Oxidative stress in juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum), were held in 8–11°C freshwater, starved for 3 days and subjected to a low‐water stressor to determine the relationship between the general stress response and oxidative stress. Lipid peroxidation (LPO) levels (lipid hydroperoxides) were measured in kidney, liver and brain samples taken at the beginning of the experiment (0‐h unstressed controls) and at 6, 24 and 48 h after application of a continuous low‐water stressor. Tissue samples were also taken at 48 h from fish that had not been exposed to the stressor (48‐h unstressed controls). Exposure to the low‐water stressor affected LPO in kidney and brain tissues. In kidney, LPO decreased 6 h after imposition of the stressor; similar but less pronounced decreases also occurred in the liver and brain. At 48 h, LPO increased (in comparison with 6‐h stressed tissues) in the kidney and brain. In comparison with 48‐h unstressed controls, LPO levels were higher in the kidney and brain of stressed fish. Although preliminary, results suggest that stress can cause oxidative tissue damage in juvenile chinook salmon. Measures of oxidative stress have shown similar responses to stress in mammals; however, further research is needed to determine the extent of the stress–oxidative stress relationship and the underlying physiological mechanisms in fish.     

Adapting bioenergetic factorial modelling to understand the implications of heat stress on barramundi (Lates calcarifer) growth,feed utilisation and optimal protein and energy requirements – potential strategies for dealing with climate change?

The implications of temperature on bioenergetics for barramundi (Lates calcarifer) were defined in an improved factorial model that encompassed revised parameters accounting for effects over the temperature range of 16–39 °C and size range of 10–3000 g. A revised growth function describing weight gain by barramundi as a function of fish weight and temperature was derived from farm and laboratory data and included a term for a shift in optimal temperature with fish size: Gain (g fish^?1 day^?1) = (K + xT + yT ² + zT³) * (weight)^ax+b. Maintenance energy and protein demand functions were also derived on a similar form, and all three functions combined to form the basis of a factorial model for energy and protein demand. Using this model, optimal iterative feed specifications were defined for a range of fish sizes at temperatures of 25, 30 and 35 °C. A feed demand model was also developed based on the demand for digestible energy (DE) at each of these temperatures. The model shows that at high temperatures (35 °C), there is an increase in digestible protein (DP) to DE demand, and that with increasing size, there is a decrease in the DP to DE demand.     

有机铬对热应激蛋鸡血清生化指标的影响

目的,研究有机铬对热应激蛋鸡血清中某些生化指标的影响。方法,将处于热应激条件下的192只35周龄新罗曼褐壳蛋鸡随机分为6组,分别饲喂不添加铬的玉米-豆粕型基础饲粮和添加10、20、40及50mg/kg铬的试验饲粮49d结果:试验组中血糖、胆固醇、总蛋白、球蛋白平均含量均低于对照组,并且添加20mg/kg铬组的血糖、胆固醇、白蛋白含量与对照组相比差异显著(P<0.05)。对于AKP活性,添加40mg/kg铬组与对照组相比差异显著(P<0.05),其它各组间相比均为差异不显著(P>0.05)。对于尿酸含量,添加50mg/kg铬组与对照组相比为差异显著。结论:有机铬具有缓解热应激的作用。     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.     

团头鲂(Megahbrama amblycephala) Caspase 9 基因全长cDNA的克隆及在氨氮胁迫下的表达分析

为了解团头鲂(Megahbrama amblycephala) Caspase 9基因序列特征及其在氨氮胁迫过程中的作用,应用末端快速扩增(Rapid amplification of cDNA ends,RACE)技术克隆得到团头鲂Caspase 9基因全长1613 bp的cDNA序列,包括185 bp的5 '末端非翻译区(Untranslated regions,UTR)、96 bp的3'UTR、1332 bp的开放阅读框(Open Reading Frame,ORF).氨基酸多序列比对显示,平均同源性为77.74％,表明团头鲂Caspase9基因具有较高的保守性;系统进化分析显示,该基因氨基酸序列与其他鱼类Caspase 9聚为一支,并与锦鲤(Cyprinus carpio) Caspase 9亲缘关系较近;荧光定量PCR结果显示,Caspase 9基因在团头鲂各组织中均有表达,在脑中表达量最高,在肌肉中表达量最低,同时,在胁迫和恢复过程中,该基因在肝和脑中的表达规律相似,均在胁迫期间出现表达量上调,整体呈类似波浪形表达谱.研究结果表明,氨氮胁迫下,Caspase 9基因参与团头鲂的免疫防御,并与细胞凋亡分子过程有关.本研究结果可为进一步了解团头鲂应答氨氮胁迫分子机制提供理论依据.     

盐胁迫对雨生红球藻虾青素累积、虾青素合成相关酶基因表达和抗氧化指标的影响

为探讨盐胁迫对雨生红球藻(Haematococcus pluvialis)虾青素合成的影响与机理,以及雨生红球藻各抗氧化机制之间的关系,本研究采用生化和分子生物学方法研究了不同浓度(0.04 mol/L、0.08 mol/L、0.12 mol/L和0.16 mol/L)和不同时间(3 d、6 d和9 d)的盐(Na Cl)胁迫对雨生红球藻生长、虾青素积累、番茄红素β-环化酶(Lcy)、β-胡萝卜素羟化酶(Crt R-B)和β-胡萝卜素酮化酶(Bkt)基因表达、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性以及丙二醛(MDA)含量的影响。结果表明,各胁迫时间的雨生红球藻的密度均随着盐胁迫浓度的增加而不断下降,在盐胁迫的第9天,雨生红球藻的死亡率和孢子比例均随着盐胁迫浓度的增加而不断升高;雨生红球藻虾青素含量、Lcy、Crt R-B和Bkt基因表达量均随着盐胁迫浓度和时间的增加而不断提高;雨生红球藻SOD、CAT和GSH-Px活性以及MDA含量在不同浓度和不同时间的盐胁迫下与对照组(0.00 mol/L Na Cl)相比均升高,且在不同时间的0.12 mol/L Na Cl胁迫下与对照组相比均显著升高(P0.05);雨生红球藻虾青素含量、Lcy、Crt R-B和Bkt基因表达量在盐胁迫的早期(第3天)和中期(第6天)阶段较低,在盐胁迫的后期(第9天)阶段较高,而SOD、CAT和GSH-Px活性以及MDA含量在盐胁迫的早期和中期阶段较高,在盐胁迫的后期阶段较低。实验结果说明了适当浓度和时间的盐胁迫能促进雨生红球藻累积虾青素,雨生红球藻在盐胁迫下主要是通过提高虾青素合成相关酶基因的转录水平来促进虾青素的合成,其虾青素和抗氧化酶的抗氧化活性可能互为补充,共同保护雨生红球藻免受盐胁迫的氧化损伤。     

热胁迫对白梭吻鲈鳃组织结构及抗氧化指标的影响

温度是导致鱼类生理变化的最重要的环境因素之一。热应激导致氧化应激,破坏抗氧化防御系统,并导致脂质和组织病理学的损害。白梭吻鲈(Sander lucioperca)属于亚冷水性鱼类,最适生长水温20～24℃。近年来,由于夏季气温极高,中国池塘出现了大量的白梭吻鲈死亡,白梭吻鲈的水产养殖业一直面临着高温的严峻挑战。鱼鳃参与鱼类许多重要功能,如呼吸,渗透调节和排泄,与外部环境保持密切接触,对水质变化特别敏感。为探究热胁迫对白梭吻鲈鳃组织结构及抗氧化指标的影响,将100尾鱼暴露于28℃,31℃,34℃,36℃热应激2h,以23℃作为对照组。收集鳃用于石蜡切片制备,并将苏木精伊红（HE）染色用于一般组织学观察。测定超氧化歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPX）活性和丙二醛（MDA）等抗氧化指标。结果显示,28 ℃组鳃小片发生充血,31 ℃组部分鳃小片末端卷曲变形、上皮细胞水肿,34 ℃组鳃小片变形加重,且有少量融合,上皮细胞水肿加重、明显隆起,鳃小片之间粘液细胞肥大、增生,36 ℃组相邻鳃小片大量融合且严重变形,卷曲混乱排列,大量上皮细胞坏死、脱落,鳃丝粘液细胞增生、肥大;随着胁迫温度升高,鳃中SOD、CAT、GPX、MDA均整体呈现先升高、后降低的变化趋势,且在36℃时显著下降。这表明在热应激后,鱼体内ROS增加,脂质过氧化增加。由于体内产生大量的ROS和H2O2,机体也增加了GPX和CAT的合成。当暴露在36℃时,胁迫强度超过白梭吻鲈的适应能力,抗氧化系统功能无法正常进行。表明热胁迫对白梭吻鲈鳃组织有损伤,且随着热胁迫温度的升高,其鳃受损情况越发严重。     

饲料溶菌酶添加水平对氨氮应激下吉富罗非鱼血清生化指标、抗菌性能和肝脏抗氧化能力的影响

为研究氨氮应激下溶菌酶对吉富罗非鱼血清生化指标、抗菌性能及肝脏抗氧化能力的影响,用6种不同溶菌酶添加水平(0、18、36、54、72和90 mg/kg)的饲料(分别记为L0、L18、L36、L54、L72和L90)饲喂初始均重为(11.35±0.08)g的罗非鱼60 d,之后应用氯化铵进行24 h氨氮应激实验。结果显示:1应激后,各组鱼的血清生化指标在组间存在较大差异(P0.05),不同溶菌酶添加水平下的鱼体对氨氮应激产生了不同的响应机制。L54组鱼主要通过激发免疫系统并调节蛋白质代谢来抵抗氨氮浓度突变;L72和L90组鱼主要通过调节高密度脂蛋白和低密度脂蛋白、胆固醇和甘油三酯之间的动态变化来缓解氨氮对机体的应激。2血清抗菌实验表明,L54和L72组鱼对大肠杆菌、金黄色葡萄球菌和嗜水气单胞菌的抑制能力最大,且显著高于对照组(P0.05);L36~L90组对溶藻弧菌的抑制作用显著高于对照组和L18组(P0.05);各溶菌酶添加组的鱼对枯草芽孢杆菌具有不同程度的保护作用。3应激后肝脏的超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶活性随溶菌酶添加水平的增加总体呈现先升后降的变化趋势,L54组显著高于对照组(P0.05);各添加组丙二醛含量均显著低于对照组(P0.05)。研究表明,54和72mg/kg溶菌酶添加水平对氨氮应激下吉富罗非鱼的血清生化指标、抗菌活性和肝脏抗氧化指标产生了最积极有效的调控,能够增强鱼体的抗应激能力。     

鱼类microRNA研究进展

microRNA是一种大小为18~25个核苷酸的非编码RNA,调控靶基因的转录后表达,广泛参与多种生理过程。本研究简要综述了microRNA生物学特性、转录后调控机制,详细讨论了第二代测序技术的相关应用、microRNA在鱼类中的表达模式、功能研究,包括渗透压调节、生殖、发育、生长、免疫、代谢等。现有的研究表明,miR-200和miR-30具有调控血浆离子浓度和渗透压平衡的功能;Dicer1是胚胎发育所必需的,在原肠胚时期,miR-20a调节头、眼、脊柱、体节的形态发生,miR-92调控内胚层的形成,miR-10影响体轴形成,miR-219影响头部和尾部细胞的凋亡;miR-122、miR-30、miR-145分别影响肝细胞成熟、肝胆管和肠道发育,miR-138和miR-143调控心脏发育,miR-126、miR-150、miR-451调控血小板、血细胞的发育和成熟,miR-10和miR-126影响血管生成,miR-200、miR-183分别影响嗅觉、听觉系统的发育,多个microRNA参与视网膜、肌肉、鱼鳍、肝脏等器官的再生;miR-133a/b、miR-206抑制肌肉细胞分裂,miR-214、miR-499、miR-199、miR-3906调控快肌纤维和慢肌纤维的生成;miR-430促进原始生殖细胞的生成;多个microRNA参与到禁食和再投喂调控的分解代谢和合成代谢平衡;此外,microRNA通过对免疫因子的调控参与到先天免疫、低氧适应、体色调控、应激反应、肌间刺形成等生理过程中。本研究通过对这些结果进行总结和讨论分析,将有助于对鱼类microRNA研究领域现状的了解和把握,促进鱼类非编码RNA研究的深入。     

碳酸盐碱度胁迫下凡纳滨对虾基因的差异表达

以广盐性养殖的凡纳滨对虾(Litopenaeus vannamei)为研究对象,采用抑制消减杂交(suppression subtractive hybridization,SSH)和定量PCR的方法,研究其在高碳酸盐碱度胁迫下转录组水平的基因表达差异,以期从基因组水平研究对虾对高碳酸盐碱度胁迫的适应机制。结果表明,以高碳酸盐碱度(20 mmol/L)胁迫第4天凡纳滨对虾鳃组织和低碱度(2 mmol/L)对照组鳃组织为材料,通过斑点杂交筛选,发现鳃组织中有158个克隆子表达上调,291个克隆子表达下调。挑选150个高表达差异的克隆子进行测序,获得100个序列,其中50个得到成功注释。经过GO分析,这些注释的差异基因主要分为8大类群,其中碳酸酐酶基因(CA)、Na+-K+-ATPase基因(NKA-α)等与离子调控相关的基因表达量上调,而溶菌酶基因等与先天免疫相关的基因表达量下调,这些结果表明高碳酸盐碱度胁迫下,凡纳滨对虾以增加离子调控的方式进行酸碱平衡的调控,同时其免疫功能受到抑制。此外,还对CA和NKA-α两个基因在鳃和触角腺中的时空表达规律进行了研究,发现高碳酸盐碱度胁迫9 d过程中,两个基因在鳃组织中的表达均呈现先高表达后回落的现象,而在触角腺中NKA-α基因则一直维持较高表达水平,说明CA和NKA-α基因在凡纳滨对虾高碳酸盐碱度适应离子调控中起着关键作用,同时还发现除了鳃组织之外,触角腺同样参与了调控。本研究从转录水平初步筛选了高碳酸盐碱度胁迫下凡纳滨对虾的表达差异基因,探索了凡纳滨对虾的耐盐碱机制,对培育适于盐碱地水产养殖的优良品种有着重要的意义。