镉、铜对非洲鲫鱼肝线粒体功能影响的初步研究

本实验以在海水中培养的非洲鲫鱼(Tilapiamossambica)为试验材料,采用氧电极法测定氯化镉、硫酸铜对鱼肝线粒体(离体)呼吸控制率(RCR)和氧化磷酸化效率(ADP/O)的影响,以探讨重金属毒物对海洋生物的致毒机理以及了解对其体內各种生化过程所起的作用。     

The action of heavy metals on the gametes of the marine mussel, Mytilus edulis (L.)—III. The effect of applied copper and zinc on sperm motilityin relation to ultrastructural damage and intracellular metal localisation

Treatment of Mytilus edulis sperm with external concentrations of copper or zinc (0-1-3-3mM) causes a decrease in motility in which zinc is more inhibitory than copper. Zinc also appeared to cause more extensive mitochondrial damage, as revealed by transmission electron microscopy, than did treatment with copper. The relationship between sperm motility and respiration in the presence of the various heavy metal concentrations used indicates that the depression of sperm motility can be explained largely on the basis of respiratory inhibition. However, zinc produces a less pronounced effect on sperm motility than on r respiration.X-ray microanalysis of thick sections of fixed treated sperm showed that copper accumulation occurs in the acrosomes, mitochondria and nuclei, whereas zinc is found in the acrosomes and in mitochondrial granules in association with calcium and phosphorus. No evidence was obtained for zinc accumulation in the nuclei. Treatment with either copper or zinc resulted in considerable reductions of bound calcium and phosphorus in both the acrosomes and mitochondria. It is suggested that the heavy metal ions cause an increase in the permeability of the organelle membranes to calcium and phosphorus.It is concluded that the less marked effect of zinc on sperm motility as compared to respiration may be due to an increase in the cytosolic free calcium concentration which, in turn, may stimulate the flagellar contractile apparatus.     

Resistance to heavy metals of heterotrophic bacteria isolated from the deep-sea hydrothermal vent polychaete, Alvinella pompejana

Fifty-two heterotrophic bacteria isolated from the deep-sea hydrothermal vent polychaete, Alvinella pompejana, were tested for susceptibility to cadmium, zinc, copper, arsenate and silver ions by an agar dilution technique. Metal resistance was exhibited 96% of the total isolates. The frequencies of resistance to heavy metals were: copper, 88%; zinc and arsenic, 58%; cadmium, 38%. However, strains demonstrated a high susceptibility to silver. 71% of the strains tested were multiresistant to metals.     

Lead, cadmium, copper, nickel and iron in limpets, mussels and snails from the coast of ras Beirut, Lebanon

Samples of Patella coerulea (Linnaeus), Brachydontes variabilis (Krauss), Monodonta turbinata (Born) and surface seawater were collected at eight locations along the coast of Ras Beirut, Lebanon, and analysed for lead, cadmium, copper, nickel and iron. With the exception of cadmium, the metal levels found in the three molluscs appear to be high in relation to levels reported by investigators from other coastal areas. The average values for lead, cadmium and nickel within these animals were quite similar. Concentrations of copper in P. coerulea were lower, but concentrations of iron were much higher than those found in both B. variabilis and M. turbinata. Further studies of metal levels in various other intertidal organisms from Ras Beirut, together with investigations into the possible effects of seasonal changes on metal concentration and distribution, are currently under way.     

浙江近岸海域贝类中重金属和贝毒污染状况研究

在浙江近岸海域采集了青蛤(Cyclina sinensis)、毛蚶(Scapharca subcrenata)、厚壳贻贝(Mytilus coruscus)、缢蛏(Sinonovacula constricta)、泥螺(Bullacta exarata)、菲律宾蛤仔(Ruditapes philippi-narum)、文蛤(Meretrix meretrix)、僧帽牡蛎(Ostrea cucullata)和栉孔扇贝(Chlamys farreri)共9种贝类计29份样品,检测其中重金属汞、砷、铜、铅、锌、镉以及麻痹性贝毒和腹泻性贝毒.结果表明,瑞安毛蚶、瑞安栉孔扇贝中有麻痹性贝类毒素检出,嵊泗文蛤、毛蚶和乐清牡蛎中存在重金属超标的情况;牡蛎对汞、铜、锌、镉有较强的富集能力.     

Organic complexation and its control of the dissolved concentrations of copper and zinc in the Scheldt estuary

Cathodic stripping voltammetry (CSV) is used to determine total (after UV-irradiation) and labile dissolved metal concentrations as well as complexing ligand concentrations in samples from the river Scheldt estuary. It was found that even at high added concentrations of catechol (1 m for copper and 0·4 m for iron) and of APDC (1 m for zinc) only part of the dissolved metal was labile (5–58% for copper, 34–69% for zinc, 10–38% for iron); this discrepancy could be explained by the low solubility of iron which is largely present as colloidal material, and by competition for dissolved copper and zinc by organic complexing ligands. Ligand concentrations varied between 28 and 206 n for copper and between 22 and 220 n for zinc; part of the copper complexing ligands could be sub-divided into strong complexing sites with concentrations between 23 and 121 n and weaker sites with concentrations between 44 and 131 n . Values for conditional stability constants varied between (logK′ values) 13·0 and 14·8 for strong and between 11·5 and 12·1 for weaker copper complexing ligands, whereas for zinc the values were between 8·6 and 10·6. The average products of ligand concentrations and conditional stability constants (a-coefficients) were 6 × 10² for zinc and 6 × 10⁶ for copper.The dissolved zinc concentration was found to co-vary with the zinc complexing ligand concentration throughout the estuary. It is argued that the zinc concentration is regulated, in this estuary at least, by interactions with dissolved organic complexing ligands. A similar relationship was apparent between the dissolved copper and the strong copper complexing ligand concentration. The total copper complexing ligand concentrations were much greater than the dissolved copper concentrations, suggesting that only strongly complexed copper is kept in solution.These results provide evidence for the first time that interactions of copper and zinc with dissolved organic complexing ligands determine the geochemical pathway of these metals.     

Transport of copper from the Bering Sea to the northwestern North Pacific

Dissolved copper concentrations in surface waters of the Bering Sea ranged from 106 to 882 ngl^–1. Higher concentrations were found in continental shelf waters. In the northwestern North Pacific dissolved copper ranged from 54 to 140 ngl^–1. Particulate copper concentrations varied regionally and seasonally from 6 to 79 ngl^–1. Regionally averaged particulate copper concentrations decreased from 175 to 33g g^–1 against an increase in suspended materials because of the dilution effects of biological fractions. Apparent sporadic increases in copper concentrations were found in the mixing area of the Kuroshio and the Oyashio waters. The feature is attributed to the lateral distribution of different water types rather than to the upwelling of deeper waters by eddies. In the same area west of 160E, waters with high concentrations of dissolved copper (96±9 ngl^–1) were found. Their origin appears to be the continental shelf of the Bering Sea. In spite of intensive biological activity, a considerable fraction of copper added to shelf waters was transported to the area off Japan via the circulation in the Bering Sea and the Oyashio current.     

Biochemical‐genetic and meristic evidence that blue and copper moki (Teleostei: Latridae: Latridopsis) are discrete species

Specimens of blue moki Latridopsis ciliaris (Forster in Bloch & Schneider, 1801) from New Zealand were shown to differ from specimens of copper moki Latridopsis forsteri (Castelnau, 1872) using two genetic methods—allozymes and muscle proteins. Allozyme techniques revealed fixed differences between blue and copper moki at 4 out of 15 loci. There was a genetic distance of 0.31–0.34 between population samples of the two colour morphs. The two morphs were also distinguished by iso‐electric focusing of muscle proteins. Meristic characters, counted in the specimens studied with genetic markers, revealed diagnostic markers in tubed lateral line scales and scales in the lower transverse series. It is concluded that specimens of blue moki from New Zealand and copper moki from New Zealand and Tasmania are valid species.     

Heavy metal resistant endophytic fungi isolated from Nypa fruticans in Kuching Wetland National Park

Heavy metal pollution is an environmental issue globally and the aim of this study was to isolate endophytic fungi from mangrove wetlands of Sarawak to assess and test their ability to grow in the presence of various heavy metals (copper (Cu), zinc (Zn), lead (Pb), and chromium (Cr)). Samples of Nypa fruticans were collected from Kuching Wetland National Park (KWNP) for subsequent endophyte isolation. Ninety-three (93) isolates were obtained and assessed and the most resistant isolates (growing at concentrations up to 1000 ppm) were identified using fungal primers ITS 1 and ITS 4. All of the endophytic fungi were identified to be closely related to Pestalotiopsis sp. and this is to our knowledge the first study reporting the ability of Pestalotiopsis sp. to grow at high concentrations of copper, lead, zinc and chromium. Our results highlight the potential of using endophytic fungi for the treatment of heavy metal pollution, for example as biosorbents.

     

铜在牙鲆(Paralichthys olivaceus)组织蓄积、分配及排放的研究

采用暴露实验方法 ,研究了海水中Cu在牙鲆 (Paralichthysolivaceus)内脏、肌肉、鳃组织内吸收、积累和排放规律 ,海水中总有机碳 (TOC)浓度、配体种类对铜吸收的影响及其与海水表观络合容量 (ACuCC)的关系。结果表明 ,Cu浓度为 0 .5mg/L时 ,各组织内Cu蓄积量随暴露时间增加而增大 ,第 1 3天均达吸收平衡 ,此时Cu蓄积量 (mg/kg)为内脏 971 .89>肌肉 2 0 4 .99>鳃 90 .0 4。染毒 1 3天牙鲆在清洁海水中排放结果表明 :随排放时间增加各组织Cu蓄积量下降 ,第 8天接近排放平衡。此时各组织Cu排出率为 :肌肉 89% >鳃 86.5 % >内脏团 83.7%。海水中TOC浓度、种类对牙鲆Cu蓄积有明显影响 ,当TOC浓度相同时 ,孔石莼分泌物比牙鲆分泌物更能降低Cu在牙鲆组织内吸收和蓄积量 ;当TOC种类相同时 ,随TOC浓度升高 ,各组织铜蓄积量均明显下降 ,表明海水TOC能降低牙鲆组织对Cu的吸收和积累。ACuCC随TOC浓度增加有明显上升趋势 ,并与TOC浓度呈线性相关。     

饲料中不同形式的铜及添加量对凡纳滨对虾(Paneaus vannamei)生长性能、血清铜蓝蛋白和生长激素水平的影响

在对照饲料中添加不同水平的硫酸铜、富马酸铜和蛋氨酸铜,使硫酸铜组铜离子含量为32mg/kg,富马酸铜和蛋氨酸铜组铜离子含量分别为16mg/kg、32mg/kg和48mg/kg,共组成8种饲料(饲料编号为对照、Sul-32、Fum-16、Fum-32、Fum48、Met-16、Met-32和Met-48),饲喂凡纳滨对虾(初重0.74±0.01g)8周。结果表明,饲料中添加三种铜源均能不同程度提高凡纳滨对虾的特定生长率(SGR)、存活率(Survival)和蛋白质效率(PER),降低饲料系数(FCE)。其中,Met-16和Fum-32组的SGR显著高于对照组(P0.05),Met-16、Met-32和Fum-32组的PER显著高于其它各组(P0.05);不同铜源对凡纳滨对虾的体组成没有显著影响(P0.05),但Met-32组的肝体比(HSI)显著下降(P0.05)。各组间血清铜蓝蛋白(CER)含量差异不显著(P0.05),Met-16和Fum-32组的生长激素(GH)含量则显著高于对照组(P0.05)。本研究结果表明,饲料中三种铜源能不同程度的提高凡纳滨对虾的生长性能,但相同浓度有机铜离子的生物利用率优于无机铜,其顺序为:蛋氨酸铜富马酸铜硫酸铜。饲料铜对凡纳滨对虾的促生长作用可能是通过提高体内相关抗氧化酶的活性及刺激生长激素的分泌来实现的。     

Production of an extracellular copper-binding compound by the heterotrophic marine bacterium Vibrio alginolyticus

The heterotrophic marine bacterium Vibrio alginolyticus was shown to produce extracellular copper-binding compound (s) when exposed to copper in a seawater medium. Fractionation and analysis of copper, and methionine incorporation in culture supernatant fractions showed that the copper-binding compound coeluted with material which was radiolabeled with ³⁵S methionine. This suggests that the copper-binding compound may be proteinaceous. Analysis of label incorporation from ¹⁴C glucose into the fraction containing the copper-binding compound indicated that it was produced actively by the cells during recovery from copper toxicity, and was not present because of non-specific release by lysed bacterial cells. Concentrated supernatants from control and copper-challenged cultures contained two compounds which could be marginally resolved by size exclusion HPLC (26 kD and 28 kD), and which were produced at about a ten-fold higher level in copper-challenged cultures than in controls. These data indicate that the bacteria respond to toxic copper levels by excretion of a ca. 28 kD compound (s), which serves to detoxify copper in the medium by formation of organic copper complexes.The data suggest a potential role for macromolecules excreted by heterotrophic bacteria in control of copper ion activity in seawater. The production of copper-complexing compounds in response to copper represents a mechanism through which bacteria may directly influence the speciation of metal ions in seawater.     

Kinetic and equilibrium studies of copper-dissolved organic matter complexation in water column of the stratified Krka River estuary (Croatia)

An interaction of dissolved natural organic matter (DNOM) with copper ions in the water column of the stratified Krka River estuary (Croatia) was studied. The experimental methodology was based on the differential pulse anodic stripping voltammetric (DPASV) determination of labile copper species by titrating the sample using increments of copper additions uniformly distributed on the logarithmic scale. A classical at-equilibrium approach (determination of copper complexing capacity, CuCC) and a kinetic approach (tracing of equilibrium reconstitution) of copper complexation were considered and compared. A model of discrete distribution of organic ligands forming inert copper complexes was applied. For both approaches, a home-written fitting program was used for the determination of apparent stability constants (K_i^equ), total ligands concentration (L_iT) and association/dissociation rate constants (k_i¹,k_i^- 1).A non-conservative behaviour of dissolved organic matter (DOC) and total copper concentration in a water column was registered. An enhanced biological activity at the freshwater–seawater interface (FSI) triggered an increase of total copper concentration and total ligand concentration in this water layer. The copper complexation in fresh water of Krka River was characterised by one type of binding ligands, while in most of the estuarine and marine samples two classes of ligands were identified. The distribution of apparent stability constants (log K₁^equ: 11.2–13.0, log K₂^equ:8.8–10.0) showed increasing trend towards higher salinities, indicating stronger copper complexation by autochthonous seawater organic matter.Copper complexation parameters (ligand concentrations and apparent stability constants) obtained by at-equilibrium model are in very good accordance with those of kinetic model. Calculated association rate constants (k₁¹:6.1–20 × 10³ (M s)^− 1, k₂¹: 1.3–6.3 × 10³ (M s)^− 1) indicate that copper complexation by DNOM takes place relatively slowly. The time needed to achieve a new pseudo-equilibrium induced by an increase of copper concentration (which is common for Krka River estuary during summer period due to the nautical traffic), is estimated to be from 2 to 4 h.It is found that in such oligotrophic environment (dissolved organic carbon content under 83 µM_C, i.e. 1 mg_CL^− 1) an increase of the total copper concentration above 12 nM could enhance a free copper concentration exceeding the level considered as potentially toxic for microorganisms (10 pM).     

汞、铅对非洲鲫鱼肝线粒体功能影响的初步研究

为了研究和探讨重金属毒物对海洋生物的致毒机理及其对生物体内各种生化过程的影响,继镉、铜对非洲鲫鱼肝线粒体功能影响的研究之后,本实验继续以非洲鲫鱼(Tilapia mossambica)为实验材料,采用氧电极法测定了氯化汞、硝酸铅对鱼肝线粒体(离体)呼吸控制率和氧化磷酸化效率的影响。     

The effect of supplementary dietary copper on copper and metallothionein levels in the fish, dab, Limanda limanda

Common dab were fed a casein based diet containing copper at a concentration of 1·4 mg copper kg⁻¹ (dry weight) or a similar diet supplemented with 200 mg copper kg⁻¹. Fish were sampled after 10 weeks and 20 weeks and hepatic and renal copper, zinc and metallothionein concentrations were measured. All hepatic cytosolic copper in test and control fish co-eluted from Sephadex G-75 with metallothionein, as did some of the cytosolic zinc. After 20 weeks there was three times as much of this metallothionein-bound copper in the livers of the test fish compared to the control fish, although metallothionein concentration was little elevated. On the basis of these results it is proposed that cytosolic liver copper concentration or heat stable liver copper concentration can be used as an indication of excess dietary copper in the dab Limanda limanda.     

The ultrastructural study of the spermatid differentiation in Cyclina sinensis

The authors have observed submicroscopically the transformation of spermatid and the sperm structure in the spermatogenesis process of Cyclma sinensis. The results show during the spermatid differentiation the nuclear morphology transforms from the elliptical into the cylindrical shape. The morphological transformation of the nuclear chromatin experiences from the small masses of fine granule aggregation to granular substructures with a granular diameter of 20 - 41 nm (containing a few masses), then to large coarsely granules with a diameter of 36. 8-52. 6 nm and at last to the homogenate with high electron density. The proacrosomal granules secreted by Golgi complex aggregate to form a large spherical granule which moves gradually to the anterior pole of the nucleus to join the acrosomal formation. The fusion of some mitochondria and the disintegration of a few ones result in the mitochondrial number decrease and volume enlargement. Finally, the mitochondrial ring which consists of five mitochondria an     

Heavy metals in some New Zealand commercial sea fishes

To establish a base line against which future pollution may be measured, eight common commercial species of New Zealand sea fish were analysed for cadmium, chromium, copper, iron, lead, manganese, nickel, and zinc. One sample of edible muscle tissue was analysed for each of 70 samples of each species. The internal organs of up to five specimens of each species were also analysed. The fish studied were: kahawai, Arripis trutta, trevally, Caranx lutescens, tarakihi, Cheilodactylis macropterus, snapper, Chrysophrys auratus, moki, Latridopsis ciliaris, hapuku, Polyprion oxygeneios, kingfish, Seriola grandis and gurnard, Trigla kumu.

Although, none of the edible parts of the fish appeared to have trace element levels likely to be a public health problem (cadmium 0.002–0.02 ppm, chromium 0.01–0.04 ppm, copper 0.04–0.95 ppm, iron 0.9–13.5 ppm, lead ‘ 0.14–0.95 ppm, manganese 0.04–2.00 ppm, nickel 0.01–0.08 ppm, zinc 2.0–36.0 ppm), some of the organs (particularly the liver) had relatively high concentrations of elements such as cadmium (up to 54 ppm). Only if some industry were to seek to exploit internal organs of fish for human consumption would such levels become important. There was some evidence for a relationship between trace element concentrations and fish size for copper in kingfish and snapper, iron in hapuku, manganese in gurnard, and for zinc in kingfish and tarakihi. Some element pairs such as copper and zinc, iron and manganese, appeared to have concentrations which were mutually related. It was assumed that the elemental concentrations reported represent natural levels and are not the result of significant man‐made pollution because there are no major industries or large urban centres adjacent to the fishing grounds.     

Solvent extraction of copper acetylacetonate in studies of copper(II) speciation in seawater

A liquid-liquid partition, ligand exchange procedure involving the formation of copper(II) complexes with acetylacetone is presented for the determination of stability constants and concentrations of copper chelators in seawater. Acetylacetone competes with natural ligands for copper, and the equilibrium concentration of the copper acetylacetonate complex is used in speciation calculations. The concentration of the complex is calculated by partitioning a fraction of it into an organic phase and determining the total Cu concentration in that phase by back extracting with acid, and analyzing by flameless atomic absorption spectroscopy. The concentration of Cu acetylacetonate in seawater in equilibrium with the organic phase is calculated from the partition coefficient. The simple, thermodynamically well characterized procedure offers several advantages over previous techniques. Studies using organic free seawater and model ligands show good agreement between experimental and calculated conditional stability constants. Studies from seawater in Biscayne Bay, Florida, indicate two ligand types are present; type 1, K₁ = 1.2 × 10¹², C_L1 = 5.1 × 10⁻⁹ M; type 2, K₂ = 2.8 × 10¹⁰, C_L2 = 1.1 × 10⁻⁷ M. Speciation is dominated by ligand type 1. Depth profiles of [Cu(II)]_free/[Cu(II)]_total measured with the procedure at ambient copper concentrations show an increase from < 5 × 10⁻⁵ at 50–60 m to > 1 × 10⁻³ at the surface at two stations off the Florida coast.     

海藻、牡蛎提取物HLK及其营养成分的分析

海藻、牡蛎于1993年6－8月采自青岛近海。经恒温水解、脱水浓缩、低温干燥等方法得到海藻、牡蛎提取物HLK。对HLK进行分析测定，结果表明，提取物含有丰富的氨基乙磺酸、多种维生素、多糖类、不饱和脂肪酸、低脂肪以及丰富而适量的钾。钙、锌、铁、铜、碘、硒等多种营养素和矿物质元素。为研制开发海藻、牡蛎保健食品提供科学依据。     

Complexation of calcium and copper with carbohydrates: Implications for seawater speciation

The extent of complexation of calcium(II) and copper(II) with 33 carbohydrates found in seawater has been determined potentiometrically using calcium and copper ion-selective electrodes. The measurements were performed in an essentially neutral aqueous medium at 25°C in 0.70 M KNO₃.Neutral sugars form very weak 1:1 complexes with Ca(II) and Cu(II) ions. Sugars with carboxylate groups from much stronger complexes. K′_Cu(II) >K′_Ca(II)) for a given sugar. The extent of complexation is dependent on the conformation of the sugar ligand.