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)     

尕海湿地植被退化过程中有机碳及相关土壤酶活性变化特征

以青藏高原东缘尕海湿地不同植被退化程度样地为研究对象,采用定位研究与室内试验相结合的方法,研究植被退化过程对土壤有机碳和土壤酶活性的影响。结果表明：植被退化显著影响土壤有机碳含量和土壤酶活性,降低了土壤有机碳的含量和酶促反应效率,且这种影响随土层深度变化有所不同。四种植被退化阶段有机碳含量和酶活性均值总体上（0~100 cm）表现为未退化>轻度退化>中度退化>重度退化。不同土层中,20~40 cm土层的有机碳含量中度退化>轻度退化;0~10 cm土层中淀粉酶和纤二糖酶中度退化阶段活性值较高;20~100 cm土层中蔗糖酶重度退化阶段活性值最高,纤二糖酶活性重度退化>中度退化。同一退化程度土壤有机碳含量、蔗糖酶和纤二糖酶活性随土层深度增加显著降低（P<0.05）;淀粉酶和纤维素酶活性在>40 cm土层中活性值有所上升,整体呈波动下降趋势。     

不同基质生物炭对厌氧处理餐厨垃圾效能及微生态的影响

探究了剩余污泥（SS）、餐厨垃圾（FW）、玉米芯（CC）、甘蔗渣（BG）4种不同基质生物炭对厌氧生物处理餐厨垃圾效能的影响,对厌氧污泥的关键酶活性、微生物群落分布以及代谢途径等微生态进行了分析.结果表明,厌氧反应器分别加入4种生物炭后,COD平均去除率分别提高了29.49%、23.16%、29.42%、40.32%;傅里叶红外分析表明,投加SS生物炭组出水中羟基、酰胺基以及C-O-C伸缩振动峰减弱.4个厌氧反应器中厌氧污泥的乙酸激酶活性分别为0.40,0.42,0.96,0.98 μmol/g,表明投加CC与BG生物炭促进了餐厨垃圾的厌氧水解酸化过程;厌氧污泥胞外聚合物的蛋白质/多糖之比分别为0.415、0.56、1.89、2.8,投加CC、BG生物炭提高了污泥的稳定性.4个厌氧反应器中拟杆菌门、变形菌门、厚壁菌门为主要菌群,投加BG生物炭促进了变形菌门与厚壁菌门的生长;对于古细菌而言,甲烷杆菌属与甲烷丝菌属为优势种群,SS组的甲烷杆菌属丰度最高（53.48%）,而BG组中甲烷丝菌属丰度最高（42.72%）.KEGG功能分析表明古菌及细菌均以碳水化合物代谢、氨基酸代谢为主;而投加BG与SS生物炭后,微生物膜运输水平得到了提高.     

构树修复对重金属污染土壤环境质量的影响

通过温室盆栽试验,研究构树（Broussonetia papyrifera）生长对重金属污染土壤酶活性和微生物群落结构的影响.结果表明,构树修复污染土壤中酶活性和微生物多样性明显提高.经270d培养后,构树生长土壤中蔗糖酶和酸性磷酸酶活性与种植土壤相比分别显著（P<0.05）提高3.12倍和2.29倍;土壤脱氢酶与有效态As、Cd、Pb、Zn和Cu含量,蔗糖酶与有效态Cd含量,以及磷酸酶与有效态Cd和Cu含量之间呈显著负相关（P<0.05）.根据16S和18S rDNA PCR-DGGE分析表明,构树修复可提高污染土壤中细菌和丛枝菌根真菌多样性.上述结果表明,构树修复可有效改善重金属污染土壤的环境质量.然而,污染土壤中重金属有效态含量下降不明显,必须辅助物理和化学措施来强化构树对重金属污染土壤的生态修复潜力.     

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。     

生物质炭对Cd污染土壤团聚体酶活性的影响

为探究生物质炭对Cd污染土壤团聚体酶活性的影响,采用盆栽试验,向模拟Cd污染土壤中添加生物质炭并测定水稻根际土壤团聚体碳循环酶与氧化还原酶活性.结果表明:Cd污染土壤团聚体酶活性对添加量为2.5%的生物质炭响应明显,氧化还原酶活性指数值介于0.522~0.792之间.在2.5mg/kg的外源Cd条件下,2.5%的生物质炭比未添加生物质炭处理显著提高蛋白酶与过氧化氢酶活性至121%与653%.在Cd污染土壤中添加生物质炭,碳循环酶与氧化还原酶及综合酶活性均在0.5~1mm中等粒径土壤团聚体中产生富集效应,其酶活性随土壤团聚体粒径的增大呈先升后降最终趋于稳定的趋势.研究显示,在2.5mg/kg的外源Cd条件下,添加2.5%的生物质炭对水稻根际土壤团聚体酶活性影响显著,土壤酶活性受酶种类、生物质炭量与团聚体粒径的综合影响;在Cd污染土壤中添加生物质炭后,土壤酶活性随团聚体粒径的增大呈“∧”形变动规律.     

Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited (C/N ratio is 56/8.8 mmol/L) medium. Several sets of shaking flask experiments were conducted. The results showed that 2 g/L glucose feeding on the first day of the culture (24 h after the inoculation) simulated both fungal biomass growth and enzyme production. The manganese peroxidase (MnP) activity was 2.5 times greater than that produced in cultures without glucose feeding. Furthermore, the glucose feeding mode in fed-batch culture was also investigated. Compared to cultures with glucose feeding every 48 h, cultures with glucose feeding of 1.5 g/L (final concentration) every 24 h produced more enzymes. The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture, respectively, and the enzyme was kept stable for 4 days with an activity of over 200 U/L. Translated from Acta Scientise Circumstantiae, 2007, 27(3): 363–368 [译自: 环境科学学报]     

镍、锌、铝对羊角月芽藻(Selenastrum capricornutum)生长及酶活性影响研究

研究了不同浓度镍，锌，铝对常见绿藻羊角月芽藻生长速度，蛋白质含量，ＡＴＰ水平，葡萄糖－６－磷酸脱氢酶，酸性磷酸酶及硝酸还原酶活性的影响。结果表明３种金属离子在所试浓度范围内对羊角月芽藻的生长速度均有抑制作用，但单位藻培养物中蛋白质含量却随着金属离子浓度的增加而增加；高浓度金属离子对酶活性有明显抑制作用；藻细胞中ＡＴＰ水平随着金属离子浓度的增加而下降。