鱼类抗菌肽研究进展（二）

<正>3 Hepcidin Hepcidin是一种富含半胱氨酸的多肽,最先在人源的hepcidin中发现其具有抗菌活性。此后,hepcidin在许多其他的脊椎动物被陆续发现,包括爬行类,两栖类和鱼类。在鱼类中,hepcidin最先从杂交条纹鲈中发现,目前至少已经在37种鱼类中鉴定到了该抗菌肽[1]。Hepcidin一般是一个由4个二硫桥形成harpin-折叠,包含8个半胱氨酸,然而鱼hepcidin序列分析表明,鱼类hepcidin只含有4,6或7个半胱氨酸[2]。鱼类hepcidin基因在进化过程中发生了重复和多样化的过程中,产生了多重基因拷贝的,最多     

暗纹东方鲀Hepcidin基因的扩增及序列分析

利用聚合酶链式反应,首次从暗纹东方纯基因组DNA中扩增Hepcidin基因的部分序列,序列分析表明,不同鱼类之间的Hepcidin基因DNA序列之间的遗传距离为0.33～1.08,所获得的Hepcidin序列,为进一步研究暗纹东方纯中Hepcidin基因的作用机制提供了基础。     

鱼类铁调素生物学功能与体外表达的研究进展

<正>近年来，细菌性疾病频发，抗生素不规范使用导致细菌耐药性问题日益严重^[1]。研发新的抗菌药物，替代抗生素是解决细菌耐药性问题的一个方向。抗菌肽是具有抗菌作用的小分子多肽，相比于抗生素，不易产生耐药性，可作为抗生素耐药细菌治疗的潜在药物^[2-3]。其中鱼类铁调素(Hepcidin)具有广谱的抗菌活性，抗菌效果明显，已成为当前抗菌肽研究的热点。     

鱼宝--鱼类高效诱食促长营养型素

鱼宝是公司2005年新推出的鱼类高效促长产品,是公司结合鱼类养殖和化工合成的专家,通过大量的生产试验而推出的新型鱼类高效促长素。一、鱼宝的设计思路鱼宝所含的有效成分SOUSA广泛分布于各种鱼类的体内,作为体内正常代谢的活性中间体,对于鱼体进行正常的脂肪、蛋白质、糖类代谢具有重要作用。SOUSA具有一种特殊的鲜味,对动物体无不良反应,对动物的健康不会产生负面影响,可长期应用,无副作用。鱼宝富含高效活性甲基供体,甲基作为机体正常代谢所大量需要的物质,机体不能合成或合成量非常少,严重制约生物的代谢和生长。在鱼体中SOUSA长…     

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。     

热应激对鱼类生命活动影响的研究进展

本文综述了热应激对鱼类生命活动的影响,包括代谢与生长、抗氧化能力、热休克蛋白以及免疫相关研究成果。鱼受到热应激时,机体会产生对自身有害的活性氧,在一定的温度范围内,鱼类通过酶联通路和非酶联通路来消除体内多余的活性氧。大多水鱼类在适宜温度内,生长率与水温成正相关,超过适宜温度,鱼类代谢紊乱,生长缓慢甚至停止,长时间的热应激会导致鱼类死亡。研究鱼类的耐高温性,对创造更高的水产养殖经济效益和选育耐高温品种具有重要的理论价值和现实意义。     

鱼类胰岛素、胰岛素受体和胰岛素受体底物的生物学特性及其对营养物质代谢的调控研究进展

为了深入解析胰岛素调控鱼类营养物质代谢的机制,本文综述了鱼类胰岛素信号系统的3个重要成员（胰岛素、胰岛素受体以及胰岛素受体底物）的结构特征和表达规律,以及胰岛素对营养物质（糖类、脂肪和蛋白质）代谢调控方面的研究进展。目前,在鱼类,虽然胰岛素信号传递系统的研究已取得了一定的进展,但仍有需要重点加强的研究：①鱼类不同亚型胰岛素、胰岛素受体和胰岛素受体底物基因的功能研究,解析鱼类胰岛素信号传递系统的调控机制;②加强胰岛素对鱼类营养物质代谢调控机制的研究,揭示不同食性鱼类胰岛素信号传递系统调控功能的异同;③深入开展鱼类胰岛素信号传导通路和调控机制的研究,全面解析胰岛素调控鱼类营养物质代谢的分子机制。     

肉食性鱼类的糖代谢——以南方鲇为例

肉食性鱼类的糖营养和代谢研究一直是国际学术界的热点之一。以典型的肉食性鱼类南方鲇为实验对象,系统研究了糖种类和添加量对鱼体摄食、消化、排泄、代谢和生长的影响。综合分析认为,对于肉食性鱼类同化的食物糖类不能很好转化为脂肪和糖原储存,也不能大量为机体提供代谢能量,但肉食性鱼类可能采取增加代谢耗能的方式消除吸收的非必需糖类,而底物循环为机体增加代谢耗能的基本方式之一。     

鱼类消化道菌群与碳水化合物代谢

碳水化合物氧化分解是鱼类能量的重要来源。由于碳水化合物来源广泛、价格相对低廉,饲料中添加适量碳水化合物,不仅可以降低饲料成本,而且可以节约蛋白原料,减少氨氮排放。然而以往研究表明,鱼类摄入超量碳水化合物时会出现抗病力受损、生长迟缓、脂肪肝、死亡率升高等问题。鱼类消化道微生物参与宿主的糖、脂类和蛋白质等代谢过程,对动物营养代谢有重要的调控作用。提高鱼类对饲料的利用率,对鱼类增产、渔民增收具有重要的现实意义。本研究综述了鱼类对碳水化合物的代谢,以鱼类消化道微生物为出发点,阐述了鱼类消化道微生物调控碳水化合物代谢的方式与可能机制,旨在为鱼类高效利用碳水化合物以及节约饲料中蛋白质提供新视角。     

鱼类胰岛素、胰岛素受体和胰岛素受体底物的生物学特性及其对营养物质代谢的调控

为了深入解析胰岛素调控鱼类营养物质代谢的机制,本文综述了鱼类胰岛素信号系统的3个重要成员(胰岛素、胰岛素受体以及胰岛素受体底物)的结构特征和表达规律,以及胰岛素对营养物质(糖类、脂肪和蛋白质)代谢调控方面的研究进展。目前,在鱼类,虽然胰岛素信号传递系统的研究已取得了一定的进展,但仍有需要重点加强的研究:(1)鱼类不同亚型胰岛素、胰岛素受体和胰岛素受体底物基因的功能研究,解析鱼类胰岛素信号传递系统的调控机制;(2)加强胰岛素对鱼类营养物质代谢调控机制的研究,揭示不同食性鱼类胰岛素信号传递系统调控功能的异同;(3)深入开展鱼类胰岛素信号传导通路和调控机制的研究,全面解析胰岛素调控鱼类营养物质代谢的分子机制。     

Comparative effects of four feed types on white spot disease susceptibility and skin immune parameters in rainbow trout, Oncorhynchus mykiss (Walbaum)

The effects on rainbow trout, Oncorhynchus mykiss (Walbaum), immune parameters by differently formulated fish feed types containing immunostimulants have been tested in a double-blind, duplicated and controlled study performed over 50 days. A total of 800 rainbow trout (10-12 g) were kept in eight duplicate fish tanks (each containing 100 fish) and fed at a daily feeding rate of 1.5% of the biomass. The feed types were (1) control feed (C) without additives, (2) feed containing beta-glucan, nucleotides, manno-oligosaccharides (MOS), vitamins C and E (GNMCE), (3) feed containing probiotic bacteria and plant extracts (PP) and (4) feed with nucleotides, manno-oligosaccharides, vitamins C and E (NMCE). Plasma lysozyme activity was increased in fish fed two feed types (GNMCE and NMCE) but slightly depressed in fish fed PP. A non-significant trend for a higher mucous cell density at days 30 and 50 was shown in all fish receiving feeds with additives compared to the control group. All fish became infected with Ichthyophthirius multifiliis when exposed, but fish fed GNMCE showed a significantly lower infection both at days 30 and 50. Expression of genes encoding C3 and MHCII was significantly up-regulated in fish fed GNMCE for 50 days, and the expression of genes coding Hepcidin was significantly down-regulated in fish fed NMCE for 50 days. Beta-glucan was the single component, when used in combination with other feed ingredients, which was found associated with increased parasite resistance, increased lysozyme and immune gene up-regulation.     

Dietary vitamin C,immunity and disease resistance in Atlantic salmon (Salmo salar)

Presmolt Atlantic salmon were fed a fish meal based experimental diet supplemented with graded levels of ascorbate-2-monophosphate (AP), equivalent to 40, 400, 2000 and 4000 mg ascorbic acid (AA)/kg for 6 months prior to a bacterial challenge experiment. The liver AA concentration reflected the dietary intake of AP, but not linearly. Growth, hematology and acid phosphatase activity in zymosan stimulated macrophages were not affected by dietary AP. Serum hemolytic complement activity was higher in fish fed the highest AP level, but the variation was not significant. Production of specific antibodies was significantly higher in fish fed the highest AP level 11 and 17 weeks after vaccination. Bacterial challenge with Aeromonas salmonicida showed increased survival in the 4000 AP group. Lysozyme activity in headkidney and serum complement activity and serum iron in fish surviving the challenge were higher in the 4000 AP group, indicating important roles of vitamin C on lysozyme, complement and iron in non-specific disease resistance. The results indicate that high dietary levels of AP favourably affect health in Atlantic salmon. Disease resistance was, however, not correlated with the AA status in the liver.     

Effects of additive iron on growth,tissue distribution,haematology and immunology of gilthead sea bream, <Emphasis Type="Italic">Sparus aurata</Emphasis>

The comparative effects of iron-supplemented levels on growth, tissue distribution, haematology and immunology of gilthead sea bream, Sparus aurata (2 g) were investigated, using four organic (50, 100, 200, 300 mg ORG/kg diet) and one inorganic iron source (200 INOR mg/kg diet). Fish were treated for 12 weeks with the experimental diets and maintained at a water temperature of 19–22°C. Growth (final weight and specific growth rate), tissue distribution (spleen, liver and muscle), haematological parameters (red blood cells, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration) and non-specific immune indexes (respiratory burst activity and antibacterial activity of serum) were analysed. No significant differences were found in growth and iron tissue distribution among the tested groups. Red blood cell counting was statistically higher in fish given 50 ORG, 100 ORG, 200 ORG and 200 INOR feeds. However, haematocrit, haemoglobin and mean corpuscular haemoglobin concentration were not significantly affected by increasing dietary iron. Fish receiving the 100 ORG diet had the best performance with respect to the respiratory burst activity and significantly higher values for antibacterial activity of serum were obtained in fish fed with the 300 ORG diet. The present findings provided no clear evidence of the optimum iron concentration. However, there was adequate indication that iron supplementation enhanced the performance of gilthead sea bream, mainly from a haematological and immunological point of view.     

Effect of dietary iron supplement on growth, haematology and microelements of juvenile grouper, Epinephelus coioides

An experiment was conducted to investigate the effect of dietary iron supplement on growth, haematology and microelements of juvenile grouper, Epinephelus coioides. Casein–gelatine‐based diets supplemented with 0, 50, 100, 150, 200 and 250 mg kg⁻¹ iron from ferrous sulphate were fed to grouper (mean initial weight: 21.0 ± 0.2 g) for 8 weeks. Weight gain was highest in fish fed the diet supplemented with 100 mg kg⁻¹ iron, intermediate in fish fed diets with 50, 150, 200 and 250 mg kg⁻¹ iron and lowest in fish fed the basal diet. Feed efficiency followed a similar trend except that the lowest value was in fish fed the basal diet and the diet supplemented with 250 mg kg⁻¹ iron. Hepatic iron was highest in fish fed diets supplemented with iron ≥100 mg kg⁻¹, followed by fish fed diet with 50 mg kg⁻¹ iron and lowest in fish fed the basal diet. The whole‐body iron was lowest in fish fed the basal diet but not significantly different from other groups, as judged by anova . Iron supplement to the basal diet had no significant effect on haematological parameters (red blood cell count, haematocrit and haemoglobin), hepatic copper concentration or manganese, zinc concentration in liver and whole body. Broken‐line analysis of hepatic iron indicated that iron supplementation of 100 mg kg⁻¹ satisfied the hepatic iron storage and that further supplementation did not expand the iron status.     

急性胁迫对团头鲂铁稳态及其相关基因表达的影响

为研究急性应激反应对鱼体铁含量及铁稳态相关基因的影响,以团头鲂为研究对象,通过腹腔注射皮质醇来模拟急性应激,在注射后不同时间点对团头鲂的血液、肠和肝脏进行取样,采用分光光度法测定血清和肝脏中的铁含量,采用实时荧光定量PCR对铁稳态相关基因的表达量进行检测。结果显示,皮质醇注射后血浆皮质醇显著升高,血浆中铁含量在4、8、10和12 h显著高于对照组,而肝脏中的铁含量在4、8、10和12 h时显著低于对照组。团头鲂肝脏中铁调素(hep)基因表达量显著增加,并分别在8和10 h达最大值,随后有所下降。肠道和肝脏中转铁蛋白基因(Tf)的表达量均呈先升高后下降的趋势。研究表明,急性应激下机体细胞内储存铁大量释放到体液中,造成病原感染和体内增殖风险增加;hep及Tf上调表达降低细胞内铁释放和促进体液铁向胞内蓄积,在鱼体内铁稳态的调控和固有免疫反应中发挥着重要的作用。     

Use of cottonseed meal supplemented with iron for detoxification of gossypol as a total replacement of fish meal in Nile tilapia, Oreochromis niloticus (L.) diets

The objective of this study was to determine the effect of total replacement of fish meal by cottonseed meal (CSM) supplemented with various levels of iron in practical diets on growth performance, feed utilization, body composition and some biological and haematological parameters of Nile tilapia, Oreochromis niloticus (L.). Juvenile fish (average weight 3.78±0.1 g) were stocked in 18 glass aquariums (80 L each) at 25 fish per aquarium. Fish meal (50% of the diet) was used as the sole source of animal protein in the control diet 1. Diets 2–6 had 100% CSM (0.145% free gossypol) protein with various levels of supplemented iron (86, 486, 972, 1458 and 1944 mg Fe kg diet^?1) in diets 2–6 respectively. Diets were fed to fish twice daily at a rate of 3% of body weight during the first 12 weeks then 2% of the total fish biomass daily until the end of the experiment (30 weeks). The results of this study revealed that, groups of fish fed diets 1, 4, 5 and 6 had significantly (P≤0.01) the higher average body weight and specific growth rate than those of fish fed diet 2 (100% CSM without iron supplementation) and diet 3 (100% CSM plus 486 mg Fe kg diet^?1). The best values for feed conversion ratio, protein efficiency ratio and condition factor (K) were recorded with groups of fish fed diet 4 (100% CSM plus 972 mg Fe kg diet^?1). Red blood cell count, haematocrit and haemoglobin were increased with increasing levels of iron and significantly affected by dietary iron. Hepatosomatic index for diets 3–6 were not significantly different (P>0.05) and superior to that of diet 1 control [100% fish meal (FM)]. The gonadosomatic index of males of Nile tilapia was not influenced by CSM diets with or without iron, while females of Nile tilapia were significantly influenced with iron and the lowest values were recorded with groups of fish fed diet 2 (100% CSM without iron supplementation). Apparent digestibility coefficients of protein, fat dry matter and energy were relatively high for most diets supplemented with iron and increased by increasing iron supplementation. There were no significant differences between groups of fish fed diet 1 (100% FM) and diets 5 and 6 which contained 100% CSM with additional 1458 and 1944 mg Fe kg diet supplemental iron^?1 respectively. Proximate composition of whole body was not influenced by diet. Adding 972 mg Fe kg diet^?1 from ferrous sulphate to the CSM‐based diets that contained 972 mg free gossypol (1:1 iron to free gossypol ratio) for Nile tilapia reduce the negative effects of gossypol and improved growth performance, feed utilization and blood parameters and can totally replace fish meal in tilapia diets.     

Haematological and biochemical analyses of Atlantic salmon, Salmo solar L., suffering from coldwater vibriosis ('Hitra disease')

Abstract. Juvenile Atlantic salmon, Salmo salar L. were sampled from a commercial Norwegian fish farm during an outbreak of 'Hitra-disease'. One group of fish subjectively judged as healthy and another as diseased were defined on the basis of the classical apathetic behaviour seen in'Hitra-disease ď salmon. Haematological and biochemical analyses were carried out from blood and organs in 10 fish from each group. The diseased fish were severely anaemic. The blood indices MVC, MCH and MCHC indicated active erythropoiesis to compensate for the loss. Alkaline phosphatase (AP) activity, total protein, albumin, creatinine, triglycerides and total cholesterol were significantly reduced in the serum of diseased fish, while the activities of aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) showed normal and significantly increased values respectively. Liver and spleen weight relative to the body weight and the content of water and lipid in the liver were significantly elevated in diseased fish. Furthermore, the iron content of the spleen was increased, whereas the zinc content showed no changes. The branched-chain amino acids valine, leucine and isoleucine were significantly higher and serine significantly lower in muscle extracts of diseased fish.     

Bioavailability and interactions with other micronutrients of three dietary iron sources in Atlantic salmon, Salmo salar, smolts

Atlantic salmon, Salmo salar L., with mean initial weight of 60 g were fed a diet based on cod muscle meal supplemented with elemental iron, iron sulphate or haem-bound iron in concentrations of 0, 25, 50, 100, 500 and 1500 mg iron kg^?1 for 8 weeks. No significant differences in growth or mortality were found, except in fish fed 1500 mg haem iron kg^?1, which showed reduced growth. In fish fed diets supplemented with elemental iron below 1500 mg iron kg^?1, blood haemoglobin and hepatic iron concentration decreased compared with fish fed the unsupplemented diet. Fish fed diets supplemented with iron sulphate showed increased blood haemoglobin and hepatic iron concentrations between 25 and 100 mg iron kg^?1. Fish fed diets supplemented with haem-bound iron showed increased hepatic iron at all dietary iron levels, while blood haemoglobin concentration decreased in the group fed 1500 mg haem iron kg^?1. The bioavailability of haem iron relative to sulphate iron was calculated by the slope ratio method to be 239% and 148% using blood haemoglobin and hepatic iron, respectively. Relative bioavailability of elemental iron was zero when dietary supplementation levels were between 25 and 500 mg iron kg^?1, while a small part was utilized when 1500 mg elemental iron kg^?1 was supplemented. Additions of 500 and 1500 mg haem-bound iron kg^?1 resulted in a complete loss of ascorbic acid in these diets. When these groups were discounted, no significant relationship between hepatic iron and hepatic ascorbic acid was found. There was no significant effect of dietary iron on whole-body manganese concentration and only a weak effect on whole-body zinc concentration. No significant correlations between dietary iron and hepatic copper concentration were found in any of the dietary treatments. This study also showed that the level of inorganic iron supplementation may be reduced by inclusion of 20 g blood meal kg^?1 in the diet.     

Partial replacement of fish meal by cottonseed meal and soybean meal with iron and phytase supplementation for parrot fish Oplegnathus fasciatus

Two consecutive feeding trials were conducted to determine the optimum dietary level of the cottonseed and soybean meal (CS) for replacement of fish meal (FM) in diets for juvenile and ongrowing parrot fish with or without iron and phytase. In experiment I, juvenile parrot fish (BW: 3.17 g) were fed one of six experimental diets for 12 weeks which were formulated to replace FM protein by equal proportion (1:1, w:w) of CS at 0, 10, 20, 30, 40, or 50% (designated as CS0, CS10, CS20, CS30, CS40, or CS50, respectively). In experiment II, ongrowing parrot fish (BW: 55 g) were fed one of five experimental diets for 9 weeks. The experimental diets were formulated to replace FM protein by CS at 0, 20, or 30% (designated as CS0, CS20, or CS30, respectively) and to include ferrous sulfate (0.1 and 0.2%) and phytase into the CS20 and CS30 diets (CS20 + Fe&P and CS30 +Fe&P, respectively). Results from the two feeding trials indicated that the CS could replace up to 20% FM protein in diets for juvenile parrot fish (3–22 g) and up to 30% in ongrowing (55–120 g) parrot fish. The supplemental effect of iron and phytase was not significant on growth performance. Total or each gossypol enantiomer concentration in the liver increased as the level of dietary cottonseed meal increased. However, gossypol in the liver of fish fed diets supplemented with iron was not detected. The dietary supplementation of CS significantly reduced the levels of plasma triacyglycerols and total cholesterol. The present study indicates that plant protein sources are better utilized in larger fish, and that up to approximately 30% FM protein could be replaced by CS with iron and phytase in the presence of supplemental lysine and methionine.     

Effect of dietary iron levels on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A 60‐day feeding trial was carried out to investigate the effect of iron on growth, body composition and digestive enzyme activities. Diets with seven levels of iron (53.9, 90.0, 115.6, 146.1, 176.0, 215.8 and 266.0 mg iron kg^?1 diet) were fed to Jian carp (initial weight 11.4 ± 0.0 g). Per cent weight gain (PWG), feed efficiency (FE) and protein efficiency ratio were the lowest in fish fed the basal diet (P < 0.05). Body protein content was increased with the increasing iron levels (P < 0.05), but moisture, lipid and ash of fish were not significantly affected by dietary iron levels (P > 0.05). Activities of trypsin, lipase, α‐amylase, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase were improved with increasing dietary iron levels. Serum iron were significantly enhanced with dietary iron levels up to 146.1 mg iron kg^?1 diet, and plateaued. In conclusion, iron improved digestive enzyme activities of juvenile Jian carp and the dietary iron requirement for serum iron of juvenile Jian carp (11.4–64.0 g) was 147.4 mg iron kg^?1 diet with ferrous fumarate as the iron source.