长江三角洲地区污水污泥与健康安全风险研究V．污泥施用对土壤微生物群落功能多样性的影响

利用BIOLOG测试方法对长江三角洲地区施污泥土壤的微生物群落功能多样性进行了初步探讨。研究结果表明,施用污泥后土壤微生物群落代谢剖面、多样性指数、动力学参数均发生了变化,说明微生物对单一碳源底物的利用能力发生了改变。微生物群落功能多样性动力学研究结果发现,供试土壤的微生物群落代谢剖面（AWCD）与培养时间之间呈非线性关系,其变化过程符合微生物种群生长动态模型（S型）。除滩潮土的杭州新鲜消化污泥和杭州风干污泥处理外,其他污泥处理均使土壤微生物群落功能多样性动力学参数K和r值降低,S值增大。土壤微生物群落功能多样性特征的变化可以较好地显示施用污泥后土壤微生物群落对碳源利用能力和模式的差异,反映施用污泥特定生境土壤微生物群落功能多样性的变化。     

长江三角洲地区污水污泥与健康安全风险研究Ⅴ.污泥施用对土壤微生物群落功能多样性的影响

利用BIOLOG测试方法对长江三角洲地区施污泥土壤的微生物群落功能多样性进行了初步探讨。研究结果表明,施用污泥后土壤微生物群落代谢剖面、多样性指数、动力学参数均发生了变化,说明微生物对单一碳源底物的利用能力发生了改变。微生物群落功能多样性动力学研究结果发现,供试土壤的微生物群落代谢剖面(AWCD)与培养时间之间呈非线性关系,其变化过程符合微生物种群生长动态模型(S型)。除滩潮土的杭州新鲜消化污泥和杭州风干污泥处理外,其他污泥处理均使土壤微生物群落功能多样性动力学参数K和r值降低,S值增大。土壤微生物群落功能多样性特征的变化可以较好地显示施用污泥后土壤微生物群落对碳源利用能力和模式的差异,反映施用污泥特定生境土壤微生物群落功能多样性的变化。     

铅锌银尾矿区土壤微生物活性及其群落功能多样性研究

通过对浙江省天台铅锌银尾矿区土壤微生物活性指标以及微生物群落功能多样性研究 ,结果表明 ,尾矿污染区土壤几种重金属含量比非矿区土壤有明显的增加。尾矿区土壤微生物特征发生了显著的变化 ,微生物生物量和可培养细菌数量显著降低 ,但土壤基础呼吸和微生物代谢商 (qCO2 )值却明显升高。Bi olog测试结果显示 ,随着重金属污染程度的加剧其土壤微生物群落结构发生了相应变化 ,尾矿区土壤微生物群落代谢剖面 (AWCD)及群落丰富度、多样性指数均显著低于非矿区土壤 ,且供试土壤间均达极显著水平差异 (p <0 .0 1) ,表明尾矿区重金属污染引起了土壤微生物群落功能多样性的下降 ,减少了能利用有关碳源底物的微生物数量、降低了微生物对单一碳源底物的利用能力     

代谢指纹评估苯噻草胺对水稻土微生物群落的短期影响

本文探讨了酰胺类除草剂苯噻草胺对水稻土微生物群落功能多样性的短期影响。本研究采用微生物群落基质利用潜力测定（Biolog法）评估生物群落。结果表明，苯噻草胺污染引起了水稻田微生物群落功能多样性的下降，降低了微生物对单-碳源底物的利用能力，但这种影响是短暂的，在试验最终没有导致土壤微生物群落功能多样性下降。多食鞘氨醇杆菌Y1（Sphingobacterium multiuorum）的添加有利于提高水稻土微生物群落的功能多样性。3个处理土壤的群落代谢剖面值与培养时间之间呈非线性关系，其变化过程符合微生物种群生长动态模型（S形）。模型模拟分析表明，动力学参数a和x0能更灵敏地表征苯噻草胺和Y1菌株处理对水稻土微生物群落功能多样性的影响。在本实验研究中，多样性指标指数Shannon（H）能灵敏而有效地指示污染环境的微生物学变化，但群落丰富度指标颜色变化孔数（S）提供的信息较片面。     

矿区侵蚀土壤的微生物活性及其群落功能多样性研究

通过对浙江省天台铅锌银矿区侵蚀土壤的微生物活性以及微生物群落功能多样性研究,结果表明:人为开矿使矿区土壤环境呈现不同层次的加速侵蚀特征。矿区侵蚀土壤的几种重金属含量比无明显侵蚀土壤有明显的增加,且其基本理化性质相应变差。矿区侵蚀土壤微生物特征发生了显著的变化,与无明显侵蚀土壤相比,微生物生物量和可培养细菌数量显著降低,但土壤基础呼吸和微生物代谢商(qCO2)值却明显升高。Biolog测试结果显示,随着土壤侵蚀程度的加剧,其微生物群落结构亦发生了相应改变,侵蚀土壤微生物群落的丰富度、多样性指数、均匀度均显著低于对照土壤,且几项指标都达极显著水平差异(P<0.001),表明矿区水土流失引起了土壤微生物群落功能多样性的下降,减少了能利用有关碳源底物的微生物数量,降低了微生物对单一碳源底物的利用能力,最终导致土壤微生物群落结构和功能多样性发生变化。     

铜和磺胺嘧啶复合污染对土壤酶活性及微生物群落功能多样性的影响

随着畜禽养殖业规模化发展以及畜禽粪便的大量农用,由此导致的土壤重金属和抗生素复合污染问题日益突出。本文以畜禽养殖常用的饲料添加剂铜(Cu)和磺胺嘧啶(Sulfadiazine, SDZ)为目标污染物,分别采用土壤酶试剂盒(微量法)和Biolog-ECO盘法,研究二者复合污染对土壤酶活性和土壤微生物群落功能多样性的影响。结果发现,低浓度Cu(200 mg/kg)复合污染可缓解SDZ对土壤脱氢酶和β-葡萄糖苷酶的抑制作用,表现为拮抗作用；但加重SDZ对磷酸酶的抑制作用,表现为协同抑制作用。高浓度Cu(500 mg/kg)和SDZ复合对所测试土壤酶均表现为协同抑制作用。Biolog分析表明,10 mg/kg SDZ添加对土壤微生物活性具有促进作用,而Cu和SDZ复合污染处理则可显著抑制土壤微生物代谢活性。多样性指数分析表明Cu和SDZ复合污染对微生物群落多性影响较小,但可显著改变微生物种群的均一性；且主成分分析也表明二者复合污染可显著改变土壤微生物群落结构组成。本研究结论如下：1)Cu和SDZ复合污染对土壤酶活性的影响与土壤酶的种类及复合浓度有关,且土壤脱氢酶是反映SDZ及其与Cu复合污染较为敏感的指标；2)Cu和SDZ复合污染可显著抑制土壤微生物群落的整体代谢活性,并改变代谢功能,进而促使土壤微生物结构的发生变化。     

重金属污染对农田土壤无脊椎动物群落结构的影响

研究了土壤无脊椎动物对土壤重金属污染程度的响应。群落多样性指数随土壤重金属污染程度的加重而降低,井灌田、荒草田、黄灌田的群落多样性最高,而长期污灌田的群落多样性最低。重金属污染田土壤无脊椎动物的优势类群是蜱螨目和弹尾目,两类无脊椎动物具有较强的耐污染能力。在同一季节土壤无脊椎动物的种群数量主要受土壤污染程度的影响,污染程度越高其种群数量越低。不同季节间土壤无脊椎动物的数量存在极显著差异,不同污染程度的农田总群落、弹尾目和蜱螨目的个体数量均以7月为最多,10月次之,4月最少。     

多氯联苯污染土壤的微生物生态效应研究

以多氯联苯（Polychlorinated biphenyls,PCBs）自然污染的农田土壤为材料,分析土壤中微生物区系组成、生物量C、N、土壤基础呼吸以及微生物群落功能多样性的变化。研究结果表明,在以4-氯、5-氯同系物为主的PCBs污染土壤中,污染程度对土壤细菌、放线菌的数量影响不明显,而真菌的数量除与土壤污染程度有关,可能还受到土壤pH等性状的影响;土壤微生物C、N与土壤基础呼吸随污染程度的加剧呈下降趋势,但微生物C／N基本没有变化;Biolog分析显示,土壤微生物代谢剖面（AWCD）及Simpson指数在污染程度相差较大的两组土壤样品中差异均达到了显著性水平,表明PCBs污染引起了土壤微生物群落功能多样性下降,降低了微生物对不同单一碳源底物的利用能力。     

多环芳烃胁迫下稻田土壤细菌及分支杆菌种群多样性研究

多环芳烃（PAHs）污染对土壤微生物群落特别是微生物功能群的影响一直备受关注。本文应用变性梯度凝胶电泳（DGGE）技术,从群落和功能群多样性两个角度,分析了PAHs胁迫条件下稻田土壤细菌遗传多样性及与PAHs降解有密切关系的分支杆菌种群多样性变化。结果表明,PAHs污染对稻田土壤细菌群落多样性指数无显著影响,但造成土壤细菌群落结构的改变,重度PAHs造成一些对污染敏感的细菌种类消失,而使一些与PAHs降解有关的细菌种类丰度增加。而对与多环芳烃降解有密切关系的分支杆菌而言,中度PAHs污染稻田土壤分支杆菌种群多样性指数较重度和轻度PAHs污染土壤的略高,不同PAHs污染程度稻田土壤的优势分支杆菌种类不尽相同,PAHs污染造成稻田土壤1种或几种分支杆菌得到富集。长期PAHs污染造成土壤细菌群落和分支杆菌种群结构的变化,将直接影响土壤生态系统功能的发挥,间接改变土壤质量。     

基于BIOLOG指纹解析土壤可培微生物对铀污染的响应

为考察铀污染对土壤微生物群落的影响,以50、100、150 mg·L-1铀处理的土壤为研究对象,未处理的土壤为对照(CK),采用BIOLOG-ECO微孔板技术考察土壤微生物群落的功能多样性和碳源利用动力学特征等的变化。根据微孔板中的孔平均颜色变化率(AWCD)可知,铀污染对土壤微生物的生理活性呈现显著的抑制作用;采用Shannon多样性指数、Simpson指数、Mc Intosh指数描述微生物功能多样性,结果表明,铀处理组与CK的微生物群落功能多样性差异在3个多样性指数上均达到显著水平;土壤微生物对各类碳源的利用能力有所不同,以氨基酸类和胺类碳源为主;铀处理后,6类碳源相对利用率均比CK显著降低,其中羧酸类和多聚物类的利用率下降50%以上。主成分分析结果表明,铀污染后的土壤微生物特异利用的碳源主要有β-甲基-D-葡萄糖苷、D-半乳糖酸-γ-内脂、L-精氨酸等。因此,铀污染改变了土壤微生物的群落结构,导致微生物生理代谢等功能特性的变化。BIOLOG-ECO技术结合数学统计分析方法可以较为直观、快捷地反映土壤微生物的生物多样性,研究结果为评估和修复铀污染生态环境提供了理论依据。     

Diversity of microorganisms from forest soils differently polluted with heavy metals

Large accumulation of heavy metals in organic layers of forest soils may adversely affect the structure and diversity of microbial communities. The objective of this study was to assess the influence of different soil chemical properties on structure and diversity of microbial communities in soils polluted with different levels of heavy metals. The soil samples were taken at ten sites located in the vicinity of the cities of Legnica and Olkusz, differently polluted with Cu, Zn and Pb. The samples were measured for pH and the contents of organic C (C_org), total N (N_t), total S (S_t) and total Zn, Cu and Pb. The measured gross microbial properties included microbial biomass (C_mic) and soil respiration (RESP). The structure of soil microbial communities was assessed using phospholipid fatty acid (PLFA) analysis and the structure of soil bacterial communities using pyrosequencing of 16S rRNA genes. To assess diversity of the bacterial communities the Chao1 index was calculated based on the pyrosequencing data. For C_mic and RESP the most important factors were N_t and C_org, respectively. The structure and diversity of soil microbial communities revealed by PLFA profiles and pyrosequencing depended mainly on soil pH. The effect of high heavy metal contents on soil microbial properties was weaker compared with other soil properties. High concentrations of heavy metals negatively affected RESP and the Chao1 diversity index. The heavy metal pollution altered the structure of microbial communities measured with PLFA analysis, but the effect of heavy metal pollution was not observed for the structure of soil bacteria measured by pyrosequencing. The obtained results indicate that the use of soil microbial properties to study heavy metal effects may be difficult due to confounding influences of other environmental factors. In large-scale studies local variability of soil properties may obscure the effect of heavy metals.     

藏中矿区重金属污染对土壤微生物学特性的影响

土壤微生物对土壤重金属污染反应敏感,是探讨矿区土壤重金属污染生态效应的有效指标之一。通过野外调查与采样和室内分析,研究了藏中矿区重金属污染对土壤蔗糖酶、脲酶、脱氢酶和酸性磷酸酶活性、微生物生物量C（MBC）、N（MBN）和P（MBP）、土壤基础呼吸、代谢商（qCO2）及可矿化N的影响。研究表明,矿区土壤重金属Cu、Zn、Pb、Cd全量和有效含量均高于对照土壤;随着矿区土壤重金属含量增加,土壤酶活性、微生物量C、N和P、可矿化N均逐渐降低,土壤基础呼吸和qCO2则逐渐升高;土壤重金属与土壤蔗糖酶活性、脲酶活性、脱氢酶活性、酸性磷酸酶活性、MBC、MBN、土壤基础呼吸、qCO2及可矿化N具有显著的线性相关;脱氢酶活性对土壤重金属污染最为敏感,表明脱氢酶活性可作为藏中矿区土壤环境质量变化的有效指标。     

重金属复合污染下红壤微生物活性及其群落结构的变化

在室内培养条件下,应用4因素5水平二次正交回归旋转组合设计方案,对重金属复合污染红壤的微生物活性及群落功能多样性进行了研究。结果表明,在Cu、Zn、Pb、Cd复合污染生境中重金属的微生物毒性效应发挥主要由Cd、Cu两元素决定,其生物毒性顺序表现为:Cd>Cu>Zn>Ph,红砂泥中表现较为突出。与单一镉、单一铅污染处理相比,重金属复合污染对供试红壤微生物生物活性及其群落功能多样性的影响并非仅仅表现出简单的加和作用,同时还存在协同作用和拮抗作用。典型相关分析显示,红壤微生物活性与重金属含量之间关系密切,第一对典型变量的相关系数r达0.885 8、0.932 8,均达极显著水平(p<0.001), 说明重金属复合污染与红壤微生物间存在较为稳定的数量反馈机制,红壤总体微生物活性指标能较好地反映供试红壤重金属复合污染状况,可作为重金属污染红壤环境质量评价及量化分类的有效指标。     

利用Illumina高通量测序对复合重金属污染下根际和非根际稻田土壤细菌的研究

There is an increasing concern about rice (Oryza sativa L.) soil microbiomes under the influence of mixed heavy metal contamination.We used the high-throughput Illumina MiSeq sequencing approach to explore the bacterial diversity and community composition of soils in four paddy fields,exhibiting four degrees of mixed heavy metal (Cd,Pb and Zn) pollution,and examined the effects of these metals on the bacterial communities.Our results showed that up to 2 104 to 4 359 bacterial operational taxonomic units (OTUs) were found in the bulk and rhizosphere soils of the paddy fields,with the dominant bacterial phyla (greater than 1％ of the overall community) including Proteobacteria,Actinobacteria,Firmicutes,Acidobacteria,Gemmatimonadetes,Chloroflexi,Bacteroidetes and Nitrospirae.A number of rare and candidate bacterial groups were also detected,and Saprospirales,HOC36,SC-I-84 and Anaerospora were rarely detected in rice paddy soils.Venn diagram analysis showed that 174 bacterial OTUs were shared among the bulk soils with four pollution degrees.Rice rhizosphere soils displayed higher bacterial diversity indices (ACE and Chao 1) and more unique OTUs than bulk soils.Total Cd and Zn in the soils were significantly negatively correlated with ACE and Chao 1,respectively,and the Mantel test suggested that total Pb,total Zn,pH,total nitrogen and total phosphorus significantly affected the community structure.Overall,these results provided baseline data for the bacterial communities in bulk and rhizosphere soils of paddy fields contaminated with mixed heavy metals.     

Changes in Microbial Functional Diversity and Activity in Paddy Soils Irrigated with Industrial Wastewaters in Bandung, West Java Province, Indonesia

Characteristics, such as microbial biomass, basal respiration, and functional diversity of the microbial communities, were investigated in paddy soils located in Bandung, West Java Province, Indonesia, that have been heavily polluted by industrial effluents for 31 years. Paddy soil samples (10?C20 cm) were taken from two sites: polluted soils and unpolluted soils (as control sites). The polluted soils contained higher salinity, higher sodicity, higher nutrient contents, and elevated levels of heavy metals (Cr, Mn, Ni, Cu, and Zn) than unpolluted soils. Soil physicochemical properties, such as maximum water holding capacity, exchangeable sodium percentage, sodium adsorption ratio, and swelling factor, in polluted soils were much greater than those in unpolluted soils (P?<?0.05). Changes in the physical and chemical soil properties were reflected by changes in the microbial communities and their activities. BIOLOG analysis indicated that the functional diversity of the microbial community of polluted soils increased and differed from that of unpolluted soils. Likewise, the average rate of color development (average well color development), microbial biomass (measured as DNA concentration), and the soil CO₂ respiration were higher in polluted soils. These results indicate that major changes in the chemical and physical properties of paddy soils following the application of industrial wastewater effluents have had lasting impacts on the microbial communities of these soils. Thus, the increased activity, biomass, and functional diversity of the microbial communities in polluted soils with elevated salinity, sodicity, and heavy metal contents may be a key factor in enhancing the bioremediation process of these heavily polluted paddy soils.     

Influence of heavy metals on the functional diversity of soil microbial communities

Three soil types-Calcaric Phaeozem, Eutric Cambisol and Dystric Lithosol-in large container pots were experimentally contaminated with heavy metals at four different levels (light pollution: 300 ppm Zn, 100 ppm Cu, 50 ppm Ni, 50 ppm V and 3 ppm Cd; medium pollution: twofold concentrations; heavy pollution: threefold concentrations; uncontaminated control). We investigated the prognostic potential of 16 soil microbial properties (microbial biomass, respiration, N-mineralization, 13 soil enzymes involved in cycling of C, N, P and S) with regard to their ability to differentiate the four contamination levels. Microbial biomass and enzyme activities decreased with increasing heavy metal pollution, but the amount of decrease differed among the enzymes. Enzymes involved in the C-cycling were least affected, whereas vartous enzyme activities related to the cycling of N, P and S showed a considerable decrease in activity. In particular, arylsulfatase and phosphatase activities were dramatically affected. Their activity decreased to a level of a few percent of their activities in the corresponding unpolluted controls. The data suggest that aside from the loss of rare biochemical capabilities-such as the growth of organisms at the expense of aromatics (Reber 1992)-heavy metal contaminated soils lose very common biochemical propertities which are necessary for the functioning of the ecosystem. Cluster analysis as well as discriminant analysis underline the similarity of the enzyme activity pattern among the controls and among the polluted soils. The trend toward a significant functional diversity loss becomes obvious already at the lowest pollution level. This implies that concentrations of heavy metals in soils near the current EC limits will most probably lead to a considerable reduction in decomposition and nutrient cycling rates. We conclude that heavy metal pollution severely decreases the functional diversity of the soil microbial community and impairs specific pathways of nutrient cycling.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

重金属复合污染农田土壤DNA的快速提取及其PCR-DGGE分析

国内首次运用FastPrep○R 核酸快速提取系统提取了重金属复合污染农田土壤的DNA ,并对其进行了聚合酶链反应—变性梯度凝胶电泳 (PCR DGGE)分析。结果表明 ,FastPrep○R核酸提取仪与相应的FastD NASPINKitforSoil试剂盒联用时 ,能有效地分离到纯度较高的重金属污染农田土壤的DNA。PCR DGGE电泳图谱表明 ,PCR产物经DGGE检测后得到的电泳条带清晰且分离效果好 ,可以明显反映出重金属复合污染导致了农田土壤微生物在基因上的损伤 ,影响到农田土壤生态系统的细菌丰富度 ,改变了土壤环境的优势菌群 ,从而使农田土壤微生物群落结构多样性发生变化。可见 ,FastPrep○R核酸提取系统同样适用于重金属污染农田土壤环境中微生物基因组DNA的快速分离和纯化 ,得到的DNA可直接用于PCR DGGE分析。