多氯联苯(PCBl254)对栉孔扇贝消化盲囊和鳃丝EROD、GST酶活力的影响

研究了4种质量浓度（0．5μg／L、1μg／L、10μg／L、50μg／L）PCB1254暴露下,栉孔扇贝（Chlamys ferrari）消化盲囊和鳃丝7-乙氧基-3-异吩唑酮-脱乙基酶（EROD）、谷胱甘肽转移酶（GST）活力的变化。于实验开始后0、0．5d、1d、3d、6d、9d、15d、21d、30d取样测定,结果显示,除鳃丝EROD活力在低质量浓度（0．5μg／L和1．0μg/L）随时间变化不明显（P〉0．05）外,其他各处理组PCB1254对栉孔扇贝消化盲囊和鳃丝的EROD都具有明显的激活作用,而且具有明显的剂量-效应关系。低浓度（0．5μg/L、1．0μg/L）处理下,消化盲囊和鳃丝的GST活力都呈现激活的趋势,而高浓度（10μg/L、50μg/L）下,GST活力都呈现先上升后下降的趋势,且鳃丝GST活力变化幅度较为明显。结果表明,EROD和GST活力的变化在一定程度上能够反应PCBs对栉孔扇贝影响的规律性,是合适的毒理学指标,栉孔扇贝是比较好的评价多氯联苯污染的指示生物,本研究也对栉孔扇贝对多氯联苯的生物转化机制作了初步的探讨。[中国水产科学,2008,15（2）：342-346]     

Effects of deltamethrin on hematological parameters and enzymatic activity in Ancistrus multispinis (Pisces, Teleostei)

Deltamethrin (DM) is a pyrethroid insecticide widely used in Brazilian crops and in pest-control programs because of its low environmental persistence and toxicity. It has been shown to exert a wide range of effects on non-targeted organisms including fishes. The aim of this study to evaluate the effects of deltamethrin through the hematological and biochemical parameters using Ancistrus multispinis as animal model. First, blood of A. multispinis was collected by cardiac puncture in order to evaluate the basal values of the total cell counts, hemoglobin and hematocrit rates. A. multispinis showed low number leukocytes, red blood cells and hemoglobin compared to other species. The intoxication with DM (0.1 or 0.3 mg kg⁻¹) induced leukocytosis and increases the number of erythrocytes and hemoglobin levels 96 h after the injection when compared to vehicle-injected animals. The injection of lipopolysaccharide (LPS) also induced a significant leukocytosis and increased the erythrocytes number 4 h after the injection. The combination of the higher dose of DM with LPS showed an additional effect in the number of leukocytes and erythrocytes. The GST and EROD activities were also evaluated in the gills and liver, respectively, 96 h after intoxication with both doses of DM. The GST activity decreased after DM intoxication in both doses while EROD activity was not significantly different. These results suggest that A. multispinis is a valuable model for ecotoxicological studies. Also, DM can affect the metabolizing system (GST) and immunologic system of A. multispinis, a condition that can be worsened in the presence of inflammation or infection.     

城市废水暴露对食蚊鱼肝脏EROD酶活性的影响

采用动力学酶标荧光法,检测了东莞市数所污水处理厂、制药厂和电子厂废水对食蚊鱼(Gambusia affinis)肝组织中7-ethoxyresorufin o-deethylase(EROD)酶活性的影响,评价了运用EROD酶活性监测水环境污染物的生物效应的可行性。结果显示,食蚊鱼分别暴露于经稀释为20%,40%,60%,80%不同梯度的废水液72 h后,肝脏EROD酶的活性分别与受试城市污水处理厂、制药厂和电子厂的废水之间存在剂量效应关系,EROD酶活性随污水浓度的增加而提高。电子厂废水的最大诱导倍数与对照组的比值可达到5.26,这表明其水体中存在的有机污染物较多,污水处理厂次之,制药厂的出水中污染物最少。研究表明,食蚊鱼肝组织EROD酶活性可以作为监测城市废水污染的理想生物标记物,后续的研究工作应使之标准化。     

Activities of biotransformation enzymes in pheasant (Phasianus colchicus) and their modulation by in vivo administration of mebendazole and flubendazole

Basal activities of certain pheasant hepatic and intestinal biotransformation enzymes and modulation of their activities by anthelmintics flubendazole (FLBZ) and mebendazole (MBZ) were investigated in subcellular fractions that were prepared from liver and small intestine of control and FLBZ or MBZ treated birds. Several oxidation, reduction and conjugation enzyme activities were assessed. In the liver, treatment of pheasants by FLBZ or MBZ caused very slight or no changes in monooxygenase activities and conjugation enzymes. More significative changes were detected in small intestine. Metyrapone and daunorubicin reductase activities were increased by both substances in the liver. This is the first evidence that certain benzimidazoles modulate reductases of carbonyl group. With respect to the relatively slight extent of the changes caused by FLBZ or MBZ we can assume that repeated administration of therapeutic doses of both FLBZ and MBZ has probably no serious influence on pheasant biotransformation enzyme system.     

Effects of the herbicide LASSO MTX (alachlor 42% W/V) on biometric parameters and liver biomarkers in the common carp (Cyprinus carpio)

The aim of the study was to evaluate the effect of subchronic exposure to the herbicide LASSO MTX (alachlor 42% W/V) on biometric parameters and important liver biomarkers in the common carp (Cyprinus carpio). One year old fish were exposed for 28 days to LASSO MTX added to the tank water at concentrations of 240 and 2400 μg L⁻¹. The exposure did not affect fish biometric parameters. Glutathione-S-tranferase (GST) activity in liver (hepatopancreas) remained unchanged in exposed fish when compared to controls. However, significant induction of total cytochrome P 450 (CYP 450), ethoxyresorufin-O-deethylase (EROD) activity and elevated glutathione (GSH) in liver of exposed fish were detected.     

The effects of flubendazole and mebendazole on cytochromes P4501A in pheasant hepatocytes

Many benzimidazoles are known inducers of cytochromes P4501A (CYP1A) in laboratory animals and cell lines. As flubendazole and mebendazole are benzimidazole anthelmintics often used in a pheasant, in the present study an effect of these drugs in primary cultures of pheasant (Phasianus colchicus) hepatocytes was investigated. After 48 h incubation of the hepatocytes with the benzimidazoles (0.2-5 microM), CYP1A activities -- ethoxyresorufin O-deethylation (EROD) and methoxyresorufin O-demethylation (MROD) activities were measured and the CYP1A protein levels were determined by Western blotting. None of the tested benzimidazoles influenced the CYP1A protein content. No pharmacologically significant enhancement of CYP1A after exposure of the hepatocytes to flubendazole and mebendazole was found. Inhibition of the EROD/MROD activities caused by both tested substances was observed only at the highest concentration (5 microM). From a point of view of CYP1A induction or inhibition, the treatment of pheasants by both anthelmintics tested seems to be safe. Our study demonstrates the inter-species differences in CYP1A inducibility and the importance of induction/inhibition studies on target animals.     

利用化学分析和生物测试方法比较研究污染土壤中芳烃受体效应物质的积累

长期使用污水或再生水灌溉的潜在生态风险已经引起普遍关注,但是少有研究持久性有机有毒物质在土壤中积累所产生的慢性毒性.采用7-乙氧基-异吩唑酮-脱乙基酶(EROD)方法测试了北京郊区某再生水灌溉土壤中的芳烃受体效应物质,并用2,3,7,8-TCDD标定出相应的二英毒性当量(TEQ_bio).同时利用化学分析得到的土壤中16种多环芳烃(PAHs)的含量,根据文献报道毒性当量因子(TEF)换算成二英的毒性当量(TEQ_PAHs).分析生物测试的结果,发现灌溉土壤中芳烃受体     

Membrane Dialysis Extraction (MDE): A Novel Approach for Extracting Toxicologically Relevant Hydrophobic Organic Compounds from Soils and Sediments for Assessment in Biotests

Goal, Scope and Background   Organic solvents are routinely used to extract toxicants from polluted soils and sediments prior to chemical analysis or bioassay. Conventional extraction methods often require the use of heated organic solvents, in some cases under high pressure. These conditions can result in loss of volatile compounds from the sample and the degradation of thermally labile target analytes. Moreover, extracts of soils and sediments also frequently contain substantial quantities of organic macromolecules which can act as sorbing phases for target analytes and in doing so interfere with both chemical analysis and bioassays. Membrane dialysis extraction (MDE) is described as a simple, passive extraction method for selectively extracting toxicologically relevant hydrophobic organic compounds (HOCs) from polluted soils and sediments and anaylzed for its applicability in ecotoxicological investigations. Methods   Toxicologically relevant hydrophobic organic compounds were extracted from wet and dry sediments by sealing replicate samples in individual lengths of pre-cleaned low-density polyethylene (LD-PE) tubing and then dialysing in n-hexane. The efficacy of the MDE method for use in ecotoxicological investigations was assessed by testing the concentrated extracts in the neutral red assay for acute cytotoxicity, in the EROD assay for the presence of dioxin-like compounds and in the Danio rerio fish egg assay for embryotoxic and teratogenic effects. Conditions of the sediment sample (with or without water content), dialysis membrane length and duration of dialysis were analyzed with respect to their impact on three endpoints. Results of the MDE investigations were compared to data obtained in samples prepared using conventional Soxhlet extraction. Results and Discussion   The membrane dialysis extraction was found to be at least as efficient as Soxhlet methodology to extract toxicologically relevant HOCs from sediment samples. In most cases, MDE-derived extracts showed a higher toxicological potential than the Soxhlet extracts. Lack of any significant effects in any MDE controls indicated these differences were not caused by contamination of the LD-PE membrane used. The elevated toxicological potential of MDE extracts is most likely the result of enhanced bioavailability of toxic compounds in consequence of lower amounts of organic macromolecules (i.e. sorbing phases) in the MDE extracts. This effect is probably the result of a size-selective restriction by the LD-PE membrane. Conclusion   Membrane dialysis extraction was found to be a simple, efficient and cost-effective method for the extraction of sediment samples. MDE can be used to extract toxicologically relevant hydrophobic organic compounds from both wet and dry sediments without the risk of loosing volatile and thermally labile target analytes. The size-selectivity of the LD-PE membrane also appears to have the capacity to increase the bioavailablity of potential target analytes in the resulting extracts by retaining much of the organic macromolecules present in the sample. Thus, results suggest that MDE may be particularly useful for the extraction of toxicologically relevant hydrophobic organic compounds from soils and sediments for bioassays and other ecotoxicological investigations. Recommendation and Perspective   Further validation of MDE has been initiated and the applicability of the methodology to other sample types will be investigated. Of particular interest is the potential application of MDE to recover hydrophobic target analytes from biological samples such as muscle, other soft tissues and blood.     

嘉庚蛸精子发生的超微结构

在研究GCL细胞中EROD催化底物反应时间的基础上分别以三种数学模型拟合，分析比较了诱导剂TCDD、PCB、PB、BaP、3-MC、BNF对GCL细胞EROD酶活的剂量．效应曲线，结果表明Log-Normal模型能较好的拟合药酶诱导前升后降的钟形效应曲线，所得方程参数值p100和EC100可用于药物代谢酶的诱导研究；Gentox模型拟合度相对较低且其EC50值显著高于半效应浓度；Logistic模型只能拟合曲线诱导上升部分而且嬲50和P100与Log-Normal模型相比显著偏高。进一步研究结果显示诱导剂TCDD对GCL细胞作用时间越长，EC100越大，P100越小，但在作用48h后即达到较大诱导效应，延长时间并无显著加强作用。