三唑磷对斑节对虾肝胰腺和鳃的氧化胁迫效应

采用半静态毒性实验方法测定了三唑磷（Triazophos,OP）对斑节对虾（Penaeusmonodon）的96hLC卯值,同时研究了低（0．3μg·L^-1）、中（0．5μg·L^-1）、高（1μg·L^-1）浓度OP胁迫1、3、7、14d和清水恢复7d后斑节对虾肝胰腺和鳃中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、丙二醛（MDA）含量和总抗氧化能力（T—AOC）的变化趋势。结果表明：在胁迫期间SOD、GPx活性都受到了显著的诱导和抑制,呈一定的变化规律;MDA含量随胁迫时间延长显著升高。肝胰腺T—AOC在低浓度OP胁迫下呈升高-下降-升高的变化趋势,高浓度则显著下降;鳃T—AOC显著升高,而随胁迫时间延长,T—AOC显著降低。经清水恢复7d后,各浓度组肝胰腺SOD活性仍然显著低于空白组,MDA含量和T-AOC显著高于空白组。低浓度组鳃SOD、T—AOC显著高于空白组,MDA含量和中、高浓度SOD能恢复到正常水平,T—AOC则显著低于空白组。肝胰腺和鳃GPx活性变化情况相同,均为低、中浓度显著升高,高浓度显著下降。上述结果显示,0P对斑节对虾的抗氧化系统有显著的影响,且具有组织特异性,肝胰腺对OP较为敏感。斑节对虾对一定浓度OP所带来的氧化损伤具有一定的自我修复能力,但在短期内无法完全修复。     

氧化鱼油对鲤肝胰脏抗氧化机能

在半纯化饲料中添加质量分数为３％的新鲜鱼油（过氧化物值ＰＯＶ为０．６４ｍｍｏｌ／ｋｇ）和ＰＯＶ分别为２９．６４、５９．４０和９４．６９ｍｍｏｌ／ｋｇ（以Ｏ２计）的氧化鱼油,投喂１００ｇ左右２龄鲤（Ｃｙｐｒｉｎｕｓ ｃａｒｐｉｏ）鱼种１５周,结果表明：鲤摄食氧化鱼油后,肝胰脏维生素Ｅ含量下降（Ｐ〈０．０５）,抗氧化酶超氧化物歧化酶（ＳＯＤ）和谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）活性减弱（Ｐ〉０．０５）,微粒体氧化稳定性降低（Ｐ〈０．０５）,鱼渔氧化程度升高,上述变化加强;肝胰脏脂肪含量增加（Ｐ〉０．０５）;肾脏和脾脏增生（Ｐ〉０．０５）;肝胰脏肝细胞纤维化、线粒体嵴降解和融合。上述系列变化表明氧化钱油可以破坏鲤肝胰脏抗氧化机能及其正常的组织结构。     

草鱼幼鱼肝胰脏抗氧化系统对MC-LR胁迫的响应

为了研究微囊藻毒素(MC-LR)对草鱼(Ctenopharygodon idella)幼鱼肝胰脏抗氧化防御系统的影响,本研究通过腹腔注射的方法(剂量为25、100μg MC-LR·kg-1,分别称为低剂量和高剂量),对草鱼幼鱼进行染毒,于胁迫24、48 h和72 h后分离其肝胰脏,随后采用分光光度法检测了草鱼幼鱼肝胰脏中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;同时采用荧光定量PCR方法分析了SOD、CAT、谷胱甘肽过氧化物酶(GPx)和谷胱甘肽还原酶(GR)基因相对表达量的变化。结果显示:高剂量MC-LR诱导24 h和48 h后,草鱼幼鱼肝胰脏SOD活性显著增加(P0.05),而SOD在低剂量组中活性没有显著变化(P0.05);低剂量MC-LR对CAT活性没有显著影响(P0.05),而高剂量MC-LR诱导24 h后可使草鱼幼鱼肝胰脏CAT活性显著增加(P0.05),随后其活性又下降,但差异不显著(P0.05)。在MC-LR胁迫过程中,SOD、CAT、GPx基因表达在两个剂量组中均显著低于对照组(P0.05),而GR基因在低剂量MC-LR胁迫24 h后相对表达量显著上调(P0.05),尽管在72 h相对表达量上调,但差异不显著(P0.05),而高剂量组中GR基因在胁迫24 h和48 h后,其表达量被抑制,但不存在显著差异(P0.05)。进行抗氧化酶活性和基因表达相关性分析发现,SOD和CAT酶活性与SOD和CAT基因表达不相关。上述研究表明MC-LR对草鱼幼鱼肝胰脏抗氧化系统产生了明显的胁迫效应,但MC-LR对机体抗氧化酶活性与基因编码调控之间的相互作用机制还有待更深入的研究。     

宝石鲈肝胰脏蛋白质双向电泳方法的摸索与优化

选取宝石鲈肝胰脏作为样品,通过优化裂解配方及摸索裂解液与样品的最适比例,设计两个等电聚焦程序,建立和优化宝石鲈鱼肝胰脏蛋白质双向电泳实验方法,为宝石鲈肝胰脏内目标蛋白的研究提供参考。结果表明,采用裂解液配方:8 M Urea,2 M Thiourea,0.5%(W:V)CHAPS,40 mMTris-Base,1%DTT,2%(W:V)Pharmalyte pH 3～10,1 mM PMSF;裂解液和样品液比为1∶3时裂解效果较好;等电聚焦程序I(S1:0～500 V,500 V.h;S2∶500 V,500 V.h;S3∶500～3 500 V,10 000V.h;S4∶3 500 V,50 000 V.h,S5∶3 500～5 000 V,8 000 V.h)具有最好的等电聚焦效果。     

Haemolymph metabolic variables in relation to eyestalk ablation and gonad development of Pacific white shrimp Litopenaeus vannamei Boone

The present study analyses some biochemical variables in haemolymph proposed as predictive indicators of the maturation capability following eyestalk ablation. Haemolymph of captive females was obtained before and 8 days after eyestalk ablation, and levels of haemocyanin, total proteins, glucose, lactate, cholesterol, triacylglycerides and vitellogenin were determined. Biochemical variables were also analysed in the hepatopancreas at the end of the experiment. Females were grouped as immature (previtellogenic stage) and mature (vitellogenic and cortical stages) based on histological analysis done 8 days after eyestalk ablation. To analyse haemolymph variables before eyestalk ablation in relation to maturation capability, immature females were classified as those with a low maturation capability and mature females as those with a high maturation capability. Females of high maturation capability had significantly higher vitellogenin levels before eyestalk ablation than females of low maturation capability. No significant differences were found for the other biochemical variables. Vitellogenin was also higher in mature than in immature females at the end of the experiment. These results indicate that vitellogenin levels in haemolymph could be used as possible predictive criteria of maturation capability, possibly because they reflect the degree of ovarian development at the time of eyestalk ablation.     

中华绒螯蟹肝胰腺白化症组织病理变化

采用常规石蜡切片技术和透射电镜技术,对患有肝胰腺白化症的中华绒螯蟹病变组织和细胞进行观察。发现病蟹肝胰腺、鳃、肌肉等组织细胞发生了不同程度的病变,其病理特征为肝细胞水肿、变性、坏死,细胞中脂肪颗粒锐减,线粒体水肿、嵴消失、形成空泡,内质网、溶酶体解体成小泡。鳃组织增厚,上皮细胞微绒毛排列疏松、不规则,部分线粒体嵴消失,出现膜性退化。坏死的肝细胞和鳃上皮细胞中出现细菌颗粒。肌细胞的病变主要表现为细胞核固缩,肌丝松驰、水肿、断裂、肌质网溶解形成小泡。中华绒螯蟹具有重要生理功能的肝胰腺、鳃组织发生了严重病变,使其功能损伤,不耐运输;而肌肉的松驰、肝胰腺脂肪的锐减影响了中华绒螯蟹的品质。同时,病变组织和细胞中未见病毒等生物病原,表明该病为非生物性疾病。图2表0参12     

致对虾肝胰腺坏死病副溶血弧菌荧光定量PCR检测方法的建立

【目的】建立一种对虾急性肝胰腺坏死病（Acute Hepatopancreatic Necrosis Disease）病原菌副溶血弧菌（Vibrio parahaemolyticus, VpAHPND）荧光定量 PCR 检测方法,对该病进行快速检测。【方法】根据副溶血弧菌基因保守序列设计特异性引物和探针,经优化反应体系及条件,建立检测 VpAHPND 的荧光定量 PCR 方法;以重组质粒为标准品制作标准曲线,并进行特异性、敏感性、重复性及临床应用试验。【结果】该方法定量范围宽,起始模板浓度范围为 2.3×101 ~2.3×107 拷贝 /μL 时,标准曲线具有良好的线性关系;最低检测限为 2.3 拷贝/μL,与对虾其他病原菌及病毒无交叉反应,重复试验变异系数为 0.45%~0.69%,可在 1 h 内完成样品检测;对临床对虾样品的检测结果表明,该方法与世界动物卫生组织（OIE）推荐的套式 PCR 法检测结果一致。【结论】应用 TaqMan 探针建立的检测 VpAHPND 的荧光定量 PCR 方法具有灵敏度高、特异性强、重复性好的特点,可有效用于临床对虾样品的 VpAHPND 检测。     

灌喂必需氨基酸模式溶液对草鱼全鱼_省略_肌肉_肝胰脏蛋白质合成代谢的影响

采用大剂量食道灌喂DL－[4-^3H]-Phe，研究不同EAA模式下，20－30g体重草鱼种的全鱼和肌肉，肝胰脏的蛋白质合成代谢，结果表明：（1）分别缺乏Lys,Met,Trp Arg的4种极端不平衡EAA模式，同草鱼肌肉EAA模式即相对平衡的基础模式相比，肌肉，肝胰脏及全鱼的蛋白质合成速率（FSR）均显著降低，其降低程度高低依次为Met,Lys,Arg,Trp;(2)肝胰脏，肌肉人的蛋白质合成代谢与整体蛋白质合成代谢对EAA模式的改变产生相同的变化趋势，并且三者蛋白质合成代谢速率为肝胰脏＞全鱼＞肌肉；（3）EAA模式的Lys,Met,Trp,Arg4因素中，对肌肉，肝胰脏蛋白质合成速率影响最大的是Met,其余依次为Lys,Arg,Trp。     

中华绒螯蟹蜕皮过程中肌肉、肝胰脏和甲壳中钙和磷含量的变动

对中华绒螯蟹蜕皮间期和蜕皮后24h、48h和96h肌肉、肝胰脏及甲壳中钙和磷的含量进行了测定。蜕皮后肌肉中钙的含量较间期低，但从24h到48h有所增加。肝胰脏中钙的含量在蜕皮后各时间内无显著变化，甲壳中钙的含量在蜕皮后24h到96h呈增加趋势。甲壳的矿化作用在蜕皮后96h已接近完成。蜕皮后肌肉中磷的含量较间期高，以后变化不明显。蜕皮后肝胰脏中磷的含量高于间期，甲壳中含量较间期低。蜕皮后肌肉中钙磷比迅速降低，肝胰脏中钙磷比在蜕皮后24h无明显变化，但在蜕皮后24h至96h迅速降低。中华绒螯蟹蜕皮后外表皮对钙的矿化速度显著快于磷的矿化速度。     

Presence and ontogeny of intestinal and pancreatic phospholipase A2-like proteins in the Red Sea bream,Pagrus major. An immunocytochemical study

We have studied the location and the ontogeny of the digestive enzyme, phospholipase A₂ (PLA₂) immunohistochemically in the adult and larvae/juvenile of the red sea breamPagrus major by using an antiserum against theNaja naja venom PLA₂. The antiserum reacts with at least one enzyme among the PLA₂s purified from the fish hepatopancreas or intestine. Although the reactivities were comparatively low, it labelled zymogen granules of the pancreatic acinar cells and secretory materials of certain epithelial cells in the depths of epithelial crypts in the pyloric caeca of the adult. The immunoreactivities of PLA₂s were investigated in the viscera of larvae and juveniles of the 0 to 85^th day after hatch. In the larvae of the 13^th day, accumulation of PLA₂-positive zymogen granules in the pancreatic acinar cells were first recognized by the immunostaining. The intensity of the labelling subsequently became stronger and dramatically increased between the 20^th and 30^th day. This increase appeared to be one of the physiological changes associated with the transition to a new benthic life as juveniles. Lack of PLA₂ in the pancreas before the 13^th day may suggest the possibility that larvae utilized exogenous PLA₂, inherent in their prey, to digest the phospholipids. On the other hand, no reactivity was found in the intestine until the 85^th day.