低剂量全氟辛酸对雄性黑斑蛙生殖毒效应及机理研究

针对新型环境有机污染物全氟辛酸(PFOA)对两栖动物的毒效应问题,以黑斑蛙(Rana nigromaculata)为实验对象,研究了低剂量PFOA暴露对雄性黑斑蛙生殖毒效应及机理.结果表明,在0.001～1mg.L-1低剂量体外暴露20d条件下,PFOA能够显著诱导黑斑蛙精子发生畸形,相对于对照组,精子畸形率呈现明显的剂量-效应关系.酶联免疫试剂盒检测发现,除0.001mg.L-1诱导组外,0.01、0.1和1mg.L-1暴露组的血清雄性激素睾酮和雌性激素雌二醇含量分别显著降低(p<0.01,F=288.7)和上升(p<0.01,F=289.0),说明PFOA作用下黑斑蛙体内性激素水平受到改变.采用荧光定量PCR技术进行进一步的分子生物学检测,发现与性激素水平密切相关的两个基因P450芳香化酶和类固醇激素合成因子(SF-1)均增强表达(0.001mg.L-1诱导组,p<0.05;0.01、0.1和1mg.L-1暴露组,p<0.01,F=140.8).全氟辛酸对两栖类动物的雄性生殖毒效应机理主要是通过介导与性激素分泌相关的两个重要基因P450芳香化酶和类固醇激素合成因子,使其表达上调,从而一方面抑制睾酮的产生,另一方面促进性激素雌二醇的分泌.在0.001mg.L-1的体外暴露剂量下,PFOA就能够明显诱导黑斑蛙精子产生畸形毒性及增强P450芳香化酶和SF-1基因的表达(p<0.05),因此,为保护两栖动物种群,在制订PFOA地表水环境质量标准时应当考虑其下限不能低于0.001mg.L-1.     

全氟辛烷磺酸盐和全氟辛酸在水环境及水生生物体内积累的研究进展

全氟辛烷磺酸盐(PFOS)和全氟辛酸(PFOA)是全氟化合物中2种最典型的化合物,在不同环境介质(水、土壤、沉积物等)及水生生物体内被广泛检出。PFOS和PFOA具有良好的化学、物理性能,被广泛地运用于工业领域,PFOS和PFOA的污染范围已涉及全球。综述了PFOS和PFOA在水环境体系的污染现状及其在水生生物体内积累特征,探讨影响水生生物积累的主要因素,为全氟化合物污染控制提供基础。     

全氟辛酸在紫贻贝体内的富集、分布与消除

以紫贻贝为受试生物,采用半静态暴露的方式,研究了全氟辛酸（PFOA）在紫贻贝体内的富集特征、器官分布与消除规律.结果表明,紫贻贝可快速富集PFOA,整贝在不同浓度PFOA暴露下,于4~6d达到平衡态,半衰期为0.34~0.44d（约8~10h）,生物富集系数（BCFs）为5.10~9.55mL/g.PFOA在紫贻贝鳃和内脏团中的富集作用大于外套膜、性腺和闭壳肌.紫贻贝中PFOA的含量在净化6~9d后趋于空白组水平,主要代谢途径可能为鳃的作用.紫贻贝对PFOA的富集作用与暴露浓度相关,浓度越低,吸收系数和生物富集系数越大.     

PFOS/PFOA环境污染行为与毒性效应及机理研究进展

全氟辛烷磺酰基化合物（PFOS）和全氟辛酸（PFOA）是一类新型的持久性有机污染物（POPs）,近年来发现在环境系统中日益广泛分布,并在生物体内蓄积或发生致毒效应.本文首先从PFOS/PFOA在环境中的污染及其水平、在野生动物体内的暴露、对人体的暴露以及污染与暴露变化趋势等4个方面,分析了PFOS/PFOA最新的环境污染与生物暴露情况;从PFOS/PFOA在大气环境中的转运转化过程、在污水污泥中转运转化过程以及在生物体内的蓄积、代谢转化与降解过程等3个方面,阐述了PFOS/PFOA在环境中的迁移转化行为;还概述了最近几年在PFOS/PFOA所导致的生态效应及其可能的机理研究进展.最后,尝试性地提出了今后在PFOS/PFOA污染生态学方面的研究重点.     

妊娠期全氟辛酸染毒对子代雄鼠发育毒性的研究

探讨妊娠期染毒全氟辛酸(perfluorooctanoic acid,PFOA)对子代雄鼠发育毒性的影响.将50只妊娠0 d小鼠随机分5组,每组10只,妊娠0~17 d每天按0、1、2.5、5和12.5 mg·kg~(-1)BW饲喂PFOA,产后仔鼠正常饲养,记录仔鼠存活数.产后21 d,检测仔鼠血清睾酮水平并计算睾丸指数;HE染色观察睾丸组织结构变化;RT-PCR法检测睾丸组织Dlk1-Dio3印记基因簇各目的基因mRNA的表达情况.结果显示,随PFOA染毒剂量增加,仔鼠存活数显著降低,血清睾酮含量极显著性降低(p0.01);睾丸指数无统计学意义;睾丸组织有不同程度的损伤并呈剂量依赖性;Dlk1-Dio3印记基因簇各目的基因的表达量都有降低趋势,其中Gtl2、Rian、Dio3均显著性降低(p0.05).结果表明,妊娠期染毒PFOA能够降低仔鼠存活数,损伤睾丸组织结构,并能够扰乱生殖激素,降低睾丸组织Dlk1-Dio3印记基因簇各目的基因mRNA的表达.     

新型消毒副产物碘代乙酸急性暴露诱导斑马鱼肝脏氧化损伤的机理研究

具有较好水溶性的碘代乙酸是新型饮用水消毒副产物,其对水生生物的毒效应及机理备受关注.肝脏是鱼类发挥解毒作用的的主要组织,极易受化学污染物影响.本研究以模式水生动物斑马鱼(Danio rerio)为研究对象,采用半静态换水法,研究了碘代乙酸急性暴露诱导鱼体解毒功能组织氧化损伤的毒效应机理.暴露96 h后,相对于对照组,不同剂量碘代乙酸(1μg·L-1、10μg·L-1和100μg·L-1)暴露导致斑马鱼肝脏组织活性氧和丙二醛含量明显增加;超氧化物歧化酶和谷胱甘肽S转移酶活性增强,而过氧化氢酶活性降低;肝脏解毒过程标志性基因CYP1A、CYP3A和GST在mRNA水平表达量不断增加;这些指标的变化都呈现明显的剂量-效应特征.上述研究表明,饮用水消毒过程形成的碘代乙酸对水生生物具有明显的氧化应激效应,鱼体肝脏组织解毒功能基因相对于其它指标更敏感,可作为生物标志物.此外,应当加强饮用水消毒工艺研究,减少碘代乙酸等消毒副产物的形成以降低其对水生生态系统的潜在危害.     

全氟辛烷磺酸(PFOS)对斑马鱼卵黄蛋白原mRNA水平的影响

为了研究环境低剂量全氟辛烷磺酸（perfluorooctane sulfonate,PFOS）对水生生物的内分泌干扰效应和可能的作用机制,测定了PFOS对斑马鱼（Brachydanio rerio）肝脏中卵黄蛋白原（vitellogenin,VTG）mRNA水平的影响.将斑马鱼暴露于4个PFOS的环境低剂量浓度组（0.1、1、10、100μg.L-1）中进行21d毒性试验,收集肝脏样品,提取RNA,采用荧光定量PCR（qRT-PCR）分别检测VTG1和VTG3的mRNA水平.结果表明：①PFOS暴露引起雄性斑马鱼肝脏VTG1和VTG3 mRNA水平升高,VTG1 mRNA水平升高与剂量呈正相关,在100μg.L-1暴露浓度处与对照组呈现显著性差异;VTG3的mRNA水平变化与剂量呈倒U型曲线,呈现典型的毒物刺激荷尔蒙效应,在10和100μg.L-1暴露浓度处与对照组呈现显著性差异;②PFOS暴露引起雌性斑马鱼肝脏中VTG1 mRNA水平升高,在10μg.L-1暴露浓度处与对照组呈现显著性差异,但在高浓度（10和100μg.L-1）处试验结果误差较大;VTG3 mRNA水平只在10μg.L-1暴露浓度处升高,但相比于对照组均没有显著性差异.试验结果表明PFOS暴露对斑马鱼的内分泌干扰作用明显,其毒性作用机制可能是类雌激素效应,而肝脏中VTG1和VTG3mRNA水平可能作为PFOS内分泌干扰效应评价的敏感生物标志物,但VTG1和VTG3 mRNA水平的响应曲线呈现基因亚型和性别差异.     

东湖表层水体中全氟辛酸和全氟辛磺酸空间分布特征

研究了东湖表层水体中全氟辛酸(PFOA)和全氟辛磺酸(PFOS)的含量及空间分布特征,总量范围为31.1～237ng.L-1,几何平均值为115 ng.L-1,表明整个湖区受到不同程度的PFOS和PFOA污染.全部水样均检出了PFOS和PFOA,峰值达到132和158 ng.L-1,几何平均值分别为60.4和55.0 ng.L-1.湖区东部污染最严重,其次是湖区南部,湖区北部和西部污染程度相对较轻.与PFOS相比,高浓度PFOA分布相对分散,PFOS和PFOA含量之间无显著相关性,表明东湖表层水体中PFOS和PFOA属于多源输入.30个(63%)取样点上PFOS含量超过可能对水生生态系统产生影响的临界数值(43 ng.L-1),20个(42%)取样点上PFOA含量超过美国新泽西洲制定的饮用水标准(40 ng.L-1).     

铅胁迫对斑马鱼抗氧化酶活性的影响

研究Pb对斑马鱼抗氧化酶活性影响,探讨重金属Pb对斑马鱼毒理学效应.以斑马鱼为模式生物,采用半静态法对斑马鱼进行驯养和试验,测定不同浓度Pb对超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性影响.Pb暴露1d胁迫斑马鱼SOD为诱导效应,胁迫CAT为诱导-抑制效应,Pb暴露7d胁迫斑马鱼SOD为诱导效应,胁迫CAT为诱导-抑制效应,且暴露1d和暴露7d酶活性变化均显著.试验结果表明,Pb胁迨斑马鱼抗氧化系统酶活性变化显著,Pb毒性显著.结果为重金属污染对水生生物毒性机理研究提供参考.     

氟对斑马鱼生殖毒性研究

为了探讨氟对鱼类的生殖毒性效应,以成年斑马鱼为试验对象,氟化钠暴露45 d和90 d后,运用组织切片观察斑马鱼性腺组织的发育情况,采用酶联免疫分析法测定性腺中雌二醇（Estradiol,E2）、卵黄蛋白原（Vitellogenin,VTG）和睾酮（Testosterone,T）的含量,并使用实时荧光定量PCR方法检测斑马鱼生殖相关基因的表达水平.结果显示：氟暴露会阻滞成年斑马鱼性腺发育,造成卵泡发育不良,精巢生精能力下降;同时降低雌性斑马鱼性腺中E2和VTG的含量,增加T的含量;雌性斑马鱼孕酮受体（Prg）和糖皮质激素受体（GR）基因相对表达量随氟浓度的增加呈先降后升的趋势,随着染毒剂量与时间的增加,雄激素受体（AR）基因相对表达量逐渐增加,而CYP19a基因相对表达量逐渐减少.雄性斑马鱼性腺中E2、VTG、T的含量及生殖相关基因表达的变化趋势则与雌性斑马鱼相反.综上所述,氟暴露可以通过影响斑马鱼体内性腺发育、E2、VTG、T含量及生殖相关基因表达来影响其生殖功能.实验结果可为进一步研究氟化物对水生动物的生殖毒性效应提供理论依据.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Sorption of 1-naphthol by plant cuticular fractions

The contribution of aliphatic-rich plant biopolymer to sorption of hydrophobic organic compounds is significantly important because of their preservation and accumulation in the soil environment,but sorption mechanism is still not fully understood.In this study, sorption of 1-naphthol by plant cuticular fractions was examined to better understand the contributions of respective fraction.Toward this end,cuticular materials were isolated from the fruits of tomato by chemical method.The tomato cuticle sheet consisted of waxes (6.5 wt%),cuticular monomer (69.5 wt%),and polysaccharide (24.0 wt%).Isotherms of l-naphthol to the cuticular fractions were nonlinear (N value (0.82-0.90)) at the whole tested concentration ranges.The KodKow ratios for bulk cuticle (TC1),dewaxed cuticle (TC2),cutin (TC4),and desugared cuticle (TC5) were larger than unity,suggested that tomato bulk cuticle and cutin are much powerful solption medium.Sorption capability of cutin (TC4) was 2.4 times higher than the nonsaponifiable fraction (TC3).The 1-naphthol interactions with tomato cuticular materials were governed by both hydrophobic-type interactions and polar (H-bonding) interactions. Removal of the wax and polysaccharide materials from the bulk tomato cuticle caused a significant increase in the sorption ability of the cuticular material.There was a linear negative trend between K_(oc) values and the amount of polysaccharides or fraction's polarities ((N O)/C);while a linear positive relationship between K_(oc) values and the content of cutin monomer (linear R~2=0.993) was observed for present in the cuticular fractions.Predominant sorbent of the hydrophobic organic compounds (HOCs) in the plant cuticular fraction was the cutin monomer,contributing to 91.7% of the total sorption of tomato bulk cuticle.