铜离子对中国花鲈幼鱼的毒性研究

为给中国花鲈(Lateolabrax maculatus)养殖提供污染生物学的理论数据,研究了中国花鲈幼鱼的铜离子中毒症状和半致死浓度(LC50);检测了幼鱼肝脏的谷胱甘肽(GSH)和丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷丙转氨酶(GPT)和谷草转氨酶(GOT)活力;分析了铜离子胁迫对...     

水环境中重金属铜暴露下青鳉鱼的行为响应

采用生物行为传感器监测青鳉鱼在重金属铜暴露下的行为数据,分析不同暴露浓度(20、10、5、1和0.1 TU)下青鳉鱼的行为响应。20、10、5、1和0.1 TU的暴露浓度下青鳉鱼对重金属污染的行为反应模式符合环境胁迫阈值模型,且不同浓度梯度重金属对青鳉鱼产生不同的行为毒性效应。利用重金属作用下青鳉鱼的行为变化来研究重金属环境胁迫导致的生物行为响应机制,从而得到重金属暴露下生物行为的实时变化过程和趋势,可对水环境生态系统质量进行综合评价。     

水环境中重金属铜暴露下青鳉鱼的行为响应

本研究采用生物行为传感器监测青鳉鱼重金属铜暴露下的行为数据，分析不同暴露浓度(20、10、5、1和0.1 TU)下青鳉鱼的行为响应。20、10、5、1和0.1 TU的暴露浓度下青鳉鱼对重金属污染的行为反应模式符合环境胁迫阈值模型，且不同浓度梯度重金属对青鳉鱼产生不同的行为毒性效应。通过本研究，利用重金属作用下青鳉鱼的行为变化来研究重金属环境胁迫导致的生物行为响应机制，从而得到重金属暴露下生物行为的实时变化过程和趋势，可对水环境生态系统质量进行综合评价。     

Molecular dynamics of stability and structures in phytochelatin complexes with Zn,Cu, Fe,Mg, and Ca: Implications for metal detoxification

Phytochelatins, or (γ-glutamyl-cysteine) _n -glycine, are specialized peptides produced by plants and algae to mitigate toxic metal exposure, for instance in response to high levels of metals such as Cu, Cd, and Zn. Stability constants and structural characterization of metal–phytochelatin complexes are lacking. This information is required to gain mechanistic insights on the metal selectivity of phytochelatins. Here, we studied structural coordination and thermodynamic stability by performing molecular dynamics simulations of a fully hydrated phytochelatin molecule complexed with Ca²⁺, Mg²⁺, Fe²⁺, Zn²⁺, and Cu²⁺. Our results predict the following decreasing order for the thermodynamic stability of the phytochelatin complexes: Zn²⁺ ≥ Cu²⁺ ≥ Fe²⁺ > Mg²⁺ > Ca²⁺. The favorable binding energies with Zn²⁺ and Cu²⁺ over the other metal cations can be explained by shorter binding distances and greater coordination from carboxylate and keto O atoms. Conformational rearrangement of phytochelatin following metal chelation was captured by monitoring changes in the solvent-accessible volume. Accessibility of solvent molecules to the phytochelatin structure was inversely proportional to the distance between the coordinated ligands and the chelated metal. These new findings demonstrate the influence of the metal–phytochelatin structure on the metal-binding thermodynamics and the phytochelatin conformation, both of which are important to evaluate the intracellular role of phytochelatin in mediating algal response to toxic heavy metal exposure.     

Modifications of photochemical efficiency,cellular viability and total phenolic content of Prosopis glandulosa leaves exposed to copper

The effects of copper (Cu²⁺) on photochemical efficiency, cellular viability and total phenolic content in Prosopis glandulosa leaves were evaluated after 4, 8 or 12 h exposure to 0.1, 10 or 100 mM Cu²⁺ using the tissue tolerance test. The results showed that significant reductions (p<0.003) of photochemical efficiency in P. glandulosa leaves were found after 8 h exposure to 100 mM Cu²⁺ compared with controls leaves. The results also showed a significant increase (p<0.05) on electrolyte leakage values in P. glandulosa leaves after 4 h exposure to 100 mM Cu²⁺ compared with control leaves. Total phenolic content in P. glandulosa leaves only differed significantly when exposed 12 h to the highest concentration of Cu²⁺. These results suggest that Cu²⁺ can produce reactive oxygen species that may form hydroxyl radicals that damage proteins and lipids, which in turn causes loss of integrity and reduce photochemical efficiency of leaves. In conclusion, the photochemical efficiency and cellular viability analyses can be used as useful physiological tools to assess early changes in photosynthetic performance of P. glandulosa in response to copper pollution.     

Acute toxicity of copper oxide nanoparticles to Daphnia magna under different test conditions

The acute toxicity of monodispersed 6 nm and <100 nm poly-dispersed copper oxide nanoparticles toward Daphnia magna was assessed using 48 h immobilization tests. CuSO₄ was used as a reference. Four different exposure conditions were tested, to study whether the toxicity of the nanoparticle suspensions changed in a way similar to what is known for dissolved Cu: first in ISO standard test conditions (pH 7.8), second with slight acidity (pH 6.5), third in the presence of citric acid, and fourth in the presence of humic acid. For all four exposure conditions, the toxicity of Cu employed in the three forms followed the same sequence, i.e., CuSO₄ > monodispersed 6 nm CuO ? poly-dispersed CuO. The toxicity of all Cu forms decreased from pH 6.5, ? pH 7.8, > pH 7.8 + citric acid, to ? pH 7.8 + humic acid. This pattern is in agreement with concentrations of Cu²⁺ calculated using the equilibrium model MINTEQ. These findings show that the acute toxicity of copper oxide nanoparticles is governed by test water composition and the chemical species Cu²⁺.     

Comparative Toxicity of Copper and Zinc to Isolated Eel Hepatocytes

Attempts were made to elucidate lethal responses of hepatocytes of Japanese eel (Anguilla japonica) exposed to Krebs-HEPES buffer at different concentrations of Cu⁺⁺ and Zn⁺⁺. It was found that the 2-hr LC₅₀ value of Cu⁺⁺ and Zn⁺⁺ on eel hepatocytes was 490 and 1576 μM for Cu⁺⁺ and Zn⁺⁺ in single solution, and 1734 and 5200 μM in mixed solution. the toxic effects of Cu⁺⁺ and Zn⁺⁺ on eel hepatocytes were characteristic of antagonistic interaction. After 2-hr exposure, the amount of accumulated metal on the hepatocytes and the mortality of hepatocytes were increased with increasing metal concentration in Krebs-HEPES buffer. the accumulated amount of both metals was significantly decreased when the eel hepatocytes were exposed to mixed metals than to single metal.     

Heavy-metal adsorption by non-living biomass

The adsorption of some heavy metals onto the walls of harvested, washed, and dried non-living biomass cells of different Pseudomonas strains was studied at optimum experimental conditions using a simplified single component system. The Langmuir adsorption model was found to be a suitable approach to describe the system via multi-step processes. Isotherms measured at 30.0°C and pH 5.5 with [M]_total?=?10–100 mM for tight, reversible Cr⁶⁺(aq), Ni²⁺(aq), Cu²⁺(aq) and Cd²⁺(aq) binding by the cell walls of the investigated biomass fit the Langmuir model and give the pH-independent stoichiometric site capacities ν _i and equilibrium constants K _i for metal binding at specific biomass sites i?=?A, B, C, and D. Tight binding sites A, B, and D of the non-living biomass are occupied by Cr^VI, sites A and C by Ni^II, sites A and D by Cd^II, and only site B by Cu^II. It is concluded that ν _i is a stoichiometric parameter that is independent of the magnitude of K _i for binding site i and that the studied heavy metals selectively and tightly bind at different biomass sites.     

Influence of arsanilic acid,Cu<Superscript>2+</Superscript>, PO<Subscript>4</Subscript><Superscript>3–</Superscript> and their interaction on anaerobic digestion of pig manure

Arsanilic acid (ASA), copper ion (Cu²⁺) and phosphate (PO₄ ^3–) are widely used as feed additives for pigs. Most of these three supplemented feed additives were excreted in feces and urine. Anaerobic digestion is often used for the management of pig manure. However, the interaction of ASAwith Cu²⁺ or PO₄ ^3– on anaerobic digestion is still not clear. In this study, the influence of ASA, Cu²⁺, PO₄ ^3– and their interaction on anaerobic digestion of pig manure and the possible mechanisms were investigated. The initial concentrations of ASA, Cu²⁺ and PO₄ ^3– were 0.46 mM, 2 mM and 2 mM in the anaerobic digester, respectively. The methanogenesis was severely inhibited in the assays with only ASA addition, only Cu²⁺ addition and ASA + PO₄ ^3– addition with the inhibition index of 97.8%, 46.6% and 82.6%, respectively, but the methanogenesis inhibition in the assay with ASA + Cu²⁺ addition was mitigated with the inhibition index of 39.4%. PO₄ ^3– had no obvious impacts on the degradation of ASA. However, Cu²⁺ addition inhibited the degradation of ASA, and mitigated the methanogenesis inhibition. The existence of ASA would inhibit methanogenesis and generate more toxic inorganic arsenic compounds during anaerobic digestion, implying the limitation of anaerobic digestion for ASA- contaminated animal manure. However, the co-existence of ASA and Cu²⁺ could mitigate the inhibition. These results could provide useful information for the management of anaerobic digestion of pig manure containing ASA and Cu²⁺.

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Sorption of cadmium(II) and copper(II) by soil humic acids: temperature effects and sorption heterogeneity

Sorption by humic acids is known to modify the bioavailability and toxicity of metals in soils and aquatic systems. The sorption of cadmium(II) and copper(II) to two soil humic acids was measured at pH 6.0 using ion-selective electrode potentiometric titration at different temperatures. Sorption reactions were studied with all components in aqueous solution, or with the humates in suspension. Adsorption reactions were described using a multiple site-binding model, and a model assuming a continuous log-normal distribution of adsorption constants. Adsorption of Cu²⁺ was more favourable than adsorption of Cd²⁺. The log-normal distribution model provided the closest fit to observations and allowed parameterisation of adsorption data using a mean adsorption constant (log K _μ). Sorption of Cd²⁺ to dissolved humic acids increased slightly in extent and sorption affinity with increasing temperature, but the effect was small (log K _μ 2.96–3.15). A slightly greater temperature effect occurred for sorption of Cd²⁺ to solid-phase humic acids (log K _μ 1.30–2.08). Sorption of copper(II) to both aqueous- and colloidal-phase humates showed more pronounced temperature dependence, with extent of sorption, and sorption affinity, increasing with increasing temperature (log K _μ 3.4–4.9 in solution and 1.4–4.5 in suspension). The weaker adsorption of Cd²⁺ than Cu²⁺, and smaller temperature effects for dissolved humates than suspended humates, suggested that the observed temperature effects had a kinetic, rather than thermodynamic, origin. For any metal-to-ligand ratio, free metal ion concentration, and by inference metal bioavailability, decreased with increasing temperature. The consistency of the data with kinetic rather than thermodynamic control of metal bioavailability suggests that equilibrium modelling approaches to estimating bioavailability may be insufficient.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Acclimation and tolerance of Artemia salina and Ophryotrocha labronica to cooper sulphate

The brine shrimp Artemia salina L. and the polychaete worm Ophryotrocha labronica La Greca and Bacci were acclimated in sea water with copper sulphate at concentrations of 0.1, 0.05, and 0.025 ppm Cu⁺⁺, for 3 and 2 generations, respectively. Both adults and larvae of A. salina showed a greater tolerance to 1 ppm Cu⁺⁺ after acclimation compared to controls of the same age, although this tolerance diminished in successive generations. The acclimation effect was less marked in O. labronica. In both species, tolerance to 10 ppm Cu⁺⁺ upwards was not enhanced. Growth-rate inhibition and an adverse effect on reproduction was observed, in some instances in direct relationship to the acclimation concentration. It is suggested that, in A. salina, a certain tolerance to copper may be acquired through exposure to low concentrations.     

水体酸化条件下Cu(II)对斑马鱼胚胎的毒性效应

为评价由酸雨、酸性矿山排水等环境污染导致的水体酸化及水体重金属联合作用对水生生物的生态毒性效应,研究了不同pH值(pH=3、4、5、6、7和7.8)条件下Cu2+对斑马鱼胚胎发育的影响。结果表明,酸性水体及Cu2+单一存在时,酸对斑马鱼胚胎24h半数效应浓度值EC50为pH=3.65,Cu2+(pH=7.8)对斑马鱼胚胎24h-EC50为0.267mg·L-1;当水体酸化及水体中的Cu2+共存时,较低的pH对Cu2+的生物毒性起协同作用,表现为随溶液pH的降低,各浓度Cu2+对斑马鱼胚胎的24h凝结率显著增高(P24h致死率=0.001),而斑马鱼胚胎96h孵化率显著降低(P96h孵化率=0.002),且不同浓度的Cu2+之间的生物毒性效应存在显著性差异(P24h致死率=0.0321;P96h孵化率=0.0028)。这说明酸性水体和Cu2+都显著影响斑马鱼胚胎的发育,且Cu2+在酸性水体中对斑马鱼胚胎的毒性显著增强。因此,在受重金属Cu2+污染的地区,如同时受到酸雨或酸性矿山排水等较低pH值和Cu2+的双重胁迫,较低浓度的Cu2+就能够对水生生物的生殖发育及水生生态系统产生严重的影响和危害。     

Effects of lead exposure on redox status,DNA and histological structures in Venus verrucosa gills and digestive gland

ABSTRACT

Lead (Pb) is one of the most toxic heavy metals that affect the physiological status of aquatic organisms. The present investigation evaluated the possible toxic effect of lead chloride (PbCl₂) on biomarkers responses, DNA damage and histological alterations in Venus verrucosa gills and digestive gland. Three concentrations of PbCl₂ (D1:1µgL^?1, D2: 10µgL^?1 and D3: 100µgL^?1) were chosen for V. verrucosa exposure during six days. At the end of the trial, it was found that Pb tended to accumulate in both gills and digestive gland in a dose-dependent manner. However, gill tissues exhibited the highest metal burden. Our results showed an increase of malondialdehyde, protein carbonyls and advanced oxidation protein product levels in both organs following PbCl₂exposure. The induction of both non-enzymatic and enzymatic antioxidant systems; as well as the decrease of the acetylcholinesterase activity and degradation of DNA structure was recorded in the gills and digestive gland. The histopathological alterations observed in gills (disruption of lamellas and cilia filaments?…) and digestive gland (lumens occlusion, necrosis and fibrosis) confirmed the aforementioned results. Our data highlighted the short-term toxicity effects of PbCl₂ on V. verrucosa and pointed out a high sensitivity of gills towards this metal.     

Alterations in serum electrolyte levels after dimethoate exposure and recovery in the freshwater air-breathing catfish,Heteropneustes fossilis (Bloch)

Dimethoate, a moderately toxic insecticide, has a wide range of agricultural and domestic applications. Like other organophosphates, dimethoate has anticholinesterase activity. Fish are non-target organisms, inadvertently exposed to pesticides and their metabolic products. The present study includes short-term (96 h) and long-term (36 d) effects of dimethoate exposure on some serum electrolytes Ca²⁺, Mg²⁺, and Pi in the freshwater air-breathing catfish Heteropneustes fossilis. The concentration of dimethoate for short-term exposure was 2.24 mg L^?1 (75% of 96 h LC₅₀) and for long-term exposure 0.75 mg L^?1 (25% of 96 h LC₅₀). The study includes the recovery pattern in serum electrolytes after placing the fish in pesticide-free water. Fish show hypocalcemia, hypermagnesemia, and hyperphosphatemia after short-term and long-term exposure to dimethoate. When placed in pesticide-free water, these electrolytes exhibit recovery towards normalization, indicating significant (p < 0.05) recovery.     

Biokinetic modeling of heavy metals in earthworms

Studies concerning bioaccumulation kinetics and bioconcentration factor (BCF) of heavy metals like zinc (Zn), lead (Pb), chromium (Cr), cadmium (Cd), and copper (Cu) in earthworm Eudrilus eugeniae tissues including integument, gizzard, clitellum, and head region were undertaken. Calculated BCF, predicted K _ow, and predicted K _oc showed a significant correlation between heavy metals in different earthworm tissues, in substrate spiked with heavy metals. The regression coefficient (r ²) between heavy metal uptake concentration and exposure time varied between 0.73 and 0.99, indicating significant correlation. The K _oc was a maximum of 13.9016 in case of Cu and integument at an exposure time of 100 days and a minimum of 0.1114 in case of Cr with respect to head at the same exposure time. Earthworms accumulated heavy metals following chronic exposure to municipal solid waste containing heavy metals. BCF and uptake rate kinetics of heavy metals were calculated and showed increased values in head tissue followed by integument.     

Evaluation of hemopoietic responses in Labeo rohita Hamilton following acute copper toxicity

Fish are often exposed to highly contaminated water, especially in areas where the dilution rate of waste water is low. Environmental pollutants or other stress may affect one or more of the immunological functions in fish. Effective control of toxic substances is essential for the success of fish culture, which requires fish health monitoring using biomarkers. This study evaluated the effects of heavy metal copper (Cu) at environmentally relevant concentrations on the hematopoiesis in Labeo rohita. L. rohita fingerlings were exposed to different Cu concentrations for 6, 24, 48, or 72 hr. Following exposure, percentage blast cells was found to be increased after 6 and decreased after 24, 48, and 72 hr both at sublethal and LC₅₀ concentrations, whereas percentage mature erythrocytes decreased after 6 and increased after 24, 48, and 72 hr. Erythropoietic and leukopoietic efficiencies were also affected upon exposure. Erythropoietic and leucopoietic efficiencies increased markedly after 6, 24, and 48 hr but decreased after 72 hr in sublethal-treated fish. Erythropoietic efficiency decreased significantly after 6, 24 and 48 hr but increased after 72 hr in LC₅₀-treated fish. Leukopoietic efficiency decreased significantly after 6 hr exposure and increased after 24 and 48 hr but decreased after 72 hr. Flow cytometric studies of head kidney cell cycle phase distribution revealed cell death and DNA synthesis. The percentage cell death increased in fish exposed to sublethal concentrations and rose further at LC₅₀ during the earlier hours of exposure. Synthesis of DNA was increased at sublethal concentrations but was significantly reduced at LC₅₀ dose.     

镉和铅对菲律宾蛤仔脂质过氧化及抗氧化酶活性的影响

为了研究亚致死浓度的重金属对海洋贝类的毒性效应,探讨其可能的作用机理,在实验生态条件下以菲律宾蛤仔(Ru-ditapes philip pinarum)为目标生物,采用半静态毒性实验方法,研究了不同浓度Cd2+(0.0948、0.237和0.474mg·L-1)和Pb2+(0.276、0.690和1.380mg·L-1)对菲律宾蛤仔鳃和消化腺组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及丙二醛(MDA)含量的影响。结果显示:(1)鳃和消化腺中的抗氧化酶及MDA的变化呈现出类似的趋势,在胁迫初期,各浓度处理组菲律宾蛤仔鳃和消化腺组织SOD和CAT与对照组相比活性都显著升高(P<0.01),呈现出明显的诱导效应,而MDA含量与对照组相比差异不显著(P>0.05);随着胁迫时间的延长,高浓度Cd2+(0.474mg·L-1)和Pb2+(1.380mg·L-1)处理组中SOD、CAT活性快速下降,与对照组相比差异显著;低浓度处理组中的抗氧化活性虽然也较对照组有所下降,但总体下降幅度不如高浓度组明显,并且所历暴露时间较长。各浓度处理组中MDA含量变化一致,均呈现出先升高后降低的趋势,且含量均高于对照组。(2)通过相关性分析,菲律宾蛤仔消化腺组织中的SOD活性与Cd2+浓度的相关性大于鳃组织,与Pb2+浓度的相关性则小于鳃组织;消化腺组织中的CAT活性与Cd2+、Pb2+浓度呈抛物线型相关,与鳃组织CAT活性相关性不十分显著。这说明消化腺组织中SOD活性对Cd2+的敏感性大于鳃组织,消化腺组织中CAT活性对Cd2+、Pb2+的敏感性大于鳃组织。因此,菲律宾蛤仔消化腺中SOD、CAT对水环境中的重金属反应敏感,且存在一定的剂量-效应关系,消化腺组织中SOD和CAT活性与其他敏感性指标一起可以作为指示早期海洋重金属污染的生物标志物。     

Effects of cadmium on intracellular cation homoeostasis in the yeast Saccharomyces cerevisiae

Previous studies have demonstrated that cadmium can induce biochemical and physiological changes in yeast Saccharomyces cerevisiae. However, studies on the influence of cadmium on the ion balance in the cell and the interaction between cadmium and other ions are still relatively few in number. By using inductively coupled plasma-atomic emission spectrometry, the contents of some cations, including Zn²⁺, Ca²⁺, Fe³⁺, Cu²⁺, Mg²⁺, K⁺, and Na⁺ were measured. The data showed that the levels of Zn²⁺ and Fe³⁺ were increased, while those of Cu²⁺, K⁺, and Na⁺ were decreased after cadmium treatment. Afterwards, using the drop test assay, the interactions between cadmium and the selected ions were investigated. The results suggested that the cytotoxicity of cadmium could be attributable to the interference of cadmium with the intracellular cation homoeostasis. Calcium channel transporter Cch1 participates in the intracellular uptake of cadmium. Additionally, Zn²⁺, Ca²⁺, Fe³⁺, Mg²⁺, and K⁺ can rescue the toxic effect of cadmium in yeast.     

Metabolic responses in zebrafish (Danio rerio) exposed to zinc and cadmium by nuclear magnetic resonance -based metabolomics

Heavy metals are common marine and soil pollutants that are mainly the result of industrial activity, and are a threat to the environment and human health. In this study, ¹H nuclear magnetic resonance (NMR)-based metabolomics was applied to adult Danio rerio to monitor the metabolic change as a response to ZnCl₂ and CdCl₂ exposure at different concentrations for 72?h. NMR spectroscopy was used to identify and quantify the metabolites extracted from D. rerio. The metabolite profiles of the control and heavy metal exposed group were classified by partial least squares – discriminant analysis (PLS-DA) analysis, and potential contaminant-specific biomarkers were suggested. For the ZnCl₂-exposed zebrafish, the levels of ATP, aspartate and NAD⁺ were increased, whereas the levels of formate, inosine, hypoxanthine and succinate decreased. In addition, the CdCl₂-exposed zebrafish showed an increase in the levels of ATP and formate and a decrease in the levels of glutamate, inosine and glutathione. Overall, Zn and Cd may lead to neurotoxicity, disturbances in the energy metabolism and oxidative stress. Our finding demonstrated that the application of NMR-based metabolomics might be useful for detecting the toxicity caused by sub-lethal concentrations of heavy metal contaminants in the environment.