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)     

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。     

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。     

商陆和烟草对锰胁迫的抗氧化响应研究

研究植物的重金属累积能力对于植物修复和植物冶金意义重大.Mn超累积植物商陆(Phytolacca americana L.)的耐性/累积机制至今还不清楚,为了研究商陆在Mn胁迫下的抗氧化酶作用,将萌发6周的商陆和烟草幼苗分别放入1 mmol·L~(-1)或3 mmol·L~(-1)的1/2 Hoagland营养液处理4 d,实验表明商陆叶片光合速率降低幅度显著低于烟草,而烟草MDA含量和电解质渗漏量增加幅度大于商陆.如1 mmol·L~(-1) Mn处理4 d后,与对照相比,商陆和烟草的光合速率分别降低13.3%和75.5%;烟草MDA含量和电解质渗漏量分别增加347.3%和120.1%,而商陆的MDA含量和电解质渗漏量没有明显增加,表明Mn胁迫对超累积植物引起的氧化损伤明显低于非累积植物.随着Mn处理浓度的提高和时间的延长,2种植物叶片SOD和POD活性均迅速增加,其中商陆SOD活性的增加幅度大于烟草;另外,商陆CAT活性持续上升,而烟草CAT活性显著下降.如在1 mmol·L~(-1) Mn处理下,商陆SOD、POD和CAT活性分别提高161.1%、 111.3%和17.5%;烟草SOD和POD提高55.5%和206.0%,而CAT下降15.6%,表明商陆的抗氧化酶,特别是CAT,能够有效地清除Mn毒产生的自由基.这些结果说明抗氧化酶的抗氧化能力是重金属累积植物商陆的Mn耐性机制之一.     

铀胁迫对香根草生理生化指标的影响

采用水培方式研究了不同质量浓度(0、0.1 mg/L、1 mg/L、5mg/L、10 mg/L、20 mg/L)铀胁迫下香根草光合色素质量比、可溶性蛋白质质量比、丙二醛(MDA)含量、抗氧化酶(超氧化物歧化酶SOD、过氧化物酶POD、过氧化氢酶CAT)活性、非酶物质(非蛋白巯基NPT、谷胱甘肽GSH和植物络合素PCs)含量的变化.结果表明,铀质量浓度为0.1 mg/L和1 mg/L时,可促进香根草光合色素和可溶性蛋白质质量比的增加,但随铀质量浓度增加,光合色素和可溶性蛋白质质量比逐渐下降.铀胁迫诱导香根草体内MDA含量、POD活性和CAT活性呈明显升高的趋势,SOD活性则表现为逐渐下降的趋势.不同质量浓度铀胁迫对香根草根部NPT、GSH和PCs含量无明显影响,铀质量浓度为0.1 mg/L时香根草根部NPT、GSH和PCs含量略有增加;随铀胁迫质量浓度增加,叶片内GSH含量逐渐下降,NPT和PCs含量则逐渐升高.     

抗生素药物诺氟沙星对锦鲤的毒性效应

为明确抗生素药物诺氟沙星对锦鲤的毒性效应,采用鱼类毒性试验方法,研究了诺氟沙星(Norfloxacin,NFLX)对锦鲤的急性毒性和亚急性毒性效应.结果表明,当NFLX暴露质量浓度达到1 000 mg/L时,NFLX对锦鲤无急性毒性.NFLK暴露15 d,在1 mg/L、5 mg/L、25mg/L和125 mg/L 4个组中,5 mg/L组锦鲤肝脏SOD活性在6d时达到最大(p＜0.01);125 mg./L组MDA含量在15 d时达到最大值(p＜0.01);25 mg/L、125 mg/L组锦鲤肝脏中MDA含量与暴露时间呈正相关,相关系数分别为0.938 1、0.953 5.各组GST活性和GSH质量比均受到不同程度的诱导,1 mg/L组NFLX可使鱼肝GST与GSH产生适应性反应;5 mg/L组诱导GST与GSH效应最为显著,呈现“钟形”变化趋势,5 mg/L很可能是NFLX对GST活性和GSH质量比影响的一个阈值或转折点.鱼肝GPT与GOT活性变化表现出一定的相似性,均呈现先升高后降低的变化规律,其中5 mg/L组被诱导效应最为显著(p＜0.01).5 mg/L很可能是锦鲤适应NFLX暴露和机体中生物酶发挥作用的一个转折点.比较上述抗氧化和转氨酶指标的敏感性和稳定性发现,在NFLX作用下,锦鲤肝脏中GST、GSH两个指标对NFLX的反应比SOD、MDA、GPT、GOT灵敏,变化规律稳定,更适合评价水环境中NFLX暴露的毒性效应.     

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.     

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（Ｔｒｉ－ｏ－ＣｒｅｓｙｌＰｈｏｓ－ｐｈａｔｅ，ＴＯＣＰ）诱导的迟发性神经毒性．试验发现，母鸡口服ＴＯＣＰ（７５０ｍｇ／ｋｇ）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，至第２１ｄ开始便发展为瘫痪，对鸡脑神经病损靶标酯酶（ＮｅｕｒｏｐａｔｈｙＴａｒｇｅｔＥｎｚｙｍｅ，ＮＴＥ）活性的测定表明，有机械诱导的迟发性神经毒性（Ｏｒｇａｎｏｐｈｏｓｐｈａｔｅ－ＩｎｄｕｃｅｄＤｅｌａｙｅｄＰｏｌｙｎｅｕｒｏｐａｔｈｙ，ＯＰＩＤＰ）与ＮＴＥ活性抑制之间的关系不平行，其毒性症状的轻重与ＮＴＥ活性被抑制的强弱也不呈正相关，很可能，在给药后２４ｈ，体内ＯＰＩＤＰ已被触发，此外，实验观察到给予ＴＯＣＰ后被试鸡的体重明显下降。     

氯氰菊酯降解菌的筛选鉴定及其降解特性研究

从农药厂废水排放口附近的污泥中分离到1株能降解氯氰菊酯的细菌LQ-3.根据其形态、生理生化特征和16S rDNA(GenBank Accession No.FJ222585)序列分析,将该菌株鉴定为Starkeya sp..LQ-3菌株只能以共代谢方式降解氯氰菊酯,在有酵母粉、蛋白胨、葡萄糖等营养物质存在的条件下,5 d内对20 mg·L-1氯氰菊酯的降解率达到72.1%.LQ-3菌株降解氯氰菊酯的最适温度为30 ℃左右,pH值为7～8.LQ-3菌株还能降解功夫菊酯、甲氰菊酯、联苯菊酯和溴氰菊酯.酶的定域试验表明,LQ-3菌株降解氯氰菊酯的酶属于胞外酶.