涕灭威、灭多威的遗传毒性研究

研究了涕灭威与灭多威的遗传毒性.在去离子水中分别加入涕灭威与灭多威标准品,涕灭威设计0.002、0.02、0.2、2、20μg/L共5个剂量组,灭多威设计0.02、0.2、2、20、200μg/L共5个剂量组.用微核试验检测其对鲤鱼血红细胞微核的诱发效应,采用Ames试验检测2种农药的致突变性,采用彗星试验检测其对人外周血淋巴细胞DNA的损伤效应,根据3种毒理学试验结果综合分析涕灭威与灭多威的遗传毒性.结果表明,所有剂量组这2种农药未诱导鲤鱼血红细胞微核率的明显上升(p0.05);2种农药各剂量组回变菌落数均未超过自发回变组的2倍,但在非代谢活化条件下,涕灭威2～20μg/L与灭多威20～200μg/L剂量组的TA97菌株回变菌落数分别达到(129.17±17.00)、(129.50±18.28)、(109.83±10.80)和(114.17±9.37)个/皿,明显高于自发回变组(p0.05,p0.01).在代谢活化条件下,灭多威200μg/L组的TA100与TA102菌株的回变菌落数为(147.83±23.29)、(275.83±20.63)个/皿,均高于自发回变组(p0.05);在彗星试验中发现,高浓度的涕灭威与灭多威均可导致人外周血淋巴细胞的DNA损伤,涕灭威20μg/L组,灭多威200μg/L组的尾部DNA(%)、尾长、Olive尾距3个指标经统计学分析,高于去离子水对照组(p0.01).研究未见涕灭威与灭多威对鲤鱼血红细胞产生明显的染色体损伤效应,虽未见其明显的致突变性,但在高浓度下存在一定的致突变的风险,并且这2种农药可能导致人外周血淋巴细胞的DNA损伤,因此涕灭威与灭多威污染对水环境和人体健康可能存在一定的远期危害.     

Cytogenotoxicity and histopathological assessment of Lekki Lagoon and Ogun River in Synodontis clarias (Linnaeus, 1758)

The cytogenotoxicity and histopathological alterations induced by xenobiotics in Lekki Lagoon and Ogun River on Synodontis clarias were investigated. Fish from these water bodies and a fish farm (control) were examined for micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood. Also gill, liver, kidney, and ovary were processed for histopathology using hematoxylin-eosin staining. Concentrations of cadmium, zinc, lead and copper in the water were determined. There was significant (p < 0.05) increase in micronucleated, binucleated, and immature erythrocytes in both gill and peripheral blood of S. clarias from the lagoon and river compared to the reference site. Loss and disorganization of the primary and secondary lamellae, multifocal degeneration, hemorrhages, cellular infiltration, congestions, vacuolations, atresia, and necrosis were common lesions in the examined tissues of fish from the lagoon and river. Cd, Zn, Pb, and Cu in water samples from the lagoon and river were higher than the reference site. Xenobiotics in Lekki Lagoon and Ogun River, mostly metals, induced deoxyribonucleic acid (DNA) and pathological damage in S. clarias.     

Chemopreventive effects of ellagic acid against genotoxicity induced by benzo(a)pyrene

Benzo(a)pyrene (B(a)P) is a commonly used indicator for polycyclic aromatic hydrocarbons (PAHs) contamination. The use of certain phytochemicals and some polyphenolic compounds proved to be effective in blocking PAH-induced effects. The aim of the present study was to examine the possible inhibitory effects of ellagic acid (EA), present in berries and nuts, against B(a)P-mediated toxicity using in vitro and in vivo test systems. In vitro method included the Ames test, using Salmonella typhimurium tester strain TA100. In vivo experiments were based on mouse bone marrow micronucleus (MN) assay and immunoblotting. EA produced a decrease in the number of revertant colonies against B(a)P in the Ames assay. Further, there was a reduction in the mean number of micronucleated polychromatic erythrocytes in the presence of EA in the co-treatment protocol of the MN test. Western blotting results showed downregulation of CYP450 isoform CYP1A1 which prevented bioactivation of B(a)P. Thus, the present study demonstrates antimutagenic role played by EA against the mutagenic activity of B(a)P in both in vitro and in vivo systems.     

全氟辛烷磺酸(PFOS)和多壁碳纳米管(MWCNTs)复合对斑马鱼外周血红细胞DNA的损伤

为了评价全氟辛烷磺酸(PFOS)和多壁碳纳米管(MWCNTs)对水环境及鱼类的影响,以斑马鱼为模式生物,研究了PFOS和MWCNTs复合对斑马鱼外周血红细胞的DNA损伤。将成年斑马鱼暴露于PFOS(0.2、0.4、0.8、1.6 mg·L-1)、MWCNTs(50 mg·L-1)、PFOS+MWCNTs(0.2+50、0.4+50、0.8+50、1.6+50 mg·L-1)和对照溶液中30 d后,断尾取血进行微核试验和彗星试验。结果表明:PFOS和MWCNTs均可造成斑马鱼外周血红细胞的DNA损伤。1.6 mg·L-1PFOS处理组的微核率、Olive尾矩及尾长分别为(36.3±0.25)‰、(87.91±14.90)μm和(250.49±34.71)μm。PFOS与MWCNTs复合后,斑马鱼外周血红细胞的DNA损伤效应明显降低。复合处理组斑马鱼外周血红细胞的微核率、Olive尾矩及尾长均低于PFOS单独处理相。1.6 mg·L-1复合处理组的微核率、Olive尾矩及尾长比PFOS单独处理组分别降低了24.7%、68.9%、52.4%。因此,在实验浓度范围内,MWCNTs可以降低PFOS对斑马鱼外周血红细胞的DNA损伤。     

稀有鮈鲫外周血红细胞微核试验方法研究

将稀有鮈鲫(Gobiocypris rarus)半静态暴露于重铬酸钾溶液中,研究发现稀有鮈鲫的本底微核率处于较低的水平,重铬酸钾在不同浓度和时间暴露后能明显观察到外周血红细胞微核增加。在一定条件下存在剂量-效应关系和时间-效应关系,表明稀有鮈鲫可用于鱼类外周血红细胞微核试验。试验中每尾鱼观察15000个细胞,能有效地减小试验偏差,保证试验结果的可靠性。暴露浓度大于等于0.01mg/L时,染毒组与空白组的外周血红细胞微核率有显著性差异,其微核率随染毒时间的延长呈先升高后下降的趋势,均在24h时出现所有测定时间微核率的峰值。与其他鱼类比较显示,稀有鮈鲫具有较高的敏感性,可用于遗传毒物诱发微核的监测。     

Toxicological characterization of the landfill leachate prior/after chemical and electrochemical treatment: A study on human and plant cells

In this research, toxicological safety of two newly developed methods for the treatment of landfill leachate from the Piškornica (Croatia) sanitary landfill was investigated. Chemical treatment procedure combined chemical precipitation with CaO followed by coagulation with ferric chloride and final adsorption by clinoptilolite. Electrochemical treatment approach included pretreatment with ozone followed by electrooxidation/electrocoagulation and final polishing by microwave irradiation. Cell viability of untreated/treated landfill leachate was examined using fluorescence microscopy. Cytotoxic effect of the original leachate was obtained for both exposure periods (4 and 24 h) while treated samples showed no cytotoxic effect even after prolonged exposure time. The potential DNA damage of the untreated/treated landfill leachate was evaluated by the comet assay and cytokinesis-block micronucleus (CBMN) assay using either human or plant cells. The original leachate exhibited significantly higher comet assay parameters compared to negative control after 24 h exposure. On the contrary, there was no significant difference between negative control and chemically/electrochemically treated leachate for any of the parameters tested. There was also no significant increase in either CBMN assay parameter compared to the negative control following the exposure of the lymphocytes to the chemically or electrochemically treated landfill leachate for both exposure periods while the original sample showed significantly higher number of micronuclei, nucleoplasmic bridges and nuclear buds for both exposure times. Results suggest that both methods are suitable for the treatment of such complex waste effluent due to high removal efficiency of all measured parameters and toxicological safety of the treated effluent.     

Micronucleus frequency in sheep lymphocytes after in vitro exposure to fungicide tolylfluanid

The fungicide tolylfluanid (N - dichlorofluoromethylthio-N′, N - dimethyl - N - p - tolylsulfamide), was investigated by cytokinesis-block micronucleus assay. Tolylfluanid at the lowest concentration (1 × 10^{? 6}mol L^{? 1})did not influence significantly the frequency of micronuclei in sheep lymphocyte cultures in comparison with control (32.33 ± 3.51/1000 binucleated cells versus 30.33 ± 2.82/1000 binucleated cells in dimethylsulfoxide (DMSO) control, P = 0.44). Higher tolylfluanid concentrations (1 × 10^{? 4} and, 1 × 10^{? 5} mol L^{? 1}) resulted in a significant dose-dependent increase in the number of micronuclei in comparison with control (74.00 ± 13.00/1000 binucleated cells and 52.67 ± 10.12/1000 binucleated cells versus 30.33 ± 2. 82/1000 binucleated cells in DMSO control, P = 0.005 and 0.02, respectively, ANOVA followed by Tukey test P < 0.05). Many of the treated cells also possessed multiple micronuclei. Tolylfluanid did not affect the nuclear division index at all treatment concentrations. Our in vitro results thus demonstrate that tolylfluanid had a significant genotoxic effect at only the highest concentration tested.     

离子液体[C16mim]Cl对泥鳅的毒性效应

以泥鳅为受试动物,采用急性毒性、微核试验及酶活分析实验,研究了离子液体[C16mim]Cl对水生生物的生物毒性作用。急性实验表明:[C16mim]Cl对泥鳅有明显的毒性,其对泥鳅24、48和96h的LC50分别为2.533、1.763和1.379mg.L-1;微核实验表明:对照组红细胞微核率为0.5‰左右,而处理组红细胞微核率最高达到12.89‰,且各处理组均明显高于对照组(P<0.05);酶活分析实验表明:处理组血清的GPT和GOT活力均明显高于对照组,且随[C16mim]Cl浓度增高而呈现上升趋势。从以上结果可以得出:[C16mim]Cl对泥鳅具有显著的遗传毒性和生理毒性效应,因而推测其大量使用可能对水生生物及生态环境具有一定的毒害作用。     

Genotoxic effects of two different classes of insecticide in developing chick embryos

Pesticides provide considerable protection against pest population; however, rampant accumulation of these chemicals into varied habitats across the globe necessitates the need for a careful screening of each chemical due to toxic manifestations. In the current study, the genotoxic potential of two different classes of commercial insecticides – chlorpyrifos and cypermethrin combination and Spinosad, a naturalyte were compared. Rhode Island Red chick embryos were exposed to different doses of either of these insecticides individually, by in ovo treatment. Genotoxicity was then evaluated through micronucleus (MN) test and Comet assay. The combination insecticide exposure at low doses of 0.05 and 0.1 μg/egg induced DNA damage as evidenced by an increased tail moment in the Comet assay. Further, the presence of micronucleated erythrocytes and also various abnormal cells including dacryocytes, microcytes, erythroplastids, squashed/notched nuclei, and spindle-shaped erythrocytes in the blood smear consolidates indicate the presence of insecticide-induced genotoxicity. Spinosad, however, was found only mildly genotoxic but at a high dose of 1.5 mg/egg. The results indicate that usage of naturalyte insecticide may be a better option to minimize the harmful effects of chemical insecticides.     

邻苯二甲酸二乙基己酯对蚕豆根尖微核及幼苗超氧化物歧化酶的影响

为了初步探讨邻苯二甲酸二乙基己酯(Di-(2-ethylhexyl)phthalate,DEHP)对植物的毒害作用,采用不同浓度的DEHP溶液对蚕豆根部进行处理(终含量0、0.2、2、20、200mg·kg-(1细沙)),检测根尖微核率和对幼苗茎、叶SOD活性的影响.结果表明:对蚕豆根尖进行短期(6h)处理后,各DEHP处理组微核率随DEHP含量的升高呈显著上升趋势(与对照比较,p<0.05或p<0.01).处理7d后,随DEHP含量的升高(0～20mg·kg-(1细沙)),蚕豆幼苗茎、叶SOD活性均呈逐渐升高趋势;DEHP含量在20～200mg·kg-(1细沙)时,SOD活性均略有下降.以上结果表明DEHP对蚕豆种子具有遗传毒性,且能够对蚕豆幼苗产生氧化损伤.