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.     

Geographic Information System as an instrument to determine suitable areas and identify suitable zones to the development of emerging marine finfish farming in Brazil

Worldwide, the availability of suitable areas for aquaculture has become a significant concern. In Brazil, the former Ministry of Fisheries and Aquaculture (MPA) has begun to promote discussions and plan meetings on Local Mariculture Development Plans (PLDMs): a planning tool for the establishment of parks and aquatic areas. However, hundreds of groups and individuals still practice aquaculture awaiting a definitive license. This study aimed to serve as a tool to identify suitable sites for the installation of cobia cage farming and also to contribute to the PLDM zoning process. We created 27 criterion maps for marine fish farming. Their individual weights were assigned based on the best judgment of technicians, researchers and other stakeholders using the pair‐wise comparison method. The criteria and their weights were combined using multi‐criteria decision rules, and a final map according to suitability was derived by reclassification. Of the total area, 54,663 ha (14%) were composed of restrictions, and 341,083 ha (86%) were found useful; of which <1% is considered top score. Even so, these are more than enough to ensure the growth of activity. In Rio de Janeiro state, two bays have suitable areas to marine fish culture development. To ensure the sustainability of marine fish farming, we suggested the adoption of limited production areas based on assimilation and production capacity. Through this methodological approach, it was possible to identify suitable areas for cobia farming development. Additionally, the results may serve as a useful tool on which the PLDM can be based on.     

利用微卫星标记分析军曹鱼养殖群体的遗传多样性

利用12个微卫星标记对北海(BH)、陵水(LS)、硇洲(NZ)、徐闻(XW)和三亚(SY) 5个军曹鱼(Rachycentron canadum)养殖群体进行遗传多样性分析。结果显示,12对微卫星引物在5个军曹鱼养殖群体中共检测到129个等位基因。各养殖群体的平均等位基因数为3.833~6.750,平均有效等位基因数为2.284~3.645,平均观测杂合度和平均期望杂合度分别为0.481~0.635和0.533~0.681,平均多态信息含量为0.463~0.630;哈迪-温伯格平衡检测结果显示,各群体在多个微卫星位点上均显著偏离平衡(P<0.05);军曹鱼养殖群体间的遗传分化指数(Fst)为0.055~0.150,遗传距离(D)为0.240~0.635,其中,BH和NZ的Fst最大(0.150),D最远(0.635);AMOVA分析表明,军曹鱼养殖群体的84%遗传变异来自于个体之间;基于Nei´s遗传距离构建的UPGMA系统进化树显示,BH和SY聚为一支,LS和XW聚为一支,两支聚为一支后与NZ聚为一支。研究结果将为军曹鱼种质资源保护和改良等提供科学的数据参考。     

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.     

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...     

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.     

饲料中添加半乳低聚糖对军曹鱼生长、部分血清免疫和生化因子的影响

以添加0、0．2％、0．4％、0．8％和1．6％半乳低聚糖（GOS）的饲料投喂军曹鱼（Rachycentron canadum）8周,研究GOS对军曹鱼生长、部分血清免疫和生化因子的影响。当饲料中GOS的添加水平低于0．4％时能促进军曹鱼生长,但影响不显著（P〉0．05）。在饲料中添加适当水平的GOS对军曹鱼血清溶菌酶和超氧化物歧化酶（SOD）活性有显著的促进作用,而对过氧化氢酶（CAT）活性有显著的抑制作用。GOS对军曹鱼血清生化因子如总蛋白、白蛋白、球蛋白、总胆红素和直接胆红素的浓度有显著影响,对间接胆红素的影响不显著;只有GOS为0．2％的试验组血清总胆固醇浓度显著高于对照组,而0．4％的试验组血清天门冬氨酸氨基转移酶（AST）浓度显著低于对照组;当添加量低于0．8％时,碱性磷酸酶（AKP）活性随饲料中GOS含量增加而降低。结果表明,在饲料中添加适当水平的GOS有利于军曹鱼的生长,并且能够改善军曹鱼的免疫功能和健康状况,其最佳添加量为0．2％～0．4％。     

Efficacy of tricaine methanesulphonate and clove oil as anaesthetics for juvenile cobia Rachycentron canadum

Six experiments were designed to determine the optimal anaesthetic dosage of tricaine methanesulphonate (TMS) and clove oil that could be used safely on juvenile cobia Rachycentron canadum of two sizes [G1=4.9±0.8 g; G2=13.9±3.1 g]. We documented the stage of anaesthesia and the acute toxicity as 96 h LC₅₀ (lethal concentration 50% population) at various exposure times of the two anaesthetics. At 10 min induction time, the TMS 96 h LC₅₀ was 93.9 mg L⁻¹ in G1 and 97.0 mg L⁻¹ in G2. Compared with clove oil, the 96 h LC₅₀ was 60.0 mg L⁻¹ in G1 and 69.8 mg L⁻¹ in G2. The difference between the two groups (G1, G2) did not influence anaesthesia safety ( P >0.05). Rachycentron canadum achieved stage 3 anaesthesia more rapidly at a lower clove oil concentration level (40 mg L⁻¹, 10 min) than TMS (60 mg L⁻¹, 10 min), but the recovery period of clove oil, was significantly longer. Clove oil was the most effective in reducing the short-term stress induced by routine biometry (20 mg L⁻¹, 10 min) and also by transporting (1 mg L⁻¹, 8 h). Whereas, for long-term exposure, 40 mg L⁻¹ TMS was found to be safe.     

军曹鱼NHE3和NKAα1a基因克隆、表达模式及相关miRNA的靶向调控

为了解军曹鱼NHE3与NKAα1a的序列特征、盐度胁迫下的基因表达模式以及相关miRNA的靶向调控作用，实验应用cDNA末端快速扩增技术克隆了NHE3与NKAα1a的全长cDNA序列；应用实时定量PCR (qPCR)技术分析了NHE3和NKAα1a的组织特异性和盐度适应性表达模式；采用双荧光素酶报告实验检测了NHE3和NKAα1a与相关miRNA的靶向调控关系。结果显示，军曹鱼NHE3与NKAα1a基因的开放阅读框长度分别为2 718和3 075 bp，分别编码905和1 024个氨基酸；NHE3、NKAα1a在鳃、肠、心脏等9种组织中均有表达，其中以鳃组织中的表达丰度最高。随着盐度升高，NHE3在鳃中表达量下降，低盐和高盐适应后均呈显著差异。NKAα1a基因表达量随着盐度升高出现不同趋势，在鳃和肠中受低盐和高盐影响后均显著上调，而高盐环境下其在体肾中的表达水平显著下调。在不同盐度适应过程中，NHE3、NKAα1a均以鳃组织中的表达水平最高。NHE3-pmirGLO-WT与miR-1335-3p共转染时，较对照组相对荧光素酶活性下降，并存在极显著差异；NKAα1a-pmirGLO-WT...