Glutathione S-transferase from an antarctic fish, Dissostichus mawsoni

Glutathione S-transferase activity was measured in the hepatic cytosol front Dissostichus mawsoni and Pagothenia borchgrevinki. Activity measures with 1-chloro-2,4-dinitrobenzene as substrate were 11·2 and 16·7 μmol/min/g tissue respectively. Little or no activity was detected when p-nitrobenzyl chloride or 3,4-dichloro-1-nitrobenzene were used as substrate. The hepatic glutathione S-transferases from D. mawsoni were partially purified using gel filtration and chromatofocusing. Three peaks of activity were resolved. The major isozyme (158-fold purification) eluting at pH7·1 appeared to be catalytically a homodimer. The isozyme was highly inhibited by triphenyltin chloride (IC₅₀ = 0·1 μ) while inhibition constants for Cibicron Blue 3GA, bromosulphophalein and hematin were 1·1, 20 and 34 μ respectively.     

Mixed-function oxidase activity and conjugating enzymes in two species of antarctic fish

Enzyme activities of phase I and phase II of the hepatic detoxication system were investigated in two species of fish from the Ross Sea in Antarctica. The fish had extremely low phase I enzyme (BPMO and aldrin epoxidase). The reductase regenerating enzyme activities were comparable to those found in fish of temperate seas. Transferases of cytosol showed highest activity in the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and little or no conjugation of other substrates. The GSSG reductase and the glutathione transferase with peroxidase activity (CuOOH substrate) were low; the Sedependent glutathione peroxidase activity was not detectable. Interspecific differences were found for most of the enzymes tested.     

Lysosomal injury and MFO activity in the liver of flounder (Platichthys flesus L.) in relation to histopathology of hepatic degeneration and carcinogenesis

Flounder (Platichthys flesus L.), 18–48 cm total length were sampled in the mouth of the Elbe River during a 3-year interdisciplinary project ‘Fish Diseases in the Wadden Sea’. Lysosomal changes (lysosomal membrane stability) and the activity of biotransforming enzymes (MFOs/EROD) were measured parallel to investigations of ultrastructural changes in liver cells, tissue pathologies and macroscopically visible changes. The aim of the study was to investigate if these cyto- and biochemical parameters were able to reflect contaminant induced biological effects. Interlinking of the results of the MFO activity to the pathological alterations observed at the electron- and light-microscopic levels as well as during macroscopic inspection of identical individuals evidenced that the activity of the biotransformation enzymes (EROD) was not induced in healthy livers, increased considerably with the onset of liver changes and dropped again in those livers with degenerative, preneoplastic and neoplastic lesions. In contrast, concurrent studies of lysosomal membrane stability measured in parallel showed a decrease with the onset and progression of liver lesions from reversible to irreversible, neoplastic change.     

Induction of EROD activity in Paralichthys olivaceus by polychlorinated biphenyl CB-28

Abstract-The test organisms (Paralichthys olivaceus) were exposed to CB-28 with differentspiked concentrations. After 72 h, EROD activities in livers of the test organisms were measured. It wasfound that EROD activities significantly mounted up with the increase of spiked quantity. Therefore,there existed a dose-response relationship between EROD activities and specific pollutant concentrations.In the meantime, the optimal conditions for the analysis of EROD activity were discussed. The limita-tion of EROD activities as a monitoring parameter was also analyzed.     

Reproductive success, xenobiotic contaminants and hepatic mixed-function oxidase (MFO) activity in Platichthys stellatus populations from San Francisco Bay

To identify some specific effects of organic contaminants on fisheries in an urbanized estuary we compared the reproductive success of starry flounder from San Francisco Bay with concentrations of tissue contaminants and hepatic mixed-function oxidase (MFO) activity. We found significantly lower (P < 0·05) sediment concentrations of total identified polynuclear aromatic hydrocarbons (PAHs) in the less urbanized San Pablo Bay (SP) area (Fig. 1) than in the more urbanized central bay (CB) stations (Table 1). For flounder in early gametogenesis (August and September) the SP fish (n = 20) had significantly lower (P < 0·01) liver concentrations of Aroclor 1260 (0·34 ± 0·14 μg/g) than those at the CB stations: Berkeley (BK, n = 20, 1·6 ± 1·6 μg/g); Oakland (OK, n = 16, 2·3 ± 2·8 μg/g); and Alameda (AL, n = 4, 2·2 ± 1 μg/g). A similar pattern existed for DDT concentrations: SP = 0·2 ± 0·16 μg/g; BK = 0·1 ± 0·34 μg/g; OK = 0·4 ± 0·53 μg/g; and AL = 0·4 ± 0·33 μg/g. Total PAHs in livers were as follows: SP = 0·14 μg/g; BK = 2·6 μg/g; OK = 1·4 μg/g; and AL = 14 μg/g. Although gonad index, liver index, and presence of fin rot are inversely related to aryl hydrocarbon hydroxylase (AHH) activity, healthy fish in a similar reproductive state have lower AHH activities in the SP area. For example, in August and September, 1984, mean AHH activities were as follows: SP = 203 ± 89, and CB = 355 ± 200 pmol 3-OH-B[a]P mg microsomal protein min. We found a log-linear relationship for AHH activity and its percent inhibition by 7,8-benzoflavone (10⁻⁴m) and only a few fish from SP showed enhanced AHH activity after addition of 7,8-benzoflavone. This suggests that most of the starry flounder in San Francisco Bay are induced.     

条斑星鲽生长与生态转化效率的初步研究

在人工养殖的条件下(自然光照,水温为11~16.3℃),对初始平均体质量为29.64g±5.98g的条斑星鲽(Verasper moseriJordan et Gilbert)幼鱼1周年的生长和生态转化效率进行定期测定。结果显示:条斑星鲽的体长和体高在实验期间皆呈直线增长;体质量的增长可以分为两个阶段:前期呈指数生长,后期为直线增长。体长和体质量呈指数函数关系:W=10-5L3.0226,r=0.9893。条斑星鲽幼鱼周年的生长率和生态转化效率都较高,属于高生长类型鱼类;生态转化效率周年波动范围是34.16%~91.07%,特定生长率的周年波动范围是0.170~1.527。     

Preliminary results of a field study on some stress-related parameters in Tapes philippinarum naturally infected by the protozoan Perkinsus sp.

The aim of the present work was to investigate the possible use of the clam Tapes philippinarum—a filter-feeding bivalve living on soft bottoms—in the monitoring of sediment pollution in the Lagoon of Venice (North-East Italy). To this end, two clam populations, exposed to different environmental conditions (a relative clean site and an industrial polluted area), have been monitored for the prevalence of the protozoan parasite Perkinsus sp. as well as for several biological parameters commonly used as determinants of contaminants exposure. First results showed a low prevalence, higher in the clean site, but a very diffuse distribution of the parasite throughout the two populations. Nevertheless, greater digestive cells height and longer lysosomal latency time were observed in clams from the clean area. As for the physiological stress-related parameters, even if clearance rates were always higher in these clams, scope for growth values indicated a more complicated biological response.     

Do endocrine disrupting chemicals threaten Mediterranean swordfish? Preliminary results of vitellogenin and Zona radiata proteins in Xiphias gladius

Endocrine Disrupting Chemicals (EDCs) have the potential to alter hormone pathways that regulate reproductive processes in wildlife and fishes. In this research the hypothesis that Mediterranean top predator species (such as large pelagic fish) are potentially at risk due to EDCs is investigated. These marine organisms tend to accumulate high concentrations of EDCs such as polyhalogenated aromatic hydrocarbons (PHAHs). The potential effects of EDCs on a fish species of commercial interest, the top predator Xiphias gladius (swordfish), were investigated using vitellogenin (Vtg) and Zona radiata proteins (Zrp) as diagnostic and prognostic biomarkers. Dramatic induction of typically female proteins (Vtg and Zrp) was detected by ELISA and Western Blot in adult males of the species. These results are the first warning of the potential risk for reproductive function of Mediterranean top predators, and suggest the need for continuous monitoring of this fragile marine environment.     

紫菜纳豆菌发酵物中醇提物和多糖生物活性的初步研究

纳豆菌(Bacillus natto)发酵后的条斑紫菜经稀释、离心后上清液用一定浓度的乙醇处理获得醇沉物和醇溶液。将醇沉物去蛋白得到粗多糖,进行抗氧化活性的研究;将醇溶液旋转蒸发得到浸膏,将浸膏按极性大小分相萃取分别得到石油醚相、乙酸乙酯相和正丁醇相浸膏,探讨分相浸膏对金黄色葡萄球菌、枯草芽孢杆菌、四联微球菌的抑菌效果。通过测定多糖和分相浸膏对羟自由基(·OH)和DPPH自由基的清除能力评价其抗氧化能力。结果表明:醇溶物对金黄色葡萄球菌具有抑制作用,且随着浓度的升高各相浸膏对羟基自由基的清除率随之升高,其中,正丁醇相浸膏在质量浓度为0.5 mg/m L时对羟自由基的清除率达到91.6%,说明条斑紫菜经纳豆菌发酵可能产生了活性化合物;与对照组相比,利用纳豆菌发酵紫菜发酵物中多糖提取率增加了0.24%,发酵得到的多糖在质量浓度为5.0 mg/m L时对羟自由基的清除率达到了95.7%。     

篮状菌DYM25胞外产物的初步鉴定及其生物学活性

本研究以西太平洋雅浦海沟分离得到的篮状菌(Talaromyces sp.)DYM25为实验菌株,通过水饱和正丁醇萃取菌株DYM25的胞外产物,利用生物碱沉淀显色法初步考察胞外产物的种类,进而研究了胞外产物的抗菌、抗氧化和抗肿瘤活性。结果表明,1 L菌株DYM25的发酵液通过水饱和正丁醇萃取旋蒸后可得到约2 g的产物;生物碱显色反应预示着胞外产物中生物碱盐类物质的存在;篮状菌DYM25胞外产物对枯萎病致病菌木贼镰刀菌(Fusarium equiseti)的生长具有显著的抑制作用,是篮状菌作为F.equiseti拮抗菌的首次报道。同时,此胞外产物还具有一定的抗氧化活性和抗肿瘤活性,其抗氧化能力随产物浓度的升高而增强,对人胰腺癌细胞PANC-1的抑制能力也呈现出一定的浓度依赖性。篮状菌DYM25多样的生物活性凸显了其良好的研究价值和应用前景。     

Cloning and sequence analysis of the self-fertilizing fish Rivulus marmoratus immediate early gene c-fos

We have cloned the proto-oncogene c-fos from a self-fertilizing fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) after screening of R. marmoratus λGEM-11 genomic DNA library, and sequenced over 12 kb including all exons, introns and the promoter region. The R. marmoratus c-fos gene consisted of one noncoding exon and four exons with high similarity to those of fugu and mammals. We sequenced ≈7 kb of the R. marmoratus c-fos gene promoter region to gain a better understanding of the molecular anatomy of the immediate response of this gene upon cellular damage. In the promoter region, R. marmoratus c-fos gene has seven xenobiotic response elements (XREs) and eight metal response elements (MREs) as well as two estradiol (E2), 4 NFκB, 2 CarG, 2 prolactin (PRL) motifs and one pit1 site, while the 3^′-UTR of this gene contains the estrogen response element (ERE). The seven XRE and eight MRE motifs raise the possibility of its regulation by exposure to environmental pollutants. In this paper, we discuss the gene structure of R. marmoratus c-fos gene and compare its promoter region with those of other organisms' c-fos genes. We propose its potential use in ecotoxicology.     

涌流式培育的大獭蛤稚贝生长的初步研究

2004年11月至2005年1月利用涌流装置和浓缩扁藻进行了大獭蛤(Lutraria(Psam-mophila)maxima)稚贝的中间培育实验。实验所采用的涌流流量为500 mL/min,扁藻密度分别为8 870个/mL±512个/mL和3 368个/mL±557个/mL,稚贝的培育密度分别为20 000,35 000,50 000粒/m2。结果表明,过高的扁藻密度不利于大獭蛤稚贝的摄食,并会造成稚贝的死亡。只有在适当的扁藻密度和涌流流量条件下,大獭蛤稚贝才能正常生长。不同的稚贝培育密度对稚贝的生长存在影响,密度越高,生长越慢;水温低于15℃时,稚贝摄食停止,生长缓慢;稚贝在游离状态下与铺沙条件下的生长基本一致。     

中国蛤蜊Mactra chinensis Philippi稚贝运动行为学初步研究

为从行为学角度解释中国蛤蜊(Mactra chinensis Philippi)的跑滩现象, 研究了规格、温度对其跳跃行为以及底质、流速和周期性干露对其潜沙行为的影响。结果表明, 中国蛤蜊运动能力与其规格和水温有关, 3 mm稚贝不具备跳跃能力, 5 mm以上具备跳跃能力; 跳跃频率和高度随水温的升高呈上升趋势。中国蛤蜊稚贝潜沙比例均随时间推移逐渐上升; 各时间节点中国蛤蜊稚贝在细沙底质的潜沙比例最高, 中沙底质次之, 粗沙底质最低。施加微弱水流后, 中国蛤蜊稚贝潜沙速度和比例显著增加; 流速增大到一定程度后部分稚贝被水流冲走, 潜沙比例随之下降。周期性干露会导致中国蛤蜊稚贝从底质中爬出, 干露时间越长, 爬出比例越高。干露后的稚贝重新放置于海水中, 部分稚贝会在水面漂浮一段时间后再下落至水底, 漂浮比例随干露时间的增加而增加。本研究从行为学角度初步查明了中国蛤蜊跑滩现象, 为解决中国蛤蜊防跑滩问题提供了思路。     

Hepatic hemosiderosis in buffalo fish (Ictiobus spp.)

Hepatocellular hemosiderosis was observed in several species of fish associated with mixed chemical pollution in a Mississippi River Basin ecosystem. Three species of buffalo fish—smallmouth buffalo (Ictiobus bubalus), bigmouth buffalo (Ictiobus cyprinellus), and black buffalo (Ictiobus niger)—were collected from a contaminated site, Devil's Swamp, and a control site, Tunica Swamp, both near Baton Rouge, Louisiana. Liver, kidney and spleen were examined histologically. Hepatocellular iron overload was observed in smallmouth buffalo and bigmouth buffalo from Devil's Swamp. Smallmouth buffalo from Tunica Swamp had no stainable iron in the hepatocytes. Bigmouth buffalo from Tunica Swamp had trace amounts of stainable iron in the hepatocytes. Black buffalo from both Devil's Swamp and Tunica Swamp had no stainable iron in the hepatocytes. Iron could accumulate in sediment where it enters the food chain through benthic organisms. Analyses of water and sediment from both study sites revealed similarly high concentrations of iron. The pollutants in the Devil's Swamp may have affected the iron absorption and metabolism in smallmouth buffalo and bigmouth buffalo. The variation in the presence of stainable iron in the liver of three species of buffalo fish may indicate significant interspecies variation in the iron absorption and metabolism under conditions of mixed pollution.     

Preliminary results of in-situ XCTD/CTD comparison test

The current status of XCTD manufactured by the Tsurumi Seiki Co. is described based on XCTD/CTD comparison tests conducted in the Pacific and Indian Ocean in November 1996 and January 1997 respectively. The falling rates of used probes are generally consistent and differences between individual probes stay within a small range of +/−5 m through the full depth range (0–1000 m), although the rate is slightly different from the depth-time equation provided by the manufacturer. Temperature and salinity accuracy is estimated to be better than 0.05°C and 0.05 psu respectively. Comparison of vertical temperature/salinity sections of XCTD/CTD constructed separately along a line on which XCTD stations locate midway of CTD stations. The hydrographic structures obtained are generally consistent, and the difference of surface dynamic height (referred to 1000 db) is 0.03 dyn m in RMS.     

夏、冬两季真鲷仔、稚、幼鱼消化酶活性的比较

通过夏、冬两季真鲷仔、稚、幼鱼蛋白酶、淀粉酶和脂肪酶活性的变化研究表明,夏、冬两季3种消化酶变化趋势基本相似。蛋白酶的活性冬季比夏季低1个数量级;淀粉酶的活性冬季比夏季低1倍左右;脂肪酶的活性冬季高于夏季,且一直维持在相对稳定的水平。结果表明,3种酶活性受温度影响外,冬季脂肪酶的高活性对鱼苗的生长、成活具有重要意义。     

副溶血弧菌tdh基因在毕赤酵母中的表达及溶血活性检测

通过PCR技术从副溶血弧菌(Vibrio Parahaemolyticus,VP)基因组DNA上扩增耐热溶血素基因tdh,双酶切后定向插入到酵母表达载体pPICZaA中,构建重组表达质粒pPICZaA-tdh,经限制性内切酶SacI线性化后电转化毕赤酵母(Pichia pastoris,P.pastoris)GS115,经PCR和测序鉴定耐热溶血素基因成功整合到毕赤酵母的基因组中。在甲醇诱导下进行tdh的表达,SDS-PAGE分析证明重组TDH(thermostable direct hemolysin,TDH)的分子质量约为23ku,经Western blotting鉴定VP抗体能与表达的TDH发生特异反应,说明tdh基因在毕赤酵母中的表达是准确的。溶血实验证明了表达的TDH具有溶血活性。本研究首次应用毕赤酵母表达耐热溶血素基因tdh,在培养基中获得分泌表达的耐热溶血素,为研究tdh基因的功能奠定了基础。     

Individual variation in patterns of benzo[a]pyrene metabolism in the marine fish scup (Stenotomus chrysops)

The metabolism at specific sites on carcinogenic hydrocarbons such as benzo[a]pyrene (BP) is responsible for activation to the ultimate mutagens and carcinogens, and patterns of metabolism can thus influence the biological effect of such compounds. Marine fish are known to efficiently metabolize BP at the benzo-ring, forming high percentages of the 9,10-dihydrodiol (DHD) and 7,8-DHD, the latter including the penultimate carcinogen.^1,2 Hydrocarbon-induced cytochrome P-450 in fish is responsible for initiating metabolism on the benzo-ring, but epoxide hydrolase (EH) activity is required for DHD formation.^3,4 Both factors could influence formation of the DHD leading to the ultimate carcinogenic diol-epoxide. In the present study, patterns of BP metabolism were evaluated in a number of individual scup Stenotomus chrysops sampled from local Woods Hole waters, and a correlation is described between variation in the DHD formation and EH activity in these feral fish.