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Feeding Energetics and Carbohydrate Digestion of the Juvenile New

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Feeding Energetics and Carbohydrate Digestion of the Juvenile New THESIS Feeding Energetics and Carbohydrate Digestion in Juvenile Jasus edwardsii ABSTRACT To enhance the on-growing of Jasus edwardsii in culture, it is important to understand the feeding physiology of juveniles. In crustaceans, there is a loss of energy and an increase in oxygen consumption (specific dynamic action (SDA)) associated with feeding. The present research measured the SDA of juvenile J. edwardsii fed either in the morning or at night held at 15°C. The present research also investigated important issues affecting the successful on­ growing of rock lobsters in culture, diet and nutrition. This was achieved by investigating carbohydrate digestion and metabolism. The present research also investigated the growth of juvenile J. edwardsii in response to three different algal carbohydrates; agar, carrageenans and alginate. Closed box respirometry was used to measure juvenile lobsters oxygen consumption (M02) and ammonia excretion. Juveniles exhibited a nocturnal rhythm in both M02 and ammonia excretion. The factorial rise in M02 (1.58±0.03 times) for lobsters fed in the morning was significantly less than lobsters fed at night (1.80±0.01 times). Lobsters fed in the morning had a significantly shorter SDA (30±1.2 h) response compared to lobsters fed at night (36±1 h). Energy loss as a result of digestion was less for lobsters fed in the morning. Therefore, if juvenile J. edwardsii are fed in the morning, they could optimise the energy content of the meal and this could result in increased growth. At 18 °C, over 80 days juvenile J. edwardsii that were fed squid in the morning had a specific growth rate (SGR) of0.92±0.07 %body weight d- 1 and grew faster than lobsters that were fed at 1 night (SGR = 0.76±0.01 %bw d- ). Morning fed lobsters had a percent weight gain (%WG) of 107.0±3.1%, which was significantly higher than lobsters fed at night (%WG = 78.4±3.1 %). Survival was also greater for morning fed lobsters (50%) than night fed lobsters (25 %). The SDA was used as a tool to determine if juvenile J. edwardsii could utilise a range of general carbohydrates from simple monosaccharides (glucose, fructose), disaccharides (maltose, sucrose) and polysaccharides (glycogen) (general carbohydrates). Three algal binding agents (polysaccharides) were also tested; agar, carrageenans and alginate (algal carbohydrates). For Feeding Energetics and Carbohydrate Digestion in Juvenile Jasus edwardsii ii lobsters fed the general carbohydrates, oxygen consumption increased to a peak followed by a slow decline back to pre-feeding levels. The more complex the carbohydrate, the greater the oxygen consumption profile. However, when lobsters were fed on algal carbohydrates they did not exhibit this trend and had similar oxygen consumption profiles to unfed lobsters. Oxygen consumption magnitudes were significantly higher in lobsters fed a meal of glycogen, sucrose, maltose and agar than unfed lobsters. If juvenile J. edwardsii were able to digest and utilise carbohydrates, there should have been a rise in haemolymph glucose concentration following feeding. The results confirm that juvenile J. edwardsii can digest all the general carbohydrates, but require more energy to digest and metabolise glycogen and sucrose. Lobsters fed agar had a significantly higher haemolymph glucose concentration than unfed lobsters. This was the only algal carbohydrate to exhibit this response as alginate and carrageenans had similar response to unfed. These results suggest that the SDA can be used as a technique for determining carbohydrate digestion in juvenile J. edwardsii and that the magnitude of the response is the most sensitive SDA parameter. Growth rates of juvenile J. edwardsii fed algal carbohydrates were measured at 18 oc, over 80 days. Growth of lobsters fed fresh blue mussels (Mytilus galloprovincialis) was higher (SGR = 1 1.64±0.02 %bw d" ) than any of those produced with the algal carbohydrate diets. Lobsters fed a diet of mussels also had higher survival rates (87 %). Growth rates were similar amongst the 1 1 algal carbohydrates (agar SGR = 0.91±0.10 %bw d" ; carrageenans SGR = 1.14±0.04 %bw d" ; 1 alginate SGR = 0.97±0.35 %bw d" ) but were superior to lobsters fed a diet of squid (SGR = 1 0.76±0.01 %bw d" ). The survival of lobsters fed an agar (50 %) diet was significantly higher than those fed carrageenans (37.5 %) and alginate (25 %) diets. Based on these results juvenile J. edwarsii can digest and utilise agar more efficiently than carrageenans and alginate. The growth experiment, oxygen consumption and haemolymph glucose results from the present research will aid in the development of a nutritional, cost effective diet for the successful aquaculture of J. edwardsii. Acknowledgements iii ACKNOWLEDGEMENTS Many thanks must go to my supervisor Associate Professor Islay D. Marsden for her support throughout this study. Your promptness in returning drafts and helpful criticism were much appreciated and made my life a lot less stressful. Thank you for always having your door open and allowing me to bounce ideas off you. Due thanks must also be given to my associate supervisors, Associate Professor Bill Davison and Associate Professor Harry H. Taylor, who also gave me constructive criticism on drafts and experimental procedures. Huge thanks must be given to Dr Andrew Jeffs of NIWA who kindly supplied me with all my animals, diet ingredients and papers that were not available in the library. Thank you for your constructive criticisms on drafts. Without your help, this research would not have been possible. I look forward to undertaking my PhD under your supervision in the near future. Thank you to the technical staff of the Zoology Department for their advice and willingness to find ways around problems. Special thanks must be given to ·Gavin Robinson for advice on holding facilities and experimental equipment. You provided unlimited help throughout this project while also meeting the needs of other research students and running laboratories for undergraduates. A HUGE BIG THANK YOU to the following people: Dr Steven Gieseg for the use of the Biochemistry laboratory and for the help with techniques and calculations of lipid and glycogen analysis. Liam Cassidy for showing me the initial procedures involved. Associate Professor Laurie Greenfield from PAMS for allowing numerous protein samples to be processed in his laboratory and Jackie Healy for freeze drying my samples. John Pirker for much appreciated advice on the grim world of statistical analysis. Acknowledgements iv Aeron Storey and Jo Alcock for feeding and taking care of my growth experiment over the Christmas break. With out your help my growth experiment would have been a disaster. Jan McKenzie for your help with my poster, which was a great success. Thanks to my office buddy Melanie Bressington for your intellectual conversions regarding the estuary and my lobsters. A big thank you to Glen Thompson for providing with valuable break time during my writing and watch out for the Chiefs - they will take out the Super 12 next year! Last but not least, a huge thank you to my wife (Hannah) who put in the hard yards with me. Without your love and support I would not have been able to do this. Thank you for providing helpful comments and proof reading all drafts. Thanks must also be given to friends and family for their love and support. Table v TABLE OF CONTENTS PAGE ABSTRACT i ACKNOWLEDGMENTS iii LIST OF FIGURES ix LIST OF TABLES xiv CHAPTER ONE: GENERAL INTRODUCTION 1.1. Spiny Lobsters 1 1.2. Status of the New Zealand Rock Lobster Fishery 3 1.3. Aquaculture 4 1.4. Nutrition 7 1.5. Feed Energetics and Specific Dynamic Action 10 1.6. Aims 11 CHAPTER TWO: SPECIFIC DYNAMIC ACTION 2.1. Introduction 13 2.2. Methodology 16 Collection and Storage 16 Experimental Design 17 Determination of Oxygen Consumption 19 Determination of Ammonia Excretion 19 Diel Rhythms 20 SDA Determination 22 Parameters and Statistical Analysis 22 2.3. Results 23 2.3.1 Effect ofTime on Oxygen Consumption 23 2.3.2 Effect of Feeding Time on Oxygen Consumption 24 2.3.3 Effect of Time on Ammonia Excretion 27 Table vi 2.3.4 Effect of Feeding Time on Ammonia Excretion 28 2.3.5 Oxygen Nitrogen Ration 29 2.4. Discussion 32 2.4.1 Oxygen Consumption 32 2.4.2 SDA Coefficient 34 2.4.3 Ammonia Excretion and O:N Ratio 34 CHAPTER THREE: OXYGEN CONSUMPTION & AMMONIA EXCRETION IN TO FEEDING DIFFERENT CARBOHYDRATES 3 .1. Introduction 37 3.2. Methodology 40 Diet Formulation 40 Experimental Design 41 Experimental Procedure 41 Determination of Oxygen Consumption 41 Determination of Ammonia Excretion 42 Statistical Analysis 42 3.3. Results 42 3.3.1 General Carbohydrates 42 Oxygen Consumption 42 Ammonia Excretion 45 3.3.2 Algae Carbohydrates 52 Oxygen Consumption 52 Ammonia Excretion 53 3.3.3 General and Algal Carbohydrates 56 Oxygen Consumption 56 Ammonia Excretion 57 3.4. Discussion 63 3.4.1 Oxygen Consumption 63 3.4.2 Ammonia Excretion 65 Table of Contents vii CHAPTER FOUR: HAEMOLYMPH GLUCOSE 4.1. Introduction 67 4.2. Methodology 68 Experimental Design 68 Experimental Procedure 69 Haemolymph Sampling 71 Glucose Determination 71 Statistical Analysis 72 4.3. Results 72 4.3.1 General Carbohydrates 72 4.3.2 Algae Carbohydrates 77 4.3 .3 General and Algal Carbohydrates 79 4.4. Discussion 84 CHAPTER FIVE: GROWTH EXPERIMENT 5 .1. Introduction 89 5.2. Methodology 90 Collection and Storage 90 Experimental System and Water Quality 90 Preparation of Diets 92 Experiment One 93 Experiment Two 93 Experimental Procedure 94 Leaching 95 Food Intake 95 Glycogen and Lipid Analysis 96 Total Protein Analysis 98 Calculations 98 Statistical Analysis 99 Table of Contents viii 5.3. Results 100 5.3.1 Diets 100 5.3.2 Experiment One 100 Lobster Growth and Feed Consumption 100 Moults and Survival 105 Biochemical Composition 107 5.3.3 Experiment Two 108 Lobster Growth and Feed Consumption 108 Moults and Survival 116 Biochemical Composition 118 5.4.
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