军曹鱼dnd基因cDNA克隆及其在性腺周年发育过程中的表达

本研究通过cDNA末端快速扩增技术(RACE)克隆了军曹鱼(Rachycentron canadum)dnd基因(Rcdnd)的cDNA序列,全长1339bp,其中,5'非编码区59bp,3'非编码区173bp,开放阅读框(ORF)1107 bp,共编码368个氨基酸.Rcdnd氨基酸序列含有1段RNA识别保守基序(R...     

Amending reduced fish‐meal feeds with marine lecithin,but not soy lecithin,improves the growth of juvenile cobia and may attenuate heightened responses to stress challenge

Sparing of marine resources in aquafeeds can be environmentally and economically advantageous; however, fish meal (FM) replacement can affect the production performance and physiological competence. Phospholipids are increasingly understood to be involved in maintaining growth and vigour in fish and may be deficient in reduced FM formulations. Accordingly, we evaluated the growth and stress tolerance of juvenile cobia fed typical (50% FM) or reduced FM feeds (12% FM) with or without phospholipid amendment [1% marine lecithin (12% FM + Marine PL) or soy lecithin (12% FM + Soy PL)] for 6 weeks in triplicate tanks (N = 3) in a recirculation aquaculture system. The 50% FM feed yielded significantly superior growth and growth efficiency in comparison with the 12% FM and 12% FM+ Soy PL feeds, but the 12% FM+ Marine PL feed yielded comparable results to 50% FM feed. A low‐water stress challenge induced elevated plasma glucose, cortisol and lactate levels in all treatments. However, a significant interaction (diet × stress) effect suggested a lesser cortisol response among fish fed the 12% FM+ Marine PL and 50% FM diets. These findings demonstrate that growth performance and, perhaps, resilience of cobia raised on reduced FM feeds may be improved by the addition of marine‐origin phospholipid to the diet.     

急性盐度胁迫下军曹鱼稚鱼应激反应的血清学指标

军曹鱼Rachycentron canadum稚鱼[体质量(25.6±4.9)g]从盐度37‰的自然海水中直接转移至盐度为0、5‰、15‰、25‰、37‰(对照)和45‰的水体中进行盐度急性胁迫试验,转移前及转移后0.5、1、2、3、6和12 h分别检测血清中皮质醇、血糖、乳酸和天门冬氨酸氨基转移酶(AST)等应激指标变化.结果显示,试验期间对照组各指标在基础值附近变幅平稳;盐度0组在6～12 h内出现100%死亡,且死亡前稚鱼皮质醇(256.7 ng·mL-1)和血糖(11.95 mmol·L-1)都显示显著高水平(P＜0.05),而乳酸和AST水平则较低;其他4个胁迫组在最初3 h内皮质醇、血糖和乳酸水平都出现不同程度升高,其中盐度25‰组变幅显著高于其他组,除盐度5‰组皮质醇和乳酸水平在后期有升高趋势外,各组3 h后都逐渐回落至基础值或对照组附近.血清AST水平前期变化较复杂,试验后期盐度5‰和25‰组水平显著高于其他3组.除在淡水中导致稚鱼死亡外,以皮质醇水平为主要应激指标,综合分析显示,稚鱼对盐度胁迫反应初期强弱依次为盐度25‰、5‰、15‰和45‰,而后期在盐度5‰下的应激强度又逐渐增强.     

硒源与硒水平对军曹鱼幼鱼生长性能、肝脏和血清抗氧化指标及组织硒含量的影响

本试验旨在研究硒源和硒水平对军曹鱼幼鱼生长性能、肝脏和血清抗氧化指标及组织硒含量的影响,以确定军曹鱼幼鱼对不同硒源的最适需要量。在基础饲料中分别添加0(对照)、0.3、0.6、0.9和1.2 mg/kg(以硒计)的亚硒酸钠(Se-S)或蛋氨酸硒(Se-Met),配制9种试验饲料(共用对照饲料),饱食投喂初始体重为(22.18±0.35)g的军曹鱼幼鱼10周。每种试验饲料投喂3个网箱(重复),每个网箱放养30尾试验鱼。结果表明:1)硒水平对特定生长率(SGR)和增重率(WGR)有极显著影响(P0.01),但对成活率(SR)和饲料系数(FCR)无显著影响(P0.05);硒源及硒源与硒水平的交互作用对SGR、W GR、FCR和SR均无显著影响(P0.05)。SGR和W GR随着硒水平的升高先升高后降低。2)硒水平极显著影响肝脏谷胱甘肽过氧化物酶(GSH-Px)、谷胱甘肽还原酶(GR)活性和丙二醛(MDA)含量及血清GSH-Px活性(P0.01),并显著影响血清GR活性(P0.05);硒源极显著影响肝脏GR、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)活性和MDA含量及血清T-SOD活性(P0.01);硒源和硒水平的交互作用显著影响肝脏CAT活性和MDA含量及血清T-SOD活性(P0.05)。肝脏和血清GSH-Px活性随着硒水平的升高呈先升高后稳定趋势,GR活性呈先下降后稳定趋势。2种硒源均在添加量为0.9 mg/kg时有最高的肝脏GSH-Px活性,最低的肝脏GR活性。肝脏CAT活性随着硒水平的升高逐渐升高。3)脊椎骨、全鱼和肝脏硒含量随硒水平的升高而增加。硒源极显著影响脊椎骨硒含量(P0.01),硒源和硒水平的交互作用极显著影响肝脏和脊椎骨硒含量(P0.01)。以蛋氨酸硒和亚硒酸钠为硒源,通过二次回归曲线分析得出饲料硒水平分别为1.29和1.46 mg/kg时军曹鱼幼鱼可以获得最大SGR。以SGR和全鱼硒含量为判据,军曹鱼幼鱼对蛋氨酸硒的生物利用率分别相当于亚硒酸钠的1.20和2.90倍。     

军曹鱼饲料中用豆粕代替鱼粉的研究

研究了饲料中用豆粕替代部分鱼粉对军曹鱼Rachycentron canadum生长的影响。将初始体长为(12.77±1.03)cm、初始体重为(7.95±2.12)g的试验鱼随机分配在15个网箱中,每箱放30尾鱼。试验分5组进行,每组设3个平行,分别投喂用豆粕替代鱼粉的基础饲料,替代比例分别为0(对照组)、5%、10%、20%和30%,日投喂量按鱼体重的3%~5%。试验历时60 d,结果表明,当基础饲料中豆粕替代鱼粉的比例不超过10%时,对军曹鱼的相对增重率、饲料系数、成活率以及肥满度没有显著影响(P>0.05);当替代比例为20%、30%时,则对军曹鱼的生长、增重率、饲料系数、成活率以及肥满度有显著影响(P<0.05);当替代比例为30%时,对内脏比才有显著影响(P<0.05)。考虑到基础饲料中已经含有20%的豆粕,建议军曹鱼饲料中豆粕与鱼粉的比例为1∶1.8。     

军曹鱼幼鱼对吡哆醇的需要量

以初始体重(3.23±0.06)g的军曹鱼为研究对象,以酪蛋白(不含维生素)、明胶、鱼肉浓缩蛋白为蛋白源,在基础饲料中分别添加0、2、4、8、16和32mgPN/kg,配制出6种实验饲料,使饲料中吡哆醇的水平分别达到0.22、1.89、3.87、7.54、14.75和29.88mgPN/kg,研究军曹鱼对吡哆醇的需要量。养殖实验在室内流水系统中(250L)进行,每个处理设3个重复,每个重复放养军曹鱼25尾。养殖实验过程中,海水盐度为30～34,水温28～32℃,溶氧>7mg/L,养殖实验持续9周。实验结果显示,在0.22～3.87mgPN/kg水平范围内,随着饲料中吡哆醇含量的升高,实验鱼特定生长率显著升高,当饲料吡哆醇水平达到或高于3.87mg/kg时,各饲料组实验鱼的特定生长率(2.68～2.71%/d)无显著差异,而都显著高于0.22和1.89mg/kg饲料组的特定生长率(1.17～2.06%/d)P<0.05)。军曹鱼肝脏吡哆醇含量、吡哆醛含量、谷丙转氨酶活力、谷草转氨酶活力均与特定生长率有相似的变化趋势,即在3.87mg/kg饲料组接近或者达到最大值,在达到或超过3.87mg/kg时达到或近似达到一...     

Cloning of Full Length cDNA and Tissue Expression Analysis of Hepcidin Gene from Cobia (Rachycentron canadum)

The techniques of homology clone and RACE were used to clone the Hepcidin gene from cobia (Rachycentron canadium).The full length cDNA of Hepcidin gene was 714 bp with a 213 bp 5'untranslated region (UTR),a 228 bp 3'UTR and a 273 bp open reading frame (ORF) encoding a polypeptide of 90 amino acid residues with a predicted molecular weight of 10.03 ku and theoretical isoelectric point of 7.54.The predicted molecular included signal peptide,prodomain peptide and mature peptide.Phylogenetic tree of Hepcidin amino acid sequences was constructed and homology compapison of amino acid sequences showed that homologies were varied from 24.4% to 85.6% with some known Hepcidin amino acids in other fishes and mammals.Quantitative Real-time PCR (qRT-PCR) analysis revealed that Hepcidin gene was expressed in all tissues with different expression levels,which expressed most in liver.The Hepcidin gene expressions in liver,head kidney and spleen were up-regulated after stimulation of LPS and formalin-inactivated Vibrio carchariae.     

微胶囊蛋氨酸或晶体蛋氨酸对军曹鱼幼鱼相关酶活性的影响

比较研究了低鱼粉饲料中添加微胶囊蛋氨酸或晶体蛋氨酸对军曹鱼(Rachycentron canadum),消化酶和蛋白质代谢酶活性的影响.将鱼粉组和减少鱼粉后蛋氨酸缺乏组分别设为正、负对照组,试验组饲料在负对照组的基础上分别添加晶体蛋氨酸(MET)、羟基蛋氨酸(MHA)、邻苯二甲酸醋酸纤维素微胶囊蛋氨酸(CAP)、丙烯酸树脂微胶囊蛋氨酸(RES)和棕榈酸甘油酯微胶囊蛋氨酸(TPA),共7组等氮等脂饲料.每个处理设3个重复,每个重复放养20尾鱼,初始体质量(5.40±0.07)g,流水养殖8周,水温29～31℃.结果显示微胶囊蛋氨酸组的军曹鱼肠道胰蛋白酶活性显著高于正对照组和MET组(P＜0.05),MET和TPA组肝脏谷草转氨酶(GOT)活性显著高于正负对照组(P＜0.05);MET组军曹鱼幼鱼摄食后0.5 h肠道Na+、K+-ATP酶活性显著高于其他各组(P＜0.05),摄食3 h后正对照组和RES组Na+、K+-ATP酶活性上升,显著高于CAP和MHA组(P＜0.05),摄食8 h后正对照组和微胶囊蛋氨酸组显著高于MHA和MET组(P＜0.05).以上结果表明,军曹鱼低鱼粉饲料中添加晶体蛋氨酸即可增进氨基酸的代谢和蛋白质的合成,微胶囊有助于蛋氨酸在肠道中的缓慢释放,提高肠道内胰蛋白酶活性.     

Dietary ascorbic acid requirement of cobia,Rachycentron canadum Linneaus

A 10‐week feeding trial was conducted to determine the optimal requirement of cobia (Rachycentron canadum Linneaus) for dietary ascorbic acid (AA). Graded levels of L‐ascorbyl‐2‐polyphosphate (LAPP) were supplemented in basal diet to formulate six semi‐purified diets containing 2.70 (the control diet), 8.47, 28.3, 80.6, 241 and 733 mg AA equivalent kg^?1 diet, respectively. Each diet was randomly fed to triplicate groups of fish in flow‐through plastic tanks (300 L), and each tank was stocked with 25 fish with average initial weight of 4.59 ± 0.36 g. Observed deficiency signs included poor growth, higher mortality and lower feeding rate (FR) in the fish of the control group. Fish fed the control diet had significantly lower weight gain (WG), lower feed efficiency ratio (FER) and lower tissue AA concentrations in fish liver and muscle. With the increase of dietary AA, the survival, WG, FER, hepatic and muscular AA concentrations of cobia significantly increased and then levelled off. The dietary AA requirement of cobia was estimated to be 44.7 mg kg^?1 based on WG, 53.9 mg kg^?1 or 104 mg kg^?1 based on either hepatic or muscular AA concentration, respectively.