利用PCR-DGGE研究膜生物反应器中微生物的群落结构

使用聚合酶链式反应-变性梯度凝胶电泳技术(PCR-DGGE)考查了天津某再生水处理厂膜生物反应器(Membrane Bioreactor,简称MBR)培养驯化直至正常运行全过程中总细菌群落结构的演替情况.结果表明,在MBR环境中,接种的传统活性污泥中的微生物群落在几天内发生了较大的变化,在进污水驯化时,微生物群落也遭受了冲击,最后经过培养驯化趋于稳定,一些菌种逐渐成长为顶级优势微生物,在反应器内占据主导地位.最终该反应器逐渐形成了自己独有的微生物群落生态系统.另外,对该微生物群落的部分优势总细菌进行了克隆测序和系统发育树分析,通过鉴定获得的10条总细菌的16S rDNA序列,它们分别与气单胞菌属、假单胞菌、亚硝酸菌属、丛毛单胞菌属和杆菌的同源性在97%以上,这些优势微生物在MBR反应器去除有机物的过程中起到了关键的作用.     

溶藻放线菌的分离鉴定及其溶藻特性

从水华发生水塘的泥土中分离到一株溶藻放线菌L74,对该菌溶解铜绿微囊藻、鱼腥藻、颤藻、水华束丝藻的效果及溶藻方式进行了研究,利用16SrDNA序列分析对其进行了鉴定。结果表明,该菌不仅对铜绿微囊藻具有良好的溶藻效果,而且对鱼腥藻、颤藻、水华束丝藻也有良好的去除作用。对铜绿微囊藻的效解作用依赖于初始菌浓度,当初始藻液中菌浓度>1×104cfu/mL时,可产生明显的溶藻作用,6d后对铜绿微囊藻的去除率达82.6%。通过溶藻方式的研究发现,该菌菌体和胞外代谢产物都具有溶藻作用,且这种胞外代谢产物对温度敏感,当温度达到110℃时会使该溶藻物质失去活性。16SrDNA序列分析表明L74菌与多株弗氏链霉菌的16SrDNA核苷酸序列的同源性达到了100%,归属于弗氏链霉菌。     

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.     

中药废水污泥群落结构解析中PCR-DGGE引物的选择与评价

在中药废水生物活性污泥群落DGGE解析过程中,为了探讨16S rDNA通用引物的扩增效率和对污泥细菌群落的表征能力,选用包括简并性引物在内的11对通用引物扩增16S rDNA序列的4个可变区,并应用DGGE图谱评价不同引物的扩增效率和微生物种群多样性.结果表明,采用不同引物对进行DGGE分析时,微生物种群多样性存在着显...     

渭河浮游细菌群落结构特征及其关键驱动因子

基于T-rflp技术和高通量测序技术,分析了渭河在平水期、枯水期和丰水期的浮游细菌群落变化特征,并采用冗余分析和典范对应分析识别了不同水文时期影响细菌群落的关键环境因子.结果表明,渭河水体浮游细菌群落空间和季节差异明显,且季节性差异比空间差异更加显著.平水期、枯水期和丰水期渭河(陕西段)干流浮游细菌群落Shannon多样性指数分别在2.13~2.82、2.05~2.84和2.61~2.91之间.平水期浮游细菌多样性指数空间差异最大(RSD=16.75%),样点间群落结构相似度最低(26.8%),流域优势T-RF片段数最多(23种);丰水期浮游细菌Shannon指数空间差异最小(RSD=9.27%),样点间群落相似度最高(62.6%),且检出的优势片段数最少(12种).咸阳-西安段是浮游细菌群落多样性最低、优势菌群结构最单一的河段.河流水体细菌群落在不同时期的关键环境驱动因子不同,而其中悬浮颗粒物(TSS)浓度是不同水文时期都不可忽视的关键影响因子.高通量分析结果表明,丰水期渭河水体浮游细菌物种涉及21个已知细菌门类和26个候选门类,其中,变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)共同的相对丰度占比达到75%以上,是最主要的细菌类群.汛期渭河干流各样点浮游细菌群落特征趋于一致,物种结构与泾河相似而与黑河存在明显差异.     

三甲胺降解细菌的分离、降解特性及其系统发育分析

分别从稻田土壤和城市污水处理厂的活性污泥中分离得到 2株三甲胺降解菌 2 2和 5 1 .分离菌株为革兰氏染色反应阴性的球菌和短杆菌 ,菌株大小为球菌直径 0 5～ 0 9μm ,短杆菌长 0 9～ 1 2 μm ,不运动 .分离菌株均能在好氧和厌氧条件下利用三甲胺作为唯一碳源 ,生成中间产物二甲胺和甲胺 ,最终将其矿化 .根据其生理生化性状和 1 6SrDNA序列分析结果表明 ,分离菌株为副球菌属 (Paracoccussp .) ,其序列与Paracoccusaminovorans菌株序列的相似性均为 98 8% .     

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.      

Characterisation and identification of carbofuran-utilising bacteria isolated from agricultural soil

A total of 96 bacterial cultures were isolated from soil. Seventeen bacterial isolates were selected following their cultivation on solid media containing 100 mg · L^?1 carbofuran as the sole source of carbon and nitrogen. Of the 17 isolates, 10F, 11M, 17N, 23B and 26M were specifically chosen because of their relatively higher growth efficiency and genetic diversity based on Box-polymerase chain reaction analysis. These bacterial cultures had 16S rRNA gene sequences that were most similar to Acinetobacter baumannii (10F), Agrobacterium tumefaciens (11M), Ochrobactrum anthropi (17N), Escherichia coli (23B) and Agrobacterium tumefaciens (26M) with 97, 95, 93, 95 and 94% similarity in their 16S rDNA gene sequence, respectively. Degradation rates of carbofuran in soil inoculated with these isolates were 1.9, 1.5, 1.6, 1.7 and 1.6 times, respectively, faster in comparison with uninoculated soil after 10 days of incubation. The maximum degradation rates of carbofuran (45 and 91%) were detected in soil inoculated with A. baumannii (10F) after 10 and 20 days’ incubation, respectively. These data indicate that these isolates may have the potential for use in bioremediation of pesticide contaminated soil.     

不对称还原丙酮酸乙酯菌株的筛选及鉴定

以丙酮酸乙酯为底物,从成都某化工厂污水池及其附近土壤中分离到36株可将丙酮酸乙酯不对称还原成(S)-乳酸乙酯的菌株.经过多次复筛,最终获得了一株具有较高催化活性的酵母菌BTY18-6.在以该菌株静息细胞为催化剂,催化不对称还原丙酮酸乙酯合成(S)-乳酸乙酯的反应中,底物浓度为60 mmol/L时,底物转化率为86.9%,产物(S)-乳酸乙酯的ee值为88.7%.通过对其形态学、生理生化特征及其26S rDNA Dl/D2区域的分析表明,BTY18-6为胶红酵母.     

Effects of two different intercropping systems on arbuscular mycorrhizal fungal colonization and polychlorinated biphenyl (PCB) dissipation北大核心CSCD

Polychlorinated biphenyls (PCBs) are typical organic contaminants in the environment. It is indicated that plants and soil microorganisms have a positive synergistic effect on the remediation of PCB-contaminated soil. To investigate the effect of intercropping on arbuscular mycorrhizal (AM) fungal colonization and PCB remediation, a pot-cultivation experiment with two intercropping treatments, corn (Zea mays L.) / ryegrass (Lolium perenne L.) and corn/alfalfa (Medicago sativa L.), and a corn monoculture was conducted in a greenhouse. All treatments were inoculated with Funneliformis mosseae M47V. Plant biomass, root mycorrhizal colonization rate, concentration of PCBs and their homologs in soil, 16S rDNA gene abundance, and community composition measured by Terminal Restriction Fragment Length Polymorphism (T-RFLP) were determined after harvesting the plants. Intercropping significantly increased the root mycorrhizal colonization rate and plant biomass of corn (P < 0.05), as well as the available N content of the soil. A significant difference of the bacterial community composition was found among different treatments (P < 0.05). Compared with corn monoculture, corn/alfalfa intercropping significantly increased soil bacteria abundance (P < 0.05). The dissipation rates of total PCBs, as well as that of penta-chloro biphenyls were significantly increased in the intercropping treatments, when compared to the corn monoculture treatment. Moreover, corn/ryegrass intercropping has a significantly positive effect on the dissipation of tri-chloro biphenyls. Non-metric multidimensional scaling (NMDS) analysis indicated that the PCBs homologues composition were significantly correlated with the relative abundance of 128 bp and 148 bp T-RFs. Corn intercropping with ryegrass or alfalfa has a positive effect on root mycorrhizal colonization rate and plant biomass of corn. Inoculation of AM fungi in intercropping treatments significantly improved the efficiency of PCB remediation by promoting bacterial abundance and shifting the bacterial community composition. In conclusion, intercropping combined with AM fungi have positive synergistic effects on the remediation of PCB-contaminated soils. © 2018 Science Press. All rights reserved.