Characterization of microbial communities from coastal waters using microarrays

Molecular methods, including DNA probes, were used to identify and enumerate pathogenic Vibrio species in the Chesapeake Bay; our data indicated that Vibrio vulnificus exhibits seasonal fluctuations in number. Our work included a characterization of total microbial communities from the Bay; development of microarrays that identify and quantify the diversity of those communities; and observation of temporal changes in those communities. To identify members of the microbial community, we amplified the 16S rDNA gene from community DNA isolated from a biofilm sample collected from the Chesapeake Bay in February, 2000. The resultant 75 sequences were 95% or more similar to 7 species including two recently described Shewanella species, baltica and frigidimarina, that have not been previously isolated from the Chesapeake. When the genera of bacteria from biofilm after culturing are compared to those detected by subcloning amplified 16S fragments from community DNA, the cultured sample exhibited a strong bias. In oysters collected in February, the most common bacteria were previously unknown. Based on our 16S findings, we are developing microarrays to detect these and other microbial species in these estuarine communities. The microarrays will detect each species using four distinct loci, with the multiple loci serving as an internal control. The accuracy of the microarray will be measured using sentinel species such as Aeromonas species, Escherichia coli, and Vibrio vulnificus. Using microarrays, it should be possible to determine the annual fluctuations of bacterial species (culturable and non-culturable, pathogenic and non-pathogenic). The data may be applied to understanding patterns of environmental change; assessing the health of the Bay; and evaluating the risk of human illness associated with exposure to and ingestion of water and shellfish.     

钒胁迫对水稻幼苗生理生化和富集特性的影响

水稻（Oryza.sativa L.）是我国最重要的粮食作物之一,水稻产量占粮食总产量的一半以上,一旦水稻受到重金属污染,将会影响水稻植株的正常生长和生理特性。目前关于钒胁迫对水稻植株生理特性指标的影响方面报道较少。通过水培实验,研究了不同钒（V）质量浓度（0、4、8、12、16、20 mg·L-1）对水稻幼苗（Oryza.sativa L）生理生化和富集特性的影响。结果表明：随着V胁迫浓度的增加,叶绿素含量、可溶性蛋白含量、过氧化氢酶（CAT）活性、过氧化物酶（POD）活性、超氧化物歧化酶（SOD）等均呈现先上升、后下降的变化趋势。当ρ（V）≤12 mg·L-1,与对照相比较,叶绿素含量、可溶性蛋白含量和酶活性增大了135.3%、104.2%、77.8%（CAT）、84.5%（POD）和273.2%（SOD）;当ρ（V）〉12 mg·L-1,则分别降低37.2%、39.4%、41.1%、24.1%和24.5%。随着 V 胁迫浓度的增加,丙二醛（MDA）含量和细胞膜透性逐渐增大,与对照相比,分别增加了38.5%~289.3%、21.2%~303.2%,根系活力下降了10.9%~82.2%。可见,低ρ（V）（≤12 mg·L-1）对水稻幼苗的生长有一定的刺激作用,水稻幼苗自身保护酶表现出较强的自我调节能力;高ρ（V）（〉12 mg·L-1）明显抑制叶绿素和蛋白合成、抗氧化酶活性和根系活力,伤害了细胞质膜系统,影响水稻幼苗的生长发育。不同V浓度胁迫下,水稻幼苗累积的V含量为：根〉茎叶。随着V胁迫浓度增加,水稻幼苗各器官V含量增大,其中根部增幅远大于茎叶,当ρ（V）从5 mg·L-1增加到40 mg·L-1,与对照相比较,根部增加了0.98~25.3倍,茎叶部增加了0.26~4.74倍。生物富集系数（BF）先增加后降低,最大值为2.8408;迁移系数（TF）下降,最低值为0.1170,说明水稻对V有较强的富集能力,但迁移能力较低,积累的V主要富集在根部,可减轻V对地上部植物的危害。     

投加聚氨酯泡沫微生物固定化载体的SBBR脱氮除磷实验研究

研究投加聚氨酯泡沫微生物固定化载体的SBBR脱氮除磷效果及机理.在SBR中填充聚氨酯泡沫微生物固定化载体,形成序批式生物膜反应器(SBBR).SBBR以实际生活污水为处理对象,在A/O运行工况下对微生物进行培养驯化,并在SBBR运行稳定时研究其脱氮除磷效果.研究结果表明,SBBR中的聚氨酯泡沫微生物固定化载体在空间上形...     

饮用水生物处理中微生物量和活性的测定方法

微生物量和微生物活性是饮用水生物处理工艺设计与运行的重要参数。总结论述了适用于表示饮用水生物处理过程生物膜中微生物数量和活性的几种主要指标的测定方法。     

Method to Partition Between Attached and Unattached E. coli in Runoff From Agricultural Lands1

Abstract: Remediation of waters impaired by bacterial indicators is usually dictated by total maximum daily load plans, which are heavily dependent on fate and transport modeling of bacterial indicators. Nonpoint source pollution models are most frequently used to assess bacterial transport to surface waters and most models typically simulate bacterial transport as a dissolved pollutant. Previous studies have found that cells preferentially attach to sediments; however, a variety of techniques have been used to assess attachment including filtration, fractional filtration, and centrifugation. In addition, a variety of chemical and physical dispersion techniques are used to release attached and bioflocculated cells from particulates. Here we developed and validated an easy‐to‐replicate laboratory procedure for separation of unattached from attached E. coli which will also identify particle sizes to which E. coli preferentially attach. Physical and chemical dispersion techniques were evaluated and a combined hand shaker treatment for 10 min followed by dilutions in 1,000 mg/l of Tween 85 significantly increased total E. coli concentrations by 31% when compared with a control. In order to separate unattached from attached fractions, two commonly used techniques, fractional filtration, and centrifugation were combined. The filtration and centrifugation treatments did not reduce E. coli concentrations when compared with a control (p > 0.05), indicating that damage was not inflicted upon the E. coli cells during the separation procedure.     

影响土壤中微生物体氮的因子

综述了影响土壤微生物体氮的因子,以及微生物体氮与可矿化氮之间的关系。着重讨论了土壤有机质、有机物料、氮肥、作物根系、土壤温度和湿度对微生物体氮的影响及研究进展;评价了土壤微生物体氮与可矿化氮的关系的研究结果。     

解磷微生物肥料的研究与进展

磷是植物生长必需的矿物质元素之一,在土壤中易被固定降低其有效性,目前,研究和生产含有解磷功能的微生物有机肥料是转化土壤中无效磷、提高磷肥利用率的有效途径之一。叙述了解磷微生物犯料中微生物的种类、生态分布规律和影响高效解磷微生物选育的外部因素,概述了解磷微生物菌肥的应用和研究进展,探讨了解磷微生物肥料的应用价值。为解磷微生物未来的研究重点提出理论依据。．     

Biofilm structure and its influence on clogging in drip irrigation emitters distributing reclaimed wastewater

Using reclaimed wastewater for crop irrigation is a practical alternative to discharge wastewater treatment plant effluents into surface waters.However,biofouling has been identified as a major contributor to emitter clogging in drip irrigation systems distributing reclaimed wastewater.Little is known about the biofilm structure and its influence on clogging in the drip emitter flow path.This study was first to investigate the microbial characteristics of mature biofilms present in the emitters and the effect of flow path structures on the biofilm microbial communities.The analysis of biofilm matrix structure using a scanning electron microscopy (SEM) revealed that particles in the matrix of the biofilm coupled extracellular polysaccharides (EPS) and formed sediment in the emitter flow path.Analysis of biofilm mass including protein,polysaccharide and phospholipid fatty acids (PLFAs) showed that emitter flow path style influenced biofilm community structure and diversity.The correlations of biofilm biomass and discharge reduction after 360 h irrigation were computed and suggest that PFLAs provide the best correlation coefficient.Comparatively,the emitter with the unsymmetrical dentate structure and shorter flow path (Emitter C) had the best anti-clogging capability.By optimizing the dentate structure,the internal flow pattern within the flow path could be enhanced as an important method to control the biofilm within emitter flow path.This study established electron microscope techniques and biochemical microbial analysis methods that may provide a framework for future emitter biofilm studies.     

垦殖对桂林会仙喀斯特湿地土壤养分与微生物活性的影响

选择桂林会仙喀斯特湿地不同垦殖方式的土壤为研究对象,通过研究原生沼泽湿地及其开垦21 a的水田和旱地0~40 cm不同深度的土壤理化性质、微生物数量、酶活性、微生物呼吸等变化趋势,以揭示垦殖对会仙喀斯特湿地土壤质量的影响.结果表明：①垦殖导致会仙喀斯特湿地土壤呈现偏酸性和养分含量降低的趋势,垦殖后土壤有机碳（SOC）、全氮（TN）、速效氮（AN）、全磷（TP）、速效磷（AP）、全钾（TK）和速效钾（AK）总体上趋于减小,并且表层土壤减小的趋势最为显著.②垦殖改变了土壤微生物数量分布的相对比例关系,水田土壤的细菌和放线菌数量最高,而土壤真菌数量则在旱地最高（P<0.001）.③垦殖降低了各土层蛋白酶、蔗糖酶、脲酶、过氧化氢酶和多酚氧化酶的活性,在旱地的数值最低.④垦殖后土壤微生物量碳（MBC）、微生物量氮（MBN）、基础呼吸（BR）、呼吸势（PR）和微生物熵（qMB）总体上均趋于减小,代谢熵（qCO₂）在湿地表现为先增大后减小的趋势,而在水田和旱地则一致表现为逐渐升高的趋势.各土层的MBC、MBN、BR、PR和qMB普遍呈现出湿地 > 水田 > 旱地的趋势.qCO₂在0~10 cm和10~20 cm土层表现为湿地 > 水田 > 旱地,而在20~30 cm和30~40 cm土层则表现为旱地 > 水田 > 湿地.综上,垦殖显著降低了土壤的微生物活性,土壤质量呈现下降趋势,对旱地土壤影响较为严重.在会仙喀斯特湿地生态恢复过程中,应减少垦殖旱地和水田的面积,退耕恢复湿地面积.     

Effect of seed sludge on characteristics and microbial community of aerobic granular sludge

Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor.The influence of seed sludge on physical and chemical properties of granular sludge was studied;the microbial community structure was probed by using scanning electron microscope and polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE).The results showed that seed sludge played an important role on the formation of aerobic granules.Seed sludge taken from beer wastewater treatment plant(inoculum A) was more suitable for cultivating aerobic granules than that of sludge from municipal wastewater treatment plant(inoculum B).Cultivated with inoculum A, large amount of mature granules formed after 35 days operation, its SVI reached 32.75 mL/g, and SOUR of granular sludge was beyond 1.10 mg/(g·min).By contrast, it needed 56 days obtaining mature granules using inoculum B.DGGE profiles indicated that the dominant microbial species in mature granules were 18 and 11 OTU when inoculum A and B were respectively employed as seed sludge.The sequencing results suggested that dominant species in mature granules cultivated by inoculum A were Paracoccus sp., Devosia hwasunensi, Pseudoxanthomonas sp., while the dominant species were Lactococcus raffinolactis and Pseudomonas sp.in granules developed from inoculum B.