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Bacterial diversity and community structure in the East China Sea by 454 sequencing of the 16S rRNA gene 被引量:8
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作者 董逸 赵苑 +7 位作者 张文燕 李艳 周锋 刘诚刚 吴莹 刘素美 张武昌 肖天 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2014年第3期527-541,共15页
The 454 sequencing method was used to detect bacterial diversity and community structure in the East China Sea. Overall, 149 067 optimized reads with an average length of 454 nucleotides were obtained from 17 seawater... The 454 sequencing method was used to detect bacterial diversity and community structure in the East China Sea. Overall, 149 067 optimized reads with an average length of 454 nucleotides were obtained from 17 seawater samples and fi ve sediment samples sourced in May 2011. A total of 22 phyla, 34 classes, 74 orders, 146 families, and 333 genera were identifi ed in this study. Some of them were detected for the fi rst time from the East China Sea. The estimated richness and diversity indices were both higher in the sediment samples compared with in the seawater samples. All the samples were divided by their diversity indices into four regions. Similarity analysis showed that the seawater samples could be classifi ed into six groups. The groups differed from each other and had unique community structure characteristics. It was found that different water masses in the sampling areas may have had some infl uence on the bacterial community structure. A canonical correspondence analysis revealed that seawater samples in different areas and at different depths were affected by different environmental parameters. This study will lay the foundation for future research on microbiology in the East China Sea. 展开更多
关键词 bacterial diversity community structure 454 sequencing water masses canonical correspondence analysis East China Sea
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宁波沿海陆源排污口假单胞菌属(Pseudomonas)分布特点 被引量:14
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作者 陈丽萍 侯付景 +7 位作者 张迪骏 何伟娜 周君 张春丹 童茜茜 王中华 李太武 苏秀榕 《海洋与湖沼》 CAS CSCD 北大核心 2013年第4期926-930,共5页
采用高通量454焦磷酸测序方法对宁波沿海2个重点排污口、8个一般排污口的20个站位水样进行分析,得到2011年3月、5月、8月、10月份各排污口假单胞菌属的分布情况。结果表明,宁波沿海陆源排污口中存在较多的假单胞菌属。在检出的假单胞菌... 采用高通量454焦磷酸测序方法对宁波沿海2个重点排污口、8个一般排污口的20个站位水样进行分析,得到2011年3月、5月、8月、10月份各排污口假单胞菌属的分布情况。结果表明,宁波沿海陆源排污口中存在较多的假单胞菌属。在检出的假单胞菌属中,维罗纳假单胞菌(Pseudomonas veronii)和莓实假单胞菌(Pseudomonas fragi)的含量相对较多,分别占56.72%、13.904%;排污口中假单胞菌属的数量存在季节性差异,3月份、5月份假单胞菌的数量相对较多,8月份和10月份较少,推测与季节性温度变化有关;假单胞菌属的含量与排污口的主要污染物有关,氨氮含量较高的排污口假单胞菌属的含量更高。 展开更多
关键词 假单胞菌属 454焦磷酸测序 陆源排污口 聚类分析
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正常胃黏膜菌群特征分析2例 被引量:3
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作者 王莉莉 董开芯 +3 位作者 周建华 于新娟 贾盛佼 董全江 《世界华人消化杂志》 CAS 北大核心 2013年第13期1250-1255,共6页
目的:分析消化不良患者的正常胃黏膜菌群特征.方法:收集2例正常胃黏膜病理学消化不良患者的胃窦黏膜,提取黏膜组织基因组DNA,应用PCR方法扩增16S rRNA,进行二代454焦磷酸测序.测序数据应用操作单元(operational taxonomic units,OTU)聚... 目的:分析消化不良患者的正常胃黏膜菌群特征.方法:收集2例正常胃黏膜病理学消化不良患者的胃窦黏膜,提取黏膜组织基因组DNA,应用PCR方法扩增16S rRNA,进行二代454焦磷酸测序.测序数据应用操作单元(operational taxonomic units,OTU)聚类分析、多样性分析和分类学分析等生物信息技术分析胃黏膜菌群的结构、多样性和丰度.结果:患者A和B胃黏膜菌群的高通量测序分析分别得到20565条和17487条优化序列,测序覆盖深度>0.98.经过97%相似度归并后分别得到638个OTUs和667个OTUs;菌群多样性分析显示正常胃黏膜菌群的chao指数和shannon指数均低于粪便菌群.患者A的胃黏膜菌群由58个属的细菌构成,归属于14个门,以变形菌门为主(99.49%),其次为放线菌门、硬壁菌门和拟杆菌门等.患者B胃黏膜菌群构成与患者A相似,由64个属的细菌构成,归属于19个门.粘质沙雷(氏)菌为正常胃黏膜菌群的优势主导菌.结论:正常胃黏膜菌群多样性丰富,以变形菌门为主,沙雷(氏)菌为优势主导菌. 展开更多
关键词 胃菌群 454焦磷酸测序 变形菌门 沙雷(氏)菌属
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玉米秸秆与猪粪混合发酵产沼气及其细菌解析 被引量:1
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作者 袁月祥 廖银章 +3 位作者 刘晓风 闫志英 曾李乐 李旭东 《太阳能学报》 EI CAS CSCD 北大核心 2015年第8期1971-1977,共7页
以总固体1:1的玉米秸秆和猪粪为混合原料,采用10L厌氧反应器进行批式全混中温沼气发酵,并利用454焦磷酸测序法测定混合发酵过程中细菌群落变化。结果表明:预处理玉米秸秆和猪粪混合沼气发酵启动迅速,在笫4天达到产气高峰,容积产... 以总固体1:1的玉米秸秆和猪粪为混合原料,采用10L厌氧反应器进行批式全混中温沼气发酵,并利用454焦磷酸测序法测定混合发酵过程中细菌群落变化。结果表明:预处理玉米秸秆和猪粪混合沼气发酵启动迅速,在笫4天达到产气高峰,容积产气率为1.17L/(L·d),46d原料沼气产率为356.55mL/gVS。该混合发酵系统中共有23~27门细菌,其中优势菌群为拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)。发酵期内的Shannon指数在4.88~5.53之间,说明发酵期内细菌多样性比较稳定。 展开更多
关键词 玉米秸秆 猪粪 混合发酵 沼气 454焦磷酸测序 细菌解析
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The Nitrogen-Cycling Network of Bacterial Symbionts in the Sponge Spheciospongia vesparium
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作者 HE Liming KARLEP Liisi LI Zhiyong 《Journal of Ocean University of China》 SCIE CAS CSCD 2021年第4期999-1012,共14页
The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen fixation.However,the whole nitrogen-cycling network has rema... The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen fixation.However,the whole nitrogen-cycling network has remained incomplete in any individual sponge holobiont.In this study,454 pyrosequencing of the 16S rRNA genes revealed that the sponge Spheciospongia vesparium from the South China Sea has a unique bacterial community(including 12 bacterial phyla),dominated particularly by the genus Shewanella(order Alteromonadales).A total of 10 functional genes,nifH,amoA,narG,napA,nirK,norB,nosZ,ureC,nrfA,and gltB,were detected in the microbiome of the sponge S.vesparium by gene-targeted analysis,revealing an almost complete nitrogen-cycling network in this sponge.Particularly,bacterial urea utilization and the whole denitrification pathway were highlighted.MEGAN analysis suggests that Proteobacteria(e.g.,Shewanella)and Bacteroidetes(e.g.,Bizionia)are probably involved in the nitrogen cycle in the sponge S.vesparium. 展开更多
关键词 Spheciospongia vesparium bacterial symbionts 454 pyrosequencing functional gene analysis nitrogen-cycling net-work
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CVTree在454高通量测序分析菌群结构中的应用 被引量:10
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作者 华蔚颖 徐昭 +3 位作者 张梦晖 李旻 张晨虹 赵立平 《中国微生态学杂志》 CAS CSCD 2010年第4期312-316,共5页
目的探究将基于短串频度的CVTree方法用于反映菌群结构的16S rRNA基因的454高通量测序数据分析的可行性,为快速分析高通量菌群结构数据提供新的方法。方法对一个四世同堂的中国家庭7名成员肠道菌群和不同基因型及饮食类型的小鼠肠道菌群... 目的探究将基于短串频度的CVTree方法用于反映菌群结构的16S rRNA基因的454高通量测序数据分析的可行性,为快速分析高通量菌群结构数据提供新的方法。方法对一个四世同堂的中国家庭7名成员肠道菌群和不同基因型及饮食类型的小鼠肠道菌群用454高通量方法获得16S rRNA基因的V3区的测序数据,用CVTree的方法进行菌群结构的比较分析。结果通过选取合适的短串长度,CVTree的方法能准确检测到各样本间的聚类关系,其结果与之前文献报道的基于Unifrac算法的结果相一致。结论CVTree能快速、有效地处理16S rRNA基因的454高通量测序数据,实现对不同菌群结构相似性的比较分析。 展开更多
关键词 CVTree 454高通量测序 菌群结构
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玉米秆与猪粪混合发酵产沼气及其古菌解析
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作者 袁月祥 曾李乐 +4 位作者 闫志英 刘晓风 廖银章 李旭东 孙太荣 《应用与环境生物学报》 CAS CSCD 北大核心 2014年第1期117-122,共6页
为了解混合发酵系统不同发酵时间的古菌组成,采用批式全混发酵工艺进行预处理后的玉米秆和猪粪混合中温发酵产沼气研究,在不同发酵时间取样,利用454焦磷酸测序法测定各样品中古菌的组成.结果表明,该混合发酵启动迅速,第4天(d 4)达到产... 为了解混合发酵系统不同发酵时间的古菌组成,采用批式全混发酵工艺进行预处理后的玉米秆和猪粪混合中温发酵产沼气研究,在不同发酵时间取样,利用454焦磷酸测序法测定各样品中古菌的组成.结果表明,该混合发酵启动迅速,第4天(d 4)达到产气高峰,料容产气率为1.46 L L-1 d-1,46 d原料产沼气率、产甲烷率分别为356.55 mL/g(VS)和208.0 mL/g(VS).454焦磷酸测序及分析表明,混合发酵系统中古菌都归属于广古菌门(Euryarchaeota,97.83%-100%)和泉古菌门(Crenarchae ota,0-2.17%);随着发酵时间的延长,甲烷杆菌纲(Methanobacteria)古菌含量由69.84%迅速下降到0.14%,杂色泉古菌(Miscellaneous Crenarchaeotic Group,MCG)和热原体纲(Thermoplasmata)古菌含量则分别由0%、1.05%逐渐上升到2.17%、20.68%,甲烷微菌纲(Methanomicrobia)古菌则从29.84%迅速上升到96.14%后保持高位.研究说明玉米秆和猪粪混合发酵可迅速启动,系统古菌多样性随发酵时间延长而逐渐上升. 展开更多
关键词 混合发酵 沼气 454焦磷酸测序 古菌 解析
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高通量测序技术在水环境微生物群落多样性中的应用 被引量:55
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作者 王绍祥 杨洲祥 +3 位作者 孙真 刘燕 汪成伟 荆毓航 《化学通报》 CAS CSCD 北大核心 2014年第3期196-203,共8页
微生物群落多样性的研究方法随着现代生物学技术经历了几代发展,基于形态学、生理学、分子生物学的传统研究方法在取得大量研究成果的同时,受对环境微生物群落多样性认识的限制,亟待获得一种能更加全面、准确、快速反映环境微生物群落... 微生物群落多样性的研究方法随着现代生物学技术经历了几代发展,基于形态学、生理学、分子生物学的传统研究方法在取得大量研究成果的同时,受对环境微生物群落多样性认识的限制,亟待获得一种能更加全面、准确、快速反映环境微生物群落结构的方法。本文首先回顾了传统研究方法,重点介绍了高通量测序技术,包括研究者所关注的测序仪分布、类型、原理、优势与制约,列举了多样性研究中的分析作图软件,最后以454的焦磷酸测序为例具体阐述了该方法在分析水环境微生物群落多样性中的应用。 展开更多
关键词 高通量测序技术 454焦磷酸测序 生物信息分析 水体微生物群落多样性
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九龙江流域潜在病原菌污染分析 被引量:5
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作者 侯丽媛 胡安谊 +1 位作者 马英 于昌平 《环境科学》 EI CAS CSCD 北大核心 2014年第5期1742-1749,共8页
近年来,由于流域经济快速发展和城镇化进程加快等原因,九龙江水污染问题日趋严重.为全面认识九龙江流域病原菌的分布状况,应用16S rRNA基因-454焦磷酸测序技术测定九龙江支流西溪、北溪水体和沉积物中细菌16S rRNA基因V1~V3高变区,共获... 近年来,由于流域经济快速发展和城镇化进程加快等原因,九龙江水污染问题日趋严重.为全面认识九龙江流域病原菌的分布状况,应用16S rRNA基因-454焦磷酸测序技术测定九龙江支流西溪、北溪水体和沉积物中细菌16S rRNA基因V1~V3高变区,共获得204 216条高质量序列.通过与病原菌参考数据库对比分析发现,九龙江分布有68个潜在病原菌属,占序列总量的6.1%.其中梭菌属(Clostridium)、分支杆菌属(Mycobacterium)和鞘氨醇单胞菌属(Sphingomonas)在所有样品中都有检出,且丰度最高,分别占病原菌属序列总量的54.5%、5.9%和5.6%.在种水平上,九龙江分布有48种病原菌,占序列总量的0.76%.其中Afipia felis、Mycobacterium asiaticum、Clostridium baratii、Brucella melitensis和Delftia tsuruhatensis是丰度最高的5种病原菌,分别占病原菌种序列总量的48.9%、20.3%、8%、2.7%和1.7%.统计分析表明,九龙江水体中分布的病原菌种类数(属或种)显著高于沉积物中,且西溪水体检出的病原菌种类(属或种)和丰度最高,说明九龙江水体,尤其是西溪水体受病原菌污染的风险较高.此外,相关分析表明,九龙江水体中病原菌的种类数(属或种)和丰度与营养盐(氮、磷)有着显著的正相关关系,说明九龙江水体中分布的病原菌与沿岸的人类活动,如养殖业、污水排放等密切相关.因此,为保证流域的公共卫生安全,需进一步加强污染源清理整顿工作,开展水环境病原菌的实时监测. 展开更多
关键词 九龙江流域 潜在病原菌 16S RRNA基因 454焦磷酸测序 生物信息分析
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Changes of microbial composition during wastewater reclamation and distribution systems revealed by high- throughput sequencing analyses 被引量:5
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作者 Yiwen LIN Dan LI +1 位作者 Siyu ZENG Miao HE 《Frontiers of Environmental Science & Engineering》 CSCD 2016年第3期539-547,共9页
This study employed 454-pyrosequencing to investigate microbial and pathogenic communities in two wastewater reclamation and distribution systems. A total of 11972 effective 16S rRNA sequences were acquired from these... This study employed 454-pyrosequencing to investigate microbial and pathogenic communities in two wastewater reclamation and distribution systems. A total of 11972 effective 16S rRNA sequences were acquired from these two reclamation systems, and then designated to relevant taxonomic ranks by using RDP classifier. The Chao index and Shannon diversity index showed that the diversities of microbial communities decreased along wastewater reclamation processes, eroteobacteria was the most dominant phylum in reclaimed water after disinfection, which accounted for 83% and 88% in two systems, respectively. Human opportunistic pathogens, including Clostridium, Escherichia, Shigella, Pseudomonas and Mycobacterium, were selected and enriched by disinfection processes. The total chlorine and nutrients (TOC, NH3-N and NO3-N) significantly affected the microbial and pathogenic communities during reclaimed water storage and distribution processes. Our results indicated that the disinfectant-resistant pathogens should be controlled in reclaimed water, since the increases in relative abundances of pathogenic bacteria after disinfec- tion implicate the potential public health associated with reclaimed water. 展开更多
关键词 wastewater reclamation systems microbial community pathogenic community 454-pyrosequencing
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Performance and microbial diversity of an expanded granular sludge bed reactor for high sulfate and nitrate waste brine treatment 被引量:7
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作者 Runhua Liao Yan Li +7 位作者 Xuemin Yu Peng Shi Zhu Wang Ke Shen Qianqian Shi Yu Miao Wentao Li Aimin Li 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2014年第4期717-725,共9页
The disposal of waste brines has become a major challenge that hinders the wide application of ion- exchange resins in the water industry in recent decades. In this study, high sulfate removal efficiency (80%-90%) w... The disposal of waste brines has become a major challenge that hinders the wide application of ion- exchange resins in the water industry in recent decades. In this study, high sulfate removal efficiency (80%-90%) was achieved at the influent sulfate concentration of 3600 mg/L and 3% NaC1 after 145 days in an expanded granular sludge bed (EGSB) reactor. Furthermore, the feasibility of treating synthetic waste brine containing high levels of sulfate and nitrate was investigated in a single EGSB reactor during an operation period of 261 days. The highest nitrate and sulfate loading rate reached 6.38 and 5.78 kg/(m3-day) at SO42--S/NO3-N mass ratio of 4/3, and the corresponding removal efficiency was 99.97% and 82.26% at 3% NaC1, respectively. Meanwhile, 454-pyrosequencing technology was used to analyze the bacterial diversity of the sludge on the 240th day for stable operation of phase X. Results showed that a total of 9194 sequences were obtained, which could be affiliated to 14 phyla, including Proteobacteria, Firmicutes, Chlorobi, Bacteroidetes, Synergistetes and so on. Proteobacteria (77.66%) was the dominant microbial population, followed by Firmicutes (12.23%) and Chlorobi (2.71%). 展开更多
关键词 454-pyrosequencing waste brine denitrification expanded granular sludge bed (EGSB) reactor
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