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)     

木质素降解酶及其调控机理研究的进展

近年来有关木质素降解微生物和木质素降解酶的产与调控机理的研究表明，腐霉和细菌在木质自然降解中起着重要作用，腐霉的降解作用发生在次级代谢阶段，而细菌发生在初级代谢阶段，影响木质素降解酶的产生酶活力高低的因素有碳氮的选择与限制；诱导物的添加；表面活性剂的使用；某些金属离子浓度水平的改变；菌丝的固定化，温度和摇床转速的变换等。     

In vitro characteristics of human fetal cells obtained from chorionic villus sampling and amniocentesis

In vitro characteristics of human fetal cells have been investigated after chorionic villus sampling at the first trimester and amniocentesis at the second trimester of pregnancy. Light microscopy revealed heterogeneous morphology of cell types in both the chorionic villus culture and the amniotic fluid cultures. Based on the experiments performed, chorionic villus cells are more sensitive to pronase, trypsin, and versene during subculture and have a higher DNA content per single cell and release more [¹²⁵I]-Beta-human chorionic gonadotropin into culture medium than those found in amniotic fluid cells. The practical applications of this study are discussed.     

有机磷酸酯诱导迟发性神经毒性及其对神经毒性酯酶(NTE)活性影响的动态研究

以成年产卵来航母鸡为试验动物，研究了三甲基苯基磷酸酯诱导的迟发性神经毒性。试验发现，母鸡口服ＴＯＣＰ后经过大约８－１０ｄ的潜伏期开始出现中毒症状，其表现为以步态失调为主要特征的运动障碍呈进行性加重，至２１ｄ开始便发展为瘫痪。     

SDS促进复配酶水解排水管网沉积物

采用SDS协同复配酶水解排水管网沉积物,分别考察了SDS投加量、反应时间、pH、温度4个影响因素对沉积物水解效果的影响,并通过 SEM、EPS、三维荧光光谱以及粒径等表征方法进行机理分析。结果表明,投加5%(w:w)SDS与8%(w:w)复配酶,在原泥pH(7.28±0.3)、25 ℃、反应时间为2 h时,沉积物水解效果最好。此时SCOD、多糖和氨氮分别由初始的5 686.9、1 913.75和87.32 mg·L⁻¹增加到8 192.9、3 561.29和153.37 mg·L⁻¹,有机质含量由56.32%下降到55.59%。在SDS+复配酶处理管网沉积物后,其内部紧实结构被破坏,TB-EPS向S-EPS转移,腐殖酸类物质荧光强度增强,表明溶解性EPS增多,且沉积物粒径向<300 μm转变。高通量测序分析结果表明,SDS+复配酶对管网沉积物微生物种类的丰富度和多样性影响不大,但在一定程度上会改变微生物的功能菌群,减少CH₄和H₂S的产生。以上研究结果可为延缓排水管网淤堵问题提供数据支持。     

曝气对潜流人工湿地中木本植物的影响

为了增加人工湿地植物的多样性,本文将花叶冬青(Ilex aquifolium)、月季(Rosa chinensis)和八角金盘(Fatsia japonica)3种木本植物引入潜流人工湿地,比较曝气前后系统对污水处理效果,植物的生物量与对TN、TP负荷减少的贡献率,根系形态与内部结构、根际微生物与基质酶活性的比较研究。结果表明,木本植物潜流人工湿地系统曝气后可以提高COD、NH4+-N、TN、TP的去除率,分别为6.99%、4.91%、10.25%、8.09%。湿地曝气有利于3种木本植物的生长,在曝气后比曝气前生物量增加了381.75、606.25和896.88 g/m2。曝气会促使木本植物长出大量须根,且曝气后的根系具有大量由薄壁细胞组成的通气组织的水生根特征。同时,曝气后增加了硝化菌数量,各处理单元中硝化菌增量分别为A:0.28 MPN×104/g,B:4.23 MPN×104/g,C:7.7 MPN×104/g。另外,曝气后植物根际基质中磷酸酶和尿酶的含量增加量大约在50%以上,这与系统中N和P的去除率提高的规律一致。     

垂直流人工湿地启动期基质酶的时空变化

为了解垂直流人工湿地系统中基质酶在去污效果中的作用及其随基质深度和运行时间的变化规律,设置3个垂直流人工湿地系统:种植皇竹草的A系统,种植象草的B系统,以及不种植物的C系统。分别监测3个系统不同基质深度与不同运行时间条件下脲酶、磷酸酶、过氧化氢酶、转化酶和纤维素酶的变化规律,以及这几种酶与TN、TP、COD、NH4+-N 和NO3--N污染物之间的相关性。实验结果表明,上述5种酶都表现出随着基质深度增加而减少的趋势,且上层0~30 cm处的基质酶活性极显著高于中层和底层的酶活性,此外,种植能源植物的A、B系统基质酶活性都高于不种植物C系统,说明湿地中污染物的去除主要集中在上层基质,种植能源植物能够有效促进微生物的活性,增加胞外酶的分泌,提高人工湿地对污染物的去除效果,可为优化人工湿地除污效果以及湿地植物的选择提供理论依据。     

东湖通道工程沙滩浴场围堰施工对水中胞外碱性磷酸酶动力学行为的影响

城镇化过程常伴随市政建设面积与强度的大幅度增加,但其与湖泊富营养化的关系尚未得到充分的研究。2014年3-5月在武汉东湖通道工程沙滩浴场围堰施工区与相关湖区（郭郑湖、汤菱湖、团湖）的19个位点分5次采集水样,分析了水中胞外碱性磷酸酶的动力学参数及其与溶解有机磷和溶解反应性磷比值的关系。结果表明,施工致使酶的最大反应速度与底物亲和力均显著降低,有机磷酶促再生为生物可利用性无机磷的周转时间明显延长。这一结果指示施工区磷营养的相对充足,施工干扰可能有效促进了沉积物磷向水柱的补给。     

纳米零价铁和零价铁对餐厨垃圾暗发酵制氢过程铁离子和酶活性的影响

通过投加不同浓度的纳米零价铁(NZVI)和零价铁(ZVI),考察了暗发酵制氢过程中铁离子组成和浓度变化、氢化酶和脱氢酶活性,研究了2种添加剂强化餐厨垃圾高温((55±1) ℃)暗发酵制氢的作用机制。结果表明：投加NZVI和ZVI均可提高餐厨垃圾暗发酵制氢性能;当投加100 mg·L⁻¹ ZVI时,产氢效果最佳,最大产氢潜力和最大产氢速率分别为425.72 mL和66.32 mL·h⁻¹,是投加NZVI实验组的1.64倍和1.34倍,代谢途径是以乙醇型发酵为主的混合型发酵;在投加NZVI和ZVI后,暗发酵制氢末端产物的Fe²⁺和Fe³⁺浓度升高,投加300 mg·L⁻¹ NZVI和100 mg·L⁻¹ ZVI实验组Fe²⁺浓度最大,是未投加实验组的2倍和1.87倍;与反应前相比,Fe²⁺显著升高,Fe³⁺由于微生物利用与转化浓度降低,同时可有效提高氢化酶活性。投加100 mg·L⁻¹ ZVI不仅可提高氢化酶活性,还可提高脱氢酶活性。以上结果可为提高餐厨垃圾等复杂有机废物的高效能源化提供参考。     

纤维素降解菌Arthrobacter oryzae HW-17的纤维素降解特性及纤维素酶学性质

采用单一碳源选择性培养基和纤维素平板水解圈法筛选到一株具有较强纤维素分解能力的菌株Arthrobacter oryzae HW-17.此外,高通量测序发现,不同驯化条件下微生物群落结构有明显差异,低温条件下优势属为类芽孢杆菌属(Paenibacillus)和伯克氏菌属(Burkholderia).本文同时对菌株Arthrobacter oryzae HW-17的微生物特性和纤维素降解特性进行了初步研究,结果发现,KNO_3、30或35℃、pH=7分别为菌株产纤维素酶的最佳氮源、温度和pH.菌株HW-17的最高纤维素酶活为18.55 U·m L~(-1),且对磨碎加工处理的纤维素样品和含鸡粪的纤维素混合样品有更好的降解效果.此外,菌株HW-17产生的纤维素酶在中温(≤50℃)和偏酸性(pH=5~7)条件下能保持较高的酶活.Cu(Ⅱ)和Pb(Ⅱ)等金属离子能够抑制该酶的活性.