Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.     

荧光染色法测定活性污泥中的活性生物量

活性污泥中活性微生物量是表征污染物去除能力的重要指标。探索了荧光染色法直接检测活性细菌的原理和测定方法,结果表明,大肠杆菌、唾液链球菌和铜绿假单胞杆菌的活菌细胞浓度与荧光光强的线性相关度都超过0.95;在惰性颗粒物浓度为20 mg/L到60 mg/L的范围内,其对荧光光强的影响可通过线性关系修正;搅拌速度为1 000 r/min且搅拌10 min时前处理效果最好。     

Monitoring of DNA damage in haemocytes of freshwater mussel Sinanodonta woodiana sampled from the Velika Morava River in Serbia with the comet assay

This study was undertaken to investigate the potential of the freshwater mussel Sinanodonta woodiana for detection of genotoxic pollution of the environment. Study was performed at two sites in the Velika Morava River, from May 2010 to February 2011. The alkaline comet assay on haemocytes was used, and the olive tail moment (OTM) was chosen as a measure of DNA damage. The specimens held on acclimation under controlled laboratory conditions for 10 d were used as a control. Chemical analysis revealed the presence of phosphates and increased concentrations of zinc, copper and nickel at both sites during the entire sampling period. The values of OTM in mussels collected from the environment, significantly correlated with the concentration of zinc (r = 0.6248), temperature (r = 0.7006) and dissolved oxygen (r = 0.7738). Seasonal variations in genotoxic response were observed, with the highest OTM values obtained during summer months. Preliminary results of the in vitro study indicated the effect of water temperature on genotoxic response to zinc and cadmium in S. woodiana suggesting that the presence of genotoxic pollutants during months with lower temperature could be under-estimated. Obtained results indicate that S. woodiana could be a valuable tool for active biomonitoring of aquatic environments and emphasizes the importance of seasonal genotoxic monitoring with this organism.     

人外周血淋巴细胞彗星试验检测饮用水水质毒性研究

应用人外周血淋巴细胞彗星试验对无锡充山水厂、中桥水厂,常州西石桥水厂进出水及金坛市拟用水源地长荡湖湖心水样的生物毒性进行了研究．结果表明,各水样的有机浓集物均可对人外周血淋巴细胞DNA产生不同程度的损伤,存在一定的遗传毒性．试验结果与各水样水质污染状况基本一致．同时,季节变化以及常规水处理过程对水样的生物毒性也有影响．研究表明,彗星试验可有效地用于检测饮用水质有机浓集物遗传毒性．     

两种兽药添加剂对蚯蚓的DNA损伤及其联合效应

采用土壤中的指示生物赤子爱胜蚓（Eisenia foetida）作为受试生物,研究了两种兽药添加剂四环素和金霉素对赤子爱胜蚓的急性毒性.结果表明：滤纸接触法染毒48h后,这两种兽药添加剂在所设最大受试剂量内没有引起蚯蚓死亡,半致死浓度LC50〉3.98×10^-2mg·cm^-2.应用彗星试验检测蚯蚓体腔细胞的DNA损...     

生物可降解材料PHBV的细胞毒性评价的研究

通过体外细胞毒性试验,对新型材料β-羟基丁酸与β-羟基戊酸共聚物(PHBV)的细胞生物相容性进行了研究.主要依据ISO10993及GB/T16886-1997的标准,应用四甲基偶氮唑盐(MTT assay)比色法对PHBV的细胞毒性进行评价.选用不同体积分数、不同温度下的PHBV材料浸提液为培养液,以及与PHBV材料直接接触培养来检测1d、3 d、5 d小鼠成纤维细胞BHK-21的相对增值率,确定PHBV材料的毒性程度.实验结果表明,PHBV材料即使在高温下其物理化学性质也很稳定;它的细胞毒性程度为0级,有着良好的细胞生物相容性.     

新型淡水发光菌(Vibrio qinghaiensis sp.——Q67)应用于环境样品毒性测试的初步研究

研究表明，新型淡水发光菌（Ｖｉｂｒｉｏｑｉｎｇｈａｉｅｎｓｉｓｓｐ．———Ｑ６７）作为环境样品毒性检验的指标生物具有快速简便的优点．对重金属污染物毒性检验比以往Ｔ３发光菌具有更高的灵敏度，对鱼体进行的感染实验未发现明显病理改变．     

鲫鱼(Carassius auratus)卵黄蛋白原的ELISA检测

采用Sephadex G-200柱层析从鲫鱼(Carassius auratus)成熟鱼卵中提取卵黄脂磷蛋白并制备抗卵黄脂磷蛋白抗血清.用DEAE-cellulose阴离子交换层析和Sephacryl S-300分子筛凝胶层析相结合的两步层析从经17b-雌二醇诱导的雌鱼血浆中提纯卵黄蛋白原.免疫印迹反应显示卵黄脂磷蛋白和卵黄蛋白原都与抗卵黄脂磷蛋白抗血清有特异的反应.以抗卵黄脂磷蛋白抗血清作为抗体,卵黄蛋白原作为竞争抗原建立了间接竞争法酶联免疫吸附反应(ELISA)方法来检测雄鱼血液中卵黄蛋白原含量.该方法可测的最低浓度为7.8ng/mL,工作范围为0.39~25mg/mL,批内和批间误差分别为6.5%和7.5%.     

石油烃对栉孔扇贝血淋巴细胞DNA损伤的初步研究

为研究石油烃对海洋生物的毒性效应,将栉孔扇贝(Chlamys farreri)暴露于0.08、0.21和0.88mg·L-1石油烃中,采用单细胞凝胶电泳实验(彗星实验)技术检测不同暴露时间扇贝血淋巴细胞的DNA损伤程度,对照组中石油烃背景浓度为0.04mg·L-1。结果显示,低浓度(0.08mg·L-1)的石油烃短期(<7d)内即可导致栉孔扇贝血淋巴细胞的DNA损伤,并且随石油烃浓度的增大和暴露时间的延长,DNA损伤程度增加,石油烃浓度达0.88mg·L-1时,DNA损伤程度已非常严重。3d恢复实验后,各浓度组DNA损伤又均有不同程度的恢复。研究表明,彗星实验是检测石油烃对海洋贝类DNA损伤的一种有效手段,贝类血淋巴细胞DNA损伤有望成为石油烃污染的一种生物标志物,用于海洋污染的早期预警监测。     

11种取代酚类内分泌干扰活性的初步筛选与评价

通过聚蔗糖-泛影葡胺密度梯度离心法，从鲫鱼(Carassius auratus)外周血中分离出淋巴细胞，体外培养24h，用MTT法测试双酚类、联苯酚类、二酚类、烷基单酚类和取代酚类等几类可疑环境内分泌干扰物对淋巴细胞增殖的影响．结果表明，双酚类、联苯酚类和烷基单酚类物质对鲫鱼淋巴细胞的增殖具有低浓度促进，高浓度抑制的作用．二酚类物质对鲫鱼淋巴细胞的增殖具有促进作用．取代酚类对鲫鱼淋巴细胞的增殖无明显作用．该结果表明，此方法可以对环境内分泌干扰物进行初步筛选，同时发现环境内分泌干扰物对淋巴细胞增殖的效应与物质的化学结构具有一定相关性。