烷基酚类化合物对斑马鱼胚胎的毒性效应

采用斑马鱼胚胎发育技术,观察邻甲酚、对甲酚、间甲酚和2,4-二叔丁基酚联合作用对斑马鱼胚胎的影响,以研究烷基酚类化合物的毒性.结果表明,4种受试物的联合对斑马鱼胚胎发育均有明显的抑制作用及不同程度的致死等效应.Ohpf染毒主要表现为:24 h后卵凝结和尾部无延展,眼点发育受抑制及无主动活动;48 h卵凝结和心率异常,无黑色素和心包囊水肿;72 h卵凝结和未孵化等效应.     

斑马鱼在环境检测领域中的应用

斑马鱼易饲养、易管理,体外受精和体外发育,可常年产卵、孵化,因此早期被作为模型动物广泛用于生物学研究中.近年来,随着环境问题日益突出,迫切需要有效、全面的环境监测手段和方法.斑马鱼或其转基因品种就是非常好的环境监测首选生物.因其对许多环境污染物十分敏感,可实时检测进入水环境的污染物.对斑马鱼在水体中重金属毒性、环境激素...     

斑马鱼在环境保护中的应用

斑马鱼易饲养、易管理,体外受精和体外发育,可常年产卵、孵化,且胚胎透明,因此早期作为模型动物广泛用于生物学研究中.近年来,随着水环境问题日益突出,迫切需要有效、全面的环境监测手段.斑马鱼或其转基因品种不失为水环境监测的首选生物.因为它们对各类水环境污染十分敏感,可实时检测进入水环境的污染物.文章对斑马鱼在环境激素、水质...     

斑马鱼对水源特定污染物的水质监测预警

选取浑浊度、石油类、亚硝酸盐、苯、砷化物、锰等不同类型的环境污染物,研究了斑马鱼在不同污染物压力下行为强度变化规律,分析了不同条件下斑马鱼的活性及在线系统报警特征。结果表明:斑马鱼的行为生态学变化与环境污染物压力存在显著的相关性。在不同类型的环境污染物下,斑马鱼行为变化存在明显差异,对砷和锰等剧毒物质更加敏感;在高浓度环境污染物压力下,斑马鱼的行为生态学特征出现剧烈变化;随着暴露时间的增加,斑马鱼的行为生态学变化会逐渐降低。斑马鱼的行为强度变化可以快速评判水质污染程度,但应结合当地的水质特征设定具体的报警阈值。     

利用转基因斑马鱼探究四溴联苯醚(BDE-47)的神经毒性作用

为探究2,2’,4,4’-四溴联苯醚（BDE-47）对斑马鱼的早期神经毒性作用，采用中枢神经特异性标记的转基因斑马鱼Tg(elavl3:EGFP)作为模式生物，研究了BDE-47短期暴露对斑马鱼幼鱼的运动行为轨迹、中枢神经发育和神经发育关键基因表达的影响。结果表明，BDE-47暴露6 d后，5和10μmol/L暴露组斑马鱼幼鱼运动行为和神经发育关键基因（elavl3和mbp）的表达受到显著抑制，同时斑马鱼幼鱼的神经细胞的发育受到显著抑制。结果提示，神经系统特异性标记的转基因斑马鱼可作为一种筛选神经毒性化合物的理想动物模型，BDE-47对斑马鱼具有早期神经发育毒性。     

水体中黑碳对斑马鱼胚胎的生长发育危害识别

黑碳(BC)是含碳物质不完全燃烧后产生的残留物,广泛分布于水体、土壤和大气中,目前对于BC对水生生物的生态效应研究还十分有限。通过燃烧煤油获得BC,以斑马鱼胚胎为模式生物,研究了质量浓度分别为0,0.05,0.5和5 mg/L的BC悬浮溶液对斑马鱼早期生命阶段的生长发育毒性。结果表明,煤油源BC对斑马鱼具有明显的毒性作用,与对照组相比,BC暴露明显增加了斑马鱼的死亡率和心率,显著降低了孵化率和体长,导致斑马鱼形态畸形并且畸形率呈浓度依赖性。5mg/L的BC对斑马鱼的各项生长发育指标影响并非是最大的,可能与BC颗粒在水溶液中的聚集有关。该研究为BC在水生环境中的危害识别提供实验依据。     

气相色谱法测定焦化厂废水中的挥发酚

焦化厂废水以苯酚、邻甲酚、对甲酚、间甲酚为主。因此,采用4—氨基安替比林比色法和引进来电子基团法都不适用焦化厂废水中挥发酚的测定。本文采用六甲基二硅胺烷和三甲基氯硅烷混合物作为硅醚化试剂,并对其操作方法进行了改进,提高了灵敏度,使之更适宜于测定焦化厂生化后外排水中的微量挥发酚。     

应用斑马鱼和凡纳对虾诊断污染场地污水的生物毒性

应用斑马鱼(Brachydanio rerio)和凡纳对虾(Penaeus vannamei)对某废弃有机污染场地中有毒有机废弃物堆积产生的污水(1号塘污水)和场地及周边地表径流积水(2号塘污水)的生物毒性进行了试验.结果表明,1号塘污水对斑马鱼和凡纳对虾96 hLC50分别为46.2%和59.6%, 2号塘污水的毒性试验中两种试验生物的死亡率皆为0.化学检测表明,1号塘污水中的主要污染物为苯酚类和酞酸脂类物质,而2号塘污水中均未检出有机污染物.化学检测结果与毒性试验结果具有较好的相关性.     

水体突发性重金属污染胁迫下斑马鱼的行为反应分析

研究了斑马鱼在不同程度突发性的Zn2＋和Cr6＋胁迫下的行为反应,采用计算机视觉技术量化研究斑马鱼游动行为数据。结果表明,在Zn2＋和Cr6＋的突发性胁迫中,斑马鱼的行为反应快速且敏感,游动行为与污染物种类、质量浓度和暴露时间直接相关,并表现出比较相似的行为变化规律;随着胁迫程度的增加,斑马鱼游动速度的变化幅度增大,反应时间缩短,且其变化曲线符合生物行为的环境压力模型。斑马鱼的行为变化远远提前于生物的病理损伤或死亡,可利用斑马鱼暴露于污染物时的行为变化实现在线水体突发性重金属污染事故的监测预警。     

四种工业固体废物的生物毒性试验研究

由于目前尚有大量的化学物质,对人体和生物的毒性未被认识,况且工业排放的废渣往往多为复合化合物,常规理化方法虽然能进行部分元素及其化合物的定量测定,但往往只能进行单项毒物含量测定和瞬间测定,不能检验出毒物联合作用的综合毒性及其长期积累性.因此本研究重视应用生物检测手段,旨在通过大型溞斑马鱼的急性毒性试验、发生细菌的生物毒性测试以及紫露草雄蕊毛突变检测等四个指标,利用生物对环境毒物具有的指示性、敏感性、蓄积性、遗     

Histopathological changes in zebrafish embryos exposed to DLPCBs extract from Zhanjiang coastal sediment

Dioxin-like polychlorinated biphenyls (DLPCBs) are ubiquitous persistent pollutants that cause adverse effects in many environmental organisms. DLPCBs in marine sediments can be absorbed by benthic organisms, bioaccumulate, and biomagnify through the food chain and threaten animal and human health. There are no reports of DLPCBs concentrations in the Zhanjiang Gulf seabed. This study was designed to investigate the concentration of DLPCBs in the Zhanjiang coastal sediment and histopathological changes in zebrafish (Diano rerio) embryos exposed to environmentally relevant concentrations of DLPCBs. Of the five sites selected, two sites TS and JSW contained DLPCBs at concentrations of 0.08 and 22.54 ng/g dry sediment, respectively. Two groups of zebrafish embryos were used. One group was exposed to 3.75, 7.5, 15, 30, and 60 mg/ml of DLPCBs extracted from the sediments sampled from the TS site and the second group to 4.375, 8.75, 17.5, 35, and 70 mg/ml of DLPCBs from JSW site from 0.75 h post-fertilization (hpf) to 96 hpf. The zebrafish exposed to 60 and 70 mg/ml of DLPCBs at 96 hpf displayed gross histopathological changes with cardiac lesions including pericardial edema being the most deleterious. Other changes observed were hydropic degeneration of gill filaments and hepatocytes, loss of intestinal folds, and uninflated swim bladder. It appears that only a few sites of the Zhanjiang gulf are contaminated with DLPCBs. This is the first report of histopathological changes in the gills, hepatocytes, intestines, heart, and the swim bladder in zebrafish embryos exposed to DLPCBs from a coastal sediment. Further studies with sampling at different stages of development are required to identify which organ/tissue is most sensitive to DLPCBs.     

Assessing water quality in a tropical lake using biomarkers in zebrafish embryos: developmental toxicity and stress protein responses

In order to achieve a more substantial appraisal of lake water quality, the assessment must not be based only on chemical measurements and analyses of the water itself, but even more so on the impact of existing conditions on aquatic biota. This is possible with the use of biotests or biomarkers, e.g. investigations of the developmental parameters (96-h embryotoxicity evaluate) or of the induction of heat shock proteins (proteotoxicity evaluate). To evaluate the suitability of these tests for environmental screening, fertilized zebrafish eggs were exposed to water samples collected from five sites of varying levels of stress from Laguna Lake, Philippines. Reconstituted water was used as laboratory control while water samples from a highly polluted freshwater source was used as positive control. Developmental parameters were noted and described within 48 and 96 h of exposure. Dilution experiments of the positive control were also done to further assess and compare toxicity potentials of Laguna Lake waters with those originating from a polluted freshwater. After the 96-h exposure, the levels of stress proteins (hsp 70) were determined in embryos from all exposure groups. Results showed 100% mortality in embryos exposed to undiluted positive control (PC) within only 12 h. Increasing dilution levels, however, resulted in lower mortality and lower abnormality rates. No detectable developmental differences were noted among embryos exposed to either the laboratory control or Laguna Lake waters at the end of 96 h, regardless of the source. Very high survival rates and high hatching success rates were observed in embryos exposed to lake waters as well as laboratory control, and the data did not differ significantly among the groups. Likewise, no significant malformations were noted among all developing embryos throughout the exposure period. However, the levels of heat shock proteins in the two sites located closest to Manila, the Philippine capital (Northern West Bay and Central West Bay), showed a pronounced elevation relative to the control, indicating that these stress proteins protect the embryos from the detrimental effects of pollutants in the water. Based on the 96-h early life stage (ELS) test, the water quality of the lake is good for fishery propagation despite the current levels of pollutants in the water. This finding is in accordance with the Class C status (i.e. suitable for fish growth and propagation) as given to the lake by the local environmental agency. On the other hand, data on proteotoxicity showed that the fish are under stress, presumably deriving from pollutants. This calls for a continuous monitoring and improvement of the lake water. The present study indicates that the two biomarker methods are very easy to use, practical, rapid, and sensitive for assessing water quality in a tropical lake and recommends for their incorporation into the future monitoring program of Laguna Lake.     

Low Levels of Sodium and Potassium in the Water from Wetlands in Minnesota that Contained Malformed Frogs Affect the Rate of Xenopus Development

Water samples were collected between 1999 and 2000 from wetlands in Minnesota that contained malformed frogs. The water samples were analyzed for 14 minerals/ions and screened for the presence of biologically active compounds using Xenopus laevis. Results indicated that water from two sites, CWB and ROI2, induced severe retardation with embryo lengths reduced 20% after 96 hr of development. The developmental delay observed with water from ROI2 was alleviated by supplementation with sodium, while both sodium and potassium alleviated the developmental delay observed with water whose mineral content mimicked that of CWB. Seasonal fluctuations in the sodium and potassium content at ROI2 and NEY correlated with changes in the rates of Xenopus development. Xenopus embryos reared on water from ROI2 for 120 hr displayed gut malformations not present in embryos reared on a synthetic media designed to mimic the mineral content of the water from ROI2. Embryos reared on water from ROI2 supplemented with minerals at levels comparable to that routinely employed in the rearing of Xenopus were neither retarded nor malformed. It is proposed that climate driven hydrology may influence the mineral composition at selected wetlands and delay development which may alter window(s) of susceptibility towards biologically active agents and the occurrence of malformed frogs.     

Environmental contaminants in nonviable eggs of the endangered Mississippi sandhill crane (Grus canadensis pulla)

Our objectives were to determine if concentrations of environmental pollutants and microbial contamination in nonviable eggs of the endangered Mississippi sandhill crane (Grus canadensis pulla) contributed to egg failure. Six eggs collected in 1990 and four in 1991 contained only background levels of organochlorine pesticides and polychlorinated biphenyls (PCBs), and tests for microbial contamination were all negative. Two eggs contained late dead embryos, but neither revealed obvious abnormalities. Three eggs contained potentially harmful concentrations (23, 39, 146 pg/g, wet mass) of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), based on 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ) for combined compounds. Because of the scarcity of material suitable for laboratory examination and the endangered status of the crane, we recommend that nonviable eggs continue to be monitored for toxic pollutants.     

Toxicity Testing of Leachate from Waste Landfills Using Medaka (Oryzias Latipes) for Monitoring Environmental Safety

To investigate the environmental safety of waste disposal landfill sites and of land reclaimed from such sites, we evaluated the toxicity of leachate from these sites by a combination of bioassays in the Japanese killifish medaka Oryzias latipes. We tested for lethal toxicity in adult and larval medaka and for hatching inhibition of embryos from eggs. As biochemical evidence of the effects of leachate exposure, CYP1A (EROD activity) and vitellogenin (Vtg) were induced. We also bioassayed water-treated leachate and downstream river water. Leachate solution was lethal to larval and adult medaka. Embryo hatchability was inhibited, and abnormal hatching, spinal deformity and anisophthalmia occurred in embryos exposed to leachate solution. CYP1A was induced by exposure to leachate solution diluted to 1.0%, and EROD activity was significantly higher than in control. Vtg and unknown proteins were induced in the sera of male medaka exposed to the diluted leachate solution. Conventional water treatments worked effectively to remove toxic compounds but did not work well to remove element ions, including heavy metals. Treated leachate produced neither lethal toxicity nor hatching abnormalities during the exposure period. Fish toxicity tests for leachate would be useful for monitoring the environmental safety of landfill sites.     

Biological effects monitoring of the North Sea employing fish embryological data

Using the reproductive capacity of fish appears to be a suitable approach for risk assessment in the aquatic environment since fish are a typical representative thereof and in addition they are of considerable societal value. Generally the early embryonic stages are considered to be one of the most sensitive parts of a fish's life cycle. A method has been developed to use the state of health of live, naturally spawned fish embryos from plankton samples for biological effects monitoring. During the years 1985–1987 in the southern North Sea and in 1991–1992 in the whole of the North Sea fish eggs were sampled from surface waters and examined for developmental abnormalities. Elevated embryo malformation rates were detected in the plume of the major rivers Elbe and Rhine as well as along the eastern coast of England. Occurring malformations are thought to be pollution-related and may be used to define areas of environmental deterioration. The method is discussed in view of its suitability for biological effects monitoring using malformations in fish embryos as biomarkers.     

Organochlorine and mercury residues in Swainson's and ferruginous hawk eggs collected in North and South Dakota, 1974–79

Residues of organochlorine pesticides, polychlorinated biphenyls (PCBs), and mercury were measured in eggs of Swainson's hawks (Buteo swainsoni) and ferruginous hawks (B. regalis) collected in North and South Dakota during 1974–79. DDE was the most common compound detected in the eggs, but residues were below levels known to have adverse effects on reproduction. Other organochlorine compounds and mercury were found at low levels. Eggs of ferruginous hawks tended to contain more compounds with higher residues than eggs of Swainson's hawks.     

Oxidative stress response in zebrafish (Danio rerio) gill experimentally exposed to subchronic microcystin-LR

The worldwide occurrence of cyanobacterial blooms makes it necessary to perform environmental risk assessment procedures to monitor the effects of microcytins on fish. Oxidative stress biomarkers are valuable tools in this regard. Considering that zebrafish (Danio rerio) is a common model species in fish toxicology and the zebrafish gill is potentially useful in screening waterborne pollutants, this study investigated the oxidative stress response in zebrafish gill exposed to subchronic microcystin-LR (MCLR) concentrations (2 or 20?μg/l) via measurement of toxin accumulation, protein phosphatase (PP) activity, and the antioxidant parameters (glutathione-S-transferase-GST; glutathione-GSH; superoxide dismutase-SOD; catalase-CAT; glutathione peroxide-GPx; glutathione reductase-GR), as well as levels of hydroxyl radical (OH) and lipid peroxidation (LPO). The results showed that after 30?days exposure, MCLR accumulated in zebrafish gill and MCLR exposure induced PP activity in gill. A linear inhibition of GST activity and GSH content was observed in the gills, revealing that they were involved in the first step of MCLR detoxification. The 2?μg/l MCLR treatment neglectably affected OH content and the antioxidant enzymes (SOD, CAT, GPx, and GR), however oxidative stress was induced under the 20?μg/l MCLR treatment in which an enhanced OH content and alterations of the antioxidant enzymes were observed in the treated gills, although both treatments exerted little effect on LPO level. The principal component analysis results indicated that the most sensitive biomarkers of MCLR exposure were GST and GSH in zebrafish gill. So, D. rerio could be regarded as a suitable bioindicator of MCLR exposure by measuring CAT, GR, GST, and GSH as biomarkers.