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Growth, Gonadal Development, and Blood Profile in African Catfish (Clarias Gariepinus, Burchell 1822) Fed Diets Containing Cocoa Bean Meal

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Growth, Gonadal Development, and Blood Profile in African Catfish (Clarias Gariepinus, Burchell 1822) Fed Diets Containing Cocoa Bean Meal i GROWTH, GONADAL DEVELOPMENT, AND BLOOD PROFILE IN AFRICAN CATFISH (CLARIAS GARIEPINUS, BURCHELL 1822) FED DIETS CONTAINING COCOA BEAN MEAL RESEARCH PROJECT REPORT SUBMITTED TO THE DEPARTMENT OF ANIMAL SCIENCE, FACULTY OF AGRICULTURE, UNIVERSITY OF NIGERIA, NSUKKA IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTERS DEGREE IN ANIMAL REPRODUCTIVE PHYSIOLOGY. BY UZOCHUKWU, IFEANYI EMMANUEL PG/M.SC/13/64916 SUPERVISOR: PROF. S.O.C UGWU JULY, 2016 ii CERTIFICATION This is to certify that this work was carried out by UZOCHUKWU, IFEANYI EMMANUEL in the Department of Animal Science, Faculty of Agriculture, University of Nigeria, Nsukka. The above statement and the entire project therefore stand approved by: __________________________ __________________________ PROF. S.O.C UGWU PROF. A.G EZEKWE (Supervisor) (Head of Department) Date………………………… Date………………………………… ____________________________________ External examiner Date……………………………………….. iii DEDICATION I dedicate this report in a special way to My Parents, Mr. and Mrs. David Uzochukwu, my uncle, Chief. Ufondu, B.O, and to my younger sister Mrs. Orji-Uzochukwu C.B. for the hope they had in me even when things seemed rough. iv ACKNOWLEDGEMENT I wish to offer my profound gratitude to God Almighty for making this programme a success. I would not have started it in the first place if not for his grace, mercies and love. I am also grateful to my parents Mr. and Mrs. David Uzochukwu for always being there for me. My hearty appreciation goes to my Uncle, Chief. Ufondu, Boniface Onyebuchi who has been the financial pillar of my academic journey right from the onset. May God bless him abundantly and keep him alive to see the great man he has trained. I wish to also say a big thanks to my siblings, Chioma, Nnamdi, Chinyere, and Uchenna for contributing enormously to the success of this programme especially during the time financial waves and tides became furious. I wish to say a very big thank you to my first academic father, Prof. S.O.C. Ugwu (Sinque) for the understanding, guidance, support and opportunity he gave me to develop myself. To all my lecturers who taught and assisted me during the course of this programme: Prof. (Emeritus) C.C.C. Nwosu, who was my statistician during the design phase of this work; my second academic father, Prof. A.G Ezekwe, who made out time to visit the farm and made crucial adjustments; Prof. S.S.I Omeje; Dr. A.E. Onyimonyi; Dr. A.O Ani, who helped greatly with the experimental feed formulation; Dr. H.N. Foleng who took the time and stress to analyze the large amount of data collected from the study at no cost whatsoever; and Mr. C.O Osita. I wish to appreciate in a very special way the assistance given to me by Mr. Ugwuoke, C.C. and Mr. Ossai, N. in designing the experiment and guiding me throughout the entire stay in the farm. Also to my field assistant, Mrs. Cordelia (Mama Fish), I say thanks for her time and understanding. I owe them a lot. Furthermore, in no little way I will forever remain grateful to the great friends I had the opportunity of knowing. My very special lady, Miss Nnamuka, C.M. My blogger brother, Ezekwe, M. Great friends, Mr. Amaefule, B.C., Mr. Egom, M.A., Miss. Umezurike, G.O., Miss. Arokesi, A., Miss. Okafor, L., Miss. Ezike, M.N., Mr. Dim, C.E., and Miss. Ozoude, L. Without this great people who devoted time and efforts to assist me through the vigorous process of data collection at no cost, I would have stopped the work long before I started. I am forever indebted to them for these great deeds. To other persons who contributed in one way or the other to the success of this work, I beg their pardon for the omission of their names. I am greatly grateful for your assistance. I pray that God will grant them opportunities even as they continue to remain blessings to other people. v TABLE OF CONTENT Title page i Certification ii Dedication iii Acknowledgment iv Table of contents v List of Table ix List of Figures xi Abstract xiv CHAPTER ONE: INTRODUCTION 1 1.2 Objectives of the Study 4 1.3 Justification of Research 4 CHAPTER TWO LITERATURE REVIEW 6 2.1 Aquaculture Production: Nigeria 6 2.2: CATFISH: Clarias species 8 2.2.1 African Catfish (Clarias gariepinus) 9 2.2.2 Catfish Production in Nigeria 10 2.3 Nutritional Requirements of Catfish 12 2.3.1 Energy Requirements 13 2.3.2 Dietary Carbohydrate 14 2.3.3 Dietary Lipids 15 2.3.4 Protein and Amino Acids Requirements 16 2.3.5 Vitamins and Minerals 18 2.4 Cocoa (Theobroma Cacao) 20 2.4.1 Origin and Taxonomy 20 vi 2.4.2 Production and Distribution 21 2.4.3 Products from cocoa 22 2.4.4 Chemical composition of the cacao fruit, cacao bean and cacao products 24 2.4.5 Polyphenols 24 2.4.5.1 Bioavailability of polyphenols 26 2.4.5.2 Cocoa Polyphenols And Health 27 2.4.5.3 Methylxanthines 30 2.4.6 Theobromine and Caffeine 31 2.4.6.1 Theobromine toxicity 33 2.4.6.2 Detheobromination methods 33 2.4.7 Proteins 33 2.4.8 Vitamins and Minerals 34 2.5 Cocoa By-Products and Their Use In Animal Production 35 2.5.2 Cocoa pod husk meal 36 2.5.3 Cocoa bean shell (CBS) 38 2.5.4 Cocoa bean meal / Cocoa meal / Cocoa bean cake (CBM/CM/CBC) 39 2.6 Reproductive Structures of the Catfish: The Gonads 39 2.6.1 The Testes 39 2.6.2 Ovary 41 2.6.3 Gonadal development and maturation of African Catfish 41 2.6.3.1 Ovarian development 42 2.6.3.2 Developmental Stages of the Oocytes 45 2.6.4 Testicular Development 49 2.6.5 Phases of Spermatogenesis during Catfish Development 53 2.7 Factors Affecting Gonadal Development 54 vii 2.7.1 Season 54 2.7.2 Temperature 54 2.7.3 Feeding levels 55 2.7.4 Age and size 55 2.7.5 Others factors 56 2.8 Gonadosomatic Index 56 2.9 Condition Factor (K) 57 CHAPTER THREE: MATERIALS AND METHODS 3.1 Research Location and Climatic Distribution 58 3.2 Duration of Study 58 3.3 Procurement and management of fish 58 3.4 Procurement of cocoa bean meal (cocoa bean cake) 59 3.5 Experimental layout/design 59 3.6 The experimental diets 60 3.7 Data collection 61 3.8 Experimental Design 67 3.9 Statistical Analysis 67 CHAPTER FOUR RESULT AND DISCUSSION 4.1: Growth Performance 68 4.2 Ovarian Development 69 4.2.1 Gross morphology and description of ovarian stages 69 4.2.2 Effect of varying dietary levels of cocoa bean meal on the ovarian development of C. gariepinus catfish. 73 4.2.3 Oocyte Development 77 viii 4.24 Effect of varrying dietary inclusions of cocoa bean meal (cbm) on oocyte distribution in C. gariepinus 84 4.2.5 Histomorphometry 91 4.3 Testicular Development 94 4.3.1 Description and classification of the testes into stages 94 4.3.2 Effect of varying dietary levels of cocoa bean meal on testicular development of C. gariepinus catfish. 96 4.3.3 Histomorphology of the C. gariepinus catfish testes 101 4.3.4 Histomorphometrical Studies 111 4.4 Effect of dietary inclusion of cocoa bean meal on haematology of C. gariepinus catfish 113 4.5 Effect of dietary inclusion of cocoa bean meal on blood lipid profile of C. gariepinus catfish 115 CHAPTER FIVE 5.1 Conclusion 117 5.2 Recommendation 117 REFERENCES 118 ix LIST OF TABLES Table 1: Projected Human Population, Fish Demand and Supply in Nigeria (2000–2015) 8 Table 2: Zoological classification of Clariasgariepinus (Burchel 1822) 10 Table 3: Estimated Catfish production (metric tonnes) and job creation from catfish in Nigeria (2001 -2012) 11 Table 4: Recommended dietary nutrient levels (in DM) for African catfish (Clariasgariepinus) 17 Table 5: Nutrient requirement of C. gariepinus fish as reported by several researchers 18 Table 6: Dietary protein and energy requirements for African catfish (Clariasgariepinus) 18 Table 7: Scientific Classification of Cocoa 20 Table 8: Global cocoa bean production from 2011-2013 21 Table 9: Nutritional Properties of Cocoa 35 Table 10: The proximate composition (% dry matter) of by-products from the manufacture of cocoa 36 Table 11: Stages of ovarian development using macroscopic and microscopic descriptions 44 Table 12: Phases of oocyte development in Clariasgariepinus, based on histological criteria 47 Table 13: Testicular development stages of the C. gariepinus testes based on Histomorphology 51 Table 14: Phases of spermatogenesis in Clariasgariepinus, based on histological criteria 53 Table 15: The Layout of the study 59 Table 16: Sampling of the experimental fish for histology studies 60 Table 17: percentage composition of the experimental diet 61 Table 18: Effect of dietary inclusion of cocoa bean meal on growth performance x of C. gariepinus catfish 68 Table 19: Ovarian development of C. gariepinus catfish fed varying dietary levels of cocoa bean meal 73 Table 20: Oocyte distribution for the different treatment groups at the 3rd, 5th, 7th, and 9th week post treatment. 84 Table 21: Means of oocyte diameters of the different developmental stages at weeks 3, 5, 7 and 9 post treatment 91 Table 22: Effect of varying dietary levels of cocoa bean meal on body weight and testicular growth C. gariepinus catfish 96 Table 23: Tubular diameter and epithelial height of C. gariepinus catfish fed diets containing varying inclusion levels of CBM.
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