Examination of in vivo mutagenicity of sodium arsenite and dimethylarsinic acid in gpt delta rats

Arsenic is a well-known human bladder and liver carcinogen, but its exact mechanism of carcinogenicity is not fully understood. Dimethylarsinic acid(DMAV) is a major urinary metabolite of sodium arsenite(i As~Ⅲ) and induces urinary bladder cancers in rats. DMAVand i As~Ⅲare negative in in vitro mutagenicity tests. However, their in vivo mutagenicities have not been determined. The purpose of present study is to evaluate the in vivo mutagenicities of DMAVand i As~Ⅲin rat urinary bladder epithelium and liver using gpt delta F344 rats.Ten-week old male gpt delta F344 rats were randomized into 3 groups and administered 0,92 mg/L DMAV, or 87 mg/L i As~Ⅲ(each 50 mg/L As) for 13 weeks in the drinking water. In the mutation assay, point mutations are detected in the gpt gene by 6-thioguanine selection(gpt assay) and deletion mutations are identified in the red/gam genes by Spi-selection(Spi-assay). Results of the gpt and Spi-assays showed that DMAVand i As~Ⅲhad no effects on the mutant frequencies or mutation spectrum in urinary bladder epithelium or liver. These findings indicate that DMAVand i As~Ⅲare not mutagenic in urinary bladder epithelium or liver in rats.     

河流表层沉积物活体毒性甄别和毒性因子初探

本文选择辽河流域6个支流表层沉积物作为研究对象,利用2种底栖生物(摇蚊幼虫和河蚬)活体毒性测试方法评价了辽河流域表层沉积物毒性.研究结果显示,所有表层沉积物对摇蚊幼虫有毒性效应,显著降低了摇蚊幼虫的存活率(p<0.05);河蚬存活率有所降低,但差异不显著.沉积物活体毒性甄别结果发现,长沟子以有机物污染为主,付家窝堡和柴河以重金属污染为主,一统河和潮沟河以有机物和氨氮污染为主,而柳河以重金属和氨氮污染为主.综上所述,沉积物活体毒性甄别方法能有效甄别致毒污染物类别,同时表明不同种类底栖生物对沉积物毒性存在敏感性差异.     

利用氧化亚氮还原酶基因 (nosZ) 评价人工湿地系统中的反硝化菌

应用人工潜流湿地净化微污染地表水,出水用于补给人工景观河.利用定量PCR测定了湿地植物根际土壤和景观河底泥中16S rDNA和nosZ的丰度,并采用PCR-DGGE技术考察了样品中含nosZ基因的群落结构及其相似性.定量PCR结果表明,潜流湿地及人工景观河16S rDNA、nosZ平均绝对丰度(以DNA计)分别为1.91E+07、1.26E+06和2.68E+07、8.37E+05 copies.ng-1,以干土计时分别为1.45E+11、9.31E+09和5.31E+11、1.45E+10 copies.g-1.景观河底泥中微生物总量和反硝化菌数量要高于湿地根际土壤,但是后者nosZ相对丰度(3.8%～10.1%)则明显高于前者(1.7%～4.1%).根际土壤和底泥样品聚类分析相似度低,根际土壤优势菌种大部分与红杆菌目(Rhodobacearales)、根瘤菌目(Rhizobiales)和伯克氏菌目(Burkholderiales)的细菌相似,而底泥的优势菌种均为不可培养的微生物.     

模拟自然水体中藻-溞-草间的相互作用研究

为了解氮磷浓度对生物操纵效果及同时恢复水生植被的影响,以小球藻、大型溞和金鱼藻分别作为浮游植物、浮游动物和大型沉水植物的代表,模拟自然条件研究了不同氮磷浓度对三者生长和相互作用的影响.结果表明:金鱼藻和小球藻共培养时,在氮浓度介于2.92～12.60mg·L-1、磷浓度介于0.06 ～0.85 mg·L-1,金鱼藻增长不明显,甚至出现负增长;而小球藻的增长率则远高于金鱼藻.小球藻和大型溞共培养时,氮、磷浓度分别为1.26～ 10.53 mg·L-1和0.04～1.16 mg·L-1时均占据优势,对小球藻的抑制效果显著,抑藻及氮磷去除效果明显好于金鱼藻和小球藻共培养,而且磷的去除效果优于氮.三者共培养时,在氮浓度介于3.15 ～23.92 mg·L-1、磷浓度介于0.07 ～0.64 mg·L-1时,大型溞与金鱼藻的增长率都较高,而小球藻则维持在较低的增长水平,水质改善效果较好;当氮、磷浓度分别升至25.95 mg·L-1和1.18 mg·L-1时,大型潘和金鱼藻的增长率均下降,水质变差.大型溞和金鱼藻的联合控藻效果好于其单一的控制效果,该效果明显受到氮磷浓度的影响.     

农业废物好氧堆肥中环境因子对nirK、nirS和nosZ数量的影响

应用定量聚合酶链式反应(real-time PCR)技术对农业废物好氧堆肥过程中参与反硝化过程的功能基因(nirK、nirS和nosZ)丰度在堆体不同位置处随时间的变化情况进行了研究.结果表明,随着堆肥进程,3种基因数量整体呈现出先升后降的变化规律,且不同位置处的反硝化基因数量之间存在着显著的差异性.使用Canoco 4.5软件对获得的反硝化功能基因丰度数据与不同时期不同层次堆体温度、pH、含水率、NH4+-N、NO3--N和水溶性有机碳(WSC)等环境因子的相关性进行冗余分析(redundancy analysis,RDA).基于手动选择的RDA分析结果表明,WSC、NH4+-N和堆体温度对反硝化基因丰度有着显著的影响(P<0.05),且前2个因子达到了极显著水平(P<0.01).应用t-value回归分析方法单独分析每种环境因子与3种基因的相关性,其中nirK与温度和pH显著正相关(P<0.05),nirS与温度显著正相关(P<0.05),nosZ与NH4+-N显著正相关(P<0.05)、与WSC显著负相关(P<0.05).     

摇蚊幼虫与霍普水丝蚓扰动对河流沉积物中氮、磷、氧释放的影响

定量研究河床沉积物中的生物扰动对污染物的影响效应,对于维护河流健康具有重要的理论指导作用.基于室内实验模拟摇蚊和霍普水丝蚓对河床沉积物的扰动过程,研究两种生物扰动作用对沉积物中氮、磷释放以及生物扰动对上覆水中溶解氧的影响.在相同的沉积物环境中分别放入单一物种,实验结果显示,相对于空白组,摇蚊幼虫组的上覆水中氨氮、总氮、总磷平均净增量分别为2.32、0.787、0.105 mg · L^-1,霍普水丝蚓组的氨氮、总氮、总磷平均净增量分别为0.72、0.462、0.063 mg · L^-1,表明摇蚊幼虫和霍普水丝蚓的扰动作用均能促进沉积物中氨氮、总氮、总磷向上覆水中的释放,且摇蚊幼虫对污染物释放的作用效果比水丝蚓更明显;当摇蚊幼虫和水丝蚓混合放入后,摇蚊幼虫对沉积物中氨氮、总氮、总磷释放起主要促进作用;在混合实验组(摇蚊23条,霍普水丝蚓47条)中,氨氮、总氮、总磷的平均净增量均低于单一物种的摇蚊组和水丝蚓组,表明在该组中,摇蚊幼虫和霍普水丝蚓在沉积物中共同扰动作用对氮、磷释放的促进效果减弱;对上覆水中溶解氧浓度而言,摇蚊所占比例越高,溶解氧浓度就越低,且上覆水中的溶解氧浓度变化,会影响生物扰动对沉积物氮、磷释放的效果.     

氮负荷升高对螺-附着藻-苦草生态关系的影响

本文设计了双因子(氮负荷升高、有无螺类)受控实验,研究了氮负荷升高(太湖正常氮负荷的3倍)对螺-附着藻-苦草关系的影响.结果表明,氮负荷升高处理明显提高了水体总氮、总溶解氮、叶绿素a等指标的含量,显著增加了附着生物干重,进而抑制了苦草生长;与正常氮负荷处理组相比,3倍氮负荷处理组中苦草的相对生长率、株高和叶片数分别降低了33%、25%和13%.环棱螺降低了附着生物干重和水体叶绿素a含量,明显促进了苦草生长.氮负荷升高和环棱螺存在对水体各项理化指标、附着生物干重和苦草的各项生长指标均无交互作用.此外,氮负荷升高还降低了环棱螺的生长速率.分析认为,氮负荷升高对沉水植物生长的抑制机理主要体现在浮游藻类与附着生物生物量增加的抑制效应;环棱螺的存在虽然在一定程度上减弱了这种效应,但由于氮负荷升高还同时使环棱螺生长率降低,削弱了其对附着生物的牧食压力,从而使得附着生物对苦草生长的抑制作用加强.因此,氮负荷升高使螺-附着藻-沉水植物之间的生态关系失衡,也可能是氮浓度较高的富营养湖泊中沉水植被稀少的重要原因.     

Sediment pollution characteristics and in situ control in a deep drinking water reservoir

Sediment pollution characteristics, in situ sediment release potential, and in situ inhibition of sediment release were investigated in a drinking water reservoir. Results showed that organic carbon (OC), total nitrogen (TN), and total phosphorus (TP) in sediments increased from the reservoir mouth to the main reservoir. Fraction analysis indicated that nitrogen in ion exchangeable form and NaOH-extractable P (Fe/Al-P) accounted for 43% and 26% of TN and TP in sediments of the main reservoir. The Risk Assessment Code for metal elements showed that Fe and Mn posed high to very high risk. The results of the in situ reactor experiment in the main reservoir showed the same trends as those observed in the natural state of the reservoir in 2011 and 2012; the maximum concentrations of total OC, TN, TP, Fe, and Mn reached 4.42 mg/L, 3.33 mg/L, 0.22 mg/L, 2.56 mg/L, and 0.61 mg/L, respectively. An in situ sediment release inhibition technology, the water-lifting aerator, was utilized in the reservoir. The results of operating the water-lifting aerator indicated that sediment release was successfully inhibited and that OC, TN, TP, Fe, and Mn in surface sediment could be reduced by 13.25%, 15.23%, 14.10%, 5.32%, and 3.94%, respectively.     

黄连根茎浸提物对隆线溞的急性毒性作用

黄连根茎和制剂具有抗菌等作用,广泛用于水产养殖.试验以隆线溞单克隆生物株为供试生物,研究了不同浓度的黄连根茎浸提物(Coptis chinensis root extracts,CRE)对隆线溞泳动能力、存活率及趋光性的影响.结果表明,黄连根茎浸提物对水溞具有急性毒性,半致死浓度半抑制浓度趋光指数显著改变的浓度,后者仅4.27 mg·L-1(以CRE浓度计),远远低于防治鱼病黄连进入水体的浓度.因此,水产养殖使用黄连药物对水溞存在生物毒性.此外,导致隆线溞趋光指数显著改变的CRE浓度比半致死浓度低30.62~36.51倍,测试时间从48 h或24 h缩短到3 h,且容易观察,利用隆线溞趋光性的改变可快速监测CRE对其的毒害作用.在水产养殖中,如果滥用黄连制剂,会影响水生食物链,导致水生态系统失衡.     

巢湖沉积物有效态磷与硫的DGT原位同步分析研究

沉积物有效态磷(P)与硫(S)的含量分布是影响湖泊营养元素循环和水体富营养化的重要因子.本文利用Zr O-Ag I薄膜梯度扩散技术(Zr O-Ag I DGT)原位同步获取了南淝河-巢湖沉积物中有效态P和S的高分辨分布信息.结果发现:绝大部分沉积物有效P和S自界面向下呈急剧升高趋势,并在50 mm深度内出现峰值;河道内沉积物有效P和S的含量高于湖体,且P在界面的表观释放通量明显高于湖体,表明河道沉积物是湖体P的输入源.部分采样点出现有效P与S同步变化的现象,两者具有显著相关性,证实沉积物内部P和S存在同步释放的规律.     

对苯二酚抑制铜绿微囊藻生长下藻毒素的产生与释放

选用对苯二酚,根据其抑藻的剂量效应关系,研究了铜绿微囊藻在不同抑制条件下,即投加剂量为EC20、EC50、EC70、EC90、EC99时,藻毒素Microcystin-LR(MCLR)的产生与释放,并通过测定总有机碳和三维荧光光谱,研究了细胞内有机物的释放状况.结果表明,EC20作用下,藻毒素的产生受到抑制,总MCLR约为对照样的72.4%~83.0%;在EC50~EC99作用下,藻细胞受到刺激,毒素产量明显增加,总MCLR可升至对照样的1.77~3.13倍,且在EC70~EC99作用下,胞内毒素大部分释放于胞外.细胞释放的其它有机物主要是类腐殖酸和类富里酸,但并不稳定,6 d后,发生了明显的降解与转化.     

基于PCR-DGGE和拟杆菌(Bacteroides)16S rRNA的岩溶地下水粪便污染来源示踪研究:以重庆南山老龙洞地下河系统为例

岩溶地下水的微生物污染日益严重,其来源的研究得到国际学术界的广泛关注.本研究以重庆南山老龙洞岩溶地下河系统为对象,采用滤膜法监测地下水中的总细菌、大肠杆菌、粪大肠菌及粪链球菌等微生物指标,以拟杆菌(Bacteriodes)为指示细菌,采用PCR-DGGE示踪地下水中大肠杆菌/粪大肠杆菌的来源.结果表明,老龙洞地下河流域各类细菌含量严重超标,总细菌数为10~2.9×10~7CFU·m L~(-1),大肠菌群总数达4.3~4.0×10~5CFU·m L~(-1),其中粪大肠菌群(FC)和粪链球菌(FS)分别最高达到1.1×10~6CFU·(100 m L)~(-1)、1.1×10~5CFU·(100 m L)~(-1);FC/FS多数为2以上,暗示流域地下水受人类粪便影响为主.地下水样和粪便样品的拟杆菌群落的PCR-DGGE比对分析表明地下水与人粪之间相似性为7.1%~69.1%,其中地下河出口处达到69.1%.地下水与猪粪之间相似性为1.1%~53.4%,地下河出口处仅为1.5%.因此,人类粪便为地下河污染的主要来源,猪粪污染为动物粪便污染的一部分,还存在其他动物粪便污染来源.此外,PCR-DGGE产物切胶测序发现大部分Bacteroides为人类肠道或粪便来源的细菌.     

陇东黄土高原石油污染土壤环境因子对金盏菊(Calendula officinalis)-微生物联合修复的响应

为筛选适合陇东黄土高原石油污染土壤修复植物,选取当地"适生"植物金盏菊(Calendula officinalis)联合微生物菌剂开展了为期5个月的原位修复实验.采用常规方法和PCR-DGGE技术分析了修复过程中土壤理化性质、酶活性及土壤微生物群落遗传多样性等15项土壤环境因子的动态变化情况及其对金盏菊生态修复的响应.结果表明,随着原位修复的进行,1土壤脲酶、多酚氧化酶、过氧化氢酶及脱氢酶活性均不同程度呈增加的趋势(p0.05).2土壤TPHs、p H、有机质、含盐率的变化情况不尽相同,但总体呈下降的趋势(p0.05);土壤碱解氮、速效磷及速效钾含量总体呈上升趋势(p0.05).3土壤微生物群落遗传多样性结果显示,Shannon-Wiener多样性指数和Pielou均匀度指数整体呈逐步增加的趋势.4PCA分析结果显示,土壤脲酶、脱氢酶及碱解氮等环境因子对金盏菊修复表现为促进作用,而有机质含量过高将不利于金盏菊的植物修复.金盏菊和微生物菌剂联合修复过程中,土壤多酚氧化酶、速效磷、脱氢酶及速效钾发挥主要作用,而土壤TPHs和含盐率过高则影响了联合修复效果.本研究旨在分析金盏菊用于陇东地区油污土壤的生态响应,结果有助于筛选出适合陇东黄土高原土壤类别-适生植物-土著降解菌群的组合,利用乡土物种的适生性降低修复成本,为陇东地区油污土壤生态修复提供新的技术方案和背景资料.     

Assessment of Bisphenol A (BPA) neurotoxicity in vitro with mouse embryonic stem cells

The adverse effects of environmental pollution on our well-being have been intensively studied with many in vitro and in vivo systems. In our group, we focus on stem cell toxicology due to the multitude of embryonic stem cell (ESC) properties which can be exerted in toxicity assays. In fact, ESCs can differentiate in culture to mimic embryonic development in vivo, or specifically to virtually any kind of somatic cells. Here, we used the toxicant Bisphenol A (BPA), a chemical known as a hazard to infants and children, and showed that our stem cell toxicology system was able to efficiently recapitulate most of the toxic effects of BPA previously detected by in vitro system or animal tests. More precisely, we demonstrated that BPA affected the proper specification of germ layers during our in vitro mimicking of the embryonic development, as well as the establishment of neural ectoderm and neural progenitor cells.     

物理和摇蚊幼虫组合扰动对内源磷再生和形态转化的协同作用

为探讨物理和摇蚊幼虫组合扰动对内源磷再生和形态转化的协同作用,选取了苏州某富营养化河道为研究对象.利用室内静态培养试验和Rhizon间隙水采样技术,着重对比了单纯的摇蚊幼虫扰动和组合扰动下,上覆水、间隙水、沉积物中磷形态数量分布的变化规律.结果表明,组合扰动下,上覆水中不同形态磷含量(TP、PP、DTP、DIP)均处于较高水平,明显高于摇蚊幼虫扰动.这可归因于微生物活性的显著增加.与摇蚊幼虫扰动相比,组合扰动下,间隙水中DIP和Fe2+浓度的降低幅度和降低范围明显更大.这主要是由于物理扰动和底栖生物扰动产生的叠加效应,导致溶解氧渗透深度增加.另外,0~2 cm沉积物中,NH4Cl-P含量明显降低,Fe/Al-P明显增加,并且,NH4Cl-P和Fe/Al-P的变化幅度均在组合扰动下最大.这暗示了物理扰动和底栖生物扰动对磷再生和迁移产生的叠加效应.     

不同处理水芹浮床对城市河道黑臭污水的脱氮效果及其机理研究

通过对冬季不同留茬高度水芹（Oenanthe Javanica）浮床对城市河道污水氮循环细菌数量和脱氮效果的影响研究,试图阐明浮床植物去除污染水体氮素的可能途径.研究结果表明：不同处理水芹浮床对城市河道黑臭污水硝氮和氨氮的去除效果分别达到了52.5%～69.8%和68.1%～76.7%,19cm留茬高度水芹浮床对脱氮效...     

Arsenic methylation by an arsenite S-adenosylmethionine methyltransferase from Spirulina platensis

Arsenic-contaminated water is a serious hazard for human health. Plankton plays a critical role in the fate and toxicity of arsenic in water by accumulation and biotransformation.Spirulina platensis(S. platensis), a typical plankton, is often used as a supplement or feed for pharmacy and aquiculture, and may introduce arsenic into the food chain, resulting in a risk to human health. However, there are few studies about how S. platensis biotransforms arsenic. In this study, we investigated arsenic biotransformation by S. platensis. When exposed to arsenite(As(Ⅲ)), S. platensis accumulated arsenic up to 4.1 mg/kg dry weight.After exposure to As(Ⅲ), arsenate(As(Ⅴ)) was the predominant species making up 64% to86% of the total arsenic. Monomethylarsenate(MMA(Ⅴ)) and dimethylarsenate(DMA(Ⅴ))were also detected. An arsenite S-adenosylmethionine methyltransferase from S. platensis(Sp Ars M) was identified and characterized. Sp Ars M showed low identity with other reported Ars M enzymes. The Escherichia coli AW3110 bearing Spars M gene resulted in As(Ⅲ) methylation and conferring resistance to As(Ⅲ). The in vitro assay showed that Sp Ars M exhibited As(Ⅲ) methylation activity. DMA(Ⅴ) and a small amount of MMA(Ⅴ) were detected in the reaction system within 0.5 hr. A truncated Sp Ars M derivative lacking the last 34 residues still had the ability to methylate As(Ⅲ). The three single mutants of Sp Ars M(C59S, C186 S, and C238S) abolished the capability of As(Ⅲ) methylation, suggesting the three cysteine residues are involved in catalysis. We propose that Sp Ars M is responsible for As methylation and detoxification of As(Ⅲ) and may contribute to As biogeochemistry.     

A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone (TH) signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A (TBBPA), a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3, whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.     

苦草（Vallisneria natans）根系对沉积物中各形态磷的影响

采用磷分级提取的方法研究苦草(Vallisneria natans)生长过程中根系分布与沉积物不同形态磷和总磷的变化过程.在实验开始后20、50、80 d,分别测定苦草根系在沉积物的分布及沉积物中各形态磷的垂直变化.结果表明,苦草根系在实验装置中垂直分布在0~14 cm区域内.平均每株苦草根系根数为58条,平均根长为5.86 cm.苦草根系生物量在沉积物深度0~3、4~6、7~10、11~14 cm分配的质量分数分别为45.99%、32.75%、16.03%、5.23%.沉积物中总磷(TP),NaOH提取磷(NaOH-P)和有机磷(OP)含量在苦草根系分布集中区域内显著降低(P<0.05).HCl提取磷(HCl-P)和无机磷(IP)的含量无显著差异(P>0.05).