不同原料厌氧发酵及其微生物种群的研究

使用PCR-DGGE技术,对以鸡粪、猪粪、牛粪、秸秆为发酵原料的发酵体系的微生物群落多样性进行了研究.在发酵不同时间取样,进行了日产甲烷量、日产甲烷浓度的变化分析和DGGE分析.结果表明,日产甲烷量整体趋势为猪粪>鸡粪>秸秆>牛粪,日平均产甲烷量分别为2.67、2.24、0.99、0.49L;猪粪、鸡粪、秸秆的日产甲烷浓度在整个发酵周期大多可维持在50%以上,牛粪的日产甲烷浓度大部分时间低于30%;细菌的优势菌群有拟杆菌属(Bacteroidetes)、密螺旋体属(Treponema)、厌氧绳菌科(Anaerolineaceae)等,新增优势菌群有梭菌属(Clostridium)、脱硫叶菌属(Desulfobulbus)、毛螺旋菌科(Lachnospiraceae)、醋弧菌属(Acetivibrio);古菌的优势菌群有甲烷鬃菌属(Methanosaeta)、甲烷八叠球菌属(Methanosarcina)、甲烷八叠球菌目(Methanosarcinales),新增优势菌群有斯氏甲烷球菌属(Methanosphaera stadtmanae).     

营养盐与盐双重胁迫下水体中浮游细菌的响应

为阐释干旱地区内陆湖泊微生物群落对营养盐和盐双重胁迫的响应机制,针对干旱、半干旱地区许多湖泊水体受咸化与富营养化双重影响的特点,通过454高通量测序16S rRNA基因和多元统计分析,研究水体营养盐和盐的添加对细菌群落结构、多样性变化的影响. 结果表明,盐与营养盐的输入引起了水体中细菌群落组成和多样性的显著变化, 具体表现为:①随着盐度(以w计)的升高,α-变形菌纲(Alphaproteobacteria)的相对丰度显著增加,从对照处理的16.0%±4.6%增至SO(贫营养-盐水)处理的39.8%±18.1%;但会受到水体营养水平的制约. ②富营养水平下,微咸条件(盐度为1.5‰)有利于细菌类群丰富度的增加,BE-30(微咸-富营养处理培养30 d样品)的Shannon-Wiener多样性指数(4.62)显著高于FE-30(淡水-富营养处理培养30 d样品,2.08)和SE-30(高盐-富营养处理培养30 d样品,1.69);③高盐(盐度为3.0‰)条件下,随着水体中营养盐含量的增加,喜营养的嗜盐细菌类群大量繁殖(SE-30中无法分类的Comamonadaceae科的相对丰度达到69.3%),因而降低了水体中细菌群落的多样性. 研究显示,α-变形菌纲和无法分类的Comamonadaceae科对盐与营养盐的变化较为敏感,可作为干旱区湖泊咸化和富营养化过程的参考类群.      

Bacterial community structure of Tricholoma matsutake stipe soil by high-throughput sequencing北大核心CSCD

In order to understand the interaction between microorganisms in soil attached to the stipes of Tricholoma matsutake, we utilized high-throughput sequencing technology using an Illumina HiSeq 2500 System (San Diego, CA, USA) to explore the bacterial community structure in such soils and analyzed the effects of microorganisms on the formation and growth of T. matsutake. A total of 6 730 sequences were obtained from the samples, and these sequences were clustered into 928 operational taxonomic units (OTUs) at a 97% similarity level. In addition, these OTUs were grouped into 22 bacterial phyla (including 62 classes, 90 orders, 162 families, and 275 genera) and 2 archaeal phyla (including 4 classes, 6 orders, 7 families, and 8 genera). Proteobacteria was the dominant phylum present in all the samples, and these bacteria were significantly more abundant in soils attached to stipe of T. matsutake than they were in the control samples. Similarly, Beta-proteobacteria were the dominant class, and bacteria in this class were less abundant than they were in the control samples. Moreover, the genus Burkholderia within the Proteobacteria was more abundant in the control samples than in the T. matsutake soil samples. Generally, Proteobacteria, especially the genus Burkholderia, played an important role in the growth of T. matsutake, whereas other bacteria, such as Unknown-Ellin 60, would probably inhibit the growth of T. matsutake. This study showed that bacteria might be involved in the growth of T. matsutake and the formation of its fruiting bodies, but they had no positive relationship with bacterial abundance, which may provide theoretical guidance for the cultivation of T. matsutake. © 2018 Science Press. All rights reserved.     

Soil bacterial community characteristics of rice-rice-rape crop rotation北大核心CSCD

To investigate the characteristic changes in soil bacterial community under rice-rice-rape rotation in southern China, soil samples from a 30-year crop rotation and continuous cropping system were collected. Clone library and gene sequence analysis were adopted. The PCR amplification was carried out using universal primers of the 16S rDNA gene. The amplified fragments were then used to construct a clone library. The subclones were sequenced and analyzed. The experimental analysis showed that, in July 2015, and October and April 2016, the Shannon Wiener index and richness index of bacteria from the rotation treatment soils were higher than those of the continuous cropping treatment soil. The soil bacterial diversity of the rotation treatment was higher than that of the continuous cropping treatment. The results of BLAST analysis in the GenBank showed that Proteobacteria accounted 55% of total bacteria in the rotation treatment soil, whereas it accounted for 45% of total bacteria in continuous cropping treatment soil. Gemmatimonadetes accounted for 13% of the total bacteria in rotation treatment and 10% in the continuous cropping treatment. The proportion of Acidobacteria, Firmicutes, and Planctomycetes in rotation soil was less than those in the continuous cropping soil. Proteobacteria and Gemmatimonadetes were the dominant flora in soil. The changes in the predominant bacteria affected the diversity of soil bacteria in rotation and continuous cropping. The sequence analysis showed that the dominant bacteria in the soil were Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria, which higher than those in the rice-rice continuous cropping treatment. The sequences similar to Chloroflexi were observed in the rice-rice-rape rotation soils, but not in the continuous cropping soils. The results showed that the soil microbial population is conducted by agricultural tillage, and rice-rice-rape rotation increased the soil bacterial abundance compared with that of the rice-rice continuous treatment. © 2018 Science Press. All rights reserved.     

CRISPR/Cas系统抵御细菌耐药

近年来,细菌耐药性问题在全球范围内日益严重,世界各地不断出现各种新型耐药基因及"超级细菌",以此形成的细菌耐药性污染已成为威胁全球公共卫生与环境安全的重大问题。目前除了在管理上规范合理用药和限制抗生素排放,应探讨抵御细菌耐药的分子机制,有效地从根本上遏制细菌耐药的产生。CRISPR系统作为一种天然免疫系统,可用来对抗入侵的外源性遗传物质,其结构和功能与细菌耐药及毒力因素密切相关,深入分析两者的关系有助于更好地理解细菌耐药机制,为防治细菌耐药提供了新的方向。     

污染浓度对多级人工湿地-塘系统处理微污染河水中COD的影响

针对微污染水水质不稳定,COD_Mn波动较大的问题,本研究构建了3套中试规模的以"前置生态氧化塘、两级水平潜流湿地和表流湿地为核心、后置沉水植物塘"为工艺的多级人工湿地-塘组合系统（处理水量230 L·d^-1,分别命名为S1、S2和S3）.在3种进水浓度条件下（原水、稀释2倍和稀释1.3倍后的水体,分别作为S1、S2和S3系统的进水）,考察了3套湿地-塘系统各单元对实际微污染河水中COD_Mn的处理效果,并采用高通量测序等方法解析了S1系统湿地单元微生物群落分布特征.研究结果表明,3套系统的平均出水COD较进水有着非显著的升高,从升高的程度来看,S2>S3>S1.污染浓度相对最高、运行时间最长的S1系统对COD有着更好的净化效果.相较于S2系统,S3系统更高的进水浓度有利于COD去除.S1和S2系统的二级水平潜流湿地较其他单元有着较好的COD去除作用,平均去除率分别为18.60%和26.52%.S3系统的表流湿地较其他单元有着较好的COD去除作用,平均去除率为8.92%.微生物群落分析结果表明,S1系统的一级、二级水平潜流湿地中细菌丰度和多样性沿程提高,两个单元所富集的相同优势菌群包括α-变形菌纲、γ-变形菌纲和拟杆菌纲,与COD降解有关的菌属分别包括unclassified_f__Burkholderiaceae、unclassified_c__Alphaproteobacteria、norank_f__Blastocatellaceae和norank_f__Saprospiraceae、红细菌属.表流湿地中细菌丰富度进一步升高,但生物多样性有所减少,其中蓝菌属和norank_c__Actinobacteria可能与表流湿地出水COD升高有关.本研究为大型河口湿地-塘复合生态系统处理微污染地表水时工作重心由浓度削减向污染物释放控制的转移提供了宝贵的工程经验.     

印染废水循环利用抗生素抗性基因丰度变化特性

城市污水处理系统中抗生素抗性基因（ARGs）分布研究较多,但是工业废水循环利用时ARGs丰度变化特性研究尚存不足.为此,本研究构建了印染废水循环利用系统,采用16S rDNA技术分析印染废水循环利用过程中微生物群落的变化特性,采用高通量测序技术表征ARGs丰度变化趋势.循环初期,活性污泥中共检出9大类52种ARGs,其中β-内酰胺类（β-lactam）抗性基因的相对丰度最高.循环过程中,芳烃类污染物浓度随循环次数的增加而升高,β-内酰胺类抗性基因丰度则先升高后降低再升高（第100 d,上升61.85%）.与此同时,与ARGs相关的Firmicutes、Actinobacteria和Cyanobacteria的丰度显著下降（降幅分别为84.66%、64.38%和85.15%）.超过21种ARGs受到芳烃类污染物富集的显著影响,其中,6种ARGs与芳烃类污染物的浓度变化极显著正相关,6种极显著负相关.这些结果表明,ARGs的丰度变化受微生物群落和芳烃类污染物的影响,在印染废水循环利用过程中呈现先上升后下降再上升的变化趋势.本研究揭示了印染废水循环利用过程中芳烃类污染物的富集与微生物群落的变化对ARGs的影响,对印染工业废水循环利用降低抗生素抗性基因对环境的污染提供了理论指导.     

ZONES OF NITRIFICATION1

ABSTRACT: This paper advances the hypothesis that nitrification exists in shallow streams as a result of surface activity and in estuaries due to growth in the water phase. Between these zones no significant levels of nitrification occur. Field measurements of ammonia, nitrite and nitrate nitrogen, enumerations of Nitrosomonas and Nitrobacter, and respirometry on two small streams and two moderately large rivers are presented. The appropriate mathematical models to describe nitrification must be based on the mechanisms involved - bacterial growth kinetics for estuaries and zero kinetics for the surface activity in shallow streams.     

Assessment of the Ecological Integrity of Traunsee (Austria) Via Analysis of Sediments and Benthic Microbial Communities

Since nearly one hundred years Traunsee experiences the import of tons of liquid and solid waste originating from salt and soda production. Today, the lake exhibits chloride concentrations of up to 170 mg L^-1 and 19% of the lake floor are directly or indirectly influenced by industrial deposits (ID). Based on the comparison of several microbial parameters in unaffected, directly affected and intermediate lake bottom sediments, the ecological integrity of the lake was evaluated. The highly alkaline ID, which were exclusively colonized by microorganisms, harbored a bacterial community reduced by a factor of 10 in abundance and biomass compared to undisturbed sediment areas within the lake. The bacterial community of ID was furthermore characterized by a reduced content of actively respiring cells (INT-formazan reduction), a lower frequency of dividing cells (FDC) and a significantly reduced cell and biomass production. A 80 to 90% reduction in carbon recycling is estimated for the area exclusively covered by ID. Protists, although occasionally absent from the industrial sediments, were in general found to be less sensitive to the contaminant stress. Differences in alkalinity and dissolved organic carbon (DOC) concentrations of sediment porewaters as well as the total organic content and C/N ratios of sediments partly explain the microbial pattern observed at the various sampling sites. Possible consequences of the continuous industrial tailings for the whole lake ecosystem and the validation of the ecological integrity are discussed.     

FECAL COLIFORM RELEASE FROM CATTLE FECAL DEPOSITS1

ABSTRACT: Cowpies molded to a standard configuration and size were subjected to simulated rainfall, and the fecal coliform counts were determined using the most probable number (MPN) method of enumeration. The standard cowpie deposits were exposed to simulated rainfall once at ages 2 through 100 days. The effects of rainfall intensity and recurrent rainfall were also tested. Naturally-occurring fecal deposits were also tested to compare their results with those from the standard cowpies. A log-log regression was found to describe the decline in peak fecal coliform release with fecal deposit age. The 100-day-old fecal deposits produced peak counts of 4,200 fecal coliform per 100 milliliters of water. This quantity of release is minimal compared to the release from fresher fecal material. Rainfall intensity had little effect on peak fecal coliform release from fecal deposits that were 2 or 10 days old. At age 20 days the effect of rainfall intensity was significant; the highest intensity gave the lowest peak counts, and the lowest intensity gave the highest peak counts. The effect of rainfall intensity appears to be related to the dryness of the fecal deposits. Peak fecal coliform counts were significantly lowered when the fecal deposits were rained on more than once. This decline was thought to be produced by the loss of bacteria from the fecal deposits during the previous wettings. Standard cowpies produced a peak release regression that was not significantly different from the regression for the natural fecal deposits. Apparently, grossly manipulating the fecal deposits did not significantly change the release patterns.