污泥转移对SBR工艺微生物群落结构及其除磷效能影响分析

采用污泥转移SBR工艺处理以生活污水为基础的合成废水。污泥转移能够强化厌氧生物选择器中聚磷菌的筛选,从而显著提升传统SBR工艺的除磷性能。对比传统SBR,应用荧光原位杂交(FISH)和聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)考察了污泥转移SBR中微生物种群结构的变化及除磷特性。结果表明,污泥转移SBR中磷的去除率总体呈上升趋势,稳定在93%左右;而传统SBR中磷的平均去除率为50%。DGGE试验表明,污泥转移SBR首尾样品的相似性系数为36.8%,传统SBR的则为54.7%;污泥转移SBR的香农指数下降率为5.6%,而传统SBR的香农指数下降率为4%。微生物种群在驯化过程中发生了变化,即微生物种群的优势菌在反应器启动阶段被筛选和富集。对比传统SBR,污泥转移SBR中微生物种群的这一驯化现象表现得更为明显。FISH试验表明,污泥转移SBR的聚磷菌占全菌比例在反应器稳定后可达到46%,聚糖菌则稳定在22%;传统SBR稳定后的聚磷菌比例为35%,聚糖菌比例为30%。传统SBR和污泥转移SBR的聚磷菌比例都先沿程增加最后稳定,且聚磷菌是优势菌。污泥转移SBR的聚糖菌沿程减少,聚磷菌抑制了聚糖菌的生长。因此,污泥转移SBR工艺具有筛选大量聚磷菌且抑制聚糖菌的除磷优势。     

Comparison the effects of bioaugmentation versus biostimulation on marine microbial community by PCR–DGGE: A mesocosm scale

In order to better understand the effects of biostimulation and bioaugmentation processes on a marine microbial community, three different mesocosm experiments were planned. Natural seawater(10.000 L) was artificially polluted with crude oil(1 L) and(1) inorganic nutrients(Biostimulating Mesocosm, BM),(2) inorganic nutrients and an inoculum of Alcanivorax borkumensis SK2(Single Bioaugmentation Mesocosm, SBM),(3) inorganic nutrients and inoculums of A. borkumensis SK2 and Thalassolituus oleivorans MIL-1(Consortium Bioaugmentation Mesocosm, CBM). During the experimental period(20 days), samples were taken from each mesocosm and the community structure was analyzed by PCR–DGGE. The 16 S r RNA gene DGGE banding patterns and sequence analysis demonstrated that biostimulation had the lowest effect on microbial biodiversity in the mesocosms; however, the biodiversity of the marine microbial community dramatically decreased in the CBM(Shannon index was 0.6 in T3). The community structures among the three mesocosms were also markedly different,and major bacteria derived from DGGE bands were related to uncultured Gamma Proteobacteria. The biodegradation results show that the Single Bioaugmentation Mesocosm(SBM) system had the highest percentage of degradation(95%) in comparison to the BM mesocosm(80%) and CBM(70%).     

Temporal variation of microbial population in a thermophilic biofilter for SO2 removal

The performance of a biofilter relies on the activity of microorganisms during the gas contaminant treatment process. In this study, SO_2 was treated using a laboratory-scale biofilter packed with polyurethane foam cubes(PUFC), on which thermophilic desulfurization bacteria were attached. The thermophilic biofilter effectively reduced SO_2 within 10 months of operation time, with a maximum elimination capacity of 48.29 g/m~3/hr.Temporal shifts in the microbial population in the thermophilic biofilter were determined through polymerase chain reaction-denaturing gradient gel electrophoresis and deoxyribonucleic acid(DNA) sequence analysis. The substrate species and environmental conditions in the biofilter influenced the microbial population. Oxygen distribution in the PUFC was analyzed using a microelectrode. When the water-containing rate in PUFC was over 98%, the oxygen distribution presented aerobic–anoxic–aerobic states along the test route on the PUFC. The appearance of sulfate-reducing bacteria was caused by the anaerobic conditions and sulfate formation after 4 months of operation.     

Responses of activities, abundances and community structures of soil denitrifiers to short-term mercury stress

The responses of activities, abundances and community structures of soil denitrifiers to mercury (Hg) stress were investigated through a short-term incubation experiment. Four soil treatments with different concentrations of Hg (CK, Hg25, Hg50, and Hg100, denoted as 0, 25, 50, and 100 mg Hg/kg dry soil, respectively) were incubated for 28 days. Soil denitrification enzyme activity (DEA) was measured at day 3, 7 and 28. The abundances and community structures of two denitrification concerning genes, nirS (cd₁-nitrite reductase gene) and nosZ (nitrous oxide reductase gene), were analyzed using real-time PCR and denaturing gradient gel electrophoresis (DGGE). Results showed that soil DEA was significantly stimulated in the treatments of Hg25 and Hg50 compared with others at day 7. Meanwhile, no difference in the abundances of soil nirS and nosZ was found between Hg spiked treatments and CK, except the lower abundance of nirS (P<0.05) in the Hg added treatments compared with that in the CK at day 28. The community structures of denitrifiers based on nirS gene presented obvious change at day 7 along with the Hg additions, however, no variation was found in all treatments based on the nosZ gene. The results indicated that Hg (Hg25 and Hg50) had a strongly short-term stimulation on soil DEA, and nirS gene is more sensitive than nosZ gene to Hg stress.     

The metabolism of neonicotinoid insecticide thiamethoxam by soil enrichment cultures,and the bacterial diversity and plant growth‐promoting properties of the cultured isolates

A soil enrichment culture (SEC) rapidly degraded 96% of 200 mg L^?1 neonicotinoid insecticide thiamethoxam (TMX) in MSM broth within 30 d; therefore, its metabolic pathway of TMX, bacterial diversity and plant growth‐promoting rhizobacteria (PGPR) activities of the cultured isolates were studied. The SEC transformed TMX via the nitro reduction pathway to form nitrso, urea metabolites and via cleavage of the oxadiazine cycle to form a new metabolite, hydroxyl CLO‐tri. In addition, 16S rRNA gene‐denaturing gradient gel electrophoresis analysis revealed that uncultured rhizobacteria are predominant in the SEC broth and that 77.8% of the identified bacteria belonged to uncultured bacteria. A total of 31 cultured bacterial strains including six genera (Achromobacter, Agromyces, Ensifer, Mesorhizobium, Microbacterium and Pseudoxanthomonas) were isolated from the SEC broth. The 12 strains of Ensifer adhaerens have the ability to degrade TMX. All six selected bacteria showed PGPR activities. E. adhaerens TMX‐23 and Agromyces mediolanus TMX‐25 produced indole‐3‐acetic acid, whereas E. adhaerens TMX‐23 and Mesorhizobium alhagi TMX‐36 are N₂‐fixing bacteria. The six‐isolated microbes were tolerant to 200 mg L^?1 TMX, and the growth of E. adhaerens was significantly enhanced by TMX, whereas that of Achromobacter sp. TMX‐5 and Microbacterium sp.TMX‐6 were enhanced slightly. The present study will help to explain the fate of TMX in the environment and its microbial degradation mechanism, as well as to facilitate future investigations of the mechanism through which TMX enhances plant vigor.     

重金属镉对水田土壤微生物基因多样性的影响

采用16S rDNA DGGE分析技术,探讨了不同处理时间、不同浓度镉胁迫条件下水田土壤微生物种群的基因多样性.研究发现,随着处理时间的不同,各处理间的土壤微生物基因多样性差异存在明显不同,培养初期镉对土壤微生物种群毒害作用非常明显,土壤微生物基因多样性下降,但培养后期不同处理之间DNA基因型差异很小,丰度差异较大.在土壤培养初期、含镉3～10 mg/kg的培养基上出现一条新增条带,为对照及低镉浓度土壤所没有,镉形态稳定后会消失,说明土壤中可能存在适官镉浓度下良好生长的微生物种,经序列测定及与网上比对分析.与一株Uncultured bacterium sp.具有96%的相似率,初步确定为一不可培养细菌种,以其作为淹水条件下镉污染初期的报告基因有一定的可行性.     

生活垃圾堆肥过程中细菌群落演替规律

应用PCR-DGGE技术研究生活垃圾堆肥过程中的细菌群落演替规律,对堆肥不同时期的宏基因组DNA进行提取,扩增16S rDNA的V3区,分析生活垃圾堆肥过程中细菌群落的变化. DGGE图谱表明,随着堆体温度的升高,DNA条带表现出了明显的动态变化,降温期出现了新的优势条带并趋于稳定,说明堆肥不同时期的细菌群落发生了更替. 对条带分布进行聚类分析,结果表明:以55 ℃为界,将14个堆肥样品划分为2个族,族间的相似性仅为13%,说明堆肥过程中常温期(<55 ℃)和高温期(>55 ℃)微生物群落结构差别较大. 对优势条带回收测序的结果表明:在升温期,堆肥堆体中检测到H. obtusa和人类排泄物中的细菌;但随着温度的升高,具有纤维素降解功能的嗜热微生物Clostridium thermocellum成为堆肥高温期的优势细菌;当堆体温度小于55 ℃时出现了大量的未培养微生物.      

Current molecular biologic techniques for characterizing environmental microbial community

Microbes are vital to the earth because of their enormous numbers and instinct function maintaining the natural balance. Since the microbiology was applied in environmental science and engineering more than a century ago, researchers desire for more and more information concerning the microbial spatio-temporal variations in almost every fields from contaminated soil to wastewater treatment plant (WWTP). For the past 30 years, molecular biologic techniques explored for environmental microbial community (EMC) have spanned a broad range of approaches to facilitate the researches with the assistance of computer science: faster, more accurate and more sensitive. In this feature article, we outlined several current and emerging molecular biologic techniques applied in detection of EMC, and presented and assessed in detail the application of three promising tools.     

内蒙古河套灌区不同盐碱程度的土壤细菌群落多样性

采用变性梯度凝胶电泳(DGGE)技术对内蒙古河套灌区三种不同盐碱程度土壤(盐土,强度盐化土,轻度盐化土)不同深度(0~20cm和20~30cm)土壤细菌16S rDNA V3~V6可变区扩增片段进行分析,并对土壤理化性质进行了测定.结果表明:细菌群落多样性随土壤盐碱化程度的加深而减少(轻度盐化土 > 强度盐化土 > 盐土),随土壤深度的增加而降低(细菌群落多样性0~20cm土层大于20~30cm土层).细菌Shannon-Wiener指数在轻度盐化土中最大为3.36,在强度盐化土和盐土分别为3.05和2.49.不同盐碱程度土壤以细菌相似系数聚类,分为0~20cm层与20~30cm层两大族群,土壤细菌群落Shannon-Wiener指数在0~20cm层中(盐土为3.04,强度盐化土为3.29,轻度盐化土为3.36)均大于在20~30cm层(盐土为2.49,强度盐化土为3.05,轻度盐化土为3.14).相关性分析和典范对应分析表明,土壤w(EC)、pH值、w(SOC)、w(TP)是土壤细菌群落结构多样性的显著影响因素,不同盐碱程度土壤中细菌群落的Shannon-Wiener指数与土壤w(EC)(r=-0.542,P < 0.05)、pH(r=-0.526,P < 0.05)呈显著负相关,与土壤w(SOC)(r=0.700,P < 0.01)和w(TP)(r=0.805,P < 0.01)呈极显著正相关.w(EC)和pH对盐碱土壤细菌群落结构的影响力最大. 回收DGGE图谱中20个优势条带进行测序分析,结果显示,变形菌纲(α-变形菌纲、β-变形菌纲、γ-变形菌纲和δ-变形菌纲)是盐碱土壤的主要类群.     

生物炭添加对秸秆还田土壤细菌群落结构和多样性影响

为探讨生物炭对秸秆还田土壤细菌群落结构和多样性的影响,采用变性梯度凝胶电泳(PCR-DGGE)技术结合克隆测序技术,对未添加(SW)和添加(SBW)生物炭秸秆还田土壤细菌群落结构和多样性进行表征.DGGE谱图和多样性分析结果表明,生物炭添加增加了秸秆还田土壤细菌DGGE图谱条带位置和灰度值及均匀度的变化,但两个处理土壤细菌群落Shannon-Wiener指数和丰富度没有显著差异(p0.05).聚类分析结果表明,SBW处理土壤细菌群落聚为5类,SW处理土壤细菌群落可聚为6类.DGGE条带测序和结构分析结果表明,生物炭添加导致秸秆还田土壤中Actinobacteria、Nitrospira消失和Spirochaetes、Gemmatimonadetes、Chloroflexi、WS3出现,并导致Firmicutes、Proteobacteria所占比例升高和Acidobacteria、Bacteroides所占比例降低,说明生物炭添加促进了秸秆还田土壤细菌群落组成结构变化.