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Thermal and Dietary Optimization in the Hatchery Culture Of THERMAL AND DIETARY OPTIMIZATION IN THE HATCHERY CULTURE OF JUVENILE PACIFIC GEODUCK CLAMS (PANOPEA GENEROSA, GOULD 1850) by Bianca Danielle Arney B.Sc. (Hons.), Hawaii Pacific University, 2009 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in The Faculty of Graduate and Postdoctoral Studies (Applied Animal Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) September 2013 © Bianca Danielle Arney, 2013 Abstract This research is the first to examine thermal and dietary optimization in the hatchery culture of juvenile Pacific geoduck (Panopea generosa). Chapter 2 investigated temperature and feed ration optimization; chapter 3 examined live algae substitution with the spray-dried species, Schizochytrium spp. or Spirulina (Arthrospira platensis). Geoduck growth and survival were measured to quantify treatment success. The temperature trial tested four temperatures (7, 11, 15, 19 °C) in juvenile and post-larval culture. Temperature promoted a significant growth effect in both sizes. The 19 °C culture elicited a delayed growth benefit in juveniles, and suppressed ash- free dry weight (AFDW), which recommends utilization of 15 °C. In contrast, geoduck post- larvae displayed immediate (post 7 d) shell growth acceleration at 19 °C. The 19 °C temperature shortened the rearing period by 2.9 d, suggesting its application in post-larval culture. The ration experiment examined the feed ration requirements of four geoduck juvenile size classes. Ration quantities between 0.0 - 128.0x10⁶ equivalent Isochrysis cells individual⁻¹ day⁻¹ were tested. All treatments received Chaetoceros muelleri and Isochrysis sp. mixed by AFDW. Following shell length/wet weight optimization, the following rations (10⁶ equivalent Isochrysis cells individual⁻¹ day⁻¹) should be applied between week 1 and 4 of the tested geoduck culture: 4.0 (1); 8.0 (2); 16.0 or 32.0 (shell length or wet weight optimum, respectively; 3); and 32.0 (4). The algae substitution trial replaced the above bi-algae diet with variant levels (0, 25, 50, 75, 100%) of Schizochytrium or Spirulina in two size classes. Between spray-dried diets, Spirulina treatments enhanced geoduck growth. However, overall growth displayed a general decline with elevated spray-dried inclusion and maximal growth occurred between 0 – 25% substitution. The superior bi-algae diet contained a polyunsaturated fatty acid Σn – 3/n – 6 ratio ii of 2.9, a value conserved in the bivalve literature. This may indicate its importance in future bivalve studies. Tissue analysis suggested potential limited digestion of spray-dried derived protein or carbohydrate, but indicated general lipid/fatty acid sequestration. These findings do not recommend dietary incorporation of Schizochytrium or Spirulina, but culture refinement may enhance juvenile geoduck hatchery production. iii Preface This thesis was completed under the Aquaculture Collaborative Research and Development Program, project number: P-10-01-007, under the supervision of Dr. R. Scott McKinley. Research design was formulated with the assistance of Dr. McKinley, Dr. Christopher M. Pearce, and Dr. Wenshan Liu, the project’s postdoctoral fellow and primary investigator. Live animal experimentation occurred at the Pacific Biological Station (PBS) under the supervision of Dr. Pearce, with the approval of the DFO Pacific Region Animal Care Committee (PRACC) and the guidelines of the Canadian Council on Animal Care (CCAC). The DFO PRACC Animal Use Protocol approval number was: 10-019. Dr. Liu maintained geoduck broodstock and larvae at PBS, and produced the geoduck post-larvae and juveniles used in experimentation. I performed the experimental husbandry and data collection, with assistance from Dr. Liu. Protein, lipid, carbohydrate, and fatty acid analyses were conducted at the Center for Aquaculture and Environmental Research, under the supervision of Dr. McKinley, and followed the recommendations of Dr. Ian Forster and Dr. Liu. I performed the analyses and data collection with assistance from Dr. Liu. Statistical analyses, data interpretation, and preparation of this thesis manuscript was completed by myself, with critical input provided by my graduate committee (Dr. McKinley, Dr. Pearce, Dr. Forster, Dr. Marina von Keyserlingk) and Dr. Liu. Chapters 2 and 3 of this thesis will be submitted for potential publication [Chapter 2: Arney, B., Liu, W., Forster, I., McKinley, R.S., Pearce, C.M. 2013. Temperature and feed ration optimization in the hatchery culture of juvenile Pacific geoduck clams (Panopea generosa Gould, 1850). Chapter 3: Arney, B., Liu, W., Forster, I., McKinley, R.S., Pearce, C.M. 2013. iv Feasibility of live algae dietary substitution with spray-dried Schizochytrium sp. or Spirulina (Arthrospira platensis) in the hatchery culture of juvenile Pacific geoduck clams (Panopea generosa Gould, 1850)]. Funding was provided by the Aquaculture Collaborative Research and Development Program. v Table of Contents Abstract ........................................................................................................................................... ii Preface............................................................................................................................................ iv Table of Contents ........................................................................................................................... vi List of Tables .................................................................................................................................. ii List of Figures ................................................................................................................................. ii List of Abbreviations and Symbols................................................................................................. ii Acknowledgements ........................................................................................................................ iv Dedication ...................................................................................................................................... vi 1 Introduction ............................................................................................................................. 1 1.1 The Pacific geoduck .............................................................................................................. 1 1.2 Commercial fisheries............................................................................................................. 2 1.3 Aquaculture ........................................................................................................................... 5 1.4 Requirements for geoduck hatchery culture .......................................................................... 9 1.5 Temperature .......................................................................................................................... 9 1.5.1 Objective 1 .................................................................................................................... 11 1.5.2 Hypothesis 1 ................................................................................................................. 11 1.6 Feed ration ........................................................................................................................... 11 1.6.1 Objective 2 .................................................................................................................... 13 1.6.2 Hypothesis 2 ................................................................................................................. 13 1.6.3 Hypothesis 3 ................................................................................................................. 13 1.7 Limitations in live microalgae............................................................................................. 14 1.8 Bivalve nutritional requirements ......................................................................................... 15 1.8.1 Total protein ................................................................................................................. 15 1.8.2 Total carbohydrate ........................................................................................................ 16 1.8.3 Total lipid ..................................................................................................................... 17 1.8.4 Fatty acid composition.................................................................................................. 18 1.9 Spray-dried live algae substitution ...................................................................................... 21 1.9.1 Objective 3 .................................................................................................................... 23 vi 1.9.2 Hypothesis 4 ................................................................................................................. 23 1.9.3 Hypothesis 5 ................................................................................................................. 24 1.9.4 Objective 5 .................................................................................................................... 24 1.9.5 Hypothesis 6 ................................................................................................................. 24 1.10 Summary ........................................................................................................................... 25 2 Temperature
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