多氯联苯在两种海洋微藻中毒性效应及富集效应的研究

通过分析2,2',3,4,4',5'-六氯联苯(PCB_(138))对牟氏角毛藻(Chaetoceros muelleri)和亚心形扁藻(Platymonas subcordiformis)生长的影响,以及PCB_(138)对两种微藻超氧化物歧化酶(Superoxide dismutase,SOD)活性的影响,研究多氯联苯对海洋微藻的毒性效应,并采用气相色谱法研究了两种微藻对PCB_(138)的生物富集效应。结果表明,PCB_(138)对牟氏角毛藻和亚心形扁藻均有明显的毒性效应作用,对2种藻类的96 h半抑制质量浓度(96hEC_(50))分别为0.057 mg/和0.354 mg/L,与亚心形扁藻相比,牟氏角毛藻对PCB_(138)更敏感,耐受性更差;低浓度的PCB_(138)对两种微藻SOD酶活性有诱导作用,但当PCB_(138)浓度过高时,SOD酶活性下降;两种微藻对PCB_(138)均有明显的富集作用,随着培养时间的延长,牟氏角毛藻中富集的PCB_(138)含量呈上升趋势,而亚心形扁藻中PCB_(138)含量呈现先上升后下降趋势。     

外源一氧化氮和金属铜对海洋微藻抗氧化系统的影响

以亚心形扁藻为研究对象,通过添加不同浓度一氧化氮(NO)(1.4×10-10～1.4×10-5 mol/L)培养,探讨NO对微藻生长、H2O2和MDA含量以及SOD活性的影响;通过对只添加金属Cu、同时添加Cu与NO实验,对微藻的H2O2和MDA含量以及SOD活性的影响进行了对比研究。结果表明,藻细胞活性氧的代谢有随NO浓度增加的趋势,同时细胞内的抗氧化酶SOD活性和MDA含量都有相应的变化,NO作为环境的"刺激物"提高了细胞内的活性氧水平,对抗氧化系统起到激活或伤害作用。在Cu胁迫下,藻细胞活性氧代谢明显提高,SOD活性提高,并导致细胞膜脂过氧化;但外源NO能明显降低活性氧的水平,并恢复SOD活性和MDA含量,对抗氧化系统起到保护作用,提高微藻的抗氧化能力。研究表明,NO在海洋浮游植物中扮演着"双重角色"——氧化剂和抗氧化剂,从而对微藻的生长起到重要的调节作用。     

2-(N-芳基氨甲基)苯并异噻唑啉酮类化合物对海洋浮游生物的生长抑制活性研究

本文研究了13个2-(N-芳基氨甲基)苯并异噻唑啉-3-酮类化合物对球等鞭金藻Isochrysis galbana、亚心形扁藻Platymonas subcordiformis及舟形藻Navicula的生长抑制性能,并测试了其对藤壶二期无节幼虫的生物活性。结果表明:2-(N-芳基氨甲基)苯并异噻唑啉-3-酮类化合物对球等鞭金藻、亚心形扁藻和舟形藻都有良好的生长抑制活性,其中对球等鞭金藻和亚心形扁藻的抑制作用较强;对藤壶幼虫的生物活性测试表明,化合物最低质量浓度为2.0 mg/L时,除化合物g和l,都具有良好的生长抑制活性。化合物的结构与浮游生物的生长抑制活性关系表现为间位取代优于对位取代。     

重金属胁迫对两种海洋饵料微藻的急性毒性效应研究

在模拟生态环境条件下研究了铜(Cu2+)、镉(Cd2+)和锌(Zn2+)3种重金属离子对2种海洋饵料微藻(青岛大扁藻(Platymonas helgolandica)和等鞭金藻8701(Isochrysis galbana 8701))的急性毒性效应,分析其对种群增长的影响,并利用透射电镜对微藻亚显微结构的变化进行分析。在实验浓度范围内,2种微藻的种群增长对3种重金属离子的急性胁迫表现出类似的变化趋势,随着胁迫浓度的增加,微藻的相对增长率均呈现出不断下降的趋势;通过比较2种微藻的48hEC50和96h-EC50发现:3种重金属离子的毒性程度为Cu2+Cd2+Zn2+,等鞭金藻8701对重金属胁迫的敏感性高于青岛大扁藻。2种微藻的亚显微结构观察显示,二者的膜结构及细胞核对Cu2+胁迫并不太敏感;与对照组相比等鞭金藻8701的色素体及贮能物质受影响较严重,而青岛大扁藻的叶绿体及淀粉粒受影响较大。     

模拟电厂温排水温升和游离余氯对亚心形扁藻生长影响的研究

以亚心形扁藻(Platymonas subcordiformis)为受试藻种,模拟温升和游离余氯条件,研究浮游植物对核电厂温排水的生态响应。结果表明,春秋季温升对其生长影响较小,而在夏季,其生长受到显著抑制。亚心形扁藻对游离余氯非常敏感,0.1 mg/L游离余氯既能抑制其生长,抑制作用随着游离余氯浓度的升高而增强。温升会加剧游离余氯的抑制作用,4℃温升和0.1 mg/L游离余氯联合作用对亚心形扁藻的抑制效果明显增加。由于温升和游离余氯的联合作用,即使在离排口较远的水域,仍然会导致亚心形扁藻数量减少。     

蒽与苯并[a]芘对海洋微藻致毒影响的比较研究

新月菱形藻 ,球等鞭金藻 870 1和亚心形扁藻 ,分别用不同浓度的蒽和苯并 [a]芘胁迫72 h,测定三种藻的生长速率 ,叶绿素 a含量。实验表明 ,三种微藻对蒽的敏感性由高到低依次为新月菱形藻、球等鞭金藻 870 1和亚心形扁藻 ,它们 72 h的半抑制浓度 EC50 分别是 0 .0 6 0 mg/L、0 .0 6 5mg/L和 0 .0 94 mg/L。而对苯并 [a]芘的敏感性由高到低依次为亚心形扁藻、新月菱形藻和球等鞭金藻 870 1,它们的 72 h· EC50 分别是 0 .0 76 mg/L、0 .0 88mg/L 和 >0 .10 mg/L(在本实验范围内未测到 )。叶绿素 a含量与各自的相对增长率呈一定的正相关。不同的海洋微藻对蒽和苯并 [a]芘的敏感性不同。     

UV-B辐射对3种海洋微藻的种间竞争性平衡的影响

为探讨UV-B辐射对海洋生态系统的影响，研究了短期(2d)UV-B辐射对金藻8701、小新月菱形藻和亚心形扁藻3种藻单养的敏感性比较以及长期(21d)UV-B辐射对3种藻混养的种间竞争性平衡的影响。实验表明，(1)UV-B条件下。单养和混养情况下，最敏感的藻是金藻8701，小新月菱形藻次之，亚心形扁藻最不敏感；(2)增强的UV-B为对UV-B具有高耐受力的亚心形扁藻提供了竞争优势，使种间竞争平衡向着有利于亚心形扁藻的方向发展，到第21天，UV-B辐射(2．88J／m^2，5．76J／m^2)的处理组中，亚心形扁藻成为优势种。     

富勒醇减轻蒽对亚心形扁藻伤害的作用研究

采用新型抗氧化剂———富勒醇((C60(OH)n),探讨了其减轻蒽对亚心形扁藻(Plattymonassubcordiformis)伤害的作用。亚心形扁藻在蒽的半致死浓度0.06mg/L胁迫下分别加入n=14,浓度为2.29×103mg/L的C60(OH)n20μL和n≥20浓度同样为为2.29×103mg/L的C60(OH)n200μL后,分别测定了对照组和胁迫组在0、12、24、36、、8、60和72h的相对增长率K 并作图。结果分析表明富勒醇(C60(OH)n)有减轻亚心形扁藻蒽伤害的作用 ,并且它的这种减轻作用与剂量和羟基的数目呈正相关     

对二甲苯和苯乙烯对海洋微藻生长和光合能力的影响

为了探讨对二甲苯和苯乙烯对海洋微藻的毒性效应,本研究以三角褐指藻和亚心形扁藻为材料,分别比较了不同浓度的对二甲苯和苯乙烯胁迫对微藻生长以及叶绿素含量的影响,并计算对二甲苯和苯乙烯对三角褐指藻和亚心形扁藻96hEC50。研究表明,对二甲苯和苯乙烯对三角褐指藻和亚心形扁藻生长的抑制呈显著的剂量-效应关系,对二甲苯和苯乙烯对三角褐指藻96hEC50分别为60.20mg/L和47.82mg/L,对亚心形扁藻的96hEC50值分别为26.49mg/L和85.62mg/L,相比而言,三角褐指藻对苯乙烯敏感,而亚心形扁藻对对二甲苯更为敏感。同时研究还发现,对二甲苯和苯乙烯胁迫下,试验微藻的叶绿素含量均随处理浓度以及处理时间而显著降低。这些结果为评估对二甲苯和苯乙烯对海洋微藻的毒性效应提供了依据。     

Cu,Zn和Cd对5种单细胞藻的酶基因表达调控的研究

报道了在Cu2 +、Cd2 +和Zn2 +的毒害作用下 ,三角褐指藻 (PhaeodactylumtricornutumBohlin) ,湛江叉鞭藻 (IsochrysiszhanjiangensisHu &Lui) ,绿色巴夫藻 (PaviovaviridisTsengChenetZhang) ,青岛大扁藻 (Platymonashelgolandicavar.TsingtaoensisTsengetT.J.Chang) ,小球藻(Chlorellasp.)等单细胞藻的苹果酸脱氢酶发生了明显的变化。其中 ,不同种类重金属离子对该酶的基因表达作用不同 ,即使是同种离子但浓度不同 ,这种作用也不同。一般情况下高离子浓度抑制了酶基因的表达。低离子浓度促进了表达。但也有例外 ,如Cd2 +(24.53μg/ml)、Zn2 +(24.31,16.21μg/ml)促进三角褐指藻MDH 1座位等位基因的表达量 ;Cd2 +(24.53,12.26μg/ml)促进小球藻MDH 1座位等位基因的表达量。由此可推出 ,重金属离子可以干扰等位基因的表达 ,并在等位酶的活性上表现出来     

Response of antioxidant systems to copper in the gills of the clam Ruditapes decussatus

Copper (Cu) is an essential element for biological systems, however, when present in excess, is toxic. Metallothioneins can play an important role in Cu homeostasis and detoxification. Moreover, Cu can catalyse the production of toxic hydroxyl radicals that cause lipid peroxidation but defence systems in the cells can limit the oxidative damage. The present study was performed to investigate the effect of three Cu concentrations (0.5, 2.5 and 25 microg l(-1)) on the response of antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathion peroxidase and total glutathion peroxidase), total proteins, metallothioneins (MT), malondialdehyde (MDA) concentrations in the gills of the clam, Ruditapes decussatus. The activity of antioxidant enzymes and total proteins, MT and MDA concentrations were measured in the gills of the clams after 1, 3, 7, 14, 21 and 28 days of Cu exposure. Results indicate that Cu only induces an imbalance in the oxygen metabolism during the first week of Cu exposure due to a decrease in mitochondrial SOD and CAT, selenium-dependent and total glutathion peroxidase activities. Cu also causes lipid peroxidation, measured by the MDA formation, that was Cu dependent. In the gills of clams exposed to 25 microg Cu/l, the excess of Cu triggers the induction of MT synthesis after 3 days of exposure.     

Response of antioxidant systems to copper in the gills of the clam Ruditapes decussatus

Copper (Cu) is an essential element for biological systems, however, when present in excess, is toxic. Metallothioneins can play an important role in Cu homeostasis and detoxification. Moreover, Cu can catalyse the production of toxic hydroxyl radicals that cause lipid peroxidation but defence systems in the cells can limit the oxidative damage. The present study was performed to investigate the effect of three Cu concentrations (0.5, 2.5 and 25 μg l⁻¹) on the response of antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathion peroxidase and total glutathion peroxidase), total proteins, metallothioneins (MT), malondialdehyde (MDA) concentrations in the gills of the clam, Ruditapes decussatus. The activity of antioxidant enzymes and total proteins, MT and MDA concentrations were measured in the gills of the clams after 1, 3, 7, 14, 21 and 28 days of Cu exposure. Results indicate that Cu only induces an imbalance in the oxygen metabolism during the first week of Cu exposure due to a decrease in mitochondrial SOD and CAT, selenium-dependent and total glutathion peroxidase activities. Cu also causes lipid peroxidation, measured by the MDA formation, that was Cu dependent. In the gills of clams exposed to 25 μg Cu/l, the excess of Cu triggers the induction of MT synthesis after 3 days of exposure.     

Temporal variation in the antioxidant defence system and lipid peroxidation in the gills and mantle of hydrothermal vent mussel Bathymodiolus azoricus

Hydrothermal vent mussels are exposed continually to toxic compounds, including high metal concentrations and other substances like dissolved sulphide, methane and natural radioactivity. Fluctuations in these parameters appear to be common because of the characteristic instability of the hydrothermal environment. Temporal variation in the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidases (Total GPx), selenium dependent glutathione peroxidases (Se-GPx)), metallothioneins and lipid peroxidation (LPO) in the gills and mantle of the mussel Bathymodiolus azoricus from Menez–Gwen hydrothermal vent site was evaluated and related to the accumulated metal concentrations (Ag, Cu, Cd, Fe, Mn and Zn) in the tissues. Maximum antioxidant enzyme activities in the gills were detected in the beginning of summer, followed by a gradual decrease throughout the following months. One year after, the levels of antioxidant enzyme activities were similar to those reported one year before. LPO in this tissue exhibited a similar temporal variation trend. A different pattern of temporal variation in antioxidant enzyme activities was observed in the mantle, with a gradual increase from summer to the end of autumn (November). LPO in the mantle exhibited an almost reverse trend of temporal variation to that of antioxidant enzyme activities in this tissue. Antioxidant defences in the gills of B. azoricus were significantly enhanced with increasing concentrations of Ag, Cu and Mn, while negative relationships between antioxidant enzymes and Cd, Cu, Mn and Zn concentrations in the mantle were observed, suggesting different pathways of reactive oxygen species (ROS) production and that these tissues responded differently to the metal accumulation. However, temporal variation in biomarkers of defence and damage were in general similar to coastal bivalve species and can be associated with temporal variations of the physiological status due to reproduction. These variations might also be linked to the highly unstable nature of the hydrothermal environment.     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se-GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 microM), Cu (0.4 microM) and Hg (0.1 microM) with different times of exposure. The experiments were performed in pressurized containers at 9+/-1 degrees C and 85 bars. Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.     

活性氧对3种海洋微藻生长的影响

以Fe2+/Vc为活性氧诱生系统,对活性氧作用下3种海洋微藻的生长情况进行了研究.结果表明,低浓度的活性氧可以促进3种海洋微藻的生长,使藻细胞的脂质过氧化程度升高,同时刺激细胞的SOD活性使其略有上升,加入抗氧化剂可以抑制其促进作用;高浓度的活性氧能抑制3种海洋微藻细胞的生长,使细胞的脂质过氧化程度过量增高,抑制细胞的SOD活性,抗氧化剂可以缓解其对藻细胞的毒性作用.不同种类的微藻对活性氧的敏感性存在差异.     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se–GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 μM), Cu (0.4 μM) and Hg (0.1 μM) with different times of exposure. The experiments were performed in pressurized containers at 9 ± 1 °C and 85 bars.Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.     

Metallothioneins and cytosolic metals in Neomysis integer exposed to cadmium at different salinities

In the present study the induction of metallothioneins (MTs) and its relation to cytosolic metal concentrations (Zn, Cu and Cd) in the euryhaline crustacean Neomysis integer exposed to Cd at different salinities was studied. N. integer was exposed to the same free cadmium ion activity of 5.74 x 10(-9) mol l(-1) (i.e. 1/5 of the 96 h LC(50) value expressed as cadmium activity) in hypo-osmotic (5 psu), isosmotic (16 psu) and hyper-osmotic media (25 psu) for 7 days. In this way, the effect of salinity on cadmium speciation was eliminated and therefore the physiological effect of salinity on Cd accumulation and MT induction could be studied. The accumulation of cytosolic Cd in N. integer changed with salinity from 1.11+/-0.05 micromol l(-1) at 5 psu up to 1.43+/-0.17 micromol l(-1) at 25 psu. This could indicate that the physiological response of euryhaline estuarine invertebrates like N. integer to salinity changes can influence the rate of trace metal uptake from solution. While the salinity changes did not cause significant differences in cytosolic Zn concentrations (mean value of all tested salinities: 34.4+/-2.8 micromol l(-1)), an inverse relationship between salinity and cytosolic Cu concentration was observed. The highest concentration of 15.7+/-2.3 micromol Cul(-1) was determined at 5 psu and the lowest 10.9+/-1.4 micromol Cul(-1) at 25 psu. This could point to a possible relationship between the copper concentration and the hemocyanin metabolism in N. integer. This is the first time that differential pulse voltammetry method was applied to MT assays with N. integer. Although the exposure to Cd resulted in a higher Cd cytosolic concentration, no subsequent MT increase was detected. The significant positive correlation between MT levels and cytosolic Cu concentrations (Spearman correlation coefficient r(s)=0.356, p=0.009) implies a strong relationship between MT and Cu in N. integer.     

Photosynthetic activity and antioxidative response of seagrass Thalassia hemprichii to trace metal stress

This study concerned the accumulation of trace metals in tissues of seagrass (Thalassia hemprichii) exposed to various concentrations of Zn²⁺+, Cd²⁺+, Pb²⁺+ and Cu²⁺+ for 10 d, and the effect of excessive metals on quantum yield (ΔF=F_m'), photosynthetic pigments and antioxidative enzymes like superoxide dismutase (SOD), guaiacol peroxidase (POD) were also examined. Cadmium was the most highly accumulated metal. Meanwhile, high metals levels led to a remarkable breakdown of photosynthetic parameters. Especially, ΔF=F_m', chlorophyll and carotenoid were significantly low during prolonged Cu exposure. Besides, ΔF=F_m' was more severely depressed by Cu and Zn than Pb and Cd. However, T. hemprichii had positive response by increasing the activity of SOD and POD. The results indicate that T. hemprichii is the most sensitive to Cu, and the antioxidative protection mechanisms of T. hemprichii are more efficiently activated to avoid damage of Zn, Cd and Pb stress. Finally, due to the high Cd-accumulation and strong Cd-tolerance capacity, T. hemprichii can be used for phytoremediation in Cd-contaminated areas.     

Metallothionein-like proteins induced by cadmium stress in the scallop Mizuhopecten yessoensis

Organisms have evolved a cellular response called stress protein response that increases their tolerance in adverse environmental conditions. Well known stress proteins that bind essential and toxic metals are metallothionein (MT). The scallop Mizuhopecten yessoensis is the most interesting organism because it is able to accumulate toxic cadmium in its digestive gland. However, in the tissue of the digestive gland of Mizuhopecten yessoensis MT (metallothioneins) have not been found. Eastern scallops, Mizuhopecten yessoensis, were collected from two locations ?? one clean and one polluted site. The concentrations of cadmium (Cd), copper (Cu) and zinc (Zn) were measured in the digestive gland. There was a significant increase in Cd concentrations in this studied tissue. We found that in the presence of cadmium Mizuhopecten yessoensis can induce high molecular proteins. The results of experiments have shown that Cd-binding ligands have a number of properties similar to MT: acetone and temperature stability; the ability to bind some metals, including Cd, Cu and Zn. Protein chromatography (FPLC, Superosa 12) from the digestive gland of scallop M. yessoensis has shown that cadmium is associated with high molecular weight Cd-binding proteins (72 kDa and 43 kDa). The major cadmium-binding protein 72 kDa is glycoprotein. In experiments we have demonstrated that Cd-binding proteins can be induced when there is cadmium exposure. The results of this study strongly suggest that the far eastern scallop Mizuhopecten yessoensis has a unique and well-developed system for the detoxification of heavy metals and it allows for biochemical systems to be maintained in a relatively stable manner in the presence of heavy metals.     

镉对中华绒螯蟹(Eriocheir sinensis)抗氧化酶活性的影响

将中华绒螯蟹(河蟹Eriocheir sinensis)浸泡在2．Omg/L氯化镉(CdCl2)溶液中，分别于第0、6、12、24、48、72、96小时提取河蟹的肝胰腺，测定抗氧化酶：超氧化物歧化酶(Superoxicle dismutase，SOD)、过氧化氢酶(Catalase，CAT)、谷胱甘肽过氧化物酶(Glutathione Peroxiclase．GPx)的活性，探讨了镉(Cd^2 )对河蟹肝胰腺抗氧化酶活性的影响。结果显示，在低浓度Cd^2 环境下，河蟹肝胰腺的抗氧化酶活性随时间发生规律性变化。暴露于Cd^2 后，河蟹肝胰脏SOD活性降低，随时间的延长，在暴露72h后，SOD活性恢复并超过未染毒时22．3％。表明Cd^2 中毒导致细胞内氧自由基大量积累从而诱导酶活性升高；CAT活性先下降后增高，随后减少，最后以低于对照的水平趋于平衡；肝胰腺中GPx酶活性的变化规律与SOD相似，在解除氧自由基毒性方面有一定的协调性。实验表明河蟹抗氧化系统酶对Cd^2 很敏感。而酶活性在短时间内的变化规律可以为镉对河蟹早期的毒性研究提供参考。