基于Miseq的好氧反硝化菌源水脱氮的种群演变

为了研究好氧反硝化菌源水脱氮过程中水体微生物群落的演变,利用Miseq高通量测序法对投菌和对照两系统水体样本的微生物信息进行统计,并对两组样品进行了优化序列统计,OTU分布统计和分类学分析的基础分析;以及细菌群落结构,PCA,Rank-Abundance,Hcluster,Specaccum和OTU分布的高级分析.结果显示,投加贫营养好氧反硝化菌的源水系统的氮素得到有效去除,脱氮效果明显;层次聚类和主成分分析显示两系统内的群落结构发生变化,投菌系统与对照系统主要表现为变形菌和拟杆菌门;细菌主要门类和水质参数的相关性分析得出,水质指标对两系统群落变化作用明显;与此同时,投菌系统中有关氮循环的细菌有上升的变化过程. Miseq高通量测序研究源水脱氮过程的微生物种群演变可行,为研究原位生物脱氮过程的水体微生物群落演变提供技术支撑.     

Impact of heavy metals on bacterial communities from mangrove soils of the Mahanadi Delta (India)

This study aimed to assess soil nutrient status and heavy metal content and their impact on the predominant soil bacterial communities of mangroves of the Mahanadi Delta. Mangrove soil of the Mahanadi Delta is slightly acidic and the levels of soil nutrients such as carbon, nitrogen, phosphorous and potash vary with season and site. The seasonal average concentrations (μg/g) of various heavy metals were in the range: 14 810–63 370 (Fe), 2.8–32.6 (Cu), 13.4–55.7 (Ni), 1.8–7.9 (Cd), 16.6–54.7 (Pb), 24.4–132.5 (Zn) and 13.3–48.2 (Co). Among the different heavy metals analysed, Co, Cu and Cd were above their permissible limits, as prescribed by Indian Standards (Co=17 μg/g, Cu=30 μ g/g, Cd=3–6 μ g/g), indicating pollution in the mangrove soil. A viable plate count revealed the presence of different groups of bacteria in the mangrove soil, i.e. heterotrophs, free-living N₂ fixers, nitrifyers, denitrifyers, phosphate solubilisers, cellulose degraders and sulfur oxidisers. Principal component analysis performed using multivariate statistical methods showed a positive relationship between soil nutrients and microbial load. Whereas metal content such as Cu, Co and Ni showed a negative impact on some of the studied soil bacteria.     

低温驯化对自养脱氮颗粒污泥功能活性与菌群结构的影响分析

低温驯化是提高部分亚硝化-厌氧氨氧化（PN/A）组合工艺低温脱氮效能和运行稳定性的有效方法.为探究低温驯化对污泥特性的具体影响,本文从温度敏感性、颗粒外观形态、胞外聚合物（EPS）组成和微生物群落结构等方面,对中高温培养（30℃）和低温驯化（15℃）PN/A颗粒污泥之间的差异性进行了分析.反应热力学的研究结果表明,驯化污泥（G_L）在低温区（10~20℃）的脱氮性能较中高温培养污泥（G_H）有了显著提高,总无机氮去除的表观反应活化能（E_a）降低了28.4%.与G_H相比,G_L的平均粒径减小了25.8%,EPS含量增长了16.6%,颗粒的沉降性能明显下降.由高通量测序结果可知,G_L具有更高的菌群多样性,同时,污泥中好氧氨氧化菌（Nitrosomonas）与厌氧氨氧化菌（Candidatus_Kuenenia）的丰度比值（0.04）远小于G_H的0.34.这意味着颗粒污泥在低温环境中对慢速生长自养菌仍具有较强的截留能力.上述发现为解析PN/A污泥在低温条件下的自适应机制,推动组合工艺的工程化应用提供了重要参考.     

钻井废弃液中石油污染物及细菌生态多样性

采用GC/MS分析方法,发现平湖油田钻井废弃液中含有烃类、酸类和酯类等23种有机污染物质,含量达到49 800 mg/L。运用16S rRNA基因分析法测得的微生物生态多样性表明,废弃液中微生物种类丰富,包括γ和β变形杆菌,γ-Proteobacteria是优势菌群,占克隆总数78.6%。Oceanospirillales、Pseudomonadales和Enterobacteriales是γ-Proteobacteria纲的主要菌群。     

Development of pretreatment protocol for DNA extraction from biofilm attached to biologic activated carbon (BAC) granules

The biologic activated carbon (BAC) process is widely used in drinking water treatments. A comprehensive molecular analysis of the microbial community structure provides very helpful data to improve the reactor performance. However, the bottleneck of deoxyribonucleic acid (DNA) extraction from BAC attached biofilm has to be solved since the conventional procedure was unsuccessful due to firm biomass attachment and adsorption capacity of the BAC granules. In this study, five pretreatments were compared, and adding skim milk followed by ultrasonic vibration was proven to be the optimal choice. This protocol was further tested using the vertical BAC samples from the full-scale biofilter of Pinghu Water Plant. The results showed the DNA yielded a range of 40 μg·g^-1 BAC (dry weight) to over 100 μg·g^-1 BAC (dry weight), which were consistent with the biomass distribution. All results suggested that the final protocol could produce qualified genomic DNA as a template from the BAC filter for downstream molecular biology researches.     

木麻黄根区土壤理化特性及酶活性与青枯病发生关联分析

为探究木麻黄(Casuarina spp.)根区土壤理化特性及酶活性与青枯病发生的关系,进而为加强生物防治提供参考.选取位于广东省潮州市饶平县大埕镇、湛江市吴川市塘尾镇和湛江市徐闻县前山镇的3个木麻黄人工林为研究对象,在每个林分下分别设置3个样地,每个样地采集木麻黄青枯病感病植株和健康植株根区土壤各3份,共54个土壤样...     

Bacterial diversity in activated sludge from a consecutively aerated submerged membrane bioreactor treating domestic wastewater

The bacterial diversity of activated sludge from submerged membrane bioreactor (SMBR) was investigated. A 16S rDNA clone library was generated, and 150 clones were screened using restriction fragment length polymorphism (RFLP). Of the screened clones, almost full-length 16S rDNA sequences of 64 clones were sequenced. Phylogenetic tree was constructed with a database containing clone sequences from this study and bacterial rDNA sequences from NCBI for identification purposes. The 90.6% of the clones were a?l...     

农村黑臭水体沉积物细菌群落结构特征

沉积物微生物是农村黑臭水体物质循环和有机物降解等过程的主要驱动者,其群落结构组成常因外界环境的微小变化而变化.以菏泽市东明县29个农村黑臭水体为研究对象,测定农村黑臭水体上覆水和沉积物氮、磷及重金属等污染物指标,结合Illumina测序结果,分析农村黑臭水体沉积物细菌群落组成和多样性特征及其与环境因子的相关性.结果表明,该区域农村黑臭水体上覆水和沉积物中污染物分布范围均较广.与农业面源相比,以农村生活污水为主要污染来源的农村黑臭水体上覆水中氮、磷污染物浓度更高,分别是农业面源的3.1倍和1.5倍.此外,该区域农村黑臭水体沉积物中重金属含量处于较低水平,普遍低于菏泽市土壤元素背景值.该区域农村黑臭水体沉积物细菌群落的优势菌门为变形菌门、放线菌门、绿弯菌门、厚壁菌门和酸杆菌门,这5种优势菌门的序列总和占全部序列的70.3%～83.6%.γ-变形菌纲、α-变形菌纲、厌氧绳菌纲和放线菌纲是沉积物细菌群落的优势菌纲;硫杆菌属和假节杆菌属是其优势菌属.Spearman相关性分析结果表明,环境因子中DO、 COD、 TN、 TP和有机质对农村黑臭水体沉积物细菌菌属有显著影响(P<0.05),沉...     

岩溶湿地与稻田土壤团聚体细菌群落结构的比较

研究湿地系统中的土地利用变化对土壤团聚体中细菌群落结构的影响,可以为阐明土地利用变化对湿地土壤团聚体稳定性的影响机制提供理论依据.以会仙岩溶湿地和稻田为研究对象,利用高通量测序技术和生态网络分析对土壤团聚体（大团聚体0.25~2 mm、微团聚体0.053~0.25 mm和粉、黏粒<0.053 mm）细菌群落结构进行研究.结果表明：①微团聚体在稻田中占29.64%,显著高于其在天然湿地中的占比（22.20%）.②门水平上,绿弯菌门在稻田土壤大团聚体、微团聚体和粉、黏粒中的相对丰度分别为7.97%、8.56%和7.40%,显著高于其在天然湿地这3种粒径团聚体中的相对丰度（4.93%、4.81%和3.76%）;目水平上,厌氧绳菌目在稻田这3种粒径团聚体中的相对丰度分别为2.35%、3.03%和2.65%,显著高于其在天然湿地这3种粒径团聚体中的相对丰度（0.92%、0.91%和0.43%）.③生态网络分析显示,稻田微团聚体和粉、黏粒网络的节点数、边数和平均邻居数均高于天然湿地,并且在这两种组分中网络的平均连通度更大,特征路径长度更短;稻田大团聚体网络具有更长的特征路径长度、更高的模块性.结果表明,天然湿地改变为稻田后,土壤团聚体细菌群落结构改变,稻田微团聚体和粉、黏粒中细菌群落具有更高的物质循环和信息传递的效率,而稻田大团聚体细菌网络紧密性低,模块性高,其细菌群落结构更稳定.     

宿根甘蔗根际土壤细菌多样性分析中培养法与非培养法比较研究

采用培养法和非培养法提取甘蔗根际土壤微生物的总DNA,基于通用引物PCR扩增构建两种方法的细菌16SrDNA文库,并进行核糖体DNA扩增片段限制性内切酶分析(ARDRA),通过部分克隆序列测定构建细菌克隆文库的系统发育树,进而对基于传统的细菌平板培养法和直接提取总细菌DNA的非培养法进行比较分析.结果显示,非培养法文库的香侬–威纳指数、辛普森指数、丰富度分别为4.94、0.998、28.91,均高于培养法文库中相应的多样性参数(分别为4.27、0.996、16.79),均一度数值都>0.95.结合统计学数据和序列测定信息,反映出甘蔗根际土壤存在着丰富的细菌多样性,但平板培养法所展现的多样性低于直接提取法,表明前者存在着很大的局限性,而非培养法文库中主要是未培养微生物的16S rDNA序列,这从一个侧面反映了土壤样品中未培养微生物占很大比例.因此,在土壤微生物群落研究中,必须结合新的分子生态学技术手段,如采用直接提取总DNA的方法进行研究,才能比较全面地认识土壤微生物多样性.图3表3参18