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)     

供水管网生物膜有机物的二氯乙腈与二氯乙酰胺生成特性

在饮用水输配系统中,来源于管壁生物膜的有机物可能耗氯并生成消毒副产物(DBPs),包括二氯乙腈(DCAN)与二氯乙酰胺(DCAcAm)等高毒性含氮DBPs(N-DBPs).研究考察管网常见的细菌与其胞外聚合物(EPS)以及模拟管壁生物膜氯化与氯胺化后DCAN与DCAcAm的生成,并与天然有机物(NOM)和水源水有机物进行比较.结果显示,铜绿假单胞菌、恶臭假单胞菌与藤黄微球菌与氯反应生成的DCAN、DCAcAm浓度分别为1.48～2.02、0.21～0.38μg·mg~(-1)(mg~(-1)以TOC计),高于同反应条件下NOM的生成量;相比于氯化反应,3株细菌细胞氯胺化生成的DCAN与DCAcAm浓度明显更低.3株菌的EPS也是氯与氯胺化反应生成DCAN与DCAcAm的前体物,且其氯胺化反应生成的DCAcAm浓度高于氯化反应生成的.与NOM、水源水相比,模拟管壁生物膜氯化后生成的N-DBPs与三氯甲烷(TCM)浓度比更高,表明生物膜有机物比NOM与水源水有机物更倾向生成DCAN与DCAcAm类N-DBPs,且模拟管壁生物膜氯胺化的DCAcAm生成量高于氯化反应的,说明管壁生物膜有机物是供水管网系统中DCAN与DCAcAm类N-DBPs的重要前体物.     

Ni2+对好氧颗粒污泥系统性能的影响

以城镇污水处理厂A²/O工艺的回流污泥作为接种污泥,在序批式反应器（SBR）中培养好氧颗粒污泥（AGS）,探究Ni²⁺对AGS系统的影响.结果表明：在25℃的条件下,50d左右培养出成熟的AGS,其形态多呈球状或椭球状,外表分布着少量丝状菌,颗粒污泥粒径主要在2~4mm,MLSS达6000mg/L,SVI维持在40~50mL/g.将Ni²⁺作用于培养出的成熟AGS,Ni²⁺在0~2mg/L的浓度范围内会提高AGS的稳定性,使MLSS上升,SVI降低;同时会促进AGS分泌胞外多聚物（EPS）,EPS组成成分中的蛋白质（PN）明显多于多糖（PS）;Ni²⁺对好氧颗粒污泥去除总氮（TN）的抑制程度要大于COD,最终COD去除率可维持在95%以上,TN去除率可维持在70%以上.     

胞外聚合物对浸没式膜-生物反应器膜过滤性能的影响

通过浸没式膜-生物反应器（SMBR）处理上海市城市内河水的试验,研究了反应器混合液中和2种疏水性不同的膜上污泥胞外聚合物（EPS）的量、组成、污泥特性及对膜过滤性能的影响.结果表明,混合液污泥中的溶解性EPS和固着性EPS比膜上污泥多,不同污泥的固着性EPS组分中蛋白质（EPSP）/多糖（EPSC）的大小顺序依次为混合液污泥〈亲水膜污泥〈疏水膜污泥.疏水膜污泥中溶解性EPS比亲水膜多,溶解性EPS和固着性EPS量及组分均有很大差异.混合液污泥的EPS流动性大,疏水性膜上污泥EPS的流动性较大.溶解性EPS对污泥特性及膜过滤性能影响较大.SMBR中亲、疏水性膜片上黏附的污泥的相对疏水性分别为53%、59%.比较2种膜片通量的变化发现,疏水性膜污染严重.     

水稻不同生育期根际与非根际土壤胞外酶对施氮的响应

与稻田土壤碳氮循环(矿化、转化等)密切相关的酶活性可以反映微生物的生长和代谢过程.为明确水稻不同生育期根际与非根际土壤胞外酶对施氮的响应,采用根际袋法区分水稻根际和非根际土壤,利用96微孔酶标板荧光分析法,测定其碳氮过程关键酶β-1,4-葡萄糖苷酶(BG)和β-1,4-N-乙酰氨基葡萄糖苷酶(NAG)活性,探讨根际效应、施氮和生育期对土壤酶活的影响及其调控机制.结果表明,施氮使拔节期土壤BG酶活性相对于不施氮处理降低了7.4~13.5 nmol·(g·h)~(-1),而使成熟期BG酶活性增大了7.0~31.4 nmol·(g·h)~(-1),同时根际与非根际土壤中BG酶活性也随水稻的生育期而发生相应的变化.与不施氮处理相比,施氮使水稻成熟期非根际土壤NAG酶活性增加了1.1倍,根际土壤降低了0.3倍.施氮和生育期显著影响土壤BG酶活性,而水稻生育期、施氮和根际效应及其交互作用均对NAG酶活性有极显著影响.RDA分析表明土壤微生物生物量碳(MBC)和可溶性有机碳(DOC)含量主要影响水稻根际土壤胞外酶活性;而非根际土壤中酶活性的变化主要受微生物生物量氮(MBN)和铵态氮(NH_4~+-N)的影响.土壤酶活性与多种因素存在复杂关系,需要综合考虑植物生理特征、土壤酶活性和土壤特征,分析N添加对微生物群落组成的影响.     

分子生物学技术在水处理中的应用研究进展

利用现代分子生物学技术对微生物进行分析和研究,以解决传统微生物分析成本高、速度慢、准确度较低和灵敏度较差等缺点。介绍分子生物学技术在水处理中的应用现状,并对未来发展方向提出建议。目前分子生物学技术在供水工程中主要用于水中细菌、病毒、原生动物、蠕虫等病原微生物的快速检测以及管网微生物种群的分析,在污水处理中主要用于脱氮除磷过程菌群数量和空间分布的剖析、污泥膨胀成因分析以及监测微生物种群多样性来优化处理工艺。分子生物学技术的深入研究将进一步促进水行业的发展。     

外源Ca2+对SBR启动期活性污泥胞外多聚物的动态影响

利用无机物如Ca~(2+),加快活性污泥反应器启动,强化活性污泥絮体性能和结构稳定性,受到越来越多的重视.采用序批式反应器,研究进水中添加Ca~(2+)对反应器启动期活性污泥沉降性能和胞外多聚物的影响.结果表明运行至28 d,与进水中不添加Ca~(2+)的反应器(对照反应器)相比,进水中添加150 mg·L~(-1)外源Ca~(2+)的反应器中活性污泥MLSS和MLVSS值分别高出了89.6%和75.6%,SVI值则降低了47.9%;活性污泥胞外多聚物总量增加了76.4%,多糖增加了28.8%,蛋白质减少了31.6%,添加150 mg·L~(-1)外源Ca~(2+)的反应器中污泥胞外多聚物中多糖/蛋白质值为68.8,对照反应器的活性污泥胞外多聚物多糖/蛋白质值仅为36.6.三维荧光光谱和红外光谱分析表明外源Ca~(2+)导致活性污泥胞外多聚物组分发生了变化.实验结果为进水中添加外源Ca~(2+)改善活性污泥沉降性能提供了基础数据.     

群体感应淬灭菌的分离及其膜污染控制性能

通过淬灭细菌的群体感应系统来抑制生物膜形成、防止膜生物污染的方法近年来受到广泛关注.本实验从实际运行污水处理厂活性污泥中分离出5株具有群体感应淬灭功能的菌株,其中菌株HG10对信号分子N-乙酰高丝氨酸环内酯(C6-HSL)分解能力最强.经16S rRNA基因序列比对,初步鉴定为蜡样芽孢杆菌(Bacillus cereus).用海藻酸钠将菌株HG10进行包埋固定,以探究其在膜过滤系统中对膜污染防治的效果.结果表明,经过8 d培养,添加细菌包埋珠(SA-HG10)的实验组B中膜通量为181.29 L·(m~2·h)~(-1),未投加包埋珠的对照组A膜通量为110.64 L·(m~2·h)~(-1),B组膜通量比A组高出63.86%;对微滤膜片上生物膜中EPS含量测定表明,实验组B中EPS多糖和蛋白质含量较对照组A分别减少了29%和48%,疏水性蛋白质含量的大量减少是造成膜污染减弱的主要原因;膜表面胞外聚合物(EPS)总含量减少了43%,表明投放SA-HG10细菌包埋珠对过滤膜片上生物膜形成具有明显抑制作用,改善了膜过滤性能.     

细菌B1胞外活性物质对球形棕囊藻的溶藻机制初探

为探讨溶藻细菌B1对球形棕囊藻的溶藻作用机理及起作用的胞外活性物质的类别,研究了从珠海香洲码头赤潮海水中分离获得的溶藻细菌BI的无菌滤液对球形棕囊藻生长过程中的叶绿素a含量、丙二醛含量(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性的影响,并对溶藻细菌B1分泌的胞外活性物质进行了初步分析.当100 mL藻液中加入1 mL的无菌滤液,除藻率达到了83％.B1无菌滤液使藻中丙二醛的含量显著上升,48 h达到最大值153％,球形棕囊藻的SOD、CAT活性在处理开始后保持下降趋势,分别在96 h、48 h达到最低值27％、33％.研究表明B1无菌滤液(不大于1 mL)对棕囊藻的溶藻效果与加入的量成正比,且主要通过降低机体中保护酶的活性和加剧膜脂质过氧化的程度等来抑制球形棕囊藻的生长;细菌B1分泌的胞外活性物质中可能含有芳香醛类且包括载有磷基团的有机物,但不含蛋白质.     

模拟干旱胁迫对诸葛菜无机碳利用的影响

为研究喀斯特适生植物的碳酸酐酶应对岩溶干旱机制,本实验以诸葛菜为研究对象,以聚乙二醇(polyethyleneglycol,PEG)浓度为控制因子,已知δ13 C值的NaHCO3为无机碳源模拟喀斯特干旱逆境,测定分析不同浓度PEG下诸葛菜叶片的碳酸酐酶活力、稳定碳同位素组成、净光合速率和叶绿素荧光等指标。结果表明,诸葛菜的碳酸酐酶活力在PEG胁迫浓度低于40g·L-1时显著性增高;碳酸氢根离子利用份额随PEG浓度变化不同,10g·L-1时利用份额最多,20g·L-1时诸葛菜的碳酸氢根离子利用份额与对照相比无显著性差异;净光合速率随着PEG浓度的增加而下降,且下降趋势成非线性关系;PEG浓度低于40g·L-1时,荧光参数显示光合系统未受到损坏。适当的渗透逆境胁迫下,诸葛菜表现出高碳酸酐酶活力应对逆境。诸葛菜可凭借碳酸酐酶作用交替利用碳酸氢根离子和大气中的二氧化碳,补充诸葛菜因渗透胁迫造成的水分和二氧化碳的不足,保护光合系统,减缓净光合速率的下降。