16SrDNA克隆文库分析含硫污水生物反应器中生物群落

利用生物脱硫技术处理炼油厂含硫污水,硫化物选择性地氧化成单质硫。在进水硫化物605.3 mg·L-1的条件下,硫化物去除率90.7%,单质硫生成率达到80.8%。提取反应器中的DNA样本,构建16S rDNA克隆文库对反应器中的生物群落进行分析。对51个阳性克隆子(长度约为1 400 bp)进行测序,测序结果在NCBI数据库利用Blast比对并构建系统发育树。测序结果表明,生物脱硫反应器细菌种群可分为10个类群,数量最多的是γ-变形菌(γ-Proteobacteria)占76.6%,其余类群共占23.4%。γ-Proteobacteria类群中数量最多的是具有脱硫功能的Halothiobacillus sp.(66.7%)、Thiovirga sp.(25.6%)以及Pseudomonas sp.(7.7%)。系统发育树分析结果表明,生物脱硫系统中形成了以化能自养型脱硫菌为主的微生物群落结构,异养菌数量较少。     

土霉素生产废水生物处理装置中微生物群落结构特征解析

抗生素对细菌具有强抑制作用,从而会影响废水生物处理系统中的微生物群落结构。在用定量PCR方法对土霉素生产废水处理装置(进水中土霉素浓度为1 662.1±248.6μg/L)中的细菌16S rRNA基因和真菌18S rRNA基因的含量进行比较的基础上,利用16S rRNA基因克隆文库方法对污泥中的细菌群落结构进行了详细解析。定量PCR结果显示,土霉素废水活性污泥中真菌(18S rRNA)/细菌(16S rRNA)基因的拷贝数比例高达1.2,明显较高于非抗生素肌苷废水活性污泥中的比例1.52×10-6,表明真菌对于土霉素废水中有机物的去除可能发挥重要作用。细菌克隆文库分析结果显示,Alpha变形菌和Beta变形菌是主要的优势菌,比例分别为23.7%和22.0%,其次是酸杆菌(17.0%)和拟杆菌(11.9%)。     

复合生物膜-活性污泥反应器同步脱氮除磷

通过实验研究比较了复合生物膜-活性污泥反应器(HY)和传统活性污泥反应器(AS)的脱氮除磷效果。结果表明,在水力停留时间(HRT)16 h、污泥龄12～15 d、水温19～21℃、pH 6.3～7.8的条件下,复合生物反应器比活性污泥反应器运行更稳定,未发生污泥膨胀。在相同运行条件下,复合生物反应器对COD、TN和TP的去除率分别为95%、91%和98%,而活性污泥反应器对COD、TN和TP的去除率分别为85%、84%和90%。稳定工况下复合生物反应器的比硝化、比反硝化速率,比吸磷、比释磷速率均高于活性污泥反应器,且微生物相更加丰富。通过建立16S rDNA克隆文库发现生物膜和活性污泥的微生物群落结构均具有高度多样性,但生物膜微生物的微生物相比活性污泥更复杂。     

自然生境中氨氧化细菌的分子生物学研究进展

摘要 硝化过程是自然界氨氮污染降解的主要生化机制,氨氧化细菌对于富营养的河流、海洋及土壤环境等自然生境具有重要作用.氨氧化细菌的群落结构很复杂,且在不同研究区的时空上存在很大差异性.随着分子生物技术的发展,各种自然环境中存在的氨氧化细菌得到了研究.目前,中国对氨氧化细菌的研究多集中于污水处理、土壤等方面,采用非培养方法对自然环境中氨氧化细菌的研究较少.首先介绍了氨氧化细菌的发现过程和生理特性,其次论述了不同的分子生物学方法对土壤和水生生态环境等自然生境中氨氧化细菌研究进展和研究方法,最后提出了自然生境中氨氧化细菌研究的问题和展望.     

污泥施用对根际和非根际石灰性土壤中细菌多样性的影响

为研究污泥施用对石灰性土壤细菌多样性的影响,在连续2年种植夏玉米大田系统中,施用不同量的腐熟污泥,采集根际和非根际土壤进行16S rDNA-V3-V4区高通量测序,分析污泥施用对石灰性土壤细菌丰富度、多样性指数、群落结构、功能基因在代谢途径的影响,同时对细菌群落和环境因素之间的相关性进行分析。结果表明：污泥用量分别在3.5～37.5 t·hm⁻²和3.5～7.5 t·hm⁻²有利于非根际和根际土壤OTU数量增加;但污泥在75 t·hm⁻²过量施用时,根际和非根际土壤细菌的Shannon多样性指数显著降低,Simpson指数显著增加,Ace,Chao丰富度指数显著降低。主成分和门水平物种丰度分析表明：污泥施用量不同会造成根际和非根际细菌群落结构差异,并且当污泥施用量为75 t·hm⁻²时,明显降低了非根际土壤放线菌门的丰度;污泥施用量在3.75～37.5 t·hm⁻²时,非根际土壤中拟杆菌门丰度会明显增加。与对照相比,当污泥用量为75 t·hm⁻²时,根际土壤拟杆菌门有明显的增加,增加丰度达1.45倍,但酸杆菌门和放线菌门丰度降低了49.74%和80.57%。冗余分析(RDA)表明,土壤TN、Cd、Cu、TP是影响土壤细菌群落最主要的因素,其中TN、Cd、Cu和细菌Shannon、Simpson多样性指数呈现显著相关性(P< 0.05)。由此可见,连续过量施入污泥会对根际和非根际石灰性土壤中细菌多样性造成不利影响,而且上述微生物多样性变化可作为污泥合理施用的判断依据。     

PCR-DGGE研究亚硝化-电化学生物反硝化全自养脱氮工艺细菌多样性

采用分子生物学手段PCR-DGGE技术对亚硝化-电化学生物反硝化全自养脱氮工艺细菌的多样性进行了研究。结果表明,亚硝化段内主要的细菌种群为相似于Nitrosomonas sp.(AJ224410)和Nitrosomonas sp.NM41(AF272421)的种群,相似性分别为97%和94%;电化学生物反硝化段细菌类群主要有β-proteobacteria类群、γ-proteobacteria类群和Chlo-roflexi类群。填料上生物膜细菌种群较底部泥水混合物丰富,两者细菌种群相似性为75%;底部泥水混合物样中存在与厌氧氨氧化菌Brocadia anammoxidans(AF375994)相似性为93%的菌种,而填料上生物膜中存在与Thioalkalivibrio sp.K90mix(EU709865)和Thiobacillus thioparus(AJ243144)相似性分别为94%、97%的菌种,其中Thiobacillus thioparus(AJ243144)是典型的硫自养反硝化菌,表明填料上生物膜中有大量的硫自养反硝化菌。     

滇池底泥微生物菌群对微囊藻毒素的生物降解

采用滇池水华蓝藻中提取提纯的微囊藻毒素(microcystins,MCs)作为微生物生长的碳源和氮源,从长期暴露于蓝藻水华的滇池底泥中,通过从含低浓度到高浓度MCs的逐步培养驯化,获得了高效降解MCs的微生物混合菌群,在初始MC-RR和LR浓度大约分别为50 mg/L和30 mg/L下,3 d内可将MCs全部降解.进一步活性研究显示,不同含碳和含氮化合物虽然能够促进混合微生物菌群的生长,但对降解MCs却无明显的促进作用,说明MCs既可以作为微生物生长的碳源,又可以作为微生物生长的氮源,在富含有机物的天然水体中并不一定能够促进微生物对MCs的生物降解.     

滇池底泥微生物菌群对微囊藻毒素的生物降解

采用滇池水华蓝藻中提取提纯的微囊藻毒素（microcystins，MCs）作为微生物生长的碳源和氮源，从长期暴露于蓝藻水华的滇池底泥中，通过从含低浓度到高浓度MCs的逐步培养驯化，获得了高效降解MCs的微生物混合菌群，在初始MC-RR和LR浓度大约分别为50mg／L和30mg／L下，3d内可将MCs全部降解。进一步活性研究显示，不同含碳和含氮化合物虽然能够促进混合微生物菌群的生长，但对降解MCs却无明显的促进作用，说明MCs既可以作为微生物生长的碳源，又可以作为微生物生长的氮源，在富含有机物的天然水体中并不一定能够促进微生物对MCs的生物降解。     

Respiration induced by substrate and bacteria diversity after application of diuron,hexazinone, and sulfometuron-methyl alone and in mixture

Abstract

After application, herbicides often reach the soil and affect non-target soil microorganisms, decreasing their population, diversity or affecting metabolic activity. Therefore, laboratory studies were performed to evaluate the effects of diuron, hexazinone and sulfometuron-methyl alone and mixed upon carbon transformation by soil microorganisms in clayey and sandy soils and the effect on bacterial diversity and structure. Control treatment without herbicide application was also performed. Sub-samples from the control and herbicide treatments (10?g – in triplicate) were collected before herbicide application and 7, 14, 28 and 42?days after treatment (DAT), then 1?mL of ¹⁴C-glucose solution was applied. The released ¹⁴CO₂ was trapped in 2?M NaOH solution and the radioactivity was analyzed by liquid scintillation counting (LSC), 12?h after glucose application. The effect of herbicides on bacterial diversity was evaluated by T-RFLP. The experiment was conducted in a complete randomized design. Hexazinone did not affect ¹⁴CO₂ evolution. Diuron showed a greater ¹⁴CO₂ evolution in sandy and clayey soil, while sulfometuron-methyl led to an increase in sandy soil, at 42 DAT. A greater evolution of carbon was observed in the treatment with herbicide mixture in sandy soil, compared with the same treatment in clayey soil or control. However, the herbicide mixture application did not affect the soil biological activity measured by the respiration rate induced by substrate. On the other hand, the herbicide mixtures affected the bacterial diversity in both soils, being the strongest effect to diuron and sulfometuron-methyl in clayey soil and hexazinone in sandy soil.     

Reduction of selenite to red elemental selenium by moderately halotolerant Bacillus megaterium strains isolated from Bhitarkanika mangrove soil and characterization of reduced product

Two Gram (+) bacterial strains, BSB6 and BSB12, showing resistance and potential for Se(IV) reduction among 26 moderately halotolerant isolates from the Bhitarkanika mangrove soil were characterized by biochemical and 16S rDNA sequence analyses. Both of them were strictly aerobic and able to grow in a wide range of pH (4-11), temperature (4-40 °C) and salt concentration (4-12%) having an optimum growth at 37 °C, pH ∼7.5 and 7% salt (NaCl). The biochemical characteristics and 16S rDNA sequence analysis of BSB6 and BSB12 showed the closest phylogenetic similarity with the species Bacillus megaterium. Both the strains effectively reduced Se(IV) and complete reduction of selenite (up to 0.25 mM) was achieved within 40 h. SEM with energy dispersive X-ray and TEM analyses revealed the formation of nano size spherical selenium particles in and around the bacterial cells which were also supported by the confocal micrograph study. The UV-Vis diffuse reflectance spectra and XRD of selenium precipitates revealed that the selenium particles are in the nanometric range and crystalline in nature. These bacterial strains may be exploited further for bioremediation process of Se(IV) at relatively high salt concentrations and green synthesis of selenium nanoparticles.     

Biodegradation of 4-aminobenzenesulfonate by Ralstonia sp. PBA and Hydrogenophaga sp. PBC isolated from textile wastewater treatment plant

A co-culture consisting of Hydrogenophaga sp. PBC and Ralstonia sp. PBA, isolated from textile wastewater treatment plant could tolerate up to 100 mM 4-aminobenzenesulfonate (4-ABS) and utilize it as sole carbon, nitrogen and sulfur source under aerobic condition. The biodegradation of 4-ABS resulted in the release of nitrogen and sulfur in the form of ammonium and sulfate respectively. Ninety-eight percent removal of chemical oxygen demand attributed to 20 mM of 4-ABS in cell-free supernatant could be achieved after 118 h. Effective biodegradation of 4-ABS occurred at pH ranging from 6 to 8. During batch culture with 4-ABS as sole carbon and nitrogen source, the ratio of strain PBA to PBC was dynamic and a critical concentration of strain PBA has to be reached in order to enable effective biodegradation of 4-ABS. Haldane inhibition model was used to fit the degradation rate at different initial concentrations and the parameters μ_max, K_s and K_i were determined to be 0.13 h⁻¹, 1.3 mM and 42 mM respectively. HPLC analyses revealed traced accumulation of 4-sulfocatechol and at least four unidentified metabolites during biodegradation. This is the first study to report on the characterization of 4-ABS-degrading bacterial consortium that was isolated from textile wastewater treatment plant.