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Microbiome analysis and-omics studies of microbial denitrification processes in wastewater treatment: recent advances

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Science China(Life Sciences) 2018年第7期61卷 753-761页

作者： Lili Miao zhipei liu State Key Laboratory of Microbial Resources Institute of Microbiology Chinese Academy of Sciences

Nitrogen pollution is an increasingly severe worldwide problem because of drainage of nitrogen-containing wastewater and intensive application of nitrogen-containing fertilizers. Denitrification, a key process in nitrogen cycles, is commonly employed for nitrogen removal in engineered wastewater treatment systems. Biological denitrification is performed by denitrifying microbes(bacteria) that use nitrate as terminal electron acceptor. Better understanding the functions of diverse microbial populations in denitrification-based wastewater treatment systems, and the interactions of these populations with operating environments, is essential for improving both treatment performance and system stability. Recent advances in "meta-omics"(e.g., genomics, transcriptomics, proteomics, metabolomics), other molecular biology tools, and microbiome analysis have greatly enhanced such understanding. This minireview summarizes recent findings regarding microbial community structure and composition, key functional microbes and their physiology, functional genes involved in nitrogen cycle, and responses of microbes and their genes to changes of environmental factors or operating parameters, in denitrification processes in wastewater treatment systems. Of particular interest are heterotrophic denitrification systems(which require alternative organic carbon sources) and the autotrophic denitrification systems(which do not require an external carbon source). Integrated microbiome and-omics approaches have great future potential for determination of optimal environmental and biotechnological parameters,novel process development, and improvement of nitrogen removal efficiency and system stability.

关键词： wastewater treatment nitrogen removal denitrification microbiome analysis

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Preliminary investigation on the role of microorganisms in the production of phosphine

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Journal of Environmental Sciences 2008年第7期20卷 885-890页

作者： liu zhipei JIA Shengfen WANG Baojun Institute of Microbiology Chinese Academy of SciencesBeijing 100101China

The relationships between the phosphine content and various microbial populations,activities of different enzymes were investigated *** results indicated that the phosphine content of samples from various environments was positively related to total anaerobic microorganisms,organic phosphate compound-dissolving bacteria,denitrifying bacteria,and the activities of alkaline phosphatase and dehydrogenase,with correlation coefficients (R^2) up to 0.93,0.90,0.69,0.79,and 0.82,*** also sh...

关键词： phosphane content microbial populations enzymatic activities sulfate reducing bacteria denitrifying bacteria

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A bioaugmentation failure caused by phage infection and weak biofilm formation ability

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Journal of Environmental Sciences 2009年第8期21卷 1153-1161页

作者： FU Songzhe FAN Hongxia liu Shuangjiang liu Ying liu zhipei State Key Laboratory of Microbial Resources Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China Graduate University of Chinese Academy of Sciences Beijing 100039 China Institute of Oceanology Chinese Academy of Sciences Qingdao 266701 China

Two biological aerated filters （BAF） were setup for ammonia removal treatment of the circulation water in a marine aquaculture. One of the BAFs was bioaugmented with a heterotrophic nitrifying bacterium, Lutimonas sp. H10, where the ammonia removal was not improved and the massive inoculation was even followed by a nitrification breakdown from day 9 to 18. The nitrification was remained stable in control BAF operated under the same conditions. Fluorescent in situ hybridization （FISH） with rRNA-targeted probes and cultivable method revealed that Lutimonas sp. H10 almost disappeared from the bioaugomented BAF within 3 d, and this was mainly due to the infection of a specific phage as revealed by flask experiment, plaque assay and transmission electron observation. Analyses of 16S rRNA gene libraries showed that bacterial groups from two reactors evolved differently and an overgrowth of protozoa was observed in the bioaugmented BAF. Therefore, phage infection and poor biofilm forming ability of the inoculated strain are the main reasons for bioaugmentation failure. In addition, grazing by protozoa of the bacteria might be the reason for the nitrification breakdown in bioaugmented BAF during day 9-18.

关键词： marine culture water circulation ammonia removal Lutimonas sp. HIO nitrification breakdown fluorescent in situhybridization (FISH)

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

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Journal of Environmental Sciences 2010年第10期22卷 1576-1585页

作者： Run Sun Jinghua Jin Guangdong Sun Ying liu zhipei liu State Key Laboratory of Microbial Resources Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China. Environmental Protection Research Institute of Light Industry Beijing 100089 China

Inoculation with efficient 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 benzo[a]pyrene 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℃, respectively; while the degradation rate of benzo[a]pyrene was only 65.1% after incubation for 28 days at 30℃. Totally, 108, 100 and 100 valid clones were randomly selected and sequenced from the libraries A, B, and E Phylogenetic analyses showed that all the clones could be divided into 5 groups, Bacteroidetes, ct-Proteobacteria, Actinobacteria, β-Proteobacteda and γ- Proteobacteria. Sequence similarity analyses showed total 39 operational taxonomic units （OTUs） in the libraries. The predominant bacterial groups were α-Proteobacteria （19 OTUs, 48.7%）, γ-Proteobacteria （90TUs, 23.1%） and β-Proteobacteria （80TUs, 20.5%）. During the transfer process, the proportions of α-Proteobacteria and β-Proteobacteria increased greatly （from 47% to 93%）, while γ-Proteobacteda decreased from 32% （library A） to 6% （library F）; and Bacteroidetes group disappeared in libraries B and F.

关键词： high-molecular-weight-PAHs bacterial consortium bacterial community structure 16S rRNA gene library

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A novel and complete gene cluster involved in the degradation of aniline by Delftia sp. AN3

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Journal of Environmental Sciences 2008年第6期20卷 717-724页

作者： ZHANG Tao ZHANG Jinglei liu Shuangjiang liu zhipei Laboratory of Environmental Microbiology Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China Graduate University of Chinese Academy of Sciences Beijing 100039 China

A recombinant strain, Escherichia coli JM109-AN1, was obtained by constructing of a genomic library of the total DNA of Delftia sp. AN3 in E. coli JM109 and screening for catechol 2,3-dioxygenase activity. This recombinant strain could grow on aniline as sole carbon, nitrogen and energy source. Enzymatic assays revealed that the exogenous genes including aniline dioxygenase （AD） and catechol 2,3-dioxygenase （C230） genes could well express in the recombinant strain with the activities of AD and C230 up to 0.31 U/mg wet cell and 1.92 U/mg crude proteins, respectively. The AD or C23O of strain AN3 could only catalyze aniline or catechol but not any other substituted substrates. This recombinant strain contained a recombinant plasmid, pKC505-AN1, in which a 29.7-kb DNA fragment from Delftia sp. AN3 was inserted. Sequencing and open reading frame （orfs） analysis of this 29.7 kb fragment revealed that it contained at least 27 orfs, among them a gene cluster （consisting of at least 16 genes, named danQTA1A2BRDCEFG1HIJKG2） was responsible for the complete metabolism of aniline to TCA-cycle intermediates. This gene cluster could be divided into two main parts, the upper sequences consisted of 7 genes （danQTA1A2BRD） were predicted to encode a multi-component aniline dioxygenase and a LysR-type regulator, and the central genes （danCEFG1HIJKG2） were expected to encode meta-cleavage pathway enzymes for catechol degradation to TCA-cycle intermediates. Unlike clusters tad from Delftia tsuruhatensis AD9 and tdn from Pseudomonas putida UCC22, in this gene cluster, all the genes were in the same transcriptional direction. There was only one set of C230 gene （danC） and ferredoxin-like protein gene （danD）. The presence of only one set of these two genes and specificity of AD and C230 might be the reason for strain AN3 could only degrade aniline. The products of danQTA1A2BRDC showed 99%-100% identity to those from Delftia acidovorans 7N, and 50%-85% identity to those of tad cluster from

关键词： aniline biodegradation Delftia sp. AN3 genomic library aniline degradative gene cluster

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Enhanced biological nutrient removal by the alliance of a heterotrophic nitrifying strain with a nitrogen removing ecosystem

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Journal of Environmental Sciences 2008年第2期20卷 216-223页

作者： Naeem ud din Ahmad XU Haiying CHEN Liping liu zhipei liu Shuangjiang State Key Laboratory of Microbial Resources Institute of Microbiology Chinese Academy of Sciences Beijing 100080 China Graduate School of Chinese Academy of Sciences Beijing 100049 China

Nitrogen removal from synthetic wastewater was investigated in an airlift bioreactor （ALB）, augmented with a novel heterotrophic nitrifier Pseudonocardia ammonioxydans H9^T under organic carbon to nitrogen ratios （Corg/N） ranging from 0 to 12. Effect of the inoculated strain was also determined on the settling properties and the removal of chemical oxygen demand （COD）. Two laboratory scale reactors were set up to achieve a stable nitrifying state under the same physicochemical conditions of hydraulic retention time （HRT）, temperature, pH and dissolved oxygen （DO）, and operated under the sequencing batch mode. The level of DO was kept at 0.5- 1.5 mg/L by periodic stirring and aeration. Each specific Corg/N ratio was continued for duration of 3 weeks. One of the reactors （BR2） was inoculated with P ammonioxydans H9^T periodically at the start of each Corg/N ratio. Sludge volumetric index （SVI） improved with the increasing Corg/N ratio, but no significant difference was detected between the two reactors. BR2 showed higher levels of nitrogen removal with the increasing heterotrophic conditions, and the ammonia removal reached to the level of 82%-88%, up to10% higher than that in the control reactor （BR1） at Corg/N ratios higher than 6; however, the ammonia removal level in experimental reactor was up to 8% lower than that in control reactor at Corg/N ratios lower than 2. The COD removal efficiency progressively increased with the increasing Corg/N ratios in both of the reactors. The COD removal percentage up to peak values of 88%-94% in BR2, up to 11% higher than that in BR1 at Corg/N ratio higher than 4. The peak values of ammonia and COD removal almost coincided with the highest number （18%-27% to total bacterial number） of the exogenous bacterium in the BR2, detected as colony forming units （CFU）. Furthermore, the removal of ammonia and COD in BR2 was closely related to the number of the inoculated strain with a coefficient index （R2） up to 0.82 and 0.85 for ammonia

关键词： air-lift bioreactor (ALB) bioaugrnentation Corg/N ratio COD and ammonia removal sequencing batch mode heterotrophic nitrifier Pseudonocardia ammonioxydans H9^T

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Denitrification characteristics of a marine origin psychrophilic aerobic denitrifying bacterium

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Journal of Environmental Sciences 2011年第11期23卷 1888-1893页

作者： Haiyan Zheng Ying liu Guangdong Sun Xiyan Gao Qingling Zhang zhipei liu State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China.

A psychrophilic aerobic denitrifying bacterium,strain S1-1,was isolated from a biological aerated filter conducted for treatment of recirculating water in a marine aquaculture *** S1-1 was preliminarily identified as Psychrobacter *** on the analysis of its 16S rRNA gene sequence,which showed 100% sequence similarity to that of Psychrobacter *** S1-1 grew well either in high nitrate or high nitrite conditions with a removal of 100% nitrate or 63.50% nitrite,and the total nitrogen removal rates could reach to 46.48% and 31.89%,*** results indicated that nitrate was mainly reduced in its logarithmic growth phase with a very low leve1 accumulation of nitrite,suggesting that the aerobic denitrification process of strain S1-1 occurred mainly in this *** GC-MS results showed that N 2 O was formed as the major intermediate during the aerobic denitrifying process of strain ***,factors affecting the growth of strain S1-1 and its aerobic denitrifying ability were also *** showed that the optimum aerobic denitrification conditions for strain S1-1 were sodium succinate as carbon source,C/N ratio15,salinity 10 g/L NaCl,incubation temperature 20°C and initial pH 6.5.

关键词： nitrogen removal aerobic denitrification marine environment low temperature Psychrobacter sp.S1-1

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Isolation and characterization of a bacterial strain that efficiently degrades sex steroid hormones

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Frontiers of Environmental Science & Engineering 2007年第3期1卷 325-328页

作者： JI Shulan liu zhipei liu Zhipeng REN Haiyan College of Environmental and Energy Engineering Beijing University of TechnologyBeijing 100022China Institute of Microbiology Chinese Academy of SciencesBeijing 100101China

A bacterial strain,ZY3,growing on sex steroid hormones as the sole source of carbon and energy was isolated from the sewage treatment plant of a prophylactic steroids ***3 degrades the 3-methoxy-17b-hydroxy-1,3,5(10),8(9)-d-4-estren(MHE).This strain was preli-minarily identified as Raoultella ***3 according to its morphology and its 16S rRNA gene *** the experimental period(72 h),the optimum temperature,pH and 3-MHE concentration for the degradation of hydride by the strain ZY3 were 35°C,10 and 10 mg/L,*** degradation rate of the sex steroid hormones increased to 87%and 85% after the addition of maltose and peptone,respectively.

关键词： sex steroid hormones 3-methoxy-17b-hydroxy-1,3,5(10) 8(9)-d-4-estren(MHE) biodegradation ZY3 strain

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1961-2020年赣江流域月尺度实际蒸发模拟及归因分析

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水土保持研究 2025年第1期32卷 195-202页

作者：姬广兴刘智培高红凯陈伟强黄珺嫦张亚丽郭宇龙陈轶楠河南农业大学资源与环境学院华东师范大学地理科学学院

[目的]探究赣江流域月时间尺度实际蒸发的变化规律和分配特征以及不同因素对实际蒸发量的贡献量，为生态环境保护和水资源高效利用提供理论依据。[方法]基于赣江流域外洲水文站1961—2020年径流量数据，结合Mann-Kendall突变检验法和Pet... 详细信息

[目的]探究赣江流域月时间尺度实际蒸发的变化规律和分配特征以及不同因素对实际蒸发量的贡献量，为生态环境保护和水资源高效利用提供理论依据。[方法]基于赣江流域外洲水文站1961—2020年径流量数据，结合Mann-Kendall突变检验法和Pettitt突变点检验法确定径流突变年份。其次利用ABCD水文模拟赣江流域基准期和突变期径流变化过程，计算得出月尺度实际蒸发量。然后利用去趋势预置白趋势检验法(TFPW-MK)分析实际蒸发的变化趋势和年内分布特征。最后采用月尺度Budyko模型对实际蒸发量变化进行归因分析。[结果](1)径流突变年份为1991年。(2)赣江流域2月、3月、4月、6月、9月的实际蒸发量呈现上升趋势，其余月份实际蒸发为下降趋势。其中2月、3月的实际蒸发量显著上升(p<0.05)。(3)赣江流域月尺度实际蒸发量呈现“单峰型”分布，在7月附近达到最大值。(4)气候变化会导致各月实际蒸发量的增加，人类活动会导致各月实际蒸发量减少。[结论]赣江流域实际蒸发量呈现先增大后减少的趋势，气候变化对实际蒸发量的贡献表现为正作用。人类活动对实际蒸发量变化表现为负作用。

关键词：实际蒸发量变化气候变化人类活动 ABCD模型月尺度Budyko模型

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Decolorization and bio degradation metabolism of azo dyes by Pseudomonas S-42

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Journal of Environmental Sciences 1991年第2期3卷 89-102页

作者： liu zhipei and Yang HaifangInstitute of Microbiology,Academis Sinica,Beijing 100080,China

A bacterial strain was isolated from activated sludge and has been identified as Pseudomonas sp. S-42 capable of decolorizing azo dyes such as Diamira Brilliant Orange RR (DBO-RR), Direct Brown M (DBM), Eriochrome Brown R (EBR) and so on. The growing cells, intact cells, cell-free extract and purified enzyme of strain S-42 could decolorize azo dyes under similar conditions at the optimum pH 7.0 and temperature of 37℃. The efficiencies of decolorization for DBO-RR, DBM, EBR with intact cells stood more than 90%. When the cell concentration was 15mg (wet)/ml and the reaction time was 5 hours, the decolorizing activities of intact cells for above three azo dyes were 1.75, 2.4, 0.95 μg dye/mg cell, respectively. Cell-free extract and purified enzyme belonged to azoreductase with molecular weight about 34000±2000 and Vmax and Km values for DBO-RR of 13μmol/mg protein/nun and 54μmol, respectively. The results from the detection of the biodegradation products of DBO-RR by spectrophotometric and NaNO2 reaction methods showed that the biodegradation of azo dyes was initiated by the reducing cleavage of azo bonds. The biodegradation metabolism path for DBO-RR by Psued. S-42 was hypothesized.

关键词： Pseudomonas sp. S-42 azo dyes decolorization biodegradation azore-ductase.

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