RAPD marker and substrate utilization pattern applied to study microbial community diversity in the soil affected by agricultural chemicals

Present analyses of random amplified polymorphic DNA (RAPD) and Biolog GN substrate utilization pattern are combined to further study the diversity of microbial communities in four soils affected by agricultural chemicals. The results showed that the four soil microbial communities were apparently distinguishable in the diversity at RAPD level in terms of the richness and modified richness in the summer, which supports our previous report using the same soils in winter. A significant difference for the average well color development (AWCD) at 72 h incubation was found among the soils in winter using Biolog GN substrate utilization pattern, but this difference was not found among the soils in summer. However, Shannon-Weaver indices for microbial communities in the summer soils polluted by agricultural chemicals were significantly higher than those in winter at metabolic level; in contrast, no significant difference existed between the two seasons for microbial communities in the soil without chemical pollution. Present results suggest that the combined approach using RAPD and substrate utilization pattern could be used to effectively quantify microbial community diversity and its changes among the seasons in the soils affected by agricultural chemicals, simultaneously at molecular and physiological levels.     

RAPD Marker and Substrate Utilization Pattern Applied to Study Microbial Community Diversity in the Soil Affected by Agricultural Chemicals

Abstract

Present analyses of random amplified polymorphic DNA (RAPD) and Biolog GN substrate utilization pattern are combined to further study the diversity of microbial communities in four soils affected by agricultural chemicals. The results showed that the four soil microbial communities were apparently distinguishable in the diversity at RAPD level in terms of the richness and modified richness in the summer, which supports our previous report using the same soils in winter. A significant difference for the average well color development (AWCD) at 72 h incubation was found among the soils in winter using Biolog GN substrate utilization pattern, but this difference was not found among the soils in summer. However, Shannon-Weaver indices for microbial communities in the summer soils polluted by agricultural chemicals were significantly higher than those in winter at metabolic level; in contrast, no significant difference existed between the two seasons for microbial communities in the soil without chemical pollution. Present results suggest that the combined approach using RAPD and substrate utilization pattern could be used to effectively quantify microbial community diversity and its changes among the seasons in the soils affected by agricultural chemicals, simultaneously at molecular and physiological levels.     

Metabolic diversity of the heterotrophic microorganisms and potential link to pollution of the Rouge River

The heterotrophic microbial communities of the Rouge River were tracked using Biolog Ecoplates to understand the metabolic diversity at different temporal and spatial scales, and potential link to river pollution. Site less impacted by anthrophogenic sources (site 1), showed markedly lower metabolic diversity. The only substrates that were utilized in the water samples were carbohydrates. Sites more impacted by anthrophogenic sources (sites 8 and 9) showed higher metabolic diversity. Higher functional diversity was linked to the physico-chemical and biological properties of the water samples (i.e. higher concentrations of DO, DOC, chlorophyll, and bacterial density). Biolog analysis was found to be useful in differentiating metabolic diversity between microbial communities; in determining factors that most influence the separation of communities; and in identifying which substrates were most utilized by the communities. It can also be used as an effective ecological indicator of changes in river function attributable to urbanization and pollution.     

Assessing microbial communities for a metabolic profile similar to activated sludge.

To search for reliable testing inocula alternatives to activated sludge cultures, several model microbial consortia were compared with activated sludge populations for their functional diversity. The evaluation of the metabolic potential of these mixed inocula was performed using the Biolog EcoPlates and GN and GP MicroPlates (Biolog, Inc., Hayward, California). The community-level physiological profiles (CLPPs) obtained for model communities and activated sludge samples were analyzed by principal component analysis and hierarchic clustering methods, to evaluate the ability of Biolog plates to distinguish among the different microbial communities. The effect of different inocula preparation methodologies on the community structure was also studied. The CLPPs obtained with EcoPlates and GN MicroPlates showed that EcoPlates are suitable to screen communities with a metabolic profile similar to activated sludge. New, well-defined, standardized, and safe inocula presenting the same metabolic community profile as activated sludge were selected and can be tested as surrogate cultures in activated-sludge-based bioassays.     

Ni2＋对活性污泥活性及群落多样性的影响

通过检测活性污泥的电子传递体系活性以及生物多样性,研究Ni2＋对活性污泥微生物活性及群落多样性的影响。结果表明：与对照系统相比,5mg／L的Ni2＋对2,3,5-lriphenylteItrazoliumchloride（TTC．ETS）活性未产生显著的影响;但当Ni2＋的浓度进一步增大到10、20和40mg／L后,其对序批式反应器内活性污泥TTC—ETS活性的抑制率分别达到（36．794-11．14）％、（55．88±13．90）％和（70．97±6．78）％。低浓度Ni2＋．能增强活性污泥微生物对碳源的利用,但高于10mg／L的Ni2＋则显著抑制了活性污泥微生物对碳源的利用。各个SBR系统中微生物群落最常见的物种相近,物种丰富度和均一性则均有所不同,其中群落物种丰富度随着Ni2＋浓度的增加而逐渐减小。TTC—ETS活性、平均每孔颜色变化率、Shan—liOn指数和Simpson指数,与Ni2＋的胁迫浓度之间的显著相关性表明,它们均可有效地表征Ni2＋胁迫对活性污泥微生物活性及群落多样性的影响程度。     

Comparison of the purification performance and microbial community functional diversity in flow-directional-switching and unidirectional-flow biotrickling filters

Because of the characteristics of low operating cost and convenient operation, the biotrickling filter is extensively researched and used to treat low concentration waste gas contaminated by volatile organic compounds (VOCs) and other odors. In this paper, two laboratory-scale biotrickling filters were constructed and toluene was selected as the sole carbon source, and the effects of different waste-gas flow configuration patterns on the purification capacity and the microbial community functional diversity of biotrickling filters were evaluated. The results indicated that the flow-directional-switching (FDS) biotrickling filter had better purification performance, and the maximum elimination capacity reached 480 g·m^?3·hr^?1, which was 17.1% higher than conventional unidirectional-flow (UF) biotrickling filter. Comparing the purification capacities of different sections in two biotrickling filters, the maximum toluene elimination capacity of section III in FDS system could reach 542 g·m^?3·hr^?1, which was 2.8 times as great as that in UF system, which resulted from the difference of elimination capacity in two systems. By analyzing the metabolic activity of two systems by community-level physiological profiling (CLPP) with Biolog (Biolog Inc., Hayward, CA) ECO-plate technique, metabolic activity in three sections of FDS system was higher than that of UF system. The metabolic activity was the highest in section III of FDS system and 46.8% higher than that of UF system. Shannon index and McIntosh index of section III in FDS system were 6.2% and 31.5% higher, respectively, than those of UF system.

Implications: The flow-directional-switching (FDS) biotrickling filter had a better purification performance than unidirectional-flow (UF) biotrickling filter at high inlet loadings, because FDS produced a more uniform distribution of biomass and microbial metabolic capacity along the length of the packed bed without diminishing activity and removal capacity in the inlet section.     

印染废水接触氧化池中悬浮污泥和生物膜的微生物群落比较分析

为了揭示接触氧化池中悬浮污泥和生物膜菌群群落结构的异同,提取基因组DNA,采用PCR扩增与454高通量测序技术对两者菌群测序并进行了操作单元(OTU)聚类分析、多样性分析和分类学分析。结果表明,悬浮污泥和生物膜菌群的高通量测序分析分别得到优化序列8 256条和10 592条,测序覆盖深度都在80%左右,经过97%相似度归并后分别得到570个OTUs和694个OTUs,菌群多样性分析显示悬浮污泥内菌群的丰度和多样性都低于生物膜,悬浮污泥有356个属的细菌构成,而生物膜内则有387个,都归属于21个门。悬浮污泥和生物膜内不仅存在共有菌种,也含有各自的特定菌种,说明不同的微环境使其有了相对应的优势群落。通过这种差异的比较分析,更能准确地了解接触氧化池中微生物的群落组成情况,有利于分析其与系统功能的关系。     

混合型污水处理厂原位生物强化脱氮中试及微生物群落结构分析研究

针对混合型污水处理厂进水水质波动大、C/N低、总氮稳定达标困难等共性问题,开发出适宜于低C/N、贫营养等苛刻环境条件的高效脱氮菌剂.现场中试结果表明:经过脱氮菌强化后,生化系统脱氮效率提升8.5百分点,出水平均总氮较强化前下降3 mg/L,出水水质稳定性明显提升,生物强化效果显著.在不补加碳源条件下,深度处理出水总氮完...     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

Effects of lead and cadmium nitrate on biomass and substrate utilization pattern of soil microbial communities

A study was conducted to evaluate the effects of different concentrations of lead (Pb) and cadmium (Cd) applied as their nitrates on soil microbial biomass carbon (C(mic)) and nitrogen (N(mic)), and substrate utilization pattern of soil microbial communities. The C(mic) and N(mic) contents were determined at 0, 14, 28, 42 and 56 days after heavy metal application (DAA). The results showed a significant decline in the C(mic) for all Pb and Cd amended soils from the start to 28 DAA. From 28 to 56 DAA, C(mic) contents changed non-significantly for all other treatments except for 600 mgkg(-1) Pb and 100 mgkg(-1) Cd in which it declined significantly from 42 to 56 DAA. The N(mic) contents also decreased significantly from start to 28 DAA for all other Pb and Cd treatments except for 200 mgkg(-1) Pb which did not show significant difference from the control. Control and 200 mgkg(-1) Pb had significantly lower soil microbial biomass C:N ratio as compared with other Pb treatments from 14 to 42 DAA, however at 56 DAA, only 1000 mgkg(-1) Pb showed significantly higher C:N ratio compared with other treatments. No significant difference in C:N ratio for all Cd treated soils was seen from start to 28 DAA, however from 42 to 56 DAA, 100 mgkg(-1) Pb showed significantly higher C:N ratio compared with other treatments. On 56 DAA, substrate utilization pattern of soil microbial communities was determined by inoculating Biolog ECO plates. The results indicated that Pb and Cd addition inhibited the functional activity of soil microbial communities as indicated by the intensity of average well color development (AWCD) during 168 h of incubation. Multivariate analysis of sole carbon source utilization pattern demonstrated that higher levels of heavy metal application had significantly affected soil microbial community structure.     

Carbendazim dissipation in the biomixture of on-farm biopurification systems and its effect on microbial communities

The impact of repeated carbendazim (CARB) applications on the extent of CARB dissipation, the microbial diversity, the community level physiological profile (CLPP), and the enzymatic activity within the biomixture of an on-farm biopurification system was evaluated. After three successive CARB applications, the CARB dissipation efficiency was high; the efficiency of dissipation was 87%, 94% and 96% after each application, respectively. Although microbial enzymatic activity was affected significantly by CARB application, it could recover after each CARB pulse. Likewise, the numbers of cultivable bacteria, fungi and actinomycetes (as measured in CFUs) were slightly affected by the addition of CARB, but the inhibitory effect of the pesticide application was temporary. Denaturing gradient gel electrophoresis (DGGE) and Biolog Ecoplate assays demonstrated that the microbial populations remained relatively stable over time when compared to the control. The results obtained herein therefore demonstrate the high dissipation capacity of this biomixture and highlight the microbiological robustness of this biological system.     

Antibiotic resistance genes in an urban river as impacted by bacterial community and physicochemical parameters

Antibiotic resistance genes (ARGs) in urban rivers are a serious public health concern in regions with poorly planned, rapid development. To gain insights into the predominant factors affecting the fate of ARGs in a highly polluted urban river in eastern China, a total of 285 ARGs, microbial communities, and 20 physicochemical parameters were analyzed for 17 sites. A total of 258 unique ARGs were detected using high-throughput qPCR, and the absolute abundance of total ARGs was positively correlated with total organic carbon and total dissolved nitrogen concentrations (P < 0.01). ARG abundance and diversity were greatly altered by microbial community structure. Variation partitioning analysis showed that the combined effects of multiple factors contributed to the profile and dissemination of ARGs, and variation of microbial communities was the major factor affecting the distribution of ARGs. The disparate distribution of some bacteria, including Bacteroides from mammalian gastrointestinal flora, Burkholderia from zoonotic infectious diseases, and Zoogloea from wastewater treatment, indicates that the urban river was strongly influenced by point-source pollution. Results imply that microbial community shifts caused by changes in water quality may lead to the spread of ARGs, and point-source pollution in urban rivers requires greater attention to control the transfer of ARGs between environmental bacteria and pathogens.

     

Microbial abundance and community in subsurface flow constructed wetland microcosms: role of plant presence

In this research, the role of plants in improving microorganism growth conditions in subsurface flow constructed wetland (CW) microcosms was determined. In particular, microbial abundance and community were investigated during summer and winter in Phragmites australis-planted CW microcosms (PA) and unplanted CW microcosms (control, CT). Results revealed that the removal efficiencies of pollutants and microbial community structure varied in winter with variable microbial abundance. During summer, PA comprised more dominant phyla (e.g., Proteobacteria, Actinobacteria, and Bacteroidetes), whereas CT contained more Cyanobacteria and photosynthetic bacteria. During winter, the abundance of Proteobacteria was >40 % in PA but dramatically decreased in CT. Moreover, Cyanobacteria and photosynthetic bacterial dominance in CT decreased. In both seasons, bacteria were more abundant in root surfaces than in sand. Plant presence positively affected microbial abundance and community. The potential removal ability of CT, in which Cyanobacteria and photosynthetic bacteria were abundant during summer, was more significantly affected by temperature reduction than that of PA with plant presence.     

The effects of perennial ryegrass and alfalfa on microbial abundance and diversity in petroleum contaminated soil

Enhanced rhizosphere degradation uses plants to stimulate the rhizosphere microbial community to degrade organic contaminants. We measured changes in microbial communities caused by the addition of two species of plants in a soil contaminated with 31,000 ppm of total petroleum hydrocarbons. Perennial ryegrass and/or alfalfa increased the number of rhizosphere bacteria in the hydrocarbon-contaminated soil. These plants also increased the number of bacteria capable of petroleum degradation as estimated by the most probable number (MPN) method. Eco-Biolog plates did not detect changes in metabolic diversity between bulk and rhizosphere samples but denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified partial 16S rDNA sequences indicated a shift in the bacterial community in the rhizosphere samples. Dice coefficient matrices derived from DGGE profiles showed similarities between the rhizospheres of alfalfa and perennial ryegrass/alfalfa mixture in the contaminated soil at week seven. Perennial ryegrass and perennial ryegrass/alfalfa mixture caused the greatest change in the rhizosphere bacterial community as determined by DGGE analysis. We concluded that plants altered the microbial population; these changes were plant-specific and could contribute to degradation of petroleum hydrocarbons in contaminated soil.     

Ecosystem Response to Climatic Change: The Importance of the Cold Season

Winter climate and snow cover are the important drivers of plant community development in polar regions. However, the impacts of changing winter climate and associated changes in snow regime have received much less attention than changes during summer. Here, we synthesize the results from studies on the impacts of extreme winter weather events on polar heathland and lichen communities. Dwarf shrubs, mosses and soil arthropods were negatively impacted by extreme warming events while lichens showed variable responses to changes in extreme winter weather events. Snow mould formation underneath the snow may contribute to spatial heterogeneity in plant growth, arthropod communities and carbon cycling. Winter snow cover and depth will drive the reported impacts of winter climate change and add to spatial patterns in vegetation heterogeneity. The challenges ahead lie in obtaining better predictions on the snow patterns across the landscape and how these will be altered due to winter climate change.     

Ecosystem response to climatic change: the importance of the cold season

Winter climate and snow cover are the important drivers of plant community development in polar regions. However, the impacts of changing winter climate and associated changes in snow regime have received much less attention than changes during summer. Here, we synthesize the results from studies on the impacts of extreme winter weather events on polar heathland and lichen communities. Dwarf shrubs, mosses and soil arthropods were negatively impacted by extreme warming events while lichens showed variable responses to changes in extreme winter weather events. Snow mould formation underneath the snow may contribute to spatial heterogeneity in plant growth, arthropod communities and carbon cycling. Winter snow cover and depth will drive the reported impacts of winter climate change and add to spatial patterns in vegetation heterogeneity. The challenges ahead lie in obtaining better predictions on the snow patterns across the landscape and how these will be altered due to winter climate change.     

生物强化对MBR系统生物特性及群落结构的影响

采用高效菌强化膜生物反应器对溴氨酸废水进行处理,考察了生物强化前后系统对溴氨酸的降解能力及其内部微生物生理状态变化.并采用核糖体基因间隔序列分析技术(RISA)解析了高效菌投加前后污泥系统的群落变化.实验表明,投菌后在进水负荷增加的条件下,上清液和膜出水的溴氨酸脱色率分别约为50%和65%,相应的COD去除率分别约为25%和50%,和投菌前基本保持一致;TTC-脱氢酶活性和胞外聚合物(EPS)浓度略有波动,但是运行一段时间后即恢复到投菌前水平.群落分析表明高效菌可以在系统中稳定存在,并且不对原菌群结构造成较大影响.     

DGGE、T-RFLP 、LH-PCR对两种活性污泥的微生物种群多样性分析的比较

采用基于PCR扩增的分子生态技术DGGE、T-RFLP和LH-PCR,对2种典型污水处理系统中的活性污泥（生活污泥、焦化污泥）进行微生物种群多样性分析;并以此比较3种技术的优劣,提出不同应用条件下研究方法的选择依据。根据实验结果：DGGE得到的条带较多,但误差来源也最多;T-RFLP技术较为灵敏,但需要选择适当的内切酶,严格控制酶切条件,并且文库比对误差较大;而LH-PCR操作简单,结果稳定性较高。虽然目前尚无法判断3种方法的准确性,但LH-PCR在活性污泥的微生物种群多样性分析中已显示出潜在优势。     

曝气强度对同时硝化反硝化（SND）脱氮过程中N2O排放的影响

为了控制污水脱氮中N2O排放,在不同曝气强度下研究了好氧硝化段同时硝化反硝化（SND）系统的N2O排放特性,并采用PCR—DGGE技术分析微生物群落特征。结果发现,随着曝气强度的增强,系统总氮去除率下降,但脱氮中N2O—N所占比例则上升,实验中从低到高3个曝气强度下,总氮去除率分别为80．01％、65．28％和58．62％,脱氮中N2O—N所占的比例为1．89％、7．84％和9．20％。PCR—DGGE分析显示,和低曝气强度下相比中、高曝气强度下系统微生物群落发生明显变化,但中曝气强度和高曝气强度下系统微生物群落表现出较高相似性。这表明,不同曝气强度下系统N2O排放受到氮素转化和微生物群落变化的影响。适宜曝气强度不仅提高总氮去除率,还可有效控制N2O排放。     

Comparative assessment of air quality in two health resorts using carbon isotopes and palynological analyses

This paper describes results of applying the palynological and carbon isotopic analysis of the organic fraction of Total Suspended Particles (TSP) to discriminate distinct pollution sources and assess the anthropogenic impact for the investigated areas. The samples of atmospheric particles were collected in Czerniawa and Cieplice (two health resorts in Lower Silesia, SW Poland) twice a year in summer and winter season (from July 2006 to February 2008). The palynological spectra represent in the vast majority local plant communities without a noticeable contribution of long-transported plant particles. Palynological analysis revealed also differences in the specificity of the two sampling areas, i.e. the higher contribution of identified organic material in Czerniawa stands for more natural character of this site, but is also responsible for the higher allergic pressure when compared to Cieplice. The carbon isotopic composition of TSP varied seasonally (δ¹³C value from ?27.09‰ in summer to ?25.47‰ in winter). The increased δ¹³C value in winter (heating period) is most probably caused by uncontrolled contribution of coal soot. On the basis of isotopic mass balance the calculated contribution of anthropogenic organic particles in the atmosphere reached in winter season 72% in Czerniawa and 79% in Cieplice.