mark Jensen Date University of Tasmania 2012 ii Mark Jensen Abstract Abstract Very little is known about the metabolic and biochemical physiology of spiny lobsters as they develop. An improved understanding of the physiological responses of spiny lobsters to different environmental and culture conditions during ontogeny is essential for gaining a better understanding of environmental influences on wild populations and for their successful propagation. This study addresses important gaps in our knowledge by examining stage-specific changes in metabolic rates, ammonia-N excretion rates, thermal tolerance thresholds, and whole body and haemolymph biochemistry of larval and juvenile Sagmariasus (Jasus) verreauxi in order to observe these physiological parameters through ontogeny. Automated intermittent flow- through respirometry was used to measure the aerobic metabolism of larval and juvenile lobsters accurately. Whole body biochemical analysis was used to examine energy storage and utilisation of phyllosoma. Haemolymph biochemistry was used to determine the thermal tolerance thresholds of juvenile lobsters. The effect of temperature change on the final instar (instar 17) was examined to assess whether it could serve as a cue for metamorphosis and it was found that temperature affected routine metabolic rate ( Rr), but did not alter instar duration, and therefore is not a cue for metamorphosis. Fewer phyllosoma, however, completed metamorphosis and progressed to the puerulus stage at sub-optimal warm temperatures compared to cooler temperature. The effect of culture density was examined from hatch to pueruli. High culture density reduced growth (dry mass) and development (moult increment) of phyllosoma. There was a shift in metabolism via energy storage and utilisation of instar 17 phyllosoma in preparation for the morphological changes associated with metamorphosis. There was also an iii Mark Jensen Abstract accumulation of lipid reserves during larval development that fuelled metamorphosis and the non-feeding puerulus stage. The aerobic scope of juvenile lobsters was determined through chasing lobsters by hand until the lobster was exhausted and did not respond to further stimuli along with the effects of handling, anaesthesia, and activity on the oxygen consumption rate ( ṀO2) and ammonia-N excretion rate. Handling caused a relatively minor increase in ṀO2 and anaesthetics reduced activity of lobsters, but did not reduce ṀO2 or recovery periods from force feeding or handling. S. verreauxi juveniles have a narrow aerobic scope. Increased ṀO2 from anaesthesia and activity uses a large proportion of energy within the metabolic scope that could otherwise be utilised for other physiological functions. Thermal tolerance thresholds were examined in different sized juvenile lobsters. Sudden changes in haemolymph O 2 concentrations with water temperature indicated large lobsters have a higher optimum water temperature than small lobsters. Maximum attainable rates of standard metabolism indicated the upper critical temperature ( Tc) for juvenile S. verreauxi , characterised by the onset of anaerobic metabolism. Juveniles utilised lipid as an energy substrate at optimal temperatures, but shifted towards protein catabolism at temperatures above their thermal tolerance range. This research revealed the long larval phase of S. verreauxi is essential for accumulating lipid reserves to fuel later larval stages and provided a more complete picture of the environmental and culture requirements of spiny lobsters during ontogeny, particularly for the rarely studied late phyllosoma stages. It also established that induced stress uses a large proportion of the aerobic scope in juvenile lobsters, which limits their ability to perform aerobically and deal with additional physiological challenges. The thermal tolerance of S. verreauxi juveniles identified iv Mark Jensen Abstract the potential to expand their geographic distribution, which may have a large impact on benthic community structures and dynamics through competition for resources with existing lobster populations and other animals within the ecosystem. This may also impact local economies, particularly valuable local southern rock lobster fisheries. v Mark Jensen Acknowledgements Acknowledgements Firstly, I would like to thank Dr Arthur Ritar for his supervision during the commencement of my PhD. I am very grateful for the knowledge he imparted on rock lobsters and the opportunity to work with such a unique marine creature. I thank my supervisors, Professor Chris Carter, Dr Quinn Fitzgibbon, and Dr Louise Ward, especially Professor Chris Carter for accepting the role as primary supervisor after the commencement of my candidature. His vast years of experience in the aquaculture nutrition and physiology field have been extremely helpful in the successful completion of my PhD. Dr Quinn Fitzgibbon for providing detailed knowledge on animal physiology and continuous assistance above and beyond the call of duty and Dr Louise Ward for making herself available for phone meetings and offering assistance in the laboratory. I also wish to thank Ross Goldsmid and Anna Overweter for making their office door always open and offering support and encouragement. Thanks for the welcome distraction of our fishing trips Rosco, hopefully there are many more in the future. Much appreciation goes to Associate professor Natalie Moltschaniwskyj for her support as Graduate Research Coordinator during the first half of my PhD. Thanks to the resident “brainiac” Dr Cedric Simon for his assistance with analytical procedures and statistics and to Craig Thomas and Allan Beech for donating their time and assistance with the experimental setup and construction. Also, to the rest of the aquaculture team, staff and students at IMAS Taroona thank you for your support throughout the project. Finally, to my family for their love and support for the past twenty seven years and always believing in me. I dedicate this thesis to my Grandfather, thank you for the motivation to continue to strive for success. vi Mark Jensen Table of Contents Table of Contents Page Title page ………………………………………………………………………………………………. i Declarations ................................................................................................................... ii Statement of originality ............................................................................................. ii Authority of access .................................................................................................... ii Abstract ........................................................................................................................ iii Acknowledgements ...................................................................................................... vi Table of Contents ........................................................................................................ vii List of Figures .............................................................................................................. xi List of Tables ............................................................................................................. xvii Chapter 1. General introduction ............................................................................. 1 1.1 Introduction .......................................................................................................... 2 1.2 Lifecycle ............................................................................................................... 3 1.3 Moult cycle ........................................................................................................... 5 1.4 Density ................................................................................................................. 6 1.5 Metabolism ........................................................................................................... 7 1.5.1 Aerobic respiration ................................................................................... 7 1.5.2 Aerobic scope ......................................................................................... 12 1.5.3 Body mass .............................................................................................. 12 1.5.4 Temperature ........................................................................................... 14 1.6 Biochemical composition ..................................................................................
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