Biological Activity in a Heavily Organohalogen-Contaminated River Sediment (8 pp)

Background, Aims and Scope Sediments of the Spittelwasser creek are highly polluted with organic compounds and heavy metals due to the discharge of untreated waste waters from the industrial region of Bitterfeld-Wolfen, Germany over the course of more than one century. However, relatively few data have been published about the chloroorganic contamination of the sediment. This paper reports on the content of different (chloro)organic compounds with special emphasis on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), and chlorobenzenes. Existing concepts for the remediation of Spittelwasser sediment include the investigation of natural attenuation processes, which largely depend on the presence of an intact microbial food web. In order to gain more insight in terms of biological activity, we analyzed the capacity of sediment microflora to degrade organic matter by measuring the activities of extracellular hydrolytic enzymes involved in the biogeochemical cycling of carbon, nitrogen, phosphorus and sulfur. Furthermore, the detection of physiologically active bacteria in the sediment, particularly of those known for their capability to reductively dehalogenate organochlorine compounds, illustrates the potential for intrinsic bioremediation processes. Methods PCDD/F and chlorobenzenes were analyzed by gas chromatography(GC)/mass spectrometry and GC/flame ionization detection, respectively. The activities of hydrolytic enzymes were determined from freshly sampled sediment layers using 4-methylumbelliferyl (MUF) or 7-amino-4-methylcoumarin-conjugated model compounds and kinetic fluorescence measurements. Physiologically active bacteria from different sediment layers were microscopically visualized by fluorescence in situ hybridization (FISH). Specific bacteria were identified by 16S rRNA gene amplification and sequencing. Results and Discussion The PCDD/F congener profile was dominated by dibenzofurans. In addition, the presence of specific tetra and pentachlorinated dibenzofurans supported the assumption that extensive magnesium production was one possible source for the high contamination. A range of other chloroorganic compounds, including several isomers of chlorobenzenes, hexachlorocyclohexane and 1,1,1-trichloro-2,2-bis (p-chloro-phenyl)ethane (DDT), was present in the sediment. Activities of extracellular hydrolytic enzymes showed a strong decrease in those sediment layers that were characterized by high contents of absorbable organic halogen (AOX), indicating disturbed organic matter decay. Interestingly, an abnormal increase of cellulolytic enzyme activities below the organochlorine-rich layers was observed, possibly caused by residual cellulose from discharges of sulfite pulping wastes. FISH revealed physiologically active bacteria in most sediment layers from the surface down to the depth of about 60 cm, including members of Desulfitobacterium (D.) and Sulfurospirillum. The presence of D. dehalogenans was confirmed by its partial 16S rRNA gene sequence. Conclusions Results of chemical sediment analyses demonstrated high loads of organochlorine compounds, particularly of PCDD/F. Several years after stopping the waste water discharge to Spittelwasser creek, this sediment remains a main source for pollution of the downstream river system by way of the ongoing mobilization of sediment during high floods. As indicated by our enzyme activity measurements, the decomposition potential for organic matter is low in organochlorine-rich sediment layers. In contrast, the comparably higher enzyme activities in less organochlorine-polluted sediment layers as well as the presence of physiologically active bacteria suggest a considerable potential for natural attenuation. Recommendations and Perspectives From our data we strongly recommend to explore the degradative capacity of sediment microorganisms and the limits for in situ activity towards specific sediment pollutants in more detail. This will give a sound basis for the integration of bioremediation approaches into general concepts to reduce the risk that permanently radiates from this highly contaminated sediment. Submission Editor: Dr. Henner Hollert (Henner.Hollert@urz.uniheidelberg.de)     

Rapid detection of numerical aberrations of chromosomes 13, 18 and 21 in chorionic mesenchymal cells

We have devised and evaluated a rapid screening method for the detection of numerical aberrations of chromosomes13, 18 and 21 in chorionic villus cells. We used non-radioactive in situ hybridization (ISH) with three chromosome-specific probes on overnight-attached mesenchymal cells from chorionic villi. A blind study was performed of 47 karyotypically normal samples, one triploid sample, two samples trisomic for chromosome 21, and two samples from a fetus with putative mosaicism (46/47, +21). All samples were hybridized with the chromosome 18- and 21-specific probes. Thirty samples were additionally hybridized with the chromosome 13-specific probe. The test could be completed within 3-4 days of sampling. In samples disomic with respect to the probed chromosomes, an average of 2 per cent (range 0-9 per cent) had three hybridization signals. By contrast, in the samples trisomic for the probed chromosome(s), 57 per cent (chromosome 13), 51 per cent (chromosome 18), and an average of 74 per cent (55-86 per cent) (chromosome 21) of the nuclei exhibited three signals. In the putative mosaic samples, the number of nuclei with three chromosome 21-specific signals ranged from 41 to 69 per cent. We conclude that this technique rapidly and clearly distinguishes between normal and trisomic/triploid samples, and consequently may be of use in future prenatal diagnosis.     

聚糖原菌富集实验及其内源过程探究

细胞衰减是微生物内源过程的一个重要组成部分,可分为由细胞死亡引起的数量衰减和由细胞活性降低引起的活性衰减两部分.通过挥发性脂肪酸（VFA）吸收速率(VFAUR)测定、荧光原位杂交技术（FISH）以及LIVE/DEAD细胞染色技术,研究了富集聚糖原菌（GAOs）在序批式反应器（SBR）系统中好氧环境下的衰减特征.结果表明,当T=30℃、进料中m(COD)∶m(P)=100时,SBR系统中GAOs富集比例达94%.测定和计算表明,SBR富集系统中GAOs衰减速率和死亡速率分别为0.132 d^-1和0.034 d^-1,其数量衰减和活性衰减占其细胞总衰减比例分别为26%和74%.可见,GAOs数量衰减只占其细胞总衰减中很小一部分,而绝大部分衰减由活性衰减所引起.     

土霉素对小麦种子发芽与幼苗生长发育的生态毒性

研究了土霉素对小麦种子发芽以及早期幼苗生长发育的生态毒性效应.结果表明,土霉素对小麦种子芽长及根长的抑制效应显著(p<0.01),而且它们之间具有良好的剂量-效应关系.根据线性回归方程得出土霉素对小麦种子芽长和根长的半抑制浓度为65.5 mg/L和34.7 mg/L.然而,土霉素对小麦种子的发芽率并没有显著的影响.研究还表明,0.15~2.4 mg/L土霉素暴露21 d后,小麦叶片中的叶绿素含量降低了35.6%~47.3%,叶片及根部的可溶性蛋白含量也均呈显著下降趋势.暴露7 d后,0.15~2.4 mg/L土霉素对小麦叶片和根部SOD与POD活性的抑制效应不显著,但随着暴露时间的延长,土霉素对小麦的SOD与POD酶活性抑制率显著下降;2.4 mg/L土霉素暴露21 d后,小麦根部的SOD活性下降72.3%,说明土霉素对小麦幼苗体内的抗氧化系统具有破坏作用.该实验结果显示,低浓度土霉素长期暴露对小麦幼苗的生长发育具有不良的生态毒性效应.     

Sulfur speciation and bioaccumulation in camphor tree leaves as atmospheric sulfur indicator analyzed by synchrotron radiation XRF and XANES

Analyzing and understanding the effects of ambient pollution on plants is getting more and more attention as a topic of environmental biology.A method based on synchrotron radiation X-ray fluorescence and X-ray absorption near edge structure spectroscopy was established to analyze the sulfur concentration and speciation in mature camphor tree leaves (CTLs),which were sampled from 5 local fields in Shanghai,China.Annual SO2 concentration,SO42-concentration in atmospheric particulate,SO42-and sulfur concentration in soil were also analyzed to explore the relationship between ambient sulfur sources and the sulfur nutrient cycling in CTLs.Total sulfur concentration in mature camphor tree leaves was 766-1704 mg/kg.The mainly detected sulfur states and their corresponding compounds were +6 (sulfate,include inorganic sulfate and organic sulfate),+5.2 (sulfonate),+2.2 (suloxides),+0.6 (thiols and thiothers),+0.2 (organic sulfides).Total sulfur concentration was strongly correlated with sulfate proportion with a linear correlation coefficient up to 0.977,which suggested that sulfur accumulated in CTLs as sulfate form.Reduced sulfur compounds (organic sulfides,thiols,thioethers,sulfoxide and sulfonate) assimilation was sufficed to meet the nutrient requirement for growth at a balanced level around 526 mg/kg.The sulfate accumulation mainly caused by atmospheric sulfur pollution such as SO2 and airborne sulfate particulate instead of soil contamination.From urban to suburb place,sulfate in mature CTLs decreased as the atmospheric sulfur pollution reduced,but a dramatic increase presented near the seashore,where the marine sulfate emission and maritime activity pollution were significant.The sulfur concentration and speciation in mature CTLs effectively represented the long-term biological accumulation of atmospheric sulfur pollution in local environment.     

辽河流域与英国中部河湖水体中溶解有机质的荧光特性

运用三维荧光光谱技术对辽河流域和英国中部水体中溶解有机质(DOM)的荧光光谱特性进行了分析.结果表明,辽河流域各研究水体中DOM包含4种荧光组分,即类色氨酸、类酪氨酸、可见光区和紫外光区类腐殖质,通过对荧光特性定量指标的分析可知,辽河流域各研究断面水体中类腐殖质物质内源作用大于陆源,DOM具有较强的新近自生源,各河流水体中DOM腐殖化程度顺序为:浑河中游海城河中游蒲河下游太子河下游.相比之下,英国河湖中,除了River Tame含有以上4种荧光组分外,其它水体中均没有发现类酪氨酸荧光物质,甚至在River Tern中仅含有类腐殖质荧光物质,河湖中DOM含有较少的新近自生源组分,以陆源影响为主.另外,辽河流域水体中类蛋白与类腐殖质荧光峰强度比值大于英国各河流水体的平均值.由此可以推断,辽河流域各研究断面大量污染物的输入,是造成其严重内源污染的主要原因,从而使水体中DOM含量增加,英国各河流断面其DOM主要来自于水体本身存在的大量微生物和浮游植物的代谢活动以及土壤有机质经雨水的冲刷流入水体的贡献.     

Determination of ammonium on an integrated microchip with LED-induced fluorescence detection

A simply fabricated microfluidic device integrated with a fluorescence detection system has been developed for on-line determination of ammonium in aqueous samples. A 365-nm light-emitting diode (LED) as an excitation source and a minor band pass filter were mounted into a polydimethylsiloxane (PDMS)-based microchip for the purpose of miniaturization of the entire analytical system. The ammonium sample reacted with o-phthaldialdehyde (OPA) on-chip with sodium sulfite as reducing reagent to produce a fluorescent isoindole derivative, which can emit fluorescence signal at about 425 nm when excited at 365 nm. Effects of pH, flow rate of solutions, concentrations of OPA-reagent, phosphate and sulfite salt were investigated. The calibration curve of ammonium in the range of 0.018-1.8 μupg/mL showed a good linear relationship with R² = 0.9985, and the detection limit was (S/N = 3) 3.6 × 10^-4 μupg/mL. The relative standard deviation was 2.8% (n = 11) by calculating at 0.18 μupg/mL ammonium for repeated detection. The system was applied to determine the ammonium concentration in rain and river waters, even extent to other analytes fluorescence detection by the presented device.     

珠江广州河段着生藻类的群落结构及其与水质的关系

于2007年6月、7月和9月,采用玻璃挂片法对珠江广州河段9个断面处着生藻类的分布状况进行了调查,同时测定了叶绿素a、总氮、总磷、溶解氧、pH等环境因子.本调查共观察到着生藻类35种,主要以丝状绿藻占据优势,其中毛枝藻是绝对优势种,其最高数量百分比达97.4%;而附着在绿藻藻体的一些硅藻,如异极藻、脆杆藻也是常见优势物种.着生藻类数量丰富,为3.16×104-3.06×106cells·cm-2;种类多样性较低,Shannon-Weaver种类多样性指数为0.15～2.68.从总氮和总磷含量来看,珠江广州河段水质为Ⅲ类至Ⅴ类之间.调查期间,2007年7月12日举行了"广州市万人游珠江活动",由于活动期间采取的一些截污治理措施,6月、7月水质特别是广州市区河段水质有所好转;但活动过后,营养盐特别是总磷浓度迅速上升,叶绿素a含量和着生藻类数量急剧增加.研究结果说明,珠江广州河段水质能够得以改善,但需要采取持续而有效的措施防止水质的进一步恶化,才能保证珠江水资源的可持续发展.     

On the effect of Fe(III) on proliferation of Microcystis aeruginosa at high nitrate and low chlorophyll condition

The impact of Fe concentrations on the growth of Microcystisaeruginosa in aquatic systems under high nitrate and low chlorophyll conditions was studied. The responses of cell density, total and cell chlorophyll-a intracellular Fe content and organic elemental composition of M. aeruginosa to different concentration gradients of Fe(III) in the solutions were analysed. The results showed that the proliferation speeds of M. aeruginosa were: (1) decelerated when the Fe(III) concentration was lower than 50 μg/L in the solutions, (2) promoted and positively related to the increase of Fe(III) concentration from 100 to 500 μg/L in the solutions over the experimental period, and (3) promoted in the early stage but decelerated in later stages by excess adsorption of Fe by cells when the Fe(III) concentration was higher than 500 μg/L in the solutions. The maximum cell density, total and cell chlorophyll-a were all observed at 500 μg Fe(III)/L concentration. The organic elemental composition of M. aeruginosa was also affected by the concentration of Fe(III) in the solutions, and the molecular formula of M. aeruginosa should be expressed as C_7–7.5H₁₄O_0.8–1.3N_3.5–5 according to the functions for different Fe(III) concentrations. Cell carbon and oxygen content appeared to increase slightly, while cell nitrogen content appeared to decrease as Fe(III) concentrations increased from 100 to 500 μg/L in the solutions. This was attributed to the competition of photosynthesis and nitrogen adsorption under varying cell Fe content.     

土壤溶解性有机质荧光特征及其与铜的络合能力

选取4种不同类型土壤和腐熟鸡粪,透析提取溶解性有机质(DOM),通过凝胶色谱、三维荧光光谱测定并分析了DOM的相对分子质量(M_r)分布、化学结构特征及其与铜的结合能力.结果表明,鸡粪DOM相对分子质量积分面积远高于其他样品,表现出其具有较高的有机物含量,泥炭土次之.暗棕壤、褐土和黑土较低,仅为鸡粪积分面积的4.5%~5%.鸡粪在中分子量段积分面积约占总物质的34.1%,低分子量段和高分子量段分别约占41.7%和24.2%.荧光光谱分析表明,各样品均出现类蛋白荧光峰(E_x/E_m=240~270/300~350 nm),在中、高分子量段(M_r500)暗棕壤存在的荧光峰消失.泥炭土出现了较为独特的可见光区类富里酸荧光峰(E_x/E_m=325/420 nm),该荧光峰和黑土的荧光峰在中、高分子量段(M_r500)都产生了红移现象.鸡粪分别出现了和黑土类似的紫外区类富里酸荧光峰,以及和泥炭土类似的可见光区类富里酸荧光峰,以及一类独特的类蛋白荧光峰(E_x/E_m=280/350 nm),但这种类蛋白荧光峰在中分子量段(500M_r12 000)消失.分析表明泥炭土与Cu的络合常数(lg K)为4.13,其他3种土壤与Cu的络合常数较小且差别不大,集中在2.10~3.10之间.鸡粪与Cu的结合能力最大,络合常数达到6.66.