Evaluation of Alternative Ingredients As Fish Meal Replacers in On-Growing Diets for Red Porgy (Pagrus Pagrus)”

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Evaluation of Alternative Ingredients As Fish Meal Replacers in On-Growing Diets for Red Porgy (Pagrus Pagrus)” lm t i(�Hl� yn IYE't�I tan o ! ªAD DE LAS PALMAS DE GRAN CANARIA Sanidad Animal Scgvrid,1d Aliml'nl,inJ Anexo 1 Dª MARÍA SORAYA DÉNIZ SUÁREZ, SECRETARIA DEL INSTITUTO UNIVERSITARIO DE SANIDAD ANIMAL Y SEGURIDAD ALIMENTARIA DE LA UNIVERSIDAD DE LAS PALMAS DE GRAN CANARIA. CERTIFICA Que el Consejo de Doctores del Departamento en su sesión de fecha 21 de diciembre de 2011 tomó el acuerdo de dar el consentimiento para su tramitación, a la tesis doctoral europea titulada: "Evaluación de ingredientes alternativos a la harina de pescado en dietas de engorde para bocinegro (pagrus pagrus)", presentada por la doctoranda Dª Josefa García Romero, dirigida por la Dra. Dª. Lidia Robaina Robaina y Dr. D. Rafael Ginés Ruiz. Y para que así conste, y a efectos de lo previsto en el Artº 73.2 del reglamento de Estudios de Doctorado de esta Universidad, firmo la presente en Las Palmas de Gran Canaria, a veintidós de diciembr� de dos mil once. AnexoAnexo IIAnexo II UNIVERSIDADUNIVERSIDAD DE LAS PALMAS DE GRAN CANARIA Departamento: Instituto Universitario de Sanidad Animal y Seguridad Alimentaria Programa de Doctorado: Acuicultura Título de la Tesis “Evaluation of alternative ingredients as fish meal replacers in on-growing diets for red porgy (Pagrus pagrus)” Tesis Doctoral presentada por Josefa García Romero Dirigida por la Doctora Lidia Esther Robaina RRobainaobaina y el Doctor Rafael Ginés Ruiz La Directora El Director La Doctorando Las Palmas de Gran CanarCanariiiia,a, a 16 de Diciembre de 2011 LIDIALIDIA ESTHER ROBAINA ROBAINA, PROFESORA EN EL ÁREA DE CONOCIMIENTO DE ZOOLOGÍA DEL DEPARTAMENTO DE BIOLOGÍA DE LA UNIVERSIDAD DE LAS PALMAS DE GRAN CANARIACANARIA.... INFORMA: Que Doña JosDoña JosefaJosefa García RomeroRomero, Licenciada en Ciencias de Mar, ha realizado bajo mi dirección y asesoramiento el presente trabajo titulado: “Evaluation of alternative ingredients as fish meal replacers in onon----growinggrowing diets for red porgy (Pagrus pagrus)”. (Evaluación de ingredientes altealternativosrnativos como sustitutos de la harina de pescado en dietas de engorde para el bocinegro (Pagrus pagrus)”))”))”),)”),,, el cual considero reúne las condiciones y calidad científica para optar al Grado de Doctor. FDO.: LIDIA ESTHER ROBAINA ROBAINROBAINAAAA RAFAELRAFAEL GINÉS RUIZ, PROFESOR TITULAR DE UNIVERSIDAD EN EL ÁREA DE CONOCIMIENTO DE PRODUCCIÓN ANIMAL DE LA FACULTAD DE VETERINARIA DE LA UNIVERSIDAD DE LAS PALMAS DE GRAN CANARIACANARIA.... INFORMA: Que Doña Josefa García RomeroRomero, Licenciada en Ciencias de Mar, ha realizado bajo mi dirección y asesoramiento el presente trabajo titulado: “Evaluation of alternative ingredients as fish meal replacers in onon----growinggrowing diets for red porgy (Pagrus pagrus)”. (Evaluación de ingredientes alternativos como sssustitutossustitutosustitutosustitutos de la harina de pescado en dietas de engorde para el bocinegro (Pagrus pagrus)”))”))”),)”),,, el cual considero reúne las condiciones y calidad científica para optar al Grado de Doctor. FDO.: RAFAEL GINÉS RUIZ EEEEvvaalluuaattiioonn ooff aalltteerrnnaattiivvee iinnggrreeddiieennttss aass ffiisshh mmmmmmeeaall rreeppllaacceerrss iinn oonn--ggrroowwwwwwiinngg ddiieettss ffoorr rreedd ppoorrggyy ((PPPPaaggrruuss ppaaggrruuss)))) JJJJoosseeffaa GGaarrccííaa RRoommeerroo Grupo de Investigación en Acuicultura (GIA) Instituto Canario de Ciencias Marinas (ICCM) Instituto Universitario de Sanidad Animal y Seguridad Alimentaria (ULPGC) Thesis for the degree of Doctor of Phylosophy University of Las Palmas de Gran Canaria, 2011 Directors: Dra. Lidia Esther Robaina Robaina & Dr. Rafael Ginés Ruiz AAA mimimi fafamimilliaia,,familia, CONTENTS List of tables LListist of figures AbbreviationsAbbreviations AAcknowledgmentscknowledgments 11.... GENERAL INTRODUCTION 1 1.11.11.1.1.1... AQUACULTURE PRODUCTION………………………………………PRODUCTION……………………………………………………………………… 1 1.21.21.2.1.2... PROTEIN IN DIETS FOR FISH SPECIES………………………………...SPECIES………………………………............... 2 1.2.1. Importance of protein in compounds feed for farming finfish……………… 3 1.2.2. Fish meal as the main protein source in fish diets………………………...... 4 1.2.3. Fish meal current world production and situation………………………...... 5 1.31.31.3.1.3... ALTERNATIVE PROTEIN INGREDIENTINGREDIENTSSSS TO FISH MEAL IN FISFISHH DIETS………………………………DIETS……………………………………………………………………………………………………………………………………………………………………….................... 6 1.3.1. Alternative protein sources from terrestrial origin………………………….. 6 1.3.2. Product and by-products from marine sources as novel ingredients in aquaculture feed…………………………………………………………………... 8 1.3.2.1. Crab products and by-products………………………………………………..... 11 1.3.2.1.1. Chitin long-change polysaccharide as potential bio-component from crustacean meals………………………………………………………………….. 14 1.3.2.1.2. Dietary crab meal and fish flesh quality…………………………………….... 15 1.3.2.2. Other potential ingredients from marine sources: by-products processing from echinoderm industry……………………………………………………….. 17 1.41.41.4.1.4... THE RED PORGY (Pagrus pPagrus pagruspagrusagrusagrus; Linnaeus, 1758)…….1758)…….……………………………………………….............. 17 1.4.1. External appearance, geographical distribution, habitat and reproduction……………………………………………………………………...... 17 1.4.2. Natural diet of fish red porgy………………………………………………….... 18 1.4.3. Red porgy culture…………………………………………………………………. 19 1.4.3.1. Dietary protein and carotenoid pigments requirements……………….…... 20 222.2... OJECTIVES 25 333.3... GENERAL MATERIALS AND METHODS 29 3.13.13.1.3.1... ANIMALS…………………………………………………………………………ANIMALS…………………………………………………………………………........ 29 3.23.23.2.3.2... CULTURE CONDITIONS……………………………………………CONDITIONS……………………………………………………………….……….……….………. 30 3.2.1. Facilities……………………………………………………………………………. 30 3.2.2. Experimental tanks………………………………………………………………. 30 3.2.3. Experimental conditions…………………………………………………………. 30 3.33.33.3.3.3... EXPERIMENTAL FEED INGREDIEINGREDIENTS………NTS………NTS………………………………………………………………………………………………… 31 3.3.1. River crab, Procambarus clarkii, meal………………………………………… 31 3.3.2. Marine crab, Chaceon affinis and Paramola cuvieri, meals………………... 32 3.3.3. Sea urchin, Diadema antillarum, meal………………………………………... 34 3.3.4. Other feed ingredients……………………………………………………………. 35 3.43.43.4.3.4... FEED PROCESSING…PROCESSING…………………………………………………………………………………………………………………………………………………………………………….….….…. 35 3.4.1. Formulation………………………………………………………………………... 35 3.4.2. Diets elaboration………………………………………………………………...... 35 CONTENTS 3.53.53.5.3.5... BIOLOGICAL AND FEED UTILIZATION PARAMETERS……………PARAMETERS……………….….….…. 37 3.63.63.6.3.6... INSTRUMENTAL COLOUR DETERMINATION…………………………DETERMINATION…………………………............ 38 3.73.73.7.3.7... ANALYSIANALYSIANALYSISANALYSIS OS OFOF PF PRPRRROXIMATEOXIMATE AND FATFATFATTYFATTY ACIDS COMPOSITIONCOMPOSITION…………………….... 39 3.83.83.8.3.8... CAROTENOIDS ANALYSIS……….…ANALYSIS……….………………………………………...……………………………………...……………………………………................... 40 3.8.1. Carotenoids extraction………………………………………………………....... 40 3.8.2. Carotenoids separation and relative quantification (%) by thin layer chromatography (TLC)…………………………………………………………… 41 3.93.93.9.3.9... AMMONIA EXCRETION ASSESSMENT…………………………………ASSESSMENT…………………………………….….….…. 42 3.9.1. Water sampling for ammonia-N determination……………………………… 42 3.9.2. Analytical method………………………………………………………………… 42 3.103.103.10.3.10... DIGESTIBILITY DETERMINATION………………………………………DETERMINATION………………………………………………….... 44 3.10.1. Experimental tanks and faecal samples collection…………………………... 44 3.10.2. Chromium (III) oxide (Cr2O3) determination…………………………………. 44 3.10.3. Apparent digestibility coefficients determination (ADCs)……………….. 44 3.113.113.11.3.11... FLESH TEXTURE PROFILE ANALYSIS…………………………………ANALYSIS…………………………………….….….…. 45 3.123.123.12.3.12... SENSORY ANALYSIS…………………………………………………………ANALYSIS…………………………………………………………………… 46 3.133.133.13.3.13... FLESHFLESHFLESH LIPID OXIDATION……OXIDATION……………………………………………………………………………………………………………………………………………..…………..…………..………….. 47 3.143.143.14.3.14... STATISTICAL ANALYSIANALYSIS……………………………………………………S……………………………………………………S……………………………………………………................ 47 STUDY I:STUDY I: Marine and freshwater crab meals in diets for red 44.... porgy (porgy (Pagrus pagrus): effect on growth, fish composition and 51 skin colour 4.1. Introduction………………………………………………………………………... 53 4.2. Materials and methods…………………………………………………………… 55 4.3. Results……………………………………………………………………………… 59 4.4. Discussion………………………………………………………………………...... 64 4.5. Conclusions………………………………………………………………………... 68 STUDY IISTUDY II:II Marine and freshwater crab memealsals in diets for red 55.... porgy (porgy (Pagrus pagrus): digestibilitydigestibility,, ammonia excretion and 69 phosphophosphophosphorousphosphorous and calcium retentionretention.... 5.1. Introduction………………………………………………………………………... 70 5.2. Materials and methods…………………………………………………………… 72 5.3. Results……………………………………………………………………………… 77 5.4. Discussion………………………………………………………………………...... 82 5.5. Conclusions……………………………………………………………………….... 86 STUDY IIIIII: Marine and freshwater crab meals in diets for red 6.6.6.6. porgy (porgy (Pagrus pagrus): effect o): onon fillet fatty acid profile and flesh 87 quality parameters 6.1. Introduction………………………………………………………………………... 88 6.2. Materials and methods…………………………………………………………… 90 6.3. Results……………………………………………………………………………… 95 6.4. Discussion………………………………………………………………………...... 101 6.5. Conclusions……………………………………………………………………….... 105 CONTENTS STUDY IV: Meals of spider marine crab (Paramola cuvieri) and 7.7.7.7. sea urchin (Diadema antillarum) in diets for red porgy (PagrusPagrusPagrus
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