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OPTIMAL PROTEIN-LIPID LEVEL AND REPLACEMENT OF FISH MEAL WITH PLANT PROTEIN SOURCES IN FEEDS FORMULATED FOR Pangasianodon hypophthalmus (Sauvage, 1878) PREEDA PHUMEE UNIVERSITI SAINS MALAYSIA JUNE 2011 OPTIMAL PROTEIN-LIPID LEVEL AND REPLACEMENT OF FISH MEAL WITH PLANT PROTEIN SOURCES IN FEEDS FORMULATED FOR Pangasianodon hypophthalmus (Sauvage, 1878) by PREEDA PHUMEE Thesis submitted in fulfillments of the requirements for the degree of Philosophy doctoral June 2011 TAHAP PROTEIN-LIPID YANG OPTIMAL DAN PENGGANTIAN SERBUK IKAN DENGAN SUMBER PROTEIN TUMBUHAN DI DALAM MAKANAN FORMULA BAGI Pangasianodon hypophthalmus (Sauvage, 1878) Oleh PREEDA PHUMEE Tesis yang diserahkan untuk memenuhi keperluan bagi Ijazah Sarjana Sains Jun 2011 ACKNOWLEDGEMENTS This study has been successfully completed with a great deal of contribution from the following helpful individuals. First of all, I would like to express my sincere and heartfelt appreciation to my supervisor, Professor Roshada Hashim for her invaluable guidance, constructive comments, encouragement and support throughout my studying in USM. Similarly, I would like to express my great appreciation to all of my friends (Dr. Sarita Ramanchandan, Amalia, Khalida, Karloon, Sharifah, Afini, Ahmed, Mohamad Aliyu Paiko, Mohamad Aldohal, Nana and Aunty Anna) for sharing their knowledge and expertise in this project. Moreover, I also would like to thanks the staffs in the School of Biological Science. I am grateful to the President of Rajamangala University of Technology Srivijaya (RMUT SV), Thailand, Associate Professor Pracheep Choopunth, for his encouragement and provide me the scholarship. My appreciation also extends to Dean of Faculty of Science and Fisheries Technology, RMUT SV, Trang campus, Thailand, Assistant Professor Dr. Suwit Jitpukdee, for his permission and support regarding my pursuing for further study. Thankful to all my colleagues in RMUT SV, Trang campus for their unselfish friendship. Most of all, I would like to express my deepest love and gratitude to my family, my mother, brothers and sisters for their devoted encouragement and patience. Special thank to my sister in law for her uncountable helping me to taking care my son. To my beloved family, Tavon and my lovely son; Phum, who give me heart power and sincerely love. Preeda Phumee ii TABLE OF CONTENTS PAGE ACKNOWLEDGEMENTS ii TABLE OF CONTENTS iii LIST OF TABLES ix LIST OF FIGURES xiii LIST OF ABBREVIATIONS xiv ABSTRAK xvii ABSTRACT xix CHAPTER 1 INTRODUCTION 1.1 Background of Aquaculture 1 1.2 Problem Statement 4 1.3 Research Objectives 6 CHAPTER 2 LITERATURE REVIEW 2.1 Aquaculture Challenges 7 2.1.1 Feeds Costs and Solution 9 2.2 Sutchi catfish 11 2.2.1 Biology, Morphology and Classification 11 2.2.2 Aquaculture 13 2.2.3 Feed 2.2.3.1 Homemade Feed 14 2.2.3.2 Commercial Feed 15 2.3 Nutritional Requirements 2.3.1 Proteins 16 2.3.2 Lipids 17 iii 2.3.3 Carbohydrates 18 2.4 Proteins and Amino acids 2.4.1 Introduction 20 2.4.2 Amino Acids 21 2.4.3 Protein Requirements 22 2.4.4 Protein-Energy Ratio 24 2.5 Sources of Protein 26 2.5.1 Fish meal 26 2.5.2 Soybean meal 26 2.6 Disadvantages of soybean 28 2.7 Nutritional Improvement of Soybean Meal 31 2.7.1 Extracting 31 2.7.2 Heat treatment 32 2.7.3 Genetic engineering 32 2.7.4 Enzyme treatment 33 2.7.5 Mixing with other protein sources 34 2.8 Spirulina 35 CHAPTER 3 MATERIALS AND METHODS 3.1 Introduction 41 3.2 Proximate Analysis 41 3.3 Amino Acid Analysis 42 3.4 Fatty Acid Analysis 43 3.5 Growth Performance, Feed Utilization and Body Indices Determination 44 iv CHAPTER 4 THE OPTIMAL PROTEIN AND LIPID REQUIREMENT IN FORMULATED DIETS FOR JUVENILE SUTCHI CATFISH (Pangasianodon hypophthalmus, Sauvage 1878). 4.1 Introduction 45 4.2 Materials and Methods 47 4.2.1 Experimental Diets 47 4.2.2 Fish and Experimental Conditions 49 4.2.3 Sampling Procedure 49 4.2.4 Growth Performance Assessment 50 4.2.5 Analytical Procedure 50 4.2.6 Data and Statistical Analysis 50 4.3 Results 51 4.3.1 Nutrients Composition of Experimental Diets 51 4.3.1 (a) Chemical Composition 51 4.3.1 (b) Amino Acid Composition of Experimental Diets 51 4.3.1 (c) Fatty Acid Composition of Ingredients and Experimental Diets 52 4.3.2 Growth Performance 54 4.3.3 Feed Utilization 56 4.3.4 Body Indices 57 4.3.5 Carcass Composition 58 4.3.6 Amino Acid Content of Fish Muscle 60 4.3.7 Fatty Acid Content of Fish Muscle 62 4.4 Discussion 66 v 4.4.1 The Effects on Growth and Feed Utilization 66 4.4.2 The Effect on Body Indices 68 4.4.3 The Effect on Carcass Composition 69 4.4.4 The Effect on Muscle Amino Acid Composition 70 4.4.5 The Effect on Muscle Fatty Acid Composition 70 4.5 Conclusion 71 CHAPTER 5 FISH MEAL REPLACEMENT BY SOYBEAN MEAL IN THE FORMULATED DIETS FOR JUVENILE SUTCHI CATFISH (Pangasianodon hypophthalmus Sauvage, 1878 ) 5.1 Introduction 72 5.2 Materials and Methods 73 5.2.1 Experimental Diets 73 5.2.2 Fish and Experimental Conditions 74 5.2.3 Sample Collection 76 5.2.4 Evaluation of Growth Parameters 76 5.2.5 Analytical Procedure 76 5.2.5 (a) Proximate Analysis 5.2.6 Statistical Analysis 77 5.3 Results 78 5.3.1 Nutrient Composition of Protein Sources and Experimental Diets 78 5.3.2 Amino Acid Composition of Experimental Diets 78 5.3.3 Fatty Acid Composition of Experimental Diets 81 5.3.4 Growth Performance 83 5.3.5 Feed Utilization Efficiency 85 5.3.6 Muscle Composition and Body Indices 87 5.3.7 Apparent Digestibility Coefficients 89 vi 5.3.8 Amino Acid Content of Fish Muscle 90 5.3.9 Fatty Acid Content of Fish Muscle 92 5.3.10 Fatty Acid Content of Fish Liver 94 5.4 Discussion 96 5.4.1 Growth Performance and Feed Utilization 96 5.4.2 Muscle Composition 99 5.4.3 Body Indices 100 5.4.4 Apparent Digestibility 100 5.4.5 Amino Acid Content of Fish Muscle 101 5.4.6 Fatty Acid Content of Fish Muscle 102 5.5 Conclusion 104 CHAPTER 6 EVALUATION OF A COMBINATION OF SPIRULINA AND SOYBEAN MEAL AS A PROTEIN SOURCE FOR JUVENILE SUTCHI CATFISH, Pangasianodon hypophthalmus (Sauvage, 1878) DIETS 6.1 Introduction 105 6.2 Materials and Methods 107 6.2.1 Experimental Diets 107 6.2.2 Fish and Experimental Conditions 111 6.2.3 Sampling Procedure 111 6.2.4 Growth Performance Assessment 112 6.2.5 Analytical Procedures 112 6.2.5 (a) Proximate Analysis 112 6.2.6. Data and Statistical Analysis 112 6.3 Results 113 6.3.1 Amino Acid Composition of Ingredients and Experimental Diets 113 vii 6.3.2 Fatty Acid Composition of Ingredients and Experimental Diets 115 6.3.3 Growth Performance 120 6.3.4 Feed Utilization 121 6.3.5 Muscle Composition and Body Indices 122 6.3.6 Amino Acid Composition of Fish Muscle 125 6.3.7 Fatty Acid Composition of Fish Muscle 127 6.4 Discussion 129 6.4.1 The Effects on Growth and Feed Utilization 129 6.4.2 The Effects on Muscle Composition and Body Indices 131 6.4.3 The Effects on Amino Acid Composition 133 6.4.4 The Effects on Fatty Acid Composition 134 6.5 Conclusion 136 CHAPTER 7 CONCLUSION AND RECOMMENDATIONS 7.1 Introduction 137 7.2 Conclusions 137 7.2.1 Growth Performance 137 7.2.2 Feed Utilization 138 7.2.3 Flesh Quality 139 7.3 Recommendations for Further Research 140 REFERENCES 141 APPENDICES LIST OF PUBLICATIONS viii LIST OF TABLES PAGE Table 2.1 Essential and non-essential amino acids in fish 22 Table 2.2 Summaries on the effects of Spirulina on growth and survival 37 Table 4.1 Ingredients and proximate composition of experimental diets 48 Table 4.2 Amino acid composition (% dry weight) of the experimental diets 52 Table 4.3 Fatty acid composition (% of total fatty acids) of feed ingredients 53 Table 4.4 Fatty acid composition (% total fatty acids) of experimental diets 54 Table 4.5 Growth performance of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks. 55 Table 4.6 Feed utilization (% dry matter basis) of sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks. 57 Table 4.7 Body indices of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks 58 Table 4.8 Proximate composition (% dry matter basis) of carcass of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks. 59 Table 4.9 Amino acid composition (% dry weight) of muscle of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks 61 ix Table 4.10 Fatty acid composition (% total fatty acids) of muscle of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks. 64 Table 4.11 Two-way ANOVA of some fatty acid (% total fatty acids) of muscle of juvenile sutchi catfish, Pangasianodon hypophthalmus fed experimental diets containing different protein and lipid levels for eight weeks. 64 Table 5.1 Ingredients and composition of experimental diets. 75 Table 5.2 Proximate compositions (% dry matter) of fish meal, soybean meal and experimental diets.