暗纹东方鲀心型脂肪酸结合蛋白H-FABP基因克隆与表达分析

为深入了解心型脂肪酸结合蛋白(heart-type fatty acid-binding protein,H-FABP)的结构与功能,采用RACE技术克隆得到暗纹东方鲀H-FABP的全长cDNA序列,共772 bp,开放阅读框为402 bp,编码133个氨基酸。生物信息学分析表明,暗纹东方鲀H-FABP基因具有一个脂质转运蛋白家族保守结构域和多个蛋白激酶磷酸化位点。NJ法系统进化分析显示,暗纹东方鲀与红鳍东方鲀亲缘关系最近,其次是鲈形目。经荧光定量PCR检测,H-FABP基因在暗纹东方鲀所有被检测的组织中都有表达,其中肝脏中的表达量最高,其次为肌肉和心脏,预示H-FABP主要参与脂肪酸氧化分解过程。为探究豆油作为脂肪源的可行性,采用不同比例(0、25%、50%和75%)豆油替代鱼油的饲料投喂初始体质量为(8.31±0.20)g的暗纹东方鲀8周。结果显示,全鱼油组肝脏中H-FABP基因的表达量显著高于50%豆油组和75%豆油组,但与25%豆油组无显著性差异,且随着饲料中豆油比例的增加,暗纹东方鲀肝脏中H-FABP基因的表达量呈下降趋势。研究表明,富含n-3多不饱和脂肪酸的鱼油可以促进脂肪酸氧化分解,能预防脂肪肝。因此,为不影响暗纹东方鲀肝脏脂肪代谢,饲料中豆油替代鱼油的比例应控制在50%以下。     

饲料中糖/脂肪比对暗纹东方鲀幼鱼生长、血液指标、肝代谢酶活性及PEPCK基因表达的影响

配制6种不同糖/脂肪比例（CHO∶L分别为4.29、2.79、1.86、1.19、0.73和0.41）的等氮等能饲料,饲养暗纹东方鲀幼鱼[初始体重为(11.2 ± 0.5) g] 60 d,探讨饲料中糖与脂肪比例对其生长、饲料利用、生理生化指标和PEPCK基因表达的影响。试验结果表明：随饲料中CHO∶L降低,暗纹东方鲀特定生长率（SGR）、蛋白质效率（PER）和饲料效率（FE）先升高后降低,且均在CHO∶L为1.86时达最大,显著高于CHO∶L为0.41、0.73和4.29时的值(P<0.05)。血浆甘油三酯和肝脏脂肪含量随CHO∶L降低而显著增加(P<0.05),而血糖和肝糖原含量随CHO∶L降低而显著降低(P<0.05)。随饲料中CHO∶L降低,肝脏脂肪酶和脂肪合成酶活性先增加后降低,分别在CHO∶L为1.19和1.86时活性最高。CHO∶L为1.86-4.29时PK活性显著高于其他饲料组(P<0.05)。CHO：L为0.41和0.73时,AMS活性显著低于其他饲料组(P<0.05),而PEPCK活性和mRNA相对表达量显著高于其他饲料组(P<0.05)。饲料中CHO∶L对暗纹东方鲀存活率和血浆总胆固醇含量无显著影响(P>0.05)。分别用二次多项回归模型拟合SGR、PER、FE和CHO∶L的关系,得到暗纹东方鲀幼鱼饲料中CHO∶L的适宜范围为2.01~2.16,且其对碳水化合物的利用能力要高于脂肪。     

暗纹东方鲀仔鱼对药物的耐受性研究

采用急性攻毒的方法，对暗纹东方鲀仔鱼进行硫酸铜、硫酸铜与硫酸亚铁合剂和福尔马林等3种水产养殖常用驱虫药物的毒性试验。结果表明：暗纹东方鲀仔鱼对3种药物敏感的大小顺序为：硫酸铜与硫酸亚铁合剂〉硫酸铜〉福尔马林；暗纹东方鲀仔鱼对硫酸铜合剂的敏感性最高，安全浓度小于0．05mg／L；对硫酸铜的安全浓度为0．21mg／L，24小时半致死浓度分别为1．03mg／L和1．25mg／L。暗纹东方鲀仔鱼对福尔马林的敏感性相对较低，浸洗浓度至少可达20mg／L。     

3种水产消毒剂对暗纹东方鲀稚鱼的急性毒性试验

在水温21~23℃,pH 8.20~8.50,溶解氧6.00~7.50mg/L条件下,采用半静水法研究了苯扎溴铵、戊二醛和聚维酮碘对全长(1.73±0.32)cm、体质量(0.19±0.01)g暗纹东方鲀稚鱼的急性毒性效应。试验结果表明,苯扎溴铵对暗纹东方鲀稚鱼24、48、72h和96h半致死质量浓度分别为1.04、0.95、0.72mg/L和0.53mg/L;戊二醛对暗纹东方鲀稚鱼24、48、72h和96h半致死质量浓度分别为4.27、3.55、2.94mg/L和2.40mg/L;聚维酮碘对暗纹东方鲀稚鱼24、48、72h和96h半致死质量浓度分别为269.80、200.00、158.54mg/L和151.09mg/L。苯扎溴铵、戊二醛和聚维酮碘对暗纹东方鲀安全质量浓度分别为0.24、0.74mg/L和32.97mg/L,均高于其厂家推荐使用质量浓度,可以在暗纹东方鲀生产中使用。但暗纹东方鲀对苯扎溴铵和戊二醛较为敏感,在生产过程中应严格控制使用质量浓度。     

非离子氨氮和亚硝酸盐氮对暗纹东方鲀稚鱼的急性毒性试验

在水温21~23℃,pH 8.2~8.5,溶解氧6.00~7.50mg/L的条件下,采用半静水法研究了非离子氨氮和亚硝酸盐氮对全长(1.6±0.2)cm、体质量为(0.11±0.05)g的暗纹东方鲀稚鱼的急性毒性效应。试验结果表明,暗纹东方鲀稚鱼受到非离子氨氮和亚硝酸盐氮胁迫后,先后出现鱼体体色变白、扭曲、侧游、失去平衡、昏迷等中毒症状。随着非离子氨氮和亚硝酸盐氮质量浓度的提高和胁迫时间的延长,暗纹东方鲀稚鱼死亡率逐渐升高,存在明显的剂量效应和时间效应关系。非离子氨氮和亚硝酸盐氮对暗纹东方鲀稚鱼96h半致死质量浓度分别为0.46mg/L(95%置信限0.34~0.64mg/L)和290.12mg/L(95%置信限255.16~329.87mg/L),安全质量浓度分别为0.046 mg/L和29.01mg/L。非离子氨氮和亚硝酸盐氮对暗纹东方鲀稚鱼具有一定毒性,且非离子氨氮毒性大于亚硝酸盐氮毒性。     

暗纹东方鲀仔鱼对药物的耐受性研究

采用急性攻毒的方法,进行硫酸铜、硫酸铜与硫酸亚铁合剂和福尔马林等三种水产养殖常用驱虫药物对暗纹东方纯仔鱼的毒性试验.结果表明:暗纹东方纯仔鱼对这三种药物敏感程度的大小顺序为:硫酸铜与硫酸亚铁合剂>硫酸铜>福尔马林;暗纹东方纯仔鱼对硫酸铜合剂的敏感性最强,安全浓度小于0.05 mg/L;硫酸铜的安全浓度为0.21 mg/L,24 h半致死浓度分别为1.03 mg/L和1.25 mg/L.暗纹东方鲀仔鱼对福尔马林的敏感性相对较弱,浸洗浓度至少可达20 mg/L.     

芳香化酶抑制剂对暗纹东方鲀CYP19A、DMRT1基因表达及性腺分化的影响

采用组织学和分子生物学方法,研究了投喂芳香化酶抑制剂来曲唑(LE)后暗纹东方鲀(Takifugu obscures)初孵仔鱼CYP19A、DMRT1基因表达以及性腺的组织学变化,以期进一步了解P450芳香化酶(P450arom)在鱼类早期性别分化过程中的作用。RT-PCR结果显示,对照组样品CYP19A和DMRT1表达显示性二态,雌性表达CYP19A基因,雄性表达DMRT1基因。LE处理组在性别分化期间,雄性样品单一表达DMRT1,雌性样品则同时表达CYP19A和DMRT1。qRT-PCR结果显示:LE处理组雌性仔鱼CYP19A基因表达被显著抑;虽然在仔鱼出膜后22d(dph)的表达水平高于9 dph,但仅为同日对照组的2.11%。LE处理组雌性样品22 dph时DMRT1基因表达量上调,至150 dph时达对照组雄性水平。55 dph的性腺组织学结果表明,LE处理可导致暗纹东方鲀稚鱼原始卵巢退化,并向功能性精巢发育。150 dph的LE处理组性腺均为精巢,并与对照组精巢发育同步。结论认为,暗纹东方鲀性腺分化期间P450arom是卵巢形成和维持发育所必须的,抑制P450arom活性可导致雌性暗纹东方鲀发生雄性化逆转。     

暗纹东方鲀抗苗勒氏管激素Ⅱ型受体基因的克隆、生物信息学及表达分析

为探究暗纹东方鲀(Takifugu obscures)抗苗勒氏管激素Ⅱ型受体(anti-Müllerian hormone receptorⅡ,Amhr2)基因的序列和结构信息,初步研究amhr2基因在性腺发育和性别分化过程中的作用,实验通过设计简并引物扩增及RACE技术,克隆出暗纹东方鲀amhr2完整的CDS区,分析其相应的生物信息学特征及其在不同组织和不同发育期的mRNA表达水平。结果表明:amhr2序列cDNA全长为1 868 bp(NCBI登录号:MH218814),编码513个氨基酸,与红鳍东方鲀(Takifugu rubripes)同源性最高,达99%。氨基酸多重序列比对结果显示,Amhr2氨基酸序列中的近C-末端区域序列较为保守;通过构建系统进化树发现,暗纹东方鲀Amhr2与红鳍东方鲀亲缘关系最近。荧光定量PCR分析显示,暗纹东方鲀amhr2仅在性腺中表达,且卵巢中的表达量明显高于精巢;并且在性腺早期发育过程中,amhr2在精巢和卵巢中的表达量均呈现出先降低再迅速升高的表达趋势。研究结果表明,amhr2在暗纹东方鲀中的表达具有组织特异性,且在性别分化和性别维持等过程中发挥重要的作用。     

玉米蛋白粉替代鱼粉对暗纹东方鲀溶菌酶活性及c型溶菌酶mRNA表达的影响

以初重为(41.26±1.09)g的暗纹东方鲀为实验鱼,配制5种玉米蛋白粉使用量为0%、5%、10%、15%、20%的实验饲料,玉米蛋白粉对饲料中鱼粉的替代水平分别为0%、7.4%、14.8%、22.2%和29.6%。用实验饲料饲养暗纹东方鲀60d,研究玉米蛋白粉替代鱼粉对暗纹东方鲀肝胰脏、头肾和脾脏组织的溶菌酶活性及其c-型溶菌酶mRNA相对表达量的影响。结果表明,(1)暗纹东方鲀肝胰脏、头肾和脾脏的溶菌酶比活力分别为9.14、42.12和40.49U/mgprot,头肾和脾脏组织中c-型溶菌酶mRNA相对表达量分别是肝胰脏的3.76和3.24倍。(2)玉米蛋白粉替代鱼粉显著影响暗纹东方鲀溶菌酶比活力。5%和10%组的肝胰脏和头肾组织溶菌酶比活力显著高于对照组(P<0.05),而15%和20%组肝胰脏和头肾组织溶菌酶比活力则显著低于对照组(P<0.05);5%和10%组的脾脏组织溶菌酶活力与对照组无显著性差异(P>0.05),而15%和20%组显著低于对照组和5%组(P<0.05)。(3)玉米蛋白粉对暗纹东方鲀c型溶菌酶基因mRNA相对表达量有显著影响(P<0.05)。肝胰脏组织中5%和10%组表达量显著高...     

低盐胁迫对红鳍东方鲀幼鱼肝脏的酶活性、组织结构及免疫基因表达的影响

为探讨低盐胁迫下肝脏在红鳍东方鲀(Takifugu rubripes)机体免疫中的作用,在肝脏酶活性、组织结构和基因表达水平3个方面进行了研究。结果表明,低盐(盐度4‰)胁迫下红鳍东方鲀肝脏中总超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)的酶活性显著高于对照组(P0.05);肝脏组织结构中肝细胞发生肿大,形状变形;肝脏IL15和TLR3基因的表达量显著升高(P0.05),而IRF2基因的表达量却未有显著性差异(P0.05)。     

Acute Toxicity and Sublethal Effects of Nitrite on Selected Hematological Parameters and Tissues in Dark-banded Rockfish, Sebastes inermis

Acute toxicity and sublethal effects of nitrite in dark‐banded rockfish, Sebastes inermis (83.3 ± 7.2 g), were studied under static conditions for a period of 96 h. The acute toxicity of nitrite evaluated for the 96‐h lethal concentration (LC₅₀) was 700 mg/L. The sublethal effects on selected hematological parameters of S. inermis, such as total erythrocyte count (TEC), hemoglobin, plasma glucose, and serum protein content, were measured after 0, 6, 12, 24, 48, 72, and 96 h of exposure to 0, 50, 100, 200, 400, and 700 mg/L of nitrite. Sublethal nitrite caused progressive reduction in the TEC, hemoglobin, and serum protein content in fish depending on the nitrite concentration and exposure period. The 96‐h exposure resulted in a 14–42% reduction in TEC and 25–33% reduction in hemoglobin content for 100–700 mg/L of nitrite compared to the control. A dose‐related reduction in plasma glucose (25.7–34.2%) was observed for concentrations of 200–700 mg/L of nitrite during 48 h of exposure, followed by an increase through 96 h. A significant reduction in serum protein (7.3–12.6%) was observed for 200–700 mg/L of nitrite after 96 h of exposure. Abnormal histological changes in skin, gill, liver, and kidney tissue were observed in fish exposed to 700 mg/L of nitrite after 96 h of exposure compared to the control. Although no mortality of S. inermis occurred at 500 mg/L of nitrite, all hematological parameters adversely responded to a nitrite dose of 200 mg/L for 96 h. These results showed that although acute toxicity concentration of nitrite in S. inermis is higher than 700 mg/L, sublethal concentrations of nitrite also negatively affect hematological parameters.     

Supplementing copper to a fish meal based diet fed to Atlantic salmon parr affects liver copper and selenium concentrations

The present experiment was conducted to study growth and tissue responses in Atlantic salmon, Salmo salar L., fed a fish meal based diet supplemented with copper (Cu). The findings of the experiment were used to evaluate the need for dietary Cu supplementation. Atlantic salmon parr, initially weighing ≊ 7.5 g, were randomly distributed among 10 tanks, with 300 fish in each tank. Duplicate groups of fish were fed a fish meal based diet containing 3.5 mg Cu kg⁻¹, or this diet supplemented with 5, 10, 50 or 100 mg Cu kg⁻¹ (as CuSO₄*5 H₂O) for 12 weeks. Growth was recorded and blood haemoglobin measured. The Cu concentrations in whole body, liver, serum and selected muscle samples were measured, as was liver selenium (Se) concentration.
There were no difference in growth among the dietary treatments. There were, however, significant differences among the dietary groups in liver Cu and Se concentrations. The fish fed the diet supplemented with 5 mg Cu kg⁻¹ had increased liver Cu concentration compared with the other groups. Similar trends were found for serum Cu concentration and whole-body Cu concentration, but these effects were not significant. Liver Cu and Se concentrations were positively correlated and liver Se concentrations were inversely correlated to dietary Cu concentration, confirming an interaction between these two elements in salmon.
There may be a positive effect of a modest Cu supplementation level, and we suggest that a small amount of Cu (5–10 mg kg⁻¹) should be added to fish meal based diets.     

铜对刀鲚幼鱼的急性毒性及对肝抗氧化酶活性与组织结构的影响

采用静水生物测试法研究铜(Cu²⁺)对刀鲚(Coilia nasus)幼鱼的急性毒性并进行安全性评价。根据预实验结果, 设定0.3、0.54、0.96、1.68 和3 mg/L 5个CuSO₄浓度梯度进行急性暴露实验, 以肝组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性为指标, 并通过观察肝、鳃组织病理变化来研究Cu²⁺污染对刀鲚幼鱼的毒性效应。结果表明, 刀鲚幼鱼随Cu^{2 +} 浓度升高毒性效应逐渐增强, 24、48、72和96 h LC₅₀分别为1.087、1.062、1.042和0.967 mg/L, 安全质量浓度为0.009 67 mg/L。刀鲚幼鱼暴露24 h之后, 各Cu²⁺浓度处理组(0.1, 0.2, 0.3 mg/L) CAT活性、SOD活性显著提高(P<0.05); 暴露48 h后, 各组CAT活性下降, CAT活性均低于对照组, 下降幅度与Cu^{2 +}质量浓度呈正相关, SOD活性提高显著(P<0.05); 暴露72 h后, SOD活性有所下降, SOD活性均低于对照组(P<0.05)。1.68 mg/L和3 mg/L的Cu²⁺浓度对刀鲚幼鱼的鳃和肝都造成严重的损伤, 暴露在高浓度组(3 mg/L)中的刀鲚, 其鳃上皮细胞肿大变性, 鳃小片几乎全部脱落; 肝呈黄色, 超微结构发现肝细胞明显增大, 线粒体空泡化, 胞质中出现空泡和大量脂滴, 表明Cu²⁺可诱发刀鲚出现脂肪肝综合征。SOD、CAT活性变化以及鳃、肝结构变化都反映刀鲚幼鱼受伤害的程度, 可用作安全性风险评价的参考依据。     

急性氨氮胁迫对黄颡鱼组织抗氧化酶活性及HSP70和HSP90基因mRNA表达水平的影响

为了探讨急性氨氮胁迫对黄颡鱼组织中抗氧化酶活性及HSP70和HSP90基因mRNA表达水平的影响,实验随机挑选了360尾黄颡鱼[初体质量(17.25±0.05)g],分别暴露于含有0(对照)、5.70(低浓度组)、28.50(中浓度组)和57.00(高浓度组)mg/L总氨氮浓度的水体中,进行96 h的急性胁迫实验。实验开始后,分别于0、12、24、48和96 h取样。结果显示,氨氮胁迫发生后,低、中浓度组实验鱼肝脏中超氧化物歧化酶(SOD)活性呈先升高后降低趋势,而高浓度组则持续降低;低、中、高浓度组实验鱼肝脏中丙二醛(MDA)含量在胁迫开始后显著升高;3 h时,高浓度组实验鱼肝脏中SOD活性达到最低,而MDA含量最高;24 h后,高浓度组实验鱼肝脏中过氧化氢酶(CAT)活性显著升高;低、中、高浓度组实验鱼肝脏中HSP70基因的mRNA表达量呈先降低后升高趋势,而鳃中HSP70基因表达量持续升高,但脑中HSP70基因在0 h后显著降低;氨氮胁迫3 h时,低、中、高浓度组实验鱼肝脏和脑中HSP70基因表达量显著低于对照组,而在鳃中正好相反;相比HSP70基因,高氨氮浓度组实验鱼肝脏和鳃中HSP90基因的mRNA表达量在24 h时达到最高。研究表明,不同浓度的氨氮胁迫会对黄颡鱼抗氧化酶活性造成不同程度的抑制,原因与丙二醛的积累量有关;相比HSP90基因,黄颡鱼HSP70基因的表达量在氨氮胁迫发生后迅速上调,这种生理调控机制提示HSP70在应对急性氨氮胁迫时发挥着更重要的作用。     

急性氨氮暴露对大弹涂鱼炎性反应相关基因表达的影响

为研究急性氨氮胁迫对大弹涂鱼炎性反应相关基因表达的影响,实验挑选初始体质量为(15.14±0.05) g的健康大弹涂鱼幼鱼180尾,进行96 h的急性氨氮胁迫实验。结果显示,大弹涂鱼96 h氨氮半致死浓度为8.99 mg/L总氨氮(0.11 mg/L非离子氨,T-AN);氨氮胁迫后TNF基因的mRNA表达量分别于12和96 h时显著上调,96 h时表达量达到0 h时的2倍;IL-1基因的mRNA表达量12 h时显著上调,为0 h时表达量的6倍;氨氮胁迫后IL-6基因的mRNA表达量分别于12和96 h时显著上调,表达量达到0 h时的1.5倍;氨氮胁迫后IL-8基因的mRNA表达量在24 h时出现显著下调。研究表明,大弹涂鱼96 h氨氮半致死浓度为8.99 mg/L总氨氮;半致死浓度的氨氮胁迫48 h后,TNF、IL-1、IL-6和IL-8基因的mRNA表达量持续升高,推测过度炎性应激可能是导致鱼类氨中毒死亡的原因之一。     

LAS与Cu2+单一及复合污染对泥鳅肝脏SOD、CAT活性的影响

采用暴露试验法研究了十二烷基苯磺酸钠（LAS）与Cu^2＋单一及复合污染对泥鳅肝脏超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性的影响。试验结果表明，单一LAS处理96h后，SOD活性变化不显著，CAT在LAS为20．0mg／L时被显著诱导（P〈0．05）；单一的Cu^2＋处理96h后，泥鳅肝脏SOD、CAT活性受到显著影响（P〈0．05），随着质量浓度的增加，表现为先诱导后抑制；LAS、Cu^2＋复合处理条件下，LAS为10．0mg／L时，CAT、SOD活性分别在Cu^2＋质量浓度为0．05、0．10mg／L时达到最低值，LAS为20．0mg／L时，CAT、SOD活性有所回升，但其活性仍低于对照组，且差异均达到显著水平（P〈0．01）。通过比较，发现泥鳅肝脏CAT活性变化更能反映出污染物对泥鳅的毒性作用。     

慢性氨氮胁迫对刺参摄食与消化酶活性的影响

以刺参(Apostichopus japonicas Selenka)为研究对象,探讨氨氮胁迫浓度为0 mg/L、2 mg/L、4 mg/L、6 mg/L、8 mg/L、10 mg/L时14 d内刺参摄食情况与消化道内3种不同消化酶活性变化。刺参规格分别为小[S,体重(15.13±0.02)g]、中[M,体重(28.42±0.32)g]、大[L,体重(60.14±0.88)g]。结果显示:(1)随着氨氮浓度的升高,实验刺参存活与生长逐渐与对照组出现显著差异。当氨氮浓度高于2 mg/L时,刺参末体重与特定生长率随氨氮浓度升高有显著下降趋势;当氨氮浓度为8~10 mg/L时,3种不同规格刺参均出现了吐肠、化皮,直至死亡,氨氮浓度为10 mg/L时中规格刺参存活率最低,为81.3%。(2)当氨氮浓度低于10 mg/L时,氨氮胁迫对3种规格刺参摄食率(FR)影响差异不显著,而全部实验组的食物转化率(FCE)均显著低于对照(P0.05)。(3)氨氮胁迫对3种不同规格刺参消化酶活性的影响存在差异。随氨氮浓度升高,蛋白酶活性呈降低趋势,氨氮浓度8 mg/L和10 mg/L时全部实验组刺参蛋白酶活性显著低于对照(P0.05);脂肪酶和淀粉酶活性随氨氮浓度升高呈先升后降趋势,在2 mg/L时出现峰值;当氨氮浓度为6 mg/L时,小规格刺参的消化道淀粉酶活性降低到0.30 U/mg(prot),与对照出现显著差异(P0.05);当氨氮浓度从6 mg/L到8 mg/L时,3种规格刺参脂肪酶活性发生急剧变化,显著低于对照(P0.05),中规格刺参在同一氨氮浓度胁迫下的消化酶活性变化显著高于其他规格。研究表明,氨氮胁迫会对刺参摄食、消化与生长产生不利影响,在氨氮浓度4 mg/L以下刺参消化酶活性短期可被显著诱导上调,高氨氮浓度对消化酶活性起抑制效应并可导致生理紊乱,此响应存在体重规格上的差异。     

硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响

为探讨硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响,以初始体质量为(24.85±0.10) g的大菱鲆为研究对象,通过在饲料中添加五水硫酸铜和亚硒酸钠,配制铜、硒含量分别为(0、0,1 000、0,1 000、2,1 000、4 mg/kg) 4种等氮等能的实验饲料,命名为D1、D2、D3和D4组。养殖实验持续84 d。结果显示:①各组间实验鱼成活率无显著差异;增重率、特定生长率和蛋白质效率均在D2组显著低于D1、D3和D4组;摄食率和饲料系数呈相反趋势,在D2组显著高于其他3组。②幼鱼全鱼和肝脏粗脂肪含量呈先下降后上升趋势;而背肌粗蛋白和粗脂肪含量在各组之间无显著性差异。全鱼、脊椎骨和肝脏中Cu含量在D2组达到最高值;Zn含量则在D4组达到最高值;肝脏中铁含量呈下降趋势,在D3和D4组显著低于D1和D2组。③D2组显著降低了幼鱼血清总抗氧化能力(T-AOC)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性;丙二醛(MDA)含量则呈相反趋势,在D2组显著高于D3和D4组;总超氧化物歧化酶(T-SOD)和铜—锌超氧化物歧化酶(CuZn-SOD)活性均在D4组达到最低值。④血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性均在D2组显著高于其他3组;而血糖浓度、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)活性均在D2组达到最低值,显著低于D1组。⑤肝脏金属硫蛋白(MT) mRNA相对表达量在D4组达到最高值,显著高于其他3组;谷胱甘肽转移酶(GST)和溶菌酶(LZM) mRNA相对表达量均在D2组显著低于其他3组;热休克蛋白70 (HSP70) mRNA相对表达量则在D2组显著高于其他3组。综上所述,在本实验条件下,饲料中添加硒(2～4 mg/kg)可缓解高铜(1 000 mg/kg)胁迫导致的鱼体生长缓慢等症状,并可调控鱼体的抗氧化能力、生理代谢及相关基因表达量,进而对鱼体在高铜胁迫下的内环境稳态恢复具有一定的促进作用。     

Tissue lipid peroxidative responses in Atlantic salmon (Salmo salar L.) parr fed high levels of dietary copper and cadmium

Atlantic salmon (Salmo salar L.) parr were fed for one month on fish meal based diets supplemented with Cd (0, 0.7, or 204 mg Cd kg^–1 DW) or Cu (0, 34, or 691 mg Cu kg^–1 DW) to assess the effects of non-essential (Cd) and essential (Cu) dietary metals on lipid peroxidation and the oxidative defence system. Cadmium accumulated significantly in the liver, intestine, and kidney of 204 mg Cd kg^–1exposed fish compared to controls. Copper accumulated significantly in the intestine, kidney, and liver of fish exposed to 691 mg Cu kg^–1, and in the intestine of 34 mg Cu kg^–1 exposed fish. Tissue Cu accumulation significantly increased intestinal and hepatic lipid peroxidation (as seen from thiobarbituric acid reactive substances, TBARS, levels) and subsequently decreased intestinal -tocopherol levels and increased intestinal and hepatic selenium dependent glutathione peroxidase (SeGSH-Px) activity. Dietary Cd significantly reduced SeGSH-Px activity in the intestine and liver of 204 mg Cd kg^–1 exposed fish compared to controls. No significant increase in tissue TBARS or reduction of -tocopherol levels was observed in the intestine of fish exposed to dietary Cd, with exception of the highest exposure group (204 mg Cd kg^–1). Dietary Cu caused depletion of tissue Se and glutathione levels, however the reduced availability of GSH and Se did not seem to explain the differences in SeGSH-Px activity. Dietary Cu had a direct effect on lipid peroxidation at a relatively low concentration (34 mg Cu kg^–1). Cadmium indirectly affected tissue lipid peroxidation by damaging the oxidative defence system at the highest dietary concentration (204 mg Cd kg^–1).     

Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder,Paralichthys olivaceus,and Red Sea Bream,Pagrus major

The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.