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Improvement of Yellow Perch Larvae Culture Via Live Food Enrichment with Polyunsaturated Fatty Acids THESIS Presented in Partial

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Improvement of Yellow Perch Larvae Culture Via Live Food Enrichment with Polyunsaturated Fatty Acids THESIS Presented in Partial Improvement of Yellow Perch Larvae Culture via Live Food Enrichment with Polyunsaturated Fatty Acids THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By John David Grayson Graduate Program in Environment and Natural Resources The Ohio State University 2014 Master's Examination Committee: Dr. Konrad Dabrowski, Advisor Dr. Suzanne Gray Dr. Robert Gates Dr. Ana Hill Copyrighted by John David Grayson 2014 Abstract Limited experience with live food regimes and fragmentary knowledge of nutritional requirements have been inhibitors for the indoor-intensive production of Yellow Perch Perca flavescens larvae. Live food enrichment with polyunsaturated fatty acids (PUFA) is a proven means of increasing the growth and survival of larval fish, but no studies to date have tested this method on Yellow Perch. This thesis consists of two live food enrichment experiments carried out in May/June of 2013 and 2014, as well as lipid analysis of live food and fish samples from both years. The 2013 study examined the effect of live food enrichment with docosahexaenoic acid (DHA; C22:6[n-3]) and arachidonic acid (ARA; C20:4[n-6]) on the growth, survival, and swim bladder inflation of larval Yellow Perch. The 2014 experiment was similar in design, but compared PUFA enrichments in ethyl ester (EE) and triglyceride (TG) forms. Both experiments were conducted in two phases. The first phase was carried out in a recirculating system with nine 50 L conical tanks, initially stocked at 50-70 larvae/L. Live rotifers Brachionus plicatilis were provided to larvae for the first two days of exogenous feeding, before transitioning to Artemia franciscana nauplii for the remaining eight days of this phase. The second phase was carried out in nine 60 L cylindrical flow-through tanks, initially stocked with 10 larvae/L. During this phase, fish were fed Artemia nauplii for 3 ii days, then gradually transitioned to a formulated starter diet (Otohime A®) over a 7 day period. At the end of the first phase the ARA and DHA enriched groups had significantly (p≤0.05) improved swim bladder inflation rates when compared to the control group. For the second phase, enriched groups had significantly larger mean weights and growth rates than the control. The EE-TG experiment was similar in design to the DHA-ARA experiment, except that the second phase was concluded after seven days of feeding. At the end of the first phase, the EE group had a significantly improved average weight and growth rate than the TG group. No significant trends were seen in the second phase. Following enrichment experiments, the fatty acid composition of live feeds and experimental fish were analyzed using the gas chromatography method. Fatty acid composition of zooplankton was heavily influenced by enrichments, and composition of larvae/juveniles generally reflects that of their live prey. Arachidonic acid was assimilated poorly in ARA enriched Artemia, but DHA was found in abundance. Also, rotifers tended to assimilate PUFA better in EE form, while Artemia achieved higher PUFA contents with TG enrichments. The data support that PUFA enrichment of live food can be utilized to increase the success of Yellow Perch culture by increasing growth and swim bladder inflation rates during the critical period of larval development. iii Acknowledgments I would like to first and foremost thank my advisor, Dr. Konrad Dabrowski, for his patient instruction, seemingly endless knowledge, and long hours of help. I would also like to thank the other members of my examination committee and the helpful staff within at the School of Environment and Natural Resources. I would like to thank Dr. Karolina Kwasek and Dr. Michal Wojno for their guidance with culturing larvae and Dr. Malgorzata Korzeniowska for assisting me greatly with lipid analysis. Lastly, I would like to thank Tim Parker, Nevine Shabana, Abigail King, Mackenzie Miller, Megan Kemski, and Mohammed Alam for their assistance and support. iv Vita June 2008 ............................Keystone High School, LaGrange, Ohio June 2012 ............................B.S. Environment and Natural Resources, The Ohio State University Sep. 2012 to April 2014 .... Graduate Administrative Assistant, SENR, The Ohio State University Aug. 2014 to present ..........Graduate Teaching Assistant, SENR, The Ohio State University Field of Study Major Field: Environment and Natural Resources v Table of Contents Abstract ................................................................................................................... ii Acknowledgments.................................................................................................. iv Vita .......................................................................................................................... v Field of Study .......................................................................................................... v Table of Contents ................................................................................................... vi Chapter 1: Comparison of DHA-based and ARA-based Enrichments ................... 1 Introduction ......................................................................................................... 1 Methods ............................................................................................................... 4 Facilities and Fish ............................................................................................ 4 Enrichment Procedure ..................................................................................... 8 Statistical Analysis ........................................................................................ 10 Results ............................................................................................................... 11 Discussion ......................................................................................................... 11 Culture System .............................................................................................. 11 Fish Performance ........................................................................................... 13 Chapter 2: Comparison of Enrichments in TG and EE forms .............................. 17 vi Introduction ....................................................................................................... 17 Methods ............................................................................................................. 18 Facilities and Fish .......................................................................................... 18 Enrichment Procedure. .................................................................................. 21 Sample Collection and Measurement. ........................................................... 22 Statistical Analysis. ....................................................................................... 22 Results ............................................................................................................... 24 Discussion ......................................................................................................... 24 Fish Performance ........................................................................................... 24 Chapter 3: Lipid Analysis of Yellow Perch and Enriched Live Feeds ................. 29 Introduction ....................................................................................................... 29 Methods ............................................................................................................. 30 Lipid Separation and Analysis ....................................................................... 30 Statistical Analysis ........................................................................................ 31 Results ............................................................................................................... 32 DHA-ARA Enrichment Experiment ............................................................. 32 EE-TG Enrichment Experiment .................................................................... 33 Discussion ......................................................................................................... 38 DHA-ARA Experiment ................................................................................. 38 vii EE-TG Experiment ........................................................................................ 44 Culture Success.............................................................................................. 46 Future Research ............................................................................................. 48 Literature Cited (AFS) .......................................................................................... 51 Appendix A: Weight Corrections for DHA-ARA Enrichment Experiment ......... 57 Appendix B: T Tests and ANOVA Tables ........................................................... 59 viii List of Tables Table 1. Mean (±SD) water chemistry parameters within culture tanks during the first 10 days of feeding (Phase I) in the DHA-ARA enrichment experiment. ........ 6 Table 2. Mean (±SD) weight (mg) of Yellow Perch juveniles at the end of Phase I (10 days feeding) and Phase II (24 days feeding) with unenriched (Control), DHA enriched, and ARA enriched live feeds. Larvae/juveniles are further stratified into those with inflated swim bladders (SB+) and those without
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