The influence of dietary fatty acid and fasting on the hepatic lipid metabolism of barramundi (Lates calcarifer)

For many fish species, dietary fish oil (FO) has been substituted with other oils such as poultry oil (PO) without affecting growth performance. However, in barramundi, the mechanisms by which fatty acid metabolism is regulated are poorly understood, and the effects of FO substitution are unknown. This study defined changes in the expression of genes controlling the metabolism of fatty acids in barramundi over a 24‐h time period after a single meal. From one to 12 h after a single feeding event, the expression of fatty acid synthesis genes in the liver was upregulated, while genes involved in the β‐oxidation showed minimal alteration. However, the expression of β‐oxidation genes was significantly correlated with the expression of genes regulating fatty acid synthesis. In a second experiment, the changes in liver fatty acid composition and gene expression were defined after FO was substituted with PO. Liver fatty acid profile reflected the diet composition, with some subtle exceptions supporting the enrichment of certain long‐chain polyunsaturated fatty acids in the liver. The fish from all experimental groups preferentially retained more docosahexaenoic acid than eicosapentaenoic acid in the liver, suggesting a bioconversion of this fatty acid to intermediate fatty acids. Replacement of FO with PO significantly regulated genes controlling both fatty acid synthesis and catabolism pathways, potentially related to a higher percentage of monounsaturated fatty acids, in the livers of fish fed these diets. The results demonstrated that diet composition significantly altered the lipid metabolism in barramundi and that there was a balance between direct dietary effects and endogenous synthetic capacity.     

鱼类营养代谢性疾病相关基因数据库的调研与分析

集约化养殖条件下鱼类营养代谢性疾病频发,如何防治是目前亟待解决的问题。寻找导致营养代谢性疾病的关键基因,并研究其功能及作用机制是目前研究的热点。高通量测序技术为研究者提供了海量的基因信息,如何有效的挖掘、分析和存储数据并用于疾病的防治显得尤为重要。目前,鱼类营养代谢性疾病相关基因数据库的开发和利用仍处于研究初期。为了合理高效地利用现有的组学数据信息,本研究系统地调研了鱼类代谢性疾病相关数据库,明晰了NCBI数据库中265种鱼类的基因组大小和64种鱼类的染色体数目,获取了Ensembl数据库中17个目58种鱼类的基因组数据,收集了KEGG数据库中41种鱼类的糖类、脂类和蛋白质代谢相关基因。本文可为鱼类营养代谢性疾病基因数据库的研发提供数据支撑,并为鱼类营养代谢性疾病的治疗和预防奠定坚实的基础。     

鱼类对饲料可消化糖的利用和代谢特征

糖类是鱼类三大营养物质之一,可消化糖作为一种性价比较高的能量物质,其合理利用对饲料成本的降低和饲料物理性状的维持具有重要意义,但鱼类对糖的利用有限,投喂含有一定比例可消化糖的膨化商业饲料易引起肝脏疾病的发生。文章总结了部分已发表关于鱼类糖耐受和生长试验结果的报道,发现不同鱼类对葡萄糖的代谢模式存在一定的差异,一些鱼类倾向于将过量葡萄糖以糖原的形式储存在肝脏中,而有些鱼类则既能合成糖原也能合成脂肪。糖耐量试验是了解鱼类利用可消化糖类的重要研究手段,但目前还没有统一的标准,导致研究结果的参考意义不强。作者建议将来在统一方法标准的基础上,重点研究不同鱼类的葡萄糖代谢特征以及调控方法。     

Effect of diet composition on growth performance,hepatic metabolism and antioxidant activities in Siberian sturgeon (<Emphasis Type="Italic">Acipenser</Emphasis><Emphasis Type="Italic">baerii</Emphasis>, Brandt, 1869) submitted to starvation and refeeding

Many fish species undergo natural starvation periods. Adaptation to starvation is possible through the activation of behavioral, biochemical and physiological mechanisms. Knowledge of the effect of dietary nutrients on the intermediary metabolism during starvation and refeeding can be useful to improve fish health and optimize aquaculture production. To analyze the effect of dietary nutrients on liver metabolism of Siberian sturgeon (Acipenser baerii) submitted to starvation and refeeding, four isoenergetic diets differing in nutrient composition were designed: LP-St (38 % protein, 12 % lipid, 36 % carbohydrate), HP-St (44 % protein, 10 % lipid, 30 % carbohydrate), LP-L (38 % protein, 18 % lipid, 25 % carbohydrate) and HP-L (44 % protein, 16 % lipid, 22 % carbohydrate). Four groups of fish were fed 3 weeks to satiety with the corresponding diet, starved for 2 weeks and then refeed 5 weeks to satiety on the same diet. Starvation mobilized the hepatic lipid store to a greater extent than glycogen. Starvation increased superoxide dismutase activity irrespective of the diet, while low protein diets (LP-St and LP-L) increased catalase activity. The oxidative damage decreased after 5 weeks of refeeding. Refeeding the starved fish on the HP-St diet promoted the greatest growth performance. In addition to reporting for the first time the effect of diet composition on growth, liver composition and antioxidant activities in Siberian sturgeon submitted to starvation and refeeding, our findings suggest that refeeding on HP-St diet stimulated the use of dietary carbohydrates and allowed a protein sparing effect in Siberian sturgeon.     

Dietary fish oil replacement with palm or poultry oil increases fillet oxidative stability and decreases liver glutathione peroxidase activity in barramundi (Lates calcarifer)

Complete dietary fish oil replacement with palm or poultry oil in barramundi (Lates calcarifer) had no detrimental effects on growth or hepatosomatic index of juvenile fish up to an average size of ~50 g. However, it significantly decreased the omega-3 (n-3) long-chain polyunsaturated fatty acid content of the fish muscle (fillet) lipids. This was particularly true for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which are recognised for their health beneficial effects in the human diet. As a result of their decreased EPA and DHA content, the peroxidation index of the muscle lipids was also decreased. This was associated with increased simulated retail storage shelf life as indicated by decreased thiobarbituric acid reactive substances in muscle samples from fish fed the palm or poultry oil-based diets. Concomitantly, glutathione peroxidase (GPx) activity, but not glutathione S-transferase (GST) activity or reduced glutathione concentration, was significantly reduced in the liver of barramundi fed the palm or poultry oil-based diets as compared with the fish fed the fish oil-based diet. Furthermore, GPx and GST activity were very low in muscle, much lower than in gastrointestinal tract, liver or swim bladder. Therefore, we propose that liver GPx activity may be a good predictor of fillet shelf life in barramundi and other fish species.     

Postprandial kinetics of gene expression of proteins involved in the digestive process in rainbow trout (<Emphasis Type="Italic">O. mykiss</Emphasis>) and impact of diet composition

The impact of increased incorporation of plant ingredients on diets for rainbow trout was evaluated in terms of gene expression of gastric (gastric lipase, pepsinogen) and intestinal (prolidase, maltase, phospholipase A2) digestive enzymes and nutrient transporters (peptide and glucose transporters), as well as of postprandial levels of plasma glucose, triglycerides and total free amino acids. For that purpose, trout alevins were fed from the start of exogenous feeding one of three different experimental diets: a diet rich in fish meal and fish oil (FM–FO), a plant-based diet (noFM–noFO) totally free from fish meal and fish oil, but containing plant ingredients and a Mixed diet (Mixed) intermediate between the FM–FO and noFM–noFO diets. After 16 months of rearing, all fish were left unfed for 72 h and then given a single meal to satiation. Blood, stomach and anterior intestine were sampled before the meal and at 2, 6 and 12 h after this meal. The postprandial kinetics of gene expression of gastric and intestinal digestive enzymes and nutrient transporters were then followed in trout fed the FM–FO diet. The postprandial profiles showed that the expression of almost all genes studied was stimulated by the presence of nutrients in the digestive tract of trout, but the timing (appearance of peaks) varied between genes. Based on these data, we have focused on the molecular response to dietary factors in the stomach and the intestine at 6 and 12 h after feeding, respectively. The reduction in FM and FO levels of dietary incorporation induced a significant decrease in the gene expression of gastric lipase, GLUT2 and PEPT1. The plasma glucose and triglycerides levels were also reduced in trout fed the noFM–noFO diet. Consequently, the present study suggests a decrease in digestive capacities in trout fed a diet rich in plant ingredients.     

黑斑原鮡肝脏与副肝代谢差异比较

黑斑原鮡(Glyptosternum maculatum)是我国雅鲁藏布江中上游江段特有鱼类, 为适应雅鲁藏布江急流、低温的水域环境条件, 进化出独特的组织器官——副肝。本研究采用组织学、生物化学以及转录组学的方法开展了黑斑原鮡主肝与副肝代谢差异调控机制的研究。结果表明, 黑斑原鮡主、副肝脏在结构组成、肝细胞线粒体数量及线粒体蛋白质组成上无显著差异。转录组学结果表明, 主、副肝脏共有差异表达基因 77 个。经 GO 功能注释, 筛选出羟甲基戊二酸单酰 CoA 合成酶(Hmgcl)、血栓素合成酶(Tbxas)、钙/钙调蛋白依赖性丝氨酸蛋白激酶(Cask)、甲酰甘氨酸生成酶(Sumf1)、蛋白质酪氨酸激酶(Jak1)以及甘油酸激酶(Glxk)等差异表达基因, 这些基因富集到氨基酸代谢、脂肪代谢等过程。KEGG 通路富集结果显示, 差异表达基因富集到丁酸盐代谢通路(butanoate metabolism)、 过氧化物酶体(peroxisome)、缬氨酸, 亮氨酸和异亮氨酸降解(valine, leucine and isoleucine degradation)、赖氨酸降解(lysine degradation)、色氨酸代谢(tryptophan metabolism)以及代谢途径(metabolic pathways)等信号通路。选出 4 个差异表达基因进行实时荧光定量 PCR (qRT-PCR)检测, 结果显示 qRT-PCR 与转录组测序结果基本一致。综上所述, 黑斑原鮡主、副肝脏存在氨基酸、脂肪酸及能量代谢差异, 其代谢差异可能与 Hmgcl、Ptk2b、Gba 和 Dnm1l 等基因在主、副肝脏中的差异性表达有关(P＜0.05)。本研究筛选出调节黑斑原鮡代谢差异的关键基因和信号通路, 为揭示黑斑原鮡主、副肝脏代谢差异机制奠定了理论基础。     

Lipid metabolism-related gene expression pattern of Atlantic bluefin tuna (<Emphasis Type="Italic">Thunnus thynnus</Emphasis> L.) larvae fed on live prey

The present study is the first to evaluate lipid metabolism in first-feeding Atlantic bluefin tuna (ABT; Thunnus thynnus L.) larvae fed different live prey including enriched rotifers Brachionus plicatilis and Acartia sp. copepod nauplii from 2 days after hatch. Understanding the molecular basis of lipid metabolism and regulation in ABT will provide insights to optimize diet formulations for this high-value species new to aquaculture. To this end, we investigated the effect of dietary lipid on whole larvae lipid class and fatty acid compositions and the expression of key genes involved in lipid metabolism in first feeding ABT larvae fed different live prey. Additionally, the expression of lipid metabolism genes in tissues of adult broodstock ABT was evaluated. Growth and survival data indicated that copepods were the best live prey for first feeding ABT and that differences in growth performance and lipid metabolism observed between larvae from different year classes could be a consequence of broodstock nutrition. In addition, expression patterns of lipid metabolic genes observed in ABT larvae in the trials could reflect differences in lipid class and fatty acid compositions of the live prey. The lipid nutritional requirements, including essential fatty acid requirements of larval ABT during the early feeding stages, are unknown, and the present study represents a first step in addressing these highly relevant issues. However, further studies are required to determine nutritional requirements and understand lipid metabolism during development of ABT larvae and to apply the knowledge to the commercial culture of this iconic species.     

Effects of dietary amylose/amylopectin ratio on growth performance,feed utilization,digestive enzymes,and postprandial metabolic responses in juvenile obscure puffer Takifugu obscurus

The effect of dietary amylose/amylopectin (AM/AP) ratio on growth, feed utilization, digestive enzyme activities, plasma parameters, and postprandial blood glucose responses was evaluated in juvenile obscure puffer, Takifugu obscurus. Five isonitrogenous (430 g kg^?1 crude protein) and isolipidic (90 g kg^?1 crude lipid) diets containing an equal starch level (250 g kg^?1 starch) with different AM/AP ratio diets of 0/25, 3/22, 6/19, 9/16 and 12/13 were formulated. Each experimental diet was fed to triplicate groups (25 fish per tank), twice daily during a period of 60 days. After the growth trial, a postprandial blood response test was carried out. Fish fed diet 6/19 showed best growth, feed efficiency and protein efficiency ratio. Hepatosomatic index, plasma total cholesterol concentration, liver glycogen and lipid content, and gluconokinase, pyruvate kinase and fructose-1,6-bisphosphatase activities were lower in fish fed highest AM/AP diet (12/13) than in fish fed the low-amylose diets. Activities of liver and intestinal trypsin in fish fed diet 3/22 and diet 6/19 were higher than in fish fed diet 9/16 and diet 12/13. Activities of liver and intestinal amylase and intestinal lipase, and starch digestibility were negatively correlated with dietary AM/AP ratio. Fish fed diet 3/22 and diet 6/19 showed higher plasma total amino acid concentration than fish fed the other diets, while plasma urea nitrogen concentration and activities of alanine aminotransferase and aspartate aminotransferase showed the opposite trend. Equal values were found for viscerosomatic index and condition factor, whole body and muscle composition, plasma high-density and low-density lipoprotein cholesterol concentrations, and activities of lipase and hexokinase and glucose-6-phosphatase in liver. Postprandial plasma glucose and triglyceride peak value of fish fed diet 12/13 were lower than in fish fed the low-amylose diets, and the peak time of plasma glucose was later than in fish fed the other diets. Plasma glucose and triglyceride concentrations showed a significant difference at 2 and 4 h after a meal and varied between dietary treatments. According to regression analysis of weight gain against dietary AM/AP ratio, the optimum dietary AM/AP ratio for maximum growth of obscure puffer was 0.25. The present result indicates that dietary AM/AP ratio could affect growth performance and feed utilization, some plasma parameters, digestive enzyme as well as hepatic glucose metabolic enzyme activities in juvenile obscure puffer.     

Modulation of appetite,lipid and glucose metabolism of juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) by different dietary protein levels

This study was undertaken to explore the systemic metabolic strategies of juvenile grass carp (Ctenopharyngodon idellus) to maintain growth when fed with different dietary protein levels. The optimal growth group and two growing discomfort groups were selected through the basic data, to explain the growth difference from appetite regulation and lipid and glucose metabolism perspective. Three experimental diets were formulated with three dietary protein levels at 200.3, 296.1 and 442.9 g kg^?1, named P1, P2 and P3, respectively. Juvenile grass carp (initial body weight 12.28 ± 0.14 g) were fed with three diets with 3 replications per dietary treatment in an indoor recirculation system for an 8-week feeding trial. Fish fed with diet P2 dietary group showed significantly higher WG, SGR, FI and PER than other groups. Compared with other groups, mRNA expressions of NPY, Y8a and Y8b in fish fed with P2 significantly down-regulated, while the expressions of CCK and CART in fish fed with P3 significantly down-regulated (P < 0.05). With increasing dietary protein levels, G6Pase, GK, PK and PEPCK were all significantly inhibited (P < 0.05). For lipid metabolism, the mRNA expression of ACC in P1 dietary group was significantly higher than P3 dietary group; besides, LPL expression in P3 group was significantly higher than other two groups (P < 0.05). PPARα expression in P2 was significantly lower than other groups (P < 0.05). These results suggested that grass carp fed with P2 (296.1 g kg^?1 protein level) showed highest weight gain, contributed to more balanced nutrient metabolism and appetite regulation. Too high dietary protein (442.9 g kg^?1) should be avoided because it induced lowest PER, body lipid and liver lipid, and inhibited glucose and lipid metabolism in juvenile grass carp.     

Carbohydrates in fish nutrition: effects on growth, glucose metabolism and hepatic enzymes

The utilisation of dietary carbohydrates and their effects on fish metabolism are reviewed. Details on how dietary carbohydrates affect growth, feed utilisation and deposition of nutrients are discussed. Variations in plasma glucose concentrations emphasizing results from glucose tolerance tests, and the impact of adaptation diets are interpreted in the context of secondary carbohydrate metabolism. Our focus then shifts to selected aspects of hormonal regulation of carbohydrate metabolism and dietary carbohydrates and their variable effects on glycogen and glucose turnover. We analyse the interaction of carbohydrates with other nutrients, especially protein and protein sparing, and de novo synthesis of lipids, and finish by discussing the correlation of dietary carbohydrates with fish health.     

Evaluation of mosquito fish Gambusia holbrooki (Girard) as a partial and complete pellet replacement for juvenile barramundi Lates calcarifer (Bloch)

Barramundi (Lates calcarifer, Bloch) fed live mosquito fish (Gambusia holbrooki, Girard) exhibited faster growth and a better food conversion ratio (on a dry‐weight basis) than those fed an iso‐calorific ration of commercial barramundi pellets and a 50:50 ration of mosquito fish and pellets. The improved performance of barramundi fed mosquito fish was attributed to their higher protein content, higher protein to energy ratio and lower carbohydrate content. The essential amino acid indices and fatty acid profiles of the two diets were similar, suggesting that imbalances or deficiencies in these nutrients were not the limiting factor to the slower growth of barramundi fed on pellets. The use of mosquito fish as barramundi food offers a potential use of this noxious species and may allow barramundi to be cultured without the use of conventional fish meal.     

Novel methodologies in marine fish larval nutrition

Major gaps in knowledge on fish larval nutritional requirements still remain. Small larval size, and difficulties in acceptance of inert microdiets, makes progress slow and cumbersome. This lack of knowledge in fish larval nutritional requirements is one of the causes of high mortalities and quality problems commonly observed in marine larviculture. In recent years, several novel methodologies have contributed to significant progress in fish larval nutrition. Others are emerging and are likely to bring further insight into larval nutritional physiology and requirements. This paper reviews a range of new tools and some examples of their present use, as well as potential future applications in the study of fish larvae nutrition. Tube-feeding and incorporation into Artemia of ¹⁴C-amino acids and lipids allowed studying Artemia intake, digestion and absorption and utilisation of these nutrients. Diet selection by fish larvae has been studied with diets containing different natural stable isotope signatures or diets where different rare metal oxides were added. Mechanistic modelling has been used as a tool to integrate existing knowledge and reveal gaps, and also to better understand results obtained in tracer studies. Population genomics may assist in assessing genotype effects on nutritional requirements, by using progeny testing in fish reared in the same tanks, and also in identifying QTLs for larval stages. Functional genomics and proteomics enable the study of gene and protein expression under various dietary conditions, and thereby identify the metabolic pathways which are affected by a given nutrient. Promising results were obtained using the metabolic programming concept in early life to facilitate utilisation of certain nutrients at later stages. All together, these methodologies have made decisive contributions, and are expected to do even more in the near future, to build a knowledge basis for development of optimised diets and feeding regimes for different species of larval fish.     

Nutritional regulation of hepatic glucose metabolism in fish

Glucose plays a key role as energy source in the majority of mammals, but its importance in fish appears limited. Until now, the physiological basis for such apparent glucose intolerance in fish has not been fully understood. A distinct regulation of hepatic glucose utilization (glycolysis) and production (gluconeogenesis) may be advanced to explain the relative inability of fish to efficiently utilize dietary glucose. We summarize here information regarding the nutritional regulation of key enzymes involved in glycolysis (hexokinases, 6-phosphofructo-1-kinase and pyruvate kinase) and gluconeogenesis (phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase and glucose-6-phosphatase) pathways as well as that of the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. The effect of dietary carbohydrate level and source on the activities and gene expression of the mentioned key enzymes is also discussed. Overall, data strongly suggest that the liver of most fish species is apparently capable of regulating glucose storage. The persistent high level of endogenous glucose production independent of carbohydrate intake level may lead to a putative competition between exogenous (dietary) glucose and endogenous glucose as the source of energy, which may explain the poor dietary carbohydrate utilization in fish.     

Oxidative effect of l -carnitine on energy metabolism in diploid and triploid rainbow trout ( Oncorhynchus mykiss ): impact on metabolites

l-Carnitine plays a key role in the regulation of energy metabolism and growth in fish. The aim of this study was to determine the effects of dietary l-carnitine levels on carnitine homeostasis and energy metabolism in diploid and triploid trout (Oncorhynchus mykiss). Diploid and triploid trout (29.5 ± 0.6 and 31.8 ± 0.5 g, respectively) were fed with three diets supplemented with 15, 200 or 530 mg l-carnitine/kg, respectively, for 56 days. Compared to 15 mg l-carnitine, total carnitine content increased in the liver by 182% in diploid and by 154% in triploid trout fed 530 mg l-carnitine. In muscle, total carnitine content increased by 60 and 73% in diploid and by 34 and 75% in triploid with the increase in dietary l-carnitine levels. l-Carnitine caused no significant change in the plasma concentrations of metabolites like proteins (ammonia, urea and protein), lipid (triglycerides), carbohydrate (glucose, lactate) and enzyme activities (lipase, lactate, alanine transaminase, lactic acid dehydrogenase). Triploid trout had significantly lower plasma ammonia (P = 0.003), lipase (P = 0.005) and triiodothyronine (T3; P = 0.003) levels than diploid trout. In conclusion, ploidy significantly affected the energy metabolism in rainbow trout, dietary l-carnitine levels altered the l-carnitine homeostasis, but not influence nutritional metabolism.     

鱼类的葡萄糖感知与糖代谢调节研究进展

为加深对鱼类糖的感知与代谢调控的认识,本文综述了鱼类的葡萄糖感知与摄食调控、糖代谢调控等领域的研究进展。鱼类的下丘脑不仅是中枢葡萄糖感受器所处的主要部位,同时也是食欲调节中枢。leptin、ghrelin、CCK、NPY等内分泌因子均能调控鱼类对糖的感知与摄食。另一方面,鱼类糖代谢受到胰岛素、GLP-1、ghrelin、CCK、NPY、SS等内分泌因子和糖、脂、蛋白质等营养素的双重调节。尽管鱼类对糖的利用能力低于陆生动物,但鱼体内亦存在较完善的糖的感知、摄食与代谢调节机制。因此,将来的重点工作应在于研究鱼类中枢神经系统整合营养和内分泌等信号的机制,研究草食性鱼类、杂食性鱼类在糖耐受力及糖异生调控机制上与肉食性鱼类存在的差异。     

The impact of nutritional factors on the immune system in Atlantic salmon, Salmo salar L.: a review

Abstract Compared to the extensive number of studies in nutritional immunology in humans as well as in livestock, the evaluation of nutritional impact on fish health is in its infancy. The mechanisms by which single nutrient modulate immunity are frequently not understood, even if experiments give some indication of the mode of action. With current knowledge in fish nutrition, the methods and criteria for optimal nutrient recommendations should be reevaluated to include health factors such as immunolgy and disease resistance. By increasing knowledge of the role of different nutrients in the general metabolism and in immunity, and by improving rearing conditions, it may be possible to reduce losses due to diseases by retarding the progress of diseases, improving recovery and improving protection through vaccines. This review covers research on dietary level of carbohydrate, n-3 polyunsaturated fatty acids, vitamin E and vitamin C in salmonids, of which the vitamins have been by far the most studied.