壬基酚对赤子爱胜蚓的生态毒理学研究

环境激素对生态的影响越来越受到人们的重视。通过赤子爱胜蚓对壬基酚的滤纸接触法毒性试验,从分子水平揭示环境污染物对蚯蚓组织的毒性影响。在壬基酚对赤子爱胜蚓(Eiseniafoetida)的急性毒性试验中,测得72hLC50为0.908μg/cm2。在系列浓度的壬基酚暴露中,过氧化氢酶(Catalase,CAT),超氧化物歧化酶(Superoxidedismutase,SOD)和谷胱甘肽硫转移酶(GlutathioneS-transferase,GST)的敏感性依次为CAT>SOD>GST。实验结果表明,这三种酶均可作为早期NP污染预警指标。     

Induction of glutathione reductase enzyme activity by β‐aminobutyric acid in plant leaves

Abstract

Treatment of pea and tobacco leaf discs with the resistance inducer DL‐β‐amino‐n‐butyric acid (BABA) led to a substantial induction of glutathione reductase (GR, E.C. 1.6.4.2.) enzyme activity. After exposure to 1 mM BABA for 96 hrs, the GR activities were 3.2‐fold and 2.9‐fold higher in pea and tobacco leaf discs, respectively, than GR activities in untreated controls. Elevated GR levels may contribute to the antioxidative protection of plants during pathogen attack.     

Safening activity of natural hydroxamic acids and analogous compounds against herbicide injury to maize

Abstract

Safening activities of natural compounds DIMBOA, DIBOA, and MBOA, as well as synthetic 1,4‐benzoxazin‐3‐ones were tested against acetochlor and EPTC injuries to maize. No safening activities of natural products and from low to moderate activity of synthetic benzoxazinones were observed. In order to explain inefficacy of natural compounds we studied the influence of these molecules on enzymes participating in metabolic detoxication of acetochlor and EPTC. Pretreatment with DIMBOA elevated maize cytochrome P450 levels. Pretreatments with chemicals containing 1,4‐benzoxazin‐3‐one backbone did not alter glutathione S‐transferase enzyme activities. However, all natural products inhibited glutathione S‐transferase activity of roots and shoots in vitro after addition to the enzyme. Safening ineffectiveness of natural hydroxamic acids may be explained by their inhibitory effects on GST enzymes due to their reaction with sulfhydryl groups on the enzyme.     

真鲷去毒相关基因cDNA序列的克隆与肝脏组成型表达分析

赤潮发生时产生的一些海洋生物毒素对人类和海洋动物的安全形成潜在的威胁甚至导致死亡.为从分子水平探讨鱼类中海洋藻毒素的去毒分子机理,采用RT-PCR法克隆了真鲷(Pagrus major)肝脏芳香烃受体核转位蛋白(ARNT)和I时相异生素代谢酶细胞色素P4501A(CYP1A)基因cDNA核心序列,同时,应用半定量RT-PCR方法,以β-肌动蛋白作为外参照,在其指数期增长的范围内研究了芳香烃受体(AHR)、ARNT、CYP1A、II时相异生素代谢酶alpha型谷胱甘肽S-转移酶(GSTA1、GSTA2)、rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70(HSP70)基因组成型表达水平.结果发现,真鲷ARNT、CYP1A基因cDNA核心序列片段分别长438bp和908bp,分别编码146和302个氨基酸.序列同源性分析发现,真鲷与门齿鲷(Stenotomus chrysops)、石首鱼(Micropogon undulatus)ARNT基因氨基酸序列同源性高达97.2%、95.2%,与斑马鱼(Brachydanio rerio)、人、大鼠、小鼠ARNT同源性较低(77.2%～79.3%).真鲷与门齿鲷、金头鲷(Sparus auratus)、欧洲川鲽(Rhombus maximus)、欧洲海鲈(Dicentrachus labrax)CYP1A基因氨基酸序列同源性较高,为84.8%～94.0%,与斑马鱼、人、小鼠CYP1A同源性较低,为59.6%～77.8%.真鲷肝脏AHR、ARNT、CYP1A、GSTA1、GSTA2、GSTR和HSP70基因组成型表达水平分别为(25.32±6.56)%、(26.22±4.24)%、(146.5±16.06)%、(55.42±3.75)%、(48.82±10.89)%、(79.47±3.13)%、(107.42±14.34)%.     

PCDD/Fs-induced oxidative damage and antioxidant system responses in tobacco cell suspension cultures

Polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) are ubiquitous contaminants and can be considerably accumulated by natural plants. In order to elucidate the biochemical and physiological responses of plant to PCDD/Fs, tobacco Bright Yellow-2 (BY-2) cells were selected as model plant and treated with time- and concentration-dependent PCDD/Fs. The toxic effect and oxidative stress caused by PCDD/Fs were evident, which could be indicted by the reduction in fresh mass, the increase in malondialdehyde (MDA) content, and the damage of tobacco cell ultrastructure. PCDD/Fs tolerance was correlated with changes in antioxidant system and hormones of tobacco cells. Superoxide dismutase (SOD) and peroxidase (POD) exhibited peak enzyme activities at the PCDD/Fs concentration of 1000 ng WHO₉₈-TEQ g⁻¹ fresh weight. Glutathione reductase (GR) enzyme activity increased monotonically at high level PCDD/Fs, but the activity of catalase (CAT) was only slightly affected at all treatment. Meanwhile, the exposure to PCDD/Fs resulted in the changes of hormones content. With the increase of exposure concentration of PCDD/Fs, the levels of indole-3-acetic acid (IAA) and abscisic acid (ABA) increased, whereas the concentration of jasmonates (JAs) decreased. The above results suggest that tobacco cells had the ability to cope with the oxidative stress induced by low concentration of PCDD/Fs through increasing the activities of antioxidant enzymes and alternating plant hormones levels. However, oxidative stress and toxicity would burst out when plant cells were exposed to the high levels of PCDD/Fs.     

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL^?1 for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL^?1 (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL^?1 in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.     

Growth, photosynthesis and antioxidant responses of two microalgal species, Chlorella vulgaris and Selenastrum capricornutum, to nonylphenol stress

The effect of nonylphenol (NP) on growth, photochemistry and biochemistry of two green microalgae, Chlorella vulgaris and Selenanstrum capricornutum, and their ability to degrade NP were compared. The 96 h EC₅₀ of C. vulgaris and S. capricornutum were greater than 4.0 and 1.0 mg L⁻¹ NP, respectively, suggesting that the former species was more tolerant to NP. Both microalgae acclimated to NP stress through down-regulating their photosynthetic activities, including antenna size (chlorophyll a content), maximal photochemistry (Fv/Fm) and the light absorbed by PSII (ABS/CS₀), but the dissipation of energy from reaction centres (DI₀/RC) increased with the increase of NP concentrations. In C. vulgaris, the changes of these parameters were more significant than in S. capricornutum and recovered completely after a 96 h exposure. The antioxidant responses, such as GSH content, CAT and POD activities in C. vulgaris increased with the increase of NP concentrations after a 24 h exposure, but these changes disappeared with exposure time and recovered to the control levels after 96 h. In S. capricornutum, although GSH content, CAT and POD activities also increased when exposed to low- to moderate-NP concentrations, these values were significantly reduced at a high concentration (4 mg L⁻¹) even after a 96 h exposure, indicating its antioxidant responses were significantly delayed. It is clear that the more NP-tolerant species, C. vulgaris, acclimated better with a faster recovery of its photosynthetic activity from the NP-induced damage, and exhibited more efficient and rapid responses to NP-induced oxidative stress. C. vulgaris also had a higher NP degradation ability than S. capricornutum.     

Determination of the mechanism of photoinduced toxicity of selected metal oxide nanoparticles (ZnO, CuO, Co3O4 and TiO2) to E. coli bacteria

Cytotoxicity of selected metal oxide nanoparticles(MNPs)(ZnO,CuO,Co 3 O 4 and TiO 2)was investigated in Escherichia coli both under light and dark conditions.Cytotoxicity experiments were conducted with spread plate counting and the LC 50 values were calculated.We determined the mechanism of toxicity via measurements of oxidative stress,reduced glutathione,lipid peroxidation,and metal ions.The overall ranking of the LC 50 values was in the order of ZnO < CuO < Co 3 O 4 < TiO 2 under dark condition and ZnO < CuO < TiO 2 < Co 3 O 4 under light condition.ZnO MNPs were the most toxic among the tested nanoparticles.Our results indicate depletion of reduced glutathione level and elevation of malondialdehyde level correlated with the increase in oxidative stress.Released metal ions were found to have partial effect on the toxicity of MNPs to E.coli.In summary,the dynamic interactions of multiple mechanisms lead to the toxicity of the tested MNPs to E.coli.     

Effects of insoluble Zn, Cd, and EDTA on the growth, activities of antioxidant enzymes and uptake of Zn and Cd in Vetiveria zizanioides

A root-bag experiment was conducted to study the effects of insoluble Zn, Cd and ethylenediaminetetraacetic acid (EDTA) on the plant growth, activities of antioxidant enzymes, proline, glutathione (GSH), water-soluble proteins and malondialdehyde (MDA) of Vetiveria zizanioides, and its uptake capacity of Zn and Cd.The results showed that plant growth of V.zizanioides was inhibited by Zn and Cd.The shoot dry weight (SDW) and root dry weight (RDW) decrease by 0.3%, 8.0%, 9.4% and 10.4%, 15.1%, 24.2% compared to the control without EDTA addition, respectively.After adding EDTA, shoot and root dry weight decreased over 10% and 15% compared to results in the absence of EDTA, respectively.The toxicity from insoluble Zn and Cd in soil on SDW and RDW of V.zizanioides was in order: Zn+Cd > Cd > Zn.The activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), and contents of MDA and proline significantly increased, while the contents of GSH and water-soluble proteins markedly decreased with increasing Zn and Cd toxicity.With EDTA, shoot and root Zn concentrations increased in the Zn treatment by 7.3% and 37.4% compared to the plants grown in absence of EDTA.Shoot and root Cd concentrations in the combined Zn and Cd treatment with EDTA increased by 18.6% and 391.9% compared to the treatment without EDTA.However, Zn and Cd concentrations in shoot and roots decreased in the Cd treatment compared to the plants grown in absence of EDTA, with exception of root Cd concentration in the presence of EDTA).     

Effects of copper on lipid peroxidation,glutathione, metallothionein,and antioxidative enzymes in the freshwater mussel Anodonta anatina

Copper is an essential element to all animals. At elevated concentrations, it is toxic and can participate in the formation of reactive oxygen species, leading to cellular damage. In this study, the ecotoxicological relevance of copper was investigated with freshwater mussels, Anodonta anatina. When the mussels were exposed to copper at environmentally realistic concentrations, either via the water (0.3?µmol?L^?1 Cu) or fed with Cu-loaded algae (equivalent to 0.06?µmol?L^?1 Cu), the level of thiobarbituric acid-reactive substances rose and glutathione decreased. This was associated with the induction of metallothionein and, relative to total protein, of glutathione reductase and the antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase. But, since the overall protein-synthetic capacity was hampered by the copper insult, the activities of the enzymes relative to tissue weight and copper concentrations were depressed. During depuration, most parameters started to normalize although not returning to control values within 12 days.