林丹短期暴露对斑马鱼生长发育和繁殖的影响

以斑马鱼为受试生物,研究在低浓度(0.005、0.05、0.5峭μg·L-1)林丹暴露条件下,林丹对斑马鱼体长、体质量、肝脏指数、性腺指数、产卵量和受精率的影响.结果显示,经14 d暴露后,处理组斑马鱼体长和体质量与空白对照组或溶剂对照组之间差异不显著;与空白对照组和溶剂对照组相比,0.5μg·L-1处理组雄性和雌性斑马鱼肝脏指数均显著增加(P＜0.05),雄性斑马鱼性腺指数显著降低(P＜0.05),其他处理组无明显变化;林丹对斑马鱼产卵量和受精率表现出明显的抑制作用,与空白对照组相比,0.05和0.5μg·L-1处理组斑马鱼平均产卵量和受精率均极显著降低(P＜0.01).该研究表明林丹具有环境内分泌干扰物特性,在低浓度暴露条件下可对斑马鱼的生长发育和繁殖产生一定抑制影响.     

氟对斑马鱼甲状腺内分泌功能的干扰效应

为探究氟对斑马鱼甲状腺内分泌功能的干扰效应,本研究以雌性斑马鱼成鱼为研究对象,分别以不同浓度氟(0、20、 40、80 mg·L~(-1))暴露45 d和90 d,对斑马鱼的生长发育指数进行测定,用组织学方法对斑马鱼甲状腺组织结构进行显微观察,用酶联免疫吸附法检测血浆中T3和T4激素水平,并用实时荧光定量PCR方法检测HPT轴上内分泌相关基因的表达.结果显示,与对照组相比,氟暴露组雌性斑马鱼的生长发育指数随着暴露浓度和时间的增加而呈下降趋势(p0.05);甲状腺组织病变程度随着氟暴露浓度的增大和时间的延长而加重;氟暴露45 d时,T3和T4水平呈升高趋势,90 d时,T4水平呈下降趋势且在80 mg·L~(-1)组下降显著(p0.05);氟暴露45 d时,40 mg·L~(-1)组除crh处,其他mRNA表达均显著下降(p0.05),80 mg·L~(-1)组除tg、nis、ttr外其他mRNA表达均显著下降(p0.05);暴露90 d时tg、dio1、dio2的mRNA表达水平在40 mg·L~(-1)和80 mg·L~(-1)组显著上升(p0.05),ugt1ab mRNA表达在80 mg·L~(-1)组显著下降(p0.05).综上,氟可通过影响雌性斑马鱼的生长发育、甲状腺组织结构、激素水平及HPT轴上内分泌相关基因的表达,从而对其甲状腺内分泌功能产生一定程度的干扰,进而影响机体的生长发育.本研究结果对水环境中氟的生态毒性效应及风险评估提供了理论资料.     

Toxic effect of microcystin-LR on blood vessel development

Abstract

The effect of microcystin-LR (MC-LR) at 50–1000?nmol/L on blood vessel development of zebrafish embryos has been explored. Upon exposure 72-h-post-fertilization (hpf), the hatching rate was significantly reduced and malformation and fatality rates were increased in a dose-dependent manner. MC-LR induced cell cycle arrest in the G0/G1-phase and damaged DNA structure in a dose-dependent manner. In human umbilical vein endothelial cells (HUVECs), MC-LR led to decreased activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and increased lipid peroxidation. These findings indicated that MC-LR could cause oxidative DNA damage, which mediated cell cycle arrest and inhibited cell proliferation afterwards, and retarded the blood vessel development.     

纳米二氧化硅和铅复合暴露对 斑马鱼幼鱼甲状腺内分泌系统的毒性影响

纳米颗粒(NPs)因其广泛的应用而备受关注。NPs会改变重金属(如Pb)在生物体内的富集和毒性作用。因此，研究选择国际公认的模式动物斑马鱼(Danio rerio)为研究对象，开展纳米二氧化硅(nano-SiO₂)对Pb在鱼体内的积累和甲状腺内分泌系统干扰的研究。受精2 h后的斑马鱼胚胎，单独暴露于环境相关浓度的Pb(如0, 5, 10和20 μg/L)或与无效应浓度nano-SiO₂(如25 μg/L)复合暴露至144 h。结果表明：和nano-SiO₂复合暴露后，斑马鱼幼鱼体内Pb的含量显著高于同浓度下Pb单独暴露(P<0.05)。此外，Pb或nano-SiO₂单独暴露都不会显著改变斑马鱼幼鱼体内甲状腺激素(T4和T3)的含量(P>0.05)；但当两者复合暴露后，斑马鱼幼鱼体内T4和T3含量显著低于对照组或同浓度下Pb单独暴露组 (P<0.01和P<0.05)。在Pb单独或与nano-SiO₂复合暴露下，tshβ基因的表达都显著上调，但ttr和tg基因的表达都显著下调，并且复合暴露造成基因表达水平的改变较同浓度下Pb单独暴露相比更明显(P<0.05)。上述结果指出，nano-SiO₂会增加Pb在生物体内的富集，从而对斑马鱼幼鱼的甲状腺内分泌系统造成更严重的影响。     

Heavy metals and pesticides in anuran spawn and tadpoles,water, and sediment

High concentrations of metals (cadmium, copper, lead, and zinc) and of lindane were measured in the sediment and in the spawn and tadpoles of Bufo bufo, Rana dalmatina, and Rana ridibunda from two Austrian locations. Residues in spawn suggest maternal transfer. Increase of the metal concentrations from spawn to advanced tadpole stage is explained by the larval microphaguous feeding habits which cause high exposition of the tadpoles to substances concentrating in sediments and suspended particles. Metal and pesticide concentrations in spawn and tadpoles are toxic to various other aquatic organisms and are furthermore considered to be potentially hazardous to the anurans themselves. The results are conform with world‐wide observations of contamination of anuran larvae with heavy metals, and prove the significance of non‐point source chemical exposure due to allochthonous toxicant input by wind load and precipitation.     

1H-NMR-based metabolomic study on toxicity of methomyl and methidathion in fish

A ¹H-nuclear magnetic resonance (NMR) spectroscopy with multivariate analysis was applied to detect the toxicity of antiacetylcholinesterase insecticides, methomyl (methyl (1E)-N-(methylcarbamoyloxy)ethanimidothioate) and methidathion (3-(dimethoxyphosphinothioyl sulfanylmethyl)-5-methoxy-1,3,4-thiadiazol-2-one), using zebrafish (Danio rerio) and Chinese bleak (Aphyocypris chinensis). Generally, methomyl and methidathion have been believed not to highly accumulate in fish tissues. However, these pesticides showed their toxicity by altering patterns of whole-body metabolites in neurotransmitter balance, energy metabolism, oxidative stress, and muscle maintenance in low concentrations. We used Pearson correlation analysis to contextualize the metabolic markers in pesticide treated groups. We observed that the positive correlations of choline with acetate and betaine in untreated control were shifted to null correlations showing acetylcholinesterase specific toxicity. This research demonstrated the applicability and potential of NMR metabolomics in detecting toxic effects of insecticide with a modicum of concentrations in aquatic environment.     

城市污水对斑马鱼胚胎的影响

采用静态法,在实验室控制条件下,观察处理前后的城市污水对斑马鱼胚胎的毒性效应,并探讨其对斑马鱼胚胎可能的毒性机理。结果表明,兰州市安宁区M污水处理厂处理前的污水、处理后接触池出水和接触池出水入黄河排污口水对胚胎均有较高的毒性,其72 h EC50在21.384%～108.492%之间;对处理胚胎孵化成的仔鱼毒性也很明显。上述几种污水均降低斑马鱼胚胎的孵化率,增加胚胎及其孵化仔鱼的畸形。     

nano-TiO_2与1,2,4-TCB联合作用对斑马鱼鳃组织结构及染色体损伤研究

采用组织病理切片法、微核及彗星试验研究nano-TiO_2和1,2,4-TCB联合染毒对斑马鱼鳃组织结构、细胞微核率及DNA损伤的毒性效应。结果表明,单独nano-TiO_2暴露不会导致斑马鱼鳃组织结构损伤,也不会引起明显的微核及染色体损伤。1,2,4-TCB(≥10 mg·L~(-1))会引起鳃组织和细胞染色体的损伤。nano-TiO_2与1,2,4-TCB联合作用会加重斑马鱼的鳃组织结构损伤,出现鳃小片排列严重紊乱,表皮细胞断失严重,鳃小片上皮细胞坏死脱落,鳃丝结构模糊等症状。两者联合作用会引起鳃细胞微核率增加和彗星细胞率的增加,呈现一定的剂量-效应关系,且两者间的联合作用具有协同作用。对彗星试验的尾部DNA百分比、尾长及尾距的分析发现,与对照组和单独作用相比,两者联合作用对彗星指标均有显著增强作用,这表明两者联合作用对染色体的损伤属于非特定片段损伤。     

Comparative quantitative structure–activity relationship (QSAR) study on acute toxicity of triazole fungicides to zebrafish

We employed two-dimensional quantitative structure–activity relationship (2D-QSAR) and hologram QSAR (HQSAR) methods to quantitatively investigate the mechanism and active site of toxicity for Danio rerio exposed to triazole fungicides. Our results showed that 2D-QSAR models constructed using the energy of the lowest unoccupied molecular orbit, the net C atom charges, the octanol–water partition coefficient and the molecular shape factor had higher predictive abilities. HQSAR models containing the fragment distinctions atoms (As), bonds (Bs), chirality (Ch) and donors and acceptors (D&A) had higher reliability. It was found that 2D-QSAR results explaining the toxicity mechanism were consistent with HQSAR. In summary, the hydrophobicity and shape/size of the molecule were the important factors influencing the toxic effect of these chemicals against D. rerio. In addition, electron exchange may occur between these fungicides and the target. The study provided a method to evaluate the environmental risk of chemicals with a similar structure, based on the QSAR models obtained.     

氯化镉暴露对斑马鱼幼鱼神经行为毒性作用

为探索低剂量镉持续暴露对斑马鱼胚胎和幼鱼神经发育的影响,将受精后36h(36hpf)的斑马鱼胚胎暴露于不同浓度氯化镉受试液中持续至受精后6d(6dpf)。在一般毒性观察的基础上,采用高通量视频跟踪技术检测了48孔板中幼鱼的自发运动能力和对外界刺激产生的惊恐反应及恢复能力。结果显示:在不引起外观形态明显改变的暴露剂量下幼鱼自发运动表现异常。其中,0.1～0.5μmol·L-1暴露组幼鱼自发运动增多、轨迹紊乱、刻板式转向运动增加、触壁活动减少;1.0μmol·L-1暴露组幼鱼出现运动抑制、静止和原地震颤等状态,幼鱼的自发运动距离与氯化镉浓度关系呈现剂量依赖性倒"U"型,这种行为毒性效应与乙醇急性暴露引起的运动效应相似。光照惊恐反射实验中,暴露组幼鱼对突发的光暗变化表现敏感或迟钝。在撤除光源后暗适应减慢或加快。以上结果说明,斑马鱼胚胎和幼鱼对氯化镉的神经发育毒性作用敏感,并且高通量的幼鱼神经行为学方法在外源性化学物的毒性评价中能够提高检测效率,具有良好的应用前景。