不同地理群体三疣梭子蟹mtDNA本底甲基化水平分析

利用重亚硫酸盐测序法,对适于三疣梭子蟹(Portunus trituberculatus)线粒体基因组甲基化分析的BSP(Bisulfite sequencing PCR)引物进行了筛选,在设计的21对引物中,获得了12对BSP引物。利用这12对引物分析了三疣梭子蟹海州湾群体(江苏连云港)和莱州湾群体(山东东营)线粒体基因组本底甲基化水平,结果表明:海州湾群体不存在甲基化现象,莱州湾群体发现了2个个体在ND2基因(NADH脱氢酶亚基2)位点存在甲基化现象;在存在甲基化的个体中,甲基化类型主要为CHG和CHH类型,还有少量Cp G类型。该研究结果说明,莱州湾群体和海州湾群体线粒体基因组本底水平上的甲基化特征存在一定差异,相关研究值得进一步深入探讨。     

银杏叶提取物对鲫CYP4503A和溶菌酶活性的影响

以红霉素为探针,采用NaSH法研究了银杏叶提取物对鲫细胞色素P4503A和溶菌酶活性的影响.研究结果显示,银杏提取物能诱导鲫细胞色素P4503A活性,试验组CYP4503A活性显著高于对照组,而且银杏叶提取物可以提高鲫头肾溶菌酶的活性和免疫保护率.鲫日粮中添加银杏叶提取物的适宜量为25 mg/kg.     

Dietary available phosphorus requirement for juvenile gibel carp (Carassius auratus gibelio var. CASIII)

A 57‐day growth experiment was conducted with juvenile gibel carp (13.48 ± 0.10 g) in a flow‐through system to study the effect of dietary phosphorus on growth performance, body composition, nutrition utilization, phosphorus loading and enzymes activities. Seven semipurifed diets were formulated to contain 0.07 (the basal), 2.27, 5.32, 8.10, 12.06, 15.24 and 19.48 g available phosphorus/kg diet. The results showed that specific growth rate, body length and feed efficiency significantly increased in the fish fed diets containing 0.07 to 15.24 g available P/kg diet (p < .05). Ash and P content increased in fish fed diets containing 0.07–12.06 P g/kg (p < .05) and then levelled off, while moisture, crude protein and lipid had no significant difference (p > .05). The protein retention efficiency increased in the fish fed with diets 0.07–5.32 g/kg P (p < .05) and then reached a plateau. The P content in faeces was higher in fish fed diets containing 15.24 and 19.48 g available P/kg. Total P concentration in tank water increased in fish fed 0.07–12.06 g available P per kg diet (p < .05). The plasma P was higher in the fish fed with 15.24 g available P/kg diet (p < .05), triglycerides was lower in the fish fed diet containing 15.24 and 19.48 g available P/kg (p < .05), no significant differences were observed in plasma Ca, plasma glucose and calcitonin (p > .05). Based on SGR, whole body P content and FE, dietary available P requirement for juvenile gibel carp were 13.37, 13.97 g/kg and 15.06 respectively.     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.     

Mitochondrial NAD(P)-dependent malic enzyme from herring testicular tissue: Purification,kinetic behaviour and regulatory properties

Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD⁺ oxidoreductase (decarboxylating)] was purified from herring testicular tissue to a specific activity of 26.4 μmol NADH/min/mg protein. Herring testicular tissue is one of the most abundant sources of this enzyme. The purification procedure involved differential centrifugation of mitochondria and then chromatography on DEAE-Sephacel, Red Agarose and Sephacryl S-300. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of Mn²⁺ and either NAD or NADP. Under V_max conditions the ratios for the rate of NAD/NADP reduction was 1.8. A study of the reductive carboxulation reaction indicated that this enzyme reaction is reversible; at pH 7.0 the reverse reaction exhibited 22% of the activity of forward reaction. Some kinetic characteristics of the enzyme were determined. ATP was found to be a competitive inhibitor with respect to malate. Fumarate reversed ATP inhibition. Regulation of NAD(P)-dependent malic enzyme from herring testicular tissue mitochondria could respond to changing levels of mitochondrial ATP and fumarate in vivo.     

草鱼肝微粒体的提取及CYP酶活性的测定

CYP酶代谢是药物生物转化的主要途径,其数量和活性大小直接影响药物在体内的活化与代谢。我们对草鱼肝微粒体CYP酶含量及其活性进行了初步研究,以差速离心法提取草鱼肝微粒体,以CO还原差示光谱法测得CYP酶及细胞色素b5含量分别为0.619±0.102 nmol/mg、0.264±0.042 nmol/mg。以7-乙氧异吩噁唑酮-O-脱乙基反应、苯胺-4-羟化反应、氨基比林-N-脱甲基反应作为CYP1A、CYP2E、CYP3A的探针反应,测得EROD酶活为0.043±0.004 nmol/mg/min,ANH酶活为0.028±0.002 nmol/mg/min,AMND酶活为0.207±0.035 nmol/mg/m in。结果表明草鱼肝微粒体中CYP酶发育完好,并且具有参与药物代谢的3种主要亚型活性,其含量与活性大小与其它实验动物相差较大。本实验的方法与结果为草鱼CYP酶的系统研究提供可靠手段,最终为指导水产合理用药提供理论依据。     

Effects of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus

This study was conducted to evaluate the effects of different concentrations of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus. Diets were formulated to contain 40% crude protein from solvent-extracted menhaden fish meal and 0, 7, 14 or 21% lipid from menhaden fish oil. The basal diet, without supplemental fish oil, contained lipid at 0.4% of dry weight. The diets were fed to groups of 25 juvenile red drum initially averaging 7.3 ± 0.18 g fish^–1 in a recirculating culture system for 8 weeks and weight gain was recorded. After an additional 8 weeks, 16 fish from each treatment were sacrificed and the following measurements were recorded: hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, and liver -tocopherol, malondialdehyde (MDA) formation, and cytochrome P-4501A activity (measured as 7-ethoxyresorufin O-deethylase (EROD) activity). The activity of alanine and aspartate aminotransferases and concentrations of -tocopherol also were measured in plasma.Weight gain was significantly (p<0.05) affected by dietary lipid concentration, with values ranging from 361% of initial weight for fish fed the basal diet to 527% of initial weight for fish fed the diet containing 7% lipid. The HSI and IPF ratio values also were significantly affected by lipid with the lowest values recorded for fish fed the basal diet and the highest values observed in fish fed the diet containing 21% lipid. Increasing dietary lipid significantly increased oxidative stress as reflected in reduced -tocopherol in liver and plasma and increased MDA formation in the liver, although no overt pathological signs were observed. These findings suggest that lipid concentrations between 7 and 14%, when the diet contains 60 IU vitamin E kg^–1, are likely to limit oxidative stress and result in normal physiological responses of red drum.     

中国对虾亲虾的能量代谢研究

本试验测定了中国对虾亲虾在禁食和静息状态下的耗氧率、二氧化碳排出率和氨氮排泄率,研究了它们的呼吸商、氧氮比、能源的化学本质及其消耗量之比。结果表明,体重为85.1±14.8克的亲虾,在水温19～22℃时的耗氧率为0.25±0.08毫克/克·小时,二氧化碳排出率为0.28±0.09毫克/克·小时,氨氮排泄率为8.86±3.53微克/克·小时;亲虾以蛋白质、脂肪和碳水化合物的混台物为能源,其中以蛋白质为主,脂肪次之,碳水化合物的利用最少;亲虾的耗氧率(Y)与体重(W)呈负相关,Y=1.89W~(-1.3),而与性腺发育的影响则相反,性腺发育好的亲虾代谢率高,性腺发育差的亲虾代谢率低。     

CYP2J3样基因在不同生长速率凡纳滨对虾中的差异表达

为了获得对虾生长差异的分子机制,通过比较不同生长速率凡纳滨对虾的转录组(速长组和慢长组),获得了一条编码P450的差异表达基因,经过cDNA全长验证后命名为LvCYP2J3-like,其开放阅读框为1 488 bp,编码495个aa,含有一个完整的P450功能域,功能分析表明,该基因可能参与了蜕皮激素的灭活。方差分析表明,LvCYP2J3-like在不同组织、发育阶段和蜕皮周期的表达量都存在显著性差异,并且速长群体的表达量显著性低于慢长群体。表明该基因参与了发育和蜕皮过程,其表达量与环境胁迫关系密切。在LvCYP2J3-like和其预测转录因子基因中分别发现了4和11个速长组和慢长组等位基因频率显著差异的单核苷酸多态性(SNP)标记,说明这些基因遗传差异与其活性和表达量相关,从而影响生长速率。研究结果为凡纳滨对虾遗传和育种研究提供了参考资料。     

中国对虾病原菌(鳗弧菌)的研究

1987年7～10月福建省平谭县对虾养殖场发生了一起严重的流行病。病虾活动力减弱,食欲下降,个体消瘦,头胸甲心区附近呈白色或浅桔红色,血淋巴液稀薄、混浊、不能凝固。因病虾步足、游泳肢及尾扇呈鲜红色,被称为“红腿病”。死亡率可高达90％以上。从五只垂死病虾的心脏血淋巴液中分离到细菌作纯焙养,经肌肉注射和浸泡的人工感染方法,得到了与虾池中患病虾相同的症状。菌原菌是革兰氏阴性短杆鼠。以极生单鞭毛运动。发酵葡萄糖,产酸不产气。氧化酶、过氧化氢酶阳性。能还原硝酸盐成亚硝酸盐。精氨酸脱羧;鸟氨酸、赖氨酸不脱羧。V.P.阳性.在42℃中不能生长。不能在无盐和含有8％NaCl 的胨水中生长,但在含有3％和6％NaCl的胨水中生长良好。对弧菌抑制剂0/129敏感。经鉴定为鳗弧菌(Vibrio anguillarum Bergman)。