卵形鲳鲹血细胞发生的观察

对卵形鲳鲹(Trachinotus ovatus)肝脏、脾脏、头肾和体肾4种组织器官内各种血细胞的发生和外周血液的血细胞分类组成进行了观察。结果表明:卵形鲳鲹的红细胞主要在脾脏、体肾和头肾中发生;淋巴细胞主要在体肾、头肾和脾脏中发生;粒细胞主要在头肾和体肾中发生;单核细胞在肝脏、头肾和脾脏中均有发生;而血栓细胞的发生在这4种组织中均未观察到。卵形鲳鲹在外周血液中的白细胞包括淋巴细胞、血栓细胞、嗜中性粒细胞和单核细胞。在这几种白细胞中,单核细胞数量最少,很难观察到,数量少于1%;嗜中性粒细胞稍多,有(9.25±5.64)%;血栓细胞占(38.23±21.86)%;淋巴细胞最多,达(52.52±21.65)%。     

深水网箱养殖卵形鲳鲹血液指标

对深水网箱养殖卵形鲳鲹(Trachinotus ovatus)成鱼的部分血液性状和血清生化指标进行测定比较.结果显示,雌、雄成鱼红细胞数量(RBC)、白细胞数量(WBC)、血细胞比容(Ht)、红细胞沉降率(ESR)等血液性状不存在显著性差异.血清指标中雌、雄成鱼血糖(GLU)的含量分别为(10.24±1.25)mmol/L、(17.36±1.34)mmol/L,存在极显著差异;雌鱼血清中低密度脂蛋白胆固醇(LDL-C)、钾离子(K+)、谷丙转氨酶(ALT)、谷草转氨酶(AST)浓度大于雄鱼(P<0.05);雄鱼血清中肌酐(CREA)、钠离子(Na+)浓度大于雌鱼(P<0.05);尿酸(UA)、尿素氮(BUN)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、γ-谷氨酰转肽酶(γ-GT)、总蛋白(TP)、白蛋白(ALB)、球蛋白(GLB)、白蛋白/球蛋白(ALB/GLB)、钙离子(Ca2+)、氯离子(Cl-)、无机磷(PO43-)等指标,雌雄差异不显著.卵形鲳鲹部分生化指标存在雌雄差异,可能与个体的生理状况有关.结果可为卵形鲳鲹血液生理研究提供基础资料.     

云斑尖塘鳢胚胎和早期仔鱼的发育

对云斑尖塘鳢(Oxyeleotris marmoratus)胚胎和早期仔鱼的发育进行了观察,详细描述了各发育阶段的形态特征。在28℃定水温条件下,云斑尖塘鳢的胚胎发育历时80h 30min,可分为24个发育分期。在整个发育过程中,眼、耳囊、心脏、消化道、肾脏、鳔、胸鳍和尾鳍等得到了优先发育。     

卵形鲳鲹组织蛋白酶B基因的克隆及表达分析

为研究卵形鲳鲹(Trachinotus ovatus)组织蛋白酶B(Cathepsin B, CatB)基因的表达特征和功能,应用RTPCR和RACE技术获得卵形鲳鲹CatB(TroCatB)基因的全长cDNA。TroCatB cDNA全长为2181 bp。其中5′UTR和3′UTR分别为391和797 bp, ORF为993 bp,推定编码330个氨基酸残基,推定分子质量和理论等电点分别为36.37 kD和5.73。蛋白结构预测TroCatB蛋白具有信号肽(1Met-18Ala)、前体肽(25Leu-64Gly)和一个典型木瓜蛋白酶家族半胱氨酸结构域,含107Cys、277His、297Asn 3个蛋白酶催化活性位点。同源性分析显示TroCatB蛋白与其他脊椎动物的同源性为67.0%—90.9%,成熟肽区与其他脊椎动物的同源性为73.7%—92.4%。NJ系统发育树显示卵形鲳鲹和其他鱼类聚为一支,与髙体鰤距离最近。实时荧光定量PCR检测TroCatB基因mRNA在健康卵形鲳鲹各组织中均有表达,在脾脏中表达最高;在溶藻弧菌感染后, TroCatB基因在脾脏、头肾组织中的mRNA表达水平...     

盐度、温度对卵形鲳鲹选育群体肝抗氧化酶活力的影响

通过盐度渐变和温度骤变的方法,分别研究了不同盐度(10、20、30、40)处理和不同温度(18.0℃、21.0℃、24.6℃、29.0℃、32.0℃)处理对卵形鲳鲹(Trachinotus ovatus)选育群体肝超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)活力的影响.在实验结束时,盐度10组酶活力与对照组无显著差异(P＞0.05),盐度20组SOD活力极显著低于对照组(P＜0.01);盐度40组120 h时,SOD和GPX活力极显著低于对照组(P＜0.01),CAT活力与对照组差异不显著(P＞0.05).18.0℃和21.0℃ SOD活力在1、3、6、12、24 h这5个取样时间点均高于对照组,CAT活力在实验结束时(24 h)极显著高于对照组(P ＜0.01);29.0℃SOD和CAT活力在实验结束时(24 h)显著高于对照组(P＜0.05);32.0℃ SOD和CAT活力在5个取样时间点均显著低于对照组(P＜0.05).结果表明,适当的盐度或温度变化可以改变卵形鲳鲹肝抗氧化酶活力,达到机体耐受极限时酶活力下降.     

急性和慢性低氧胁迫对卵形鲳鲹幼鱼肝组织损伤和抗氧化的影响

自然海域和养殖水体环境频现低氧,本研究针对卵形鲳鲹(Trachinotus ovatus)不耐低氧的特性,将(31.59±3.01)g(n=30)的卵形鲳鲹在(23±0.7)℃下进行3、6、12和24 h的急性和14 d的慢性低氧[溶解氧为(1.55±0.20)mg/L]胁迫。运用光学和电子显微技术,比较急、慢性低氧胁迫对卵形鲳鲹肝组织显微和超微结构的影响。通过测定肝组织中的丙二醛(MDA)含量及过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)的活性,分析低氧胁迫对卵形鲳鲹肝是否造成氧化损伤。急性低氧胁迫下,卵形鲳鲹肝组织间出现空泡、小叶结构破坏,细胞内线粒体数量减少,出现过氧化物酶体,肝细胞间血窦剧烈扩张。这些病理损伤随胁迫时间延长更趋严重,24 h时甚至出现局部肝细胞融合、坏死,慢性低氧胁迫14 d时,肝细胞局部坏死,细胞膜溶解,细胞核破裂分解,胞质内细胞器不明显,只可分辨粗面内质网,细胞内空泡体积大,细胞结构松散,血窦扩张。急性低氧胁迫下丙二醛(MDA)含量随时间先上升后下降,慢性低氧胁迫14 d时丙二醛(MDA)则显著增加。急性低氧胁迫下超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)上升后恢复,过氧化氢酶(CAT)则持续上调,慢性低氧胁迫下超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GSH)显著上调,过氧化氢酶(CAT)活性下降。结果表明,卵形鲳鲹幼鱼肝组织在低氧胁迫下病理变化明显,氧化损伤严重,且慢性低氧胁迫比急性更甚。     

卵形鲳鲹幼鱼耗氧率和排氨率的初步研究

运用封闭流水式实验方法对不同体重和放养密度的卵形鲳鲹(Trachinoms ovatus)幼鱼耗氧率和排氨率进行了初步的研究。结果表明,卵形鲳鲹幼鱼的耗氧率随着鱼体重的增加而逐渐下降,排氨率总体上也呈降低的趋势,体重对卵形鲳鲹耗氧率、排氨率的影响均达到显著水平(P0.05);光照对卵形鲳鲹幼鱼的耗氧率和排氨率影响非常显著(P0.01),幼鱼在遮光条件下的耗氧率和排氨率分别比在自然光照条件下要低25.18%～40.76%和16.28%～40.28%;随着放养密度的增加,卵形鲳鲹幼鱼的耗氧率逐渐降低,排氨率随着密度的增加呈先上升后下降的趋势,放养密度对卵形鲳鲹耗氧率、排氨率的影响均达到显著水平(P0.05)。卵形鲳鲹的耗氧率和排氨率具有明显的昼夜变化,白天的耗氧率和排氨率均高于夜间值,耗氧率的低谷值为高峰值的69.68%,排氨率的低谷值为高峰值的30.91%。卵形鲳鲹幼鱼的窒息点溶解氧含量为(0.991±0.058)mg/L。     

卵形鲳鲹对刺激隐核虫的免疫应答和免疫保护研究

用刺激隐核虫（Cryptocaryon irritans）的幼虫对卵形鲳鲹（Trachinotus ovatus）进行腹腔注射和体表感染,然后每隔1周用阻动试验（Immobilization assay）检测免疫鱼的抗血清和皮肤培养液对激刺隐核虫幼虫的阻动效价,在第14周,分别用亚致死剂量和致死剂量的刺激隐核虫幼虫对免疫鱼攻毒以检测所产生的免疫保护力。实验结果显示：两种免疫方法都能让卵形鲳鲹的血清和皮肤生成阻动刺激隐核虫幼虫的特异性抗体,并能使被免疫鱼获得明显的免疫保护,但是体表感染免疫组的血清和皮肤培养液的阻动效价都要比腹腔注射免疫组高,所获得的免疫保护力也更强。同时还发现,免疫鱼血清和皮肤培养液中的抗体存在明显的差异：两者的最初生成时间、达到峰值的时间、变化规律以及阻动效价等都不一致。因此,我们推测鱼类的系统免疫应答和皮肤粘膜免疫应答有可能是相互独立的,或者是不同步的。鱼类的体液免疫应答,特别是粘膜免疫应答对抵御刺激隐核虫的感染起了重要的作用,采用刺激隐核虫虫体疫苗可能成为预防海水鱼类白点病的一种选择。     

斑点胡子鲶的胚胎和仔鱼发育观察

斑点胡子鲶胚胎和仔鱼发育分化的次序与同属的胡子鲶相似，但胚胎发育速度较快。在水温２５－２７℃范围内，从受精到孵化出膜的时间约为２６小时，出膜仔鱼４天后卵黄囊被吸收耗尽，并在此前１天开始摄饵；经１２天左右初步完成器官分化发育，总长达１．６－１．８ｃｍ，具有成鱼的基本外观。     

粘皮鲻虎鱼胚胎及仔鱼的发育

粘皮鲻鰕虎鱼(Mugilogobius myxodermus)的卵产出时为球型,直径0.40~0.60mm,具黏性,卵颜色淡黄,半透明,油球多且数量不固定。受精5~10min后吸水膨胀成椭圆形。水温16~18℃的条件下,胚胎发育共需138h。初孵仔鱼体长2.30~3.10mm,体高0.35~0.40mm,总肌节29~30。出膜30min~24h内鳔充气,从孵出到各鳍分化完全约需要40d。     

不同盐度下人工选育卵形鲳鲹(Trachinotus ovatus)子代鳃线粒体丰富细胞结构变化

该文采用光学显微镜和透射电镜技术观察不同盐度下(5、20、30)人工选育卵形鲳鲹(Trachinotus ovatus)鳃线粒体丰富细胞的分布和超微结构变化。结果表明,线粒体丰富细胞主要分布于鳃丝和鳃小片基部,且随盐度升高而体积增大,数量增多;三个盐度组均存在由线粒体丰富细胞、扁平细胞和附细胞构成的顶端小窝,盐度5组线粒体丰富细胞顶膜面积较大,微脊发达,顶端小窝内凹,盐度20和30组线粒体丰富细胞顶膜面积相对较小,微脊不发达,顶端小窝明显内陷;盐度5和30组线粒体丰富细胞胞质内存在发达的微细小管系统,线粒体内脊丰富,盐度20组胞质内微细小管系统分布不均匀,结构松散,部分收缩成珠泡状结构,与粗面内质网相混杂。线粒体丰富细胞的结构变化与其所处的渗透压环境相适应。     

Establishment of a new fish cell line from the caudal fin of golden pompano Trachinotus ovatus and its susceptibility to iridovirus

A new cell line derived from the caudal fin of golden pompano Trachinotus ovatus (TOCF) was successfully established and characterized. TOCF cells grew well at 28° C in L‐15 medium supplemented with 10% foetal bovine serum (FBS). The cell line has been subcultured in more than 100 passages. Molecular characterization of 18S ribosomal (r)RNA and cytochrome oxidase subunit 1 (COI) confirmed that the TOCF cells were derived indeed from T. ovatus. TOCF cells have a modal chromosome number of 54. It was further showed that TOCF cells were transfected successfully with pEGFP‐N3 and pDsRED‐N1 plasmid, suggesting that TOCF cells could be used to research gene functions in vitro. Viral susceptibility tests showed that TOCF cells were susceptible to Singapore grouper iridovirus (SGIV), observed by the occurrence of the cytopathic effect (CPE) with the formation of inclusion bodies. In addition, the expression of major capsid protein (MCP) gene of SGIV changed during virus infection in TOCF cells. Thus, our present results described the characteristic of a TOCF cell line that could be a valuable tool for genetic manipulation, as well as isolation and propagation of iridovirus studies.     

Establishment and characterization of a cell line from the head kidney of golden pompano Trachinotus ovatus and its application in toxicology and virus susceptibility

A cell line derived from the head kidney of golden pompano Trachinotus ovatus (TOHK) was established and characterized in this study. The TOHK cells grew most rapidly at 28° C and the optimum foetal bovine serum concentration in L‐15 medium was 10%. The TOHK cells have a diploid chromosome number of 2N = 54. The transfection efficiency of TOHK cells was 7·5% at the 15th passage and 72% at the 40th passage. The transfection efficiency in TOHK cells was high, so these cells are suitable for foreign gene expression. The cytotoxic effects of heavy metals and extracellular products from Vibrio anguillarum and Vibrio alginolyticus were demonstrated in TOHK cells, so this TOHK cell line could also be applied in environmental monitoring of heavy metals and pathogenic bacteria. TOHK cell line showed high virus susceptibility, such as grouper nervous necrosis virus (GNNV) and Singapore grouper iridovirus (SGIV). Then, TOHK cell line could be used for the study of viral pathogenesis and the development of antiviral strategies.     

河鲈胚胎及卵黄囊期仔鱼发育

为探究河鲈(Perca fluviatilis)早期生活史和发育生物学,采用体视解剖镜、显微镜仔细观察、测量、描述、绘图的方法,连续观察了6个批次河鲈胚胎及卵黄囊期仔鱼发育状况,进行比较分析。结果显示:(1)在水温8~13℃时,胚胎期约需265h,有效积温2540~2880℃.h;水温11~13℃时,卵黄囊期约需6d,有效积温1750~2120℃.h;(2)辐射状次级卵膜将受精卵连成长带形单层网片状,每个胚胎周围有6个胚胎,排列很有规则。胚胎卵黄囊表面有一个大圆形油球。出膜前期可见眼球色素、胸鳍突起;(3)胚胎出膜的不同步主要是由于出膜前期长短不一和孵化水温较低所致。     

Individual variation in the rate of oxygen consumption by zebrafish embryos

A sensitive microsensor‐based method was used to measure oxygen consumption of individual zebrafish Danio rerio embryos at 6 h intervals from 24 to 75 h post‐fertilization. An increase in oxygen consumption rates from 4·54 to 8·29 nmol O₂ h⁻¹ was found during this period. At the individual level the differences in oxygen consumption rates caused the total oxygen consumption from 24 to 75 h post‐fertilization to vary between 0·261 and 0·462 μmol O₂ per individual with a mean of 0·379 μmol O₂ per individual. A separate carbon mass balance study corroborated the mean total oxygen consumption obtained by yielding a respiratory quotient of 0·80 for this period. These results suggest that there is significant intraspecific variation in the metabolic rate of developing zebrafish embryos, which may influence other early life‐history traits such as growth and starvation resistance.     

Cold-induced vasodilatation of finger and maximal oxygen consumption of young female athletes born in Hokkaido

To determine whether there is a direct correlation between endurance capacity and cold tolerance, maximal oxygen consumption (VO₂max), and cold-induced vasodilatation (CIVD), we measured these factors in 14 young female athletes born in Hokkaido, Japan's northernmost island. We determined the VO₂max by a standard incremental test on a cycle ergometer and measured the oxygen consumption (VO₂) by means of the Douglas-bag method. We determined the CIVD reaction by measuring the skin temperature of the left middle finger during immersion in cold water at 0°C for 20 min. The athletes showed significant positive correlations between VO₂max, expressed as l/min, and CIVD as well as other peripheral cold tolerance indexes (resistance index against frostbite and CIVD index). The body weight VO₂max (VO₂max/kg body weight) failed to correlate significantly with either the CIVD or with other cold tolerance indexes. These results suggest that CIVD in females may depend on factors other than those determined in this study, in addition to the functional spread of the vascular beds in peripheral tissues, including striated muscle; it is known that the size and the vascular bed in this tissue are affected by exercise training and that this results in the elevation of VO₂max and VO₂max/kg body weight.     

温度、盐度对强壮箭虫耗氧率和窒息点的影响

研究了不同温度、盐度下强壮箭虫的耗氧率和窒息点.结果表明:温度和盐度均对强壮箭虫的耗氧率和比耗氧率有显著影响.试验温度在5℃-25℃,强壮箭虫个体耗氧率(IO)和比耗氧率(SO)开始随温度的升高而升高,之后呈明显下降趋势.其回归方程分别为y=0.0058x3 -0.2956x2+4.415x-8.7816(R2=0.99,P＜0.05)和y=0.0011x3 -0.0546x2+0.8161x-1.6232(R2=0.99,P＜0.05),数值分别为6.30～11.71μg·ind-1·h-1和1.22 ～2.16μg· mg-1·h-1,窒息点为4.18～6.87 mg·L-1.盐度10 ～40条件下,IO和SO随盐度的升高逐渐下降,回归方程分别为y=-0.0068x2 -0.1412x+21.702(R2=0.89,P＜0.05)和y=-0.0013x2-0.0261x+4.0114( R2 =0.89,P＜0.05),数值分别为4.98～17.73μg ·ind-1·h-1和0.92～3.56 μg·mg-1 ·h-1,窒息点为4.02～6.24 mg·L-1.对强壮箭虫与其他水生动物的耗氧率和窒息点进行比较得出,强壮箭虫是一种狭氧性浮游动物.     

Metabolite availability as a window to view the early embryo microenvironment in vivo

A preimplantation embryo exists independent of blood supply, and relies on energy sources from its in vivo environment (e.g., oviduct and uterine fluid) to sustain its development. The embryos can survive in this aqueous environment because it contains amino acids, proteins, lactate, pyruvate, oxygen, glucose, antioxidants, ions, growth factors, hormones, and phospholipids—albeit the concentration of each component varies by species, stage of the estrous cycle, and anatomical location. The dynamic nature of this environment sustains early development from the one‐cell zygote to blastocyst, and is reciprocally influenced by the embryo at each embryonic stage. Focusing on embryo metabolism allowed us to identify how the local environment was deliberately selected to meet the dynamic needs of the preimplantation embryo, and helped reveal approaches to improve the in vitro culture of human embryos for improved implantation rates and pregnancy outcome.