Screening and degrading characteristics and community structure of a high molecular weight polycyclic aromatic hydrocarbon-degrading bacterial consortium from contaminated soil

Inoculation with effcient microbes had been proved to be the most important way for the bioremediation of polluted environments. For the treatment of abandoned site of Beijing Coking Chemical Plant contaminated with high level of high-molecular-weight polycyclic aromatic hydrocarbons(HMW-PAHs) ,a bacterial consortium capable of degrading HMW-PAHs,designated 1-18-1,was enriched and screened from HMW-PAHs contaminated soil.Its degrading ability was analyzed by high performance liquid chromatography(HPLC) ,and the community structure was investigated by construction and analyses of the 16S rRNA gene clone libraries(A,B and F) at different transfers.The results indicated that 1-18-1 was able to utilize pyrene,fluoranthene and benzoapyrene as sole carbon and energy source for growth.The degradation rate of pyrene and fluoranthene reached 82.8%and 96.2%after incubation for 8 days at 30°C,respectively;while the degradation rate of benzoapyrene was only 65.1%after incubation for 28 days at 30°C. Totally,108,100 and 100 valid clones were randomly selected and sequenced from the libraries A,B,and F.Phylogenetic analyses showed that all the clones could be divided into 5 groups,Bacteroidetes,α-Proteobacteria,Actinobacteria,β-Proteobacteria andγ-Proteobacteria.Sequence similarity analyses showed total 39 operational taxonomic units(OTUs) in the libraries.The predominant bacterial groups wereα-Proteobacteria(19 OTUs,48.7%) ,γ-Proteobacteria(9 OTUs,23.1%) andβ-Proteobacteria(8 OTUs,20.5%) . During the transfer process,the proportions ofα-Proteobacteria andβ-Proteobacteria increased greatly(from 47%to 93%) ,while γ-Proteobacteria decreased from 32%(library A) to 6%(library F) ;and Bacteroidetes group disappeared in libraries B and F.     

Fungal diversity in major oil-shale mines in China

As an insufficiently utilized energy resource,oil shale is conducive to the formation of characteristic microbial communities due to its special geological origins.However,little is known about fungal diversity in oil shale.Polymerase chain reaction cloning was used to construct the fungal ribosomal deoxyribonucleic acid internal transcribed spacer(r DNA ITS)clone libraries of Huadian Mine in Jilin Province,Maoming Mine in Guangdong Province,and Fushun Mine in Liaoning Province.Pure culture and molecular identification were applied for the isolation of cultivable fungi in fresh oil shale of each mine.Results of clone libraries indicated that each mine had over 50% Ascomycota(58.4%–98.9%)and 1.1%–13.5%unidentified fungi.Fushun Mine and Huadian Mine had 5.9% and 28.1% Basidiomycota,respectively.Huadian Mine showed the highest fungal diversity,followed by Fushun Mine and Maoming Mine.Jaccard indexes showed that the similarities between any two of three fungal communities at the genus level were very low,indicating that fungi in each mine developed independently during the long geological adaptation and formed a community composition fitting the environment.In the fresh oil-shale samples of the three mines,cultivable fungal phyla were consistent with the results of clone libraries.Fifteen genera and several unidentified fungi were identified as Ascomycota and Basidiomycota using pure culture.Penicillium was the only genus found in all three mines.These findings contributed to gaining a clear understanding of current fungal resources in major oil-shale mines in China and provided useful information for relevant studies on isolation of indigenous fungi carrying functional genes from oil shale.     

硫铁填料和微电流强化再生水脱氮除磷的研究

为提高再生水质量,在不同C/N和HRT条件下,对比分析硫铁复合填料和微电流作用强化再生水深度脱氮除磷效果.结果表明,硫铁复合填料和微电流作用均能够强化氮、磷的深度去除效果,且二者结合能够使反硝化系统pH值稳定在7.2~8.5之间.系统中TN主要靠异养反硝化、氢自养反硝化和硫自养反硝化作用去除,94.04%的TP是以生成磷酸铁沉淀的形式去除.分别从填料上取生物膜,进行Miseq高通量测序,构建细菌16S rRNA基因克隆文库.结果发现,在仅有海绵铁作用系统中,同时具有异养反硝化和氢自养反硝化功能的细菌所占比例达到29.47%;硫铁复合填料和硫铁微电流作用系统中,具有硫自养反硝化功能的Thiobacillus(硫杆菌属)所占比例分别达到60.47%和40.62%.因此,硫铁复合填料和微电流作用用于强化再生水深度脱氮除磷具有明显的优势.     

三唑磷作用下翡翠贻贝抑制性消减杂交文库构建与分析

采用抑制性差减杂交技术,构建了三唑磷农药急性胁迫下翡翠贻贝雌贝性腺正反向消减杂交cDNA文库.正反向文库共有117个表达序列标签（expressed sequence tags,ESTs）与NCBI数据库中已知功能蛋白具较高同源性,假定蛋白12种,另有195个ESTs未检索到同源序列.正向文库中含有细胞周期蛋白、性激素合成蛋白、DNA修复蛋白、能量代谢相关蛋白等一系列与生殖、应激适应和代谢相关的转录本信息,而反向文库中的差异ESTs主要涉及DNA复制、转录和翻译调控基因以及骨架蛋白.结果表明三唑磷对翡翠贻贝的毒性涉及到生殖调控、能量代谢、应激适应、转录翻译调控等生理代谢的多个方面,正向文库中发现的几种生殖调控基因（细胞周期蛋白、周期蛋白依赖性蛋白激酶、合子DNA复制许可因子、精子特异性蛋白、雌激素硫酸转移酶和卵黄膜卵透明带域蛋白）可能是三唑磷生殖干扰毒性作用过程中的关键基因,这为有机磷农药的生殖毒性探索提供了基础数据.     

3DBER-S反硝化脱氮性能及其菌群特征

针对污水处理厂尾水TN去除问题,采用16S rDNA克隆文库法,探究了3DBER-S(三维电极生物膜耦合硫自养脱氮工艺)的强化脱氮机制及其菌群特征. 结果表明,I(电流)和HRT(水力停留时间)对3DEBR-S中氢自养和硫自养反硝化作用所占比例的影响较大,但对脱氮效率影响不显著. 当进水C/N〔ρ(COD_Cr)/ρ(TN)〕为1、ρ(NO₃--N)为35 mg/L、I为300 mA、HRT为4 h时,NO₃--N和TN去除率可分别稳定在80%和74%以上. 16S rDNA克隆文库结果显示,反应器中β变形菌纲为优势菌群,占47.89%〔以OUT(操作单元)计〕. 在β变形菌纲中,与具有反硝化功能的陶厄氏菌属(Thauera)相似的细菌所占比例最大,为52.94%;与可分别利用硫和氢为电子供体进行反硝化脱氮的硫杆菌属(Thiobacillus)和食酸菌属(Acidovorax)相似的细菌分别占17.65%和14.71%. 3DBER-S中存在异养联合氢自养和硫自养反硝化协同去除硝酸盐氮的作用,可为反硝化脱氮提供充足的电子供体,节约了有机碳源消耗,并保证了稳定高效的脱氮效果.      

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...     

A2/O与倒置A2/O工艺低温条件下的氨氮去除能力解析

对相同进水、平行运行的A²/O与倒置A²/O工艺的冬季氨氮（NH₄⁺-N）去除能力进行了全面解析.在运行水温为14℃时,倒置A²/O工艺表现出更低的NH₄⁺-N容积去除负荷［0.13 kg·（m³·d）^-1和0.29 kg·（m³·d）^-1］和氨氧化速率（AOR）［0.07 kg·（kg·d）^-1和0.11 kg·（kg·d）^-1］,而26℃时两个工艺则相差无几.两个平行工艺中的氨氧化菌（AOB）种群定量结果几乎始终相等（倒置A²/O工艺为3.2%±0.24%,A²/O工艺为3.4%±0.31%）.克隆文库分析的结果表明,造成低温时倒置A²/O工艺中具有较低氨氮去除能力的原因是其AOB优势种属为AOR较慢的慢生型（K-生长策略）亚硝化螺菌属（Nitrosospira）,而在A²/O工艺中则为AOR较快的快生型（r-生长策略）亚硝化单胞菌属（Nitrosomonas）;而在26℃环境下两个工艺中的优势种属则均为Nitrosomonas.结合对污染物沿程去除过程的全面分析,发现尽管温度是决定AOB优势种属演替的首要原因,但由于倒置A²/O的工艺结构变化造成其好氧单元具有较高的COD负荷和高NH₄⁺-N浓度等不利于AOB生长的因素,决定了其可以在常规城市污水的条件下出现K-生长策略型的优势AOB种属.因此倒置A²/O针对异养菌（聚磷菌与反硝化菌）的工艺构造变化却通过COD负荷等间接影响到自养型氨氧化菌的种群分布与演替,并最终造成工艺在低温条件下硝化能力的减弱.     

Seasonal variation of microbial populations and biomass in Tatachia grassland soils of Taiwan

To investigate the seasonal variations of microbial ecology in grassland of Tatachia forest, soil properties, microbial populations, microbial biomass, and 16S rDNA clone library analysis were determined. The soil had temperatures 6.6–18.4°C, pH 3.6–5.1, total organic carbon 1.11–10.68%, total nitrogen 0.18–0.78%, and C/N ratios 3.46–20.55. Each gram of dry soil contained bacteria, actinomycetes, fungi, cellulolytic, phosphate-solubilizing microbes, and nitrogen-fixing microbes 4.54 × 10⁴ to 3.79 × 10⁷, 3.43 × 10² to 2.17 × 10⁵, 5.74 × 10³ to 3.76 × 10⁶, 1.97 × 10³ to 1.34 × 10⁶, 8.49 × 10² to 5.59 × 10⁵, and 3.86 × 10² to 3.75 × 10⁵ CFU, respectively. Each gram of soil contained 117–2,482 μg of microbial biomass carbon, 23–216 μg of microbial biomass nitrogen and 9–29 μg of DNA. The microbial populations, microbial biomass, and DNA decreased stepwise with the depth of soil, and they had low values in winter seasons. The microbial populations, microbial biomass carbon, microbial biomass nitrogen, and DNA at the BW2 horizon were 8.42–17.84, 19.26–64.40, 16.84–61.11, and 31.03–46.26% of those at the O horizon, respectively. When analyzing 16S rDNA library, members of Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Firmicutes, candidate division TM1, candidate division TM7, Gammatimonadetes, and Verrucomicrobia were identified. Members of Proteobacteria (44.4%) and Acidobacteria (33.3%) dominated the clone libraries. Within the phylum Proteobacteria, α-, β-, and γ-Proteobacteria were most numerous, followed by δ-Proteobacteria.     

Analytical solutions of three-dimensional contaminant transport in uniform flow field in porous media: A library

The purpose of this study is to present a library of analytical solutions for the three-dimensional contaminant transport in uniform flow field in porous media with the first-order decay, linear sorption, and zero-order production. The library is constructed using Green’s function method (GFM) in combination with available solutions. The library covers a wide range of solutions for various conditions. The aquifer can be vertically finite, semi-infinitive or infinitive, and laterally semi-infinitive or infinitive. The geometry of the sources can be of point, line, plane or volumetric body; and the source release can be continuous, instantaneous, or by following a given function over time. Dimensionless forms of the solutions are also proposed. A computer code FlowCAS is developed to calculate the solutions. Calculated results demonstrate the correctness of the presented solutions. The library is widely applicable to solve contaminant transport problems of one- or multiple- dimensions in uniform flow fields.