by Scott P. Kelly. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 168-190). / Acknowledgements --- p.iv / Chapter Chapter 1 : --- Introduction --- p.1 / Chapter Chapter 2 : --- Review of the literature / Part A : Nutritional requirements of fish and performance assessment / Chapter A2.1 --- Introduction --- p.5 / Chapter A2.2 --- Dietary protein requirement --- p.6 / Chapter A2.2.1 --- Factors influencing dietary protein requirement / Chapter A2.2.1.1 --- Protein quality / Chapter A2.2.1.2 --- Nonpotein energy / Chapter A2.2.1.3 --- Protein energy to total energy ratio / Chapter A2.2.1.4 --- Fish size/age / Chapter A2.2.1.5 --- Feeding rate / Chapter A2.2.1.6 --- Natural food / Chapter A2.2.1.7 --- Economics / Chapter A2.2.1.8 --- Environmental factors / Chapter A2.3 --- Dietary lipid --- p.14 / Chapter A2.3.1 --- Dietary lipid requirement / Chapter A2.4 --- Dietary carbohydrate --- p.16 / Chapter A2.4.1 --- Dietary carbohydrate requirement / Chapter A2.5 --- Dietary vitamins / Chapter A2.5.1 --- Dietary vitamin requirements / Chapter A2.6 --- Dietary minerals --- p.19 / Chapter A2.6.1 --- Dietary mineral requirements / Chapter A2.7 --- Evaluation criteria --- p.21 / Chapter A2.7.1 --- Introduction / Chapter A2.7.2 --- Growth and conversion efficiencies / Chapter A2.8 --- Digestion --- p.22 / Chapter A2.9 --- Metabolism in relation to nutritional status --- p.23 / Chapter A2.9.1 --- Nitrogen excretion / Chapter A2.9.2 --- Metabolic rate (Oxygen consumption) / Chapter A2.10 --- Biochemical indices of metabolic performance --- p.25 / Chapter A2.10.1 --- Tissue composition / Chapter A2.10.1.1 --- Proximate composition and organ indices / Chapter A2.10.1.2 --- Essential amino acid (EAA) profile / Chapter A2.10.2.3 --- Lipid/essential fatty acid (EFA) profile / Chapter A2.10.2.4 --- Tissue RNA/DNA ratio / Chapter A2.11 --- Haematological characteristics --- p.27 / Chapter A2.12 --- Enzymes --- p.28 / Chapter A2.12.1 --- Enzymes of the intermediate metabolism / Chapter A2.12.2 --- Intestinal enzymes / Chapter A2.13 --- Thyroid hormones --- p.30 / Part B : Teleostean salinity adaptation / Chapter B2.1 --- Introduction --- p.31 / Chapter B2.2 --- Influence of salinity on growth --- p.32 / Chapter B2.3 --- Influence of salinity on weight loss during starvation --- p.37 / Chapter B2.4 --- Metabolic rate (oxygen consumption) and salinity adaptation --- p.38 / Chapter B2.5 --- Biochemical indices of performance during salinity adaptation --- p.41 / Chapter B2.5.1 --- Carcass and tissue composition / Chapter B2.5.2 --- Haematological characteristics / Chapter B2.6 --- Influence of salinity on protein requirements of fish --- p.46 / Chapter B2.7 --- Effect of salinity on digestion and digestive enzymes --- p.48 / Chapter B2.8 --- Cortisol and osmotic adjustment --- p.50 / Chapter B2.9 --- Conclusion --- p.52 / Chapter Chapter 3: --- Materials and methods / Chapter 3.1 --- Culture conditions --- p.54 / Chapter 3.2 --- Composition of experimental diets --- p.55 / Chapter 3.2.1 --- Dietary protein / Chapter 3.2.2 --- Dietary carbohydrate / Chapter 3.2.3 --- Dietary lipid / Chapter 3.2.4 --- Dietary vitamins / Chapter 3.2.5 --- Dietary minerals / Chapter 3.2.6 --- Chromic oxide / Chapter 3.2.7 --- Binder / Chapter 3.3 --- Proximate analysis of experimental diets --- p.61 / Chapter 3.3.1 --- Moisture / Chapter 3.3.2 --- Lipid / Chapter 3.3.3 --- Protein / Chapter 3.3.4 --- Ash / Chapter 3.3.5 --- Nitrogen free extractives / Chapter 3.3.6 --- Metabolizable energy / Chapter 3.4 --- Dietary evaluation criteria --- p.65 / Chapter 3.4.1 --- Growth rate / Chapter 3.4.2 --- Protein efficiency ratio / Chapter 3.4.3 --- Food conversion ratio / Chapter 3.4.4 --- Feed conversion efficiency / Chapter 3.5 --- Digestibility --- p.66 / Chapter 3.6 --- Metabolism --- p.67 / Chapter 3.6.1 --- Ammonia excretion / Chapter 3.6.2 --- Oxygen consumption / Chapter 3.7 --- Biochemical analysis / Chapter 3.7.1 --- Organ indices / Chapter 3.7.2 --- Serum metabolites / Chapter 3.7.2.1 --- Collection of serum / Chapter 3.7.2.2 --- Serum ions / Chapter 3.7.2.3 --- Serum protein / Chapter 3.7.2.4 --- Serum α-amino acids / Chapter 3.7.2.5 --- Serum ammonia / Chapter 3.7.2.6 --- Serum glucose / Chapter 3.7.2.7 --- Serum lipids / Chapter 3.7.2.8 --- Serum hormones / Chapter 3.7.3 --- Proximate analysis of white muscle --- p.74 / Chapter 3.7.4 --- Analysis of hepatic tissue --- p.75 / Chapter 3.7.4.1 --- Proximate analysis / Chapter 3.7.4.2 --- Liver glycogen / Chapter 3.7.4.3 --- Hepatic enzymes / Chapter 3.7.4.3. --- a Glucose-6-phosphatase / Chapter 3.7.4.3. --- b Glucose-6-phosphate dehydrogenase / Chapter 3.7.4.3. --- C Hexokinase / Chapter 3.7.5 --- Intestinal enzymes --- p.80 / Chapter 3.7.5.1 --- γ-Glutamyltranspeptidase / Chapter 3.7.5.2 --- Trypsin / Chapter 3.7.5.3 --- α-Amylase / Chapter 3.7.6 --- Statistical analysis --- p.83 / Chapter Chapter 4: --- Results / Chapter 4.1 --- Growth and conversion efficiencies --- p.84 / Chapter 4.2 --- Metabolism --- p.90 / Chapter 4.2.1 --- Ammonia excretion / Chapter 4.2.2 --- Oxygen consumption / Chapter 4.3 --- Biochemical analysis --- p.93 / Chapter 4.3.1 --- Organ indices --- p.93 / Chapter 4.3.2 --- Serum metabolites --- p.98 / Chapter 4.3.2.1 --- Serum ions / Chapter 4.3.2.2 --- "Serum protein, α-amino acids and ammonia" / Chapter 4.3.2.3 --- Serum glucose / Chapter 4.3.2.4 --- Serum lipids / Chapter 4.3.2.5 --- Serum hormones / Chapter 4.3.3 --- Proximate composition of white muscle --- p.113 / Chapter 4.3.4 --- Composition of hepatic tissue --- p.117 / Chapter 4.3.4.1 --- Proximate composition and liver glycogen / Chapter 4.3.4.2 --- Hepatic enzymes / Chapter 4.3.5 --- Total digestibility --- p.126 / Chapter 4.3.6 --- Intestinal enzymes --- p.126 / Chapter Chapter 5: --- Discussion / Chapter 5.1 --- Growth and conversion efficiencies --- p.132 / Chapter 5.2 --- Metabolism --- p.135 / Chapter 5.2.1 --- Ammonia excretion / Chapter 5.2.2 --- Oxygen consumption / Chapter 5.3 --- Organ indices --- p.137 / Chapter 5.4 --- Serum metabolites --- p.140 / Chapter 5.4.1 --- Serum ions / Chapter 5.4.2 --- "Serum protein, α-amino acids and ammonia" / Chapter 5.4.3 --- Serum glucose / Chapter 5.4.4 --- Serum lipids / Chapter 5.4.5 --- Serum hormones / Chapter 5.5 --- Proximate composition of white muscle --- p.150 / Chapter 5.6 --- Proximate composition of liver --- p.152 / Chapter 5.7 --- Hepatic enzymes / Chapter 5.7.1 --- Glucose-6-phosphate dehydrogenase (G6P-DH) / Chapter 5.7.2 --- Hexokinase / Chapter 5.7.3 --- Glucose-6-phosphatase (G-6-Pase) / Chapter 5.8 --- Total digestibility --- p.159 / Chapter 5.9 --- Intestinal enzymes / Chapter 5.9.1 --- Trypsin / Chapter 5.9.2 --- γ-Glutamyltranspeptidase (γ-GT) / Chapter 5.9.3 --- α-Amylase / Chapter Chapter 6: --- Conclusion --- p.164 / References --- p.168

Thesis (Ph.D.)--Murdoch University, 2006. / Thesis submitted to the Division of Science and Engineering. Includes bibliographical references (leaves 149-162).

by Scott P. Kelly. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (p. 378-410). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web.

催乳素是一種負責廣鹽性硬骨魚低滲環境適應的重要激素。最近發現的催乳素釋放肽被報導在魚類生理活動中參與了刺激催乳素釋放和滲透調節過程。本研究以黃錫鯛為實驗動物，旨在詮釋催乳素和催乳素釋放肽在廣鹽性硬骨魚中的滲透調節作用和調控機制。 / 實驗第一部份將黃錫鯛催乳素在大腸桿菌中過量表達並用鎳離子親和層析法將之純化。此重組黃錫鯛催乳素可被抗黃錫鯛催乳素抗體特異識別。將此重組催乳素注射入活體紅箭魚可導致其血漿鈉離子濃度和滲透壓顯著提高，從而證明了此催乳素的生物活性。在第二部份的研究中，黃錫鯛催乳素受體和催乳素釋放肽受體的全長編碼基因被克隆和分析。黃錫鯛催乳素受體和硬骨魚二型催乳素受體蛋白質相似性較高（可達47%至94%），但同魚類一型催乳素受體及其他高等脊椎動物催乳素受體同源性較低，相似性只有22%至37%。此催乳素受體廣泛表達在各種組織器官中，包括腦組織、腮、心臟、腸、腎、肌肉及性腺。黃錫鯛催乳素釋放肽受體屬於G-蛋白偶聯受體家族，它具備該家族成員的各種保守序列特徵。並且它含有同其他催乳素釋放肽受體一樣的PDZ域結合位點。組織表達分佈結果顯示此受體在中樞神經系統各部份及腮、心臟、肝臟和性腺中有表達。本研究最後一部份以黃錫鯛原代培養腮細胞為實驗對象，分別探究了腮細胞對於不同滲透壓力、催乳素和催乳素釋放肽處理的反應。在這些實驗中，我們檢測了腮細胞的死亡及凋亡狀況，并用實時定量PCR分析了一些滲透調節相關基因的表達狀況。另外，我們還採用了蛋白免疫印跡法檢測了腮細胞處於不同滲透壓條件下，其熱休克蛋白70的蛋白質豐度變化。我們的結果顯示：低滲壓力會引起腮細胞死亡包括誘導凋亡。催乳素和催乳素釋放肽處理均可以顯著降低低滲壓力所誘導的細胞死亡，並且催乳素還可以阻止一部份細胞進行凋亡。腮細胞中鈉鉀泵兩個亞基、熱休克同族蛋白70及催乳素受體的基因表達均被催乳素和催乳素釋放肽處理上調。但是水通道蛋白3的基因表達并未出現明顯的變化。當腮細胞暴露于高滲透壓力環境時，水通道蛋白3、囊性纤维化跨膜转运调节因子、鈉鉀泵兩個亞基和熱休克同族蛋白70的基因表達都明顯上調。與之相反，這些基因的表達在處於低滲環境的腮細胞中均被下調。 / 本研究提供了許多關於催乳素和催乳素釋放肽在魚類腮滲透調節過程中的詳細作用，并且首次報導了催乳素釋放肽可以直接作用于魚類滲透調節組織發揮功能從而參與滲透調節過程，而不只是如以往被認為的通過旁分泌或者自分泌方式刺激催乳素的表達來發揮生理功能。本研究的探索和發現使我們更加深刻地理解催乳素和催乳素釋放肽在廣鹽性硬骨魚鹽度適應過程中的調控機制。 / Prolactin (PRL) is well known as a crucial hormone responsible for fresh (hypoosmotic) water acclimation in euryhaline teleosts. The recently discovered prolactin-releasing peptide (PrRP) has been reported to take part in stimulating PRL release and affecting osmoregulatory processes in fish. This study aims to investigate the osmoregulatory effects and regulatory mechanisms of PRL and PrRP in a euryhaline teleost, silver sea bream (Sparus sarba). / First, silver sea bream prolactin (ssPRL) was over-expressed in E. coli by IPTG induction and purified by Ni-based immobilized metal ion affinity chromatography. This recombinant silver sea bream prolactin (rssPRL) was recognized by specific antibodies against ssPRL. Subsequently, its bioactivity was confirmed by in vivo injection to swordtails, which resulted in significant increase in plasma Na⁺ level and osmolality. In the second part, full-length cDNAs of silver sea bream PRL receptor (ssPRLR) and PrRP receptor (ssPrRPR) were cloned and characterized. ssPRLR shares high amino acid identities (47% to 94%) with teleost PRLR2s but low identities (22% to 37%) with piscine PRLR1s and higher vertebrate PRLRs. It is widely distributed in brain, gill, heart, gut, kidney, muscle and gonad. ssPrRPR belongs to G protein-coupled receptor (GPCR) family with all the conserved features of GPCRs, and possesses the special PDZ domain-binding motifs of other PrRPRs. Its expression was detected in the central nervous system, gill, heart, liver and gonad. Lastly, a primary gill cell culture of silver sea bream was developed and used as a tool for studying responses of gill cells following exposure to media of different osmotic stress, rssPRL and ssPrRP, respectively. Then, cytotoxicity and apoptosis assays were performed, and effects of these treatments on expression profiles of osmoregulatory genes were analyzed by real time PCR. Influence of osmotic stresses on protein abundance of heat shock protein 70s was examined by Western blot. The results showed that hypoosmotic challenge could induce cell death including apoptosis. Both PRL and PrRP treatment markedly decreased the induced cell death, and PRL treatment prevented some gill cells from apoptosis. Expression levels of Na⁺-K⁺-ATPase alpha (NKA-α) and beta (NKA-β) subunits, heat shock cognate 70 (HSC70) and PRLR were up-regulated in PRL- and PrRP- treated gill cells, however no significant effect on expression of aquaporin3 (AQP3) mRNA was apparent. After hyperosmotic exposure, expression levels of AQP3, cystic fibrosis transmembrane conductance regulator (CFTR), NKA-α, NKA-β, and HSC70 were significantly increased. In contrast, hypoosmotic exposure considerably down-regulated expression levels of these genes. / The present study represented considerable addition of details about the precise actions of PRL and PrRP in branchial osmoregulation. Furthermore, it first reported that PrRP can exert direct effects on fish osmoregulatory epithelia, instead of its oft presumed autocrine or paracrine action. All these efforts provide new insights into the control mechanisms of PRL and PrRP in euryhaline teleost during salinity acclimation. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Qu, Zhe. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 108-142). / Abstracts also Chinese. / Title page --- p.i / Thesis committee --- p.ii / Acknowledgement --- p.iii / Abstract --- p.v / Abstract-Chinese version --- p.viii / Table of contents --- p.x / List of figures --- p.xiv / List of abbreviations --- p.xvi / Chapter Chapter 1 --- General introduction --- p.1 / Chapter Chapter 2 --- Literature review --- p.6 / Chapter 2.1 --- The role of gill in teleost osmoregulation --- p.7 / Chapter 2.1.1 --- An overview of fish osmoregulation --- p.7 / Chapter 2.1.2 --- The gill structure and epithelial ionocytes --- p.8 / Chapter 2.1.3 --- Osmoregulatory molecules in gill cells --- p.10 / Chapter 2.2 --- Piscine prolactin --- p.16 / Chapter 2.2.1 --- General features and distribution --- p.16 / Chapter 2.2.2 --- Prolactin receptor --- p.17 / Chapter 2.2.3 --- Prolactin and teleost osmoregulation --- p.18 / Chapter 2.3 --- Prolactin-releasing peptide in teleost --- p.21 / Chapter 2.3.1 --- Discovery and characteristics --- p.21 / Chapter 2.3.2 --- Distribution and its receptor --- p.22 / Chapter 2.3.3 --- Functions of PrRP --- p.23 / Chapter A. --- PrRP and prolactin release --- p.23 / Chapter B. --- PrRP and neuroendocrine stress response --- p.24 / Chapter C. --- Effects on food intake and energy metabolism --- p.26 / Chapter D. --- PrRP and cardiovascular system --- p.27 / Chapter Chapter 3 --- Production and characterization of recombinant silver sea bream (Sparus sarba) prolactin --- p.29 / Chapter Abstract --- p.30 / Chapter 3.1 --- Introduction --- p.31 / Chapter 3.2 --- Materials and methods --- p.33 / Chapter 3.2.1 --- Experimental animals --- p.33 / Chapter 3.2.2 --- Construction of recombinant silver sea bream prolactin expression vector --- p.33 / Chapter 3.2.3 --- Large-scale production and purification of rssPRL --- p.34 / Chapter 3.2.4 --- SDS polyacrylamide gel electrophoresis and Western blotting analysis of rssPRL --- p.35 / Chapter 3.2.5 --- In vivo bioassay of rssPRL --- p.36 / Chapter 3.2.6 --- Statistical analysis --- p.37 / Chapter 3.3 --- Results --- p.38 / Chapter 3.3.1 --- Overexpression and purification of rssPRL --- p.38 / Chapter 3.3.2 --- Biological activity assay of rssPRL in vivo --- p.38 / Chapter 3.4 --- Discussion --- p.43 / Chapter Chapter 4 --- Molecular cloning and characterization of prolactin receptor (PRLR) and prolactin-releasing peptide receptor (PrRPR) in silver sea bream (Sparus sarba) --- p.46 / Chapter Abstract --- p.47 / Chapter 4.1 --- Introduction --- p.48 / Chapter 4.2 --- Materials and methods --- p.51 / Chapter 4.2.1 --- Fish and tissue samples --- p.51 / Chapter 4.2.2 --- Molecular cloning of full-length ssPRLR and ssPrRPR cDNA --- p.51 / Chapter 4.2.3 --- Multiple sequence alignments and phylogenetic analysis --- p.53 / Chapter 4.2.4 --- Tissue distribution of ssPRLR and ssPrRPR --- p.53 / Chapter 4.3 --- Results --- p.54 / Chapter 4.3.1 --- Cloning and characterization of silver sea bream PRLR and PrRPR cDNA --- p.54 / Chapter 4.3.2 --- Tissue distribution of ssPRLR and ssPrRPR mRNA --- p.56 / Chapter 4.4 --- Discussion --- p.65 / Chapter Chapter 5 --- Influences of osmotic stress and hormones on gill cell of silver sea bream (Sparus sarba) --- p.69 / Chapter Abstract --- p.70 / Chapter 5.1 --- Introduction --- p.72 / Chapter 5.2 --- Methods and materials --- p.77 / Chapter 5.2.1 --- Fish and primary gill cell culture --- p.77 / Chapter 5.2.2 --- Experiments using gill cell culture --- p.78 / Chapter 5.2.3 --- Cell survival assay and apoptosis relevant detections --- p.79 / Chapter 5.2.4 --- Real time PCR analysis --- p.80 / Chapter 5.2.5 --- Western blot analysis of HSP70 --- p.82 / Chapter 5.2.6 --- Data processing and statistical analysis --- p.82 / Chapter 5.3 --- Results --- p.83 / Chapter 5.3.1 --- Hypoosmotic stress induced cell death --- p.83 / Chapter 5.3.2 --- Influences of PRL and PrRP on hypoosmotic challenge induced cell death --- p.84 / Chapter 5.3.3 --- Effects of hormones and osmotic stresses on expression levels of branchial genes --- p.84 / Chapter 5.3.4 --- Osmotic stresses induced changes in branchial protein amount of HSP70s --- p.85 / Chapter 5.4 --- Discussion --- p.94 / Chapter Chapter 6 --- General discussion and conclusions --- p.102 / References --- p.108

Leung Ling Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 117-138). / Abstracts in English and Chinese. / Abstract (English) --- p.ii / Abstract (Chinese) --- p.iv / Acknowledgements --- p.vi / List of Abbreviation --- p.vii / List of Figures and Tables --- p.viii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter Chapter 2 --- The effect of salinity on pentose phosphate pathway activity and red blood cells resistance to oxidative stress in silver seabream (Sparus sarba) --- p.4 / Abstract --- p.7 / Chapter 2.1 --- Literature review --- p.8 / Chapter 2.2 --- Materials and Methods --- p.17 / Chapter 2.3 --- Results --- p.32 / Chapter 2.4 --- Discussion --- p.54 / Chapter 2.5 --- Conclusion --- p.63 / Chapter Chapter 3 --- The effect of dietary carbohydrate level on pentose phosphate pathway activity and red blood cells resistance to oxidative stress in of silver seabream (Sparus sarba) --- p.65 / Abstract --- p.67 / Chapter 3.1 --- Literature review --- p.68 / Chapter 3.2 --- Materials and Methods --- p.77 / Chapter 3.3 --- Results --- p.80 / Chapter 3.4 --- Discussion --- p.101 / Chapter 3.5 --- Conclusion --- p.112 / Chapter Chapter 4 --- Summary --- p.113 / References --- p.117

Lipase, from porgy (Stenotomus chrysops) viscera, was purified by polyethylene glycol (PEG) 1000 precipitation, followed by dialysis and affinity chromatography on EAH-Sepharose 48. The digestive lipase from porgy showed seasonal variation in activity with high activity found in late summer and early fall compared with a spring sample. Polyethylene glycol (PEG) precipitation fraction was used to characterize this enzyme using p-nitrophenyl palmitate (pNPP) as substrate. Porgy lipase did not behave like a bile salt activated/depended lipase because it was able to hydrolyze pNPP without bile salt (e.g., sodium cholate). Porgy lipase was stable within the pH range of pH 6.0-10.0, with an optimum activity at pH 8.5. The enzyme was quite stable at temperatures below 40°C, but lost its activity rapidly at temperatures above 40°C. The optimum activity for hydrolysis pNPP was at 40°C, but the enzyme also demonstrated relatively high activity at temperatures below 40°C (i.e., 10-40°C) as well. Detergents, Triton X-100, Tween 40 and Tween 80, at final concentrations of 0.5 mM and 1 mM were found to have inhibitory effects on porgy digestive lipase activity. However, all three tested detergents appeared to increase the activity of porgy digestive lipase at elevated temperatures (i.e., 60-80°C).

No description available.

by Chow Cheuk Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1989. / Bibliography: leaves 179-192.

Hui Fong Fong Liza. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-182). / Abstracts in English and Chinese. / Chapter I --- Title page --- p.I / Chapter II --- Thesis committee --- p.II / Chapter III --- Acknowledgements --- p.III / Chapter IV --- Abstract (Chinese version) --- p.IV / Chapter V --- Abstract (English version) --- p.VII / Chapter VI --- Table of contents --- p.X / Chapter VII --- List of figures --- p.XIV / Chapter VIII --- List of table --- p.XVIII / Chapter Chapter 1: --- General introduction --- p.1 / Chapter Chapter 2: --- Literature review --- p.5 / Chapter 2.1. --- Gill --- p.5 / Chapter 2.2. --- Chloride cells (Mitochondria-rich cells) --- p.6 / Chapter 2.2.1. --- Ion extrusion by fish in seawater --- p.9 / Chapter 2.2.2. --- Ion uptake by fish in hypo-osmotic condition --- p.12 / Chapter 2.3. --- Sparus sarba (Silver seabream) --- p.14 / Chapter 2.4. --- Sodium-potassium adenosinetriphosphatase (Na+-K+-ATPase) --- p.15 / Chapter 2.4.1. --- Na+-K+-ATPase α-subunit --- p.17 / Chapter 2.4.2. --- Na+-K+-ATPase β-subunit --- p.18 / Chapter 2.4.3. --- Regulation of Na+-K+-ATPase --- p.20 / Chapter 2.5. --- Hormones --- p.21 / Chapter 2.5.1. --- Growth hormone-prolactin family --- p.21 / Chapter 2.5.2. --- Structure of hormones --- p.22 / Chapter 2.5.2.1. --- Structure of growth hormone and prolactin in fish --- p.22 / Chapter 2.5.2.2. --- Structure of insulin-like growth factors in fish --- p.26 / Chapter 2.5.2.3. --- Structure of Cortisol in fish --- p.27 / Chapter 2.5.3. --- Regulation of hormones --- p.28 / Chapter 2.5.3.1. --- Regulation of growth hormone in fish --- p.28 / Chapter 2.5.3.2. --- Regulation of prolactin in fish --- p.32 / Chapter 2.5.3.3. --- Regulation of insulin-like growth factor-I in fish --- p.33 / Chapter 2.5.3.4. --- Regulation of Cortisol in fish --- p.33 / Chapter 2.5.4. --- Functions of hormones --- p.33 / Chapter 2.5.4.1. --- Functions of growth hormone in fish --- p.33 / Chapter 2.5.4.2. --- Functions of prolactin in fish --- p.39 / Chapter 2.5.4.3. --- Functions of insulin-like growth factor-I in fish --- p.44 / Chapter 2.5.4.4. --- Functions of Cortisol in fish --- p.45 / Chapter 2.5.4.5. --- "Combined effects of GH, IGF-I, PRL and Cortisol" --- p.49 / Chapter 2.6. --- Salinity effects on Na+-K+-ATPase expression --- p.52 / Chapter Chapter 3: --- In vitro effect of hormones on branchial Na+-K+- ATPase expression in marine teleost Sparus sarba --- p.58 / Chapter 3.1. --- Abstract --- p.58 / Chapter 3.2. --- Introduction --- p.60 / Chapter 3.3. --- Materials and methods --- p.62 / Chapter 3.3.1. --- Overall experimental design --- p.62 / Chapter 3.3.2. --- Fish preparation --- p.62 / Chapter 3.3.3. --- Tissue sampling --- p.62 / Chapter 3.3.4. --- RNA extraction and dot blot analysis --- p.63 / Chapter 3.3.5. --- Protein extraction --- p.65 / Chapter 3.3.6. --- Protein quantification --- p.65 / Chapter 3.3.7. --- Na+-K+-ATPase activity --- p.65 / Chapter 3.3.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.66 / Chapter 3.3.9. --- Statistical analysis --- p.67 / Chapter 3.4. --- Results --- p.69 / Chapter 3.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.69 / Chapter 3.4.2. --- Analysis of Na+-K+-ATPase protein α-subunit --- p.81 / Chapter 3.4.3. --- Analysis of Na+-K+-ATPase activity --- p.87 / Chapter 3.5. --- Discussion --- p.92 / Chapter 3.5.1. --- Effects of rbGH and rbIGF-I on Na+-K+-ATPase expression --- p.92 / Chapter 3.5.2. --- Effects of oPRL on Na+-K+-ATPase expression --- p.102 / Chapter 3.5.3 --- Effects of Cortisol on Na+-K+-ATPase expression --- p.104 / Chapter 3.6. --- Conclusion --- p.108 / Chapter Chapter 4: --- In vivo effect of salinity on branchial Na+-K+-ATPase expression in marine teleost Sparus sarba --- p.109 / Chapter 4.1. --- Abstract --- p.109 / Chapter 4.2. --- Introduction --- p.110 / Chapter 4.3. --- Materials and methods --- p.112 / Chapter 4.3.1. --- Overall experimental design --- p.112 / Chapter 4.3.2. --- Fish preparation --- p.112 / Chapter 4.3.3. --- Tissue sampling --- p.113 / Chapter 4.3.4. --- "RNA extraction, dot blot analysis, protein extraction, quantification, Na+-K+-ATPase activity, protein gel electrophoresis and immunoblotting (Western blotting)" --- p.113 / Chapter 4.3.5. --- Statistical analysis --- p.114 / Chapter 4.4. --- Results --- p.114 / Chapter 4.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.114 / Chapter 4.4.2. --- Analysis of Na+-K+-ATPase protein a-subunit --- p.114 / Chapter 4.4.3. --- Analysis of Na+-K+-ATPase activity --- p.115 / Chapter 4.5. --- Discussion --- p.120 / Chapter 4.6. --- Conclusion --- p.125 / Chapter Chapter 5: --- General discussion and conclusion --- p.126 / References --- p.130

The aim of the current study was to determine the organic iron (Fe) requirements of gilthead sea bream (Sparus aurata). A total number of four experiments have been carried out each one for 12 weeks, in order to address and extend the knowledge on nutritional issues and challenges related with the culture of the gilthead sea bream in the Greek aquaculture industry and therefore enhance the fish health status under intensive culture conditions. These experiments aimed to determine the optimum level of organic Fe supplemented in commercial type diets of sea bream, the comparison between an organic Fe form and two inorganic Fe forms added in the diet of the fish, the effect of supplemented organic Fe on sea bream species exposed to oxygen deprivation stressors related to poor aquaculture husbandry practices and finally the interaction of organic Fe in the diet of sea bream with various levels of other trace minerals (Zn, Cu). The parameters evaluated were the growth performance of the fish, the Fe concentration in three selected tissues (the spleen, the liver and the muscle), the haematological status of the fish (the haematocrit, the red blood cell count, and the haemoglobin) and both the humoral and cellular immunology of the fish (the antibacterial activity of serum and the respiratory burst respectively). 150 mg/Kg of added organic Fe appears to be the recommended level as well as the minimum amount on fish exposed to overstocking conditions. The comparison between the two inorganic Fe forms (Ferrous Sulphate and Ferrous Carbonate) added in the diets show no significant effect on the fish. While, the fish fed the diets with 150 mg/Kg organic Fe and Cu levels lower that 5 mg/Kg had higher Hb values.