Changes in freshwater bacterial community composition during measurements of microbial and community respiration

The respiration rates of a pelagic community and of its microbialfraction (< 1.2 µm) were measured at two depths inthe oxic layer of a meromictic alpine lake (Cadagno, Switzerland)using the oxygen technique. The duration of the incubationswere 12, 24 and 55 h. Bacterioplankton abundance (DAPI counts)and composition (whole cell hybridization using 11 group-specificrRNA-targeted oligonucleotide probes) were measured during theincubations. Respiration generally increased with time, especiallyin the microbial fraction, or remained similar. This resultwas not always consistent with changes in bacterial abundanceand cell volume. The composition of the community also changedduring the incubations. The abundance of ß-Proteobacteriaincreased during the course of all the experiments. These resultsextend the previous conclusions drawn in marine environmentsto fresh waters and demonstrate that, in addition to changesin bacterial abundance, cell volume and biomass, changes inthe taxonomic composition of the bacterial community can occurduring discrete incubations of freshwater planktonic communities.     

Changes in microbial community composition and geochemistry during uranium and technetium bioimmobilization

In a previous column study, we investigated the long-term impact of ethanol additions on U and Tc mobility in groundwater (M. M. Michalsen et al., Environ. Sci. Technol. 40:7048-7053, 2006). Ethanol additions stimulated iron- and sulfate-reducing conditions and significantly enhanced U and Tc removal from groundwater compared to an identical column that received no ethanol additions (control). Here we present the results of a combined signature lipid and nucleic acid-based microbial community characterization in sediments collected from along the ethanol-stimulated and control column flow paths. Phospholipid fatty acid analysis showed both an increase in microbial biomass (approximately 2 orders of magnitude) and decreased ratios of cyclopropane to monoenoic precursor fatty acids in the stimulated column compared to the control, which is consistent with electron donor limitation in the control. Spatial shifts in microbial community composition were identified by PCR-denaturing gradient gel electrophoresis analysis as well as by quantitative PCR, which showed that Geobacteraceae increased significantly near the stimulated-column outlet, where soluble electron acceptors were largely depleted. Clone libraries of 16S rRNA genes from selected flow path locations in the stimulated column showed that Proteobacteria were dominant near the inlet (46 to 52%), while members of candidate division OP11 were dominant near the outlet (67%). Redundancy analysis revealed a highly significant difference (P = 0.0003) between microbial community compositions within stimulated and control sediments, with geochemical variables explaining 68% of the variance in community composition on the first two canonical axes.     

Changes in microbial community structure and function during vermicomposting of pig slurry

Most studies investigating the effects of earthworms on microorganisms have focused on the changes before and after vermicomposting rather than those that occur throughout the process. In the present study, we designed continuous feeding reactors in which new layers of pig slurry (1.5 and 3 kg) were added sequentially to form an age gradient inside the reactors in order to evaluate the impact of the earthworm species Eisenia fetida on microbial community structure and function. The activity of earthworms greatly reduced the bacterial and fungal biomass and microbial diversity relative to the control values. However, the pronounced presence of earthworms in the younger layers stimulated microbial activity and as such increased carbon mineralization probably due to the fact that the microorganisms may have been less resource-limited as a result of earthworm activity, as indicated by the ratio of monounsaturated to saturated PLFAs.     

Dynamics of microbial community composition and function during in situ bioremediation of a uranium-contaminated aquifer

A pilot-scale system was established to examine the feasibility of in situ U(VI) immobilization at a highly contaminated aquifer (U.S. DOE Integrated Field Research Challenge site, Oak Ridge, TN). Ethanol was injected intermittently as an electron donor to stimulate microbial U(VI) reduction, and U(VI) concentrations fell to below the Environmental Protection Agency drinking water standard (0.03 mg liter(-1)). Microbial communities from three monitoring wells were examined during active U(VI) reduction and maintenance phases with GeoChip, a high-density, comprehensive functional gene array. The overall microbial community structure exhibited a considerable shift over the remediation phases examined. GeoChip-based analysis revealed that Fe(III)-reducing bacterial (FeRB), nitrate-reducing bacterial (NRB), and sulfate-reducing bacterial (SRB) functional populations reached their highest levels during the active U(VI) reduction phase (days 137 to 370), in which denitrification and Fe(III) and sulfate reduction occurred sequentially. A gradual decrease in these functional populations occurred when reduction reactions stabilized, suggesting that these functional populations could play an important role in both active U(VI) reduction and maintenance of the stability of reduced U(IV). These results suggest that addition of electron donors stimulated the microbial community to create biogeochemical conditions favorable to U(VI) reduction and prevent the reduced U(IV) from reoxidation and that functional FeRB, SRB, and NRB populations within this system played key roles in this process.     

Changes in soil microbial community structure and function following degradation in a temperate grassland

温带草原退化后土壤微生物群落结构和功能的变化 草原退化是草原生产力维持面临的一个重大挑战,这一过程显著影响着草原生态系统的能量流动和土壤养分变化过程,进而直接或间接地影响着土壤微生物。我们的研究目的首先是调查不同草原退化程度(即未退化、中度退化和严重退化)如何影响着内蒙古温带草原的土壤微生物组成、多样性和功能,其次是阐明哪些生物和非生物因素导致了这些变化。我们的研究主要通过高通量测序技术分析土壤微生物的群落组成,并且采用FAPROTAX工具和FUNGuild工具分别预测细菌群落和真菌群落的功能。研究发现：草原退化显著降低了土壤细菌的多样性,但对真菌多样性影响不大。地下生物量、土壤有机碳和总氮与细菌的多样性变化呈显著正相关关系。草原退化显著增加了绿弯菌门的相对丰度(由2.48%提高到8.40%),降低了厚壁菌门的相对丰度(由3.62%降低到1.08%)。其次,草原退化也显著增加了球囊菌门的相对丰度(从0.17%提高到1.53%),降低了担子菌门的相对丰度(从19.30%降低到4.83%)。致病菌的相对丰度在草原退化过程中显著下降。此外,草原退化对土壤细菌群落的功能有显著的影响,尤其是与土壤碳氮循环相关的土壤细菌群落。我们的结果表明,土壤细菌群落对草原退化的响应比真菌群落更敏感。     

Changes in the microbial community during repeated anaerobic microbial dechlorination of pentachlorophenol

Pentachlorophenol (PCP) has been widely used as a pesticide in paddy fields and has imposed negative ecological effect on agricultural soil systems, which are in typically anaerobic conditions. In this study, we investigated the effect of repeated additions of PCP to paddy soil on the microbial communities under anoxic conditions. Acetate was added as the carbon source to induce and accelerate cycles of the PCP degradation. A maximum degradation rate occurred at the 11th cycle, which completely transformed 32.3 μM (8.6 mg L^?1) PCP in 5 days. Illumina high throughput sequencing of 16S rRNA gene was used to profile the diversity and abundance of microbial communities at each interval and the results showed that the phyla of Bacteroidates, Firmicutes, Proteobacteria, and Euryarchaeota had a dominant presence in the PCP-dechlorinating cultures. Methanosarcina, Syntrophobotulus, Anaeromusa, Zoogloea, Treponema, W22 (family of Cloacamonaceae), and unclassified Cloacamonales were found to be the dominant genera during PCP dechlorination with acetate. The microbial community structure became relatively stable as cycles increased. Treponema, W22, and unclassified Cloacamonales were firstly observed to be associated with PCP dechlorination in the present study. Methanosarcina that have been isolated or identified in PCP dechlorination cultures previously was apparently enriched in the PCP dechlorination cultures. Additionally, the iron-cycling bacteria Syntrophobotulus, Anaeromusa, and Zoogloea were enriched in the PCP dechlorination cultures indicated they were likely to play an important role in PCP dechlorination. These findings increase our understanding for the microbial and geochemical interactions inherent in the transformation of organic contaminants from iron rich soil, and further extend our knowledge of the PCP-transforming microbial communities in anaerobic soil conditions.     

Changes in soil microbial community composition induced by cometabolism of toluene and trichloroethylene

The effects of trichloroethylene (TCE) on microbial community composition were analyzed by reverse sample genome probing. Soil enrichments were incubated in dessicators containing an organic phase of either 1 or 10% (w/w) toluene in vacuum pump oil, delivering constant equilibrium aqueous concentrations of 16 and 143mg/l, respectively. Increasing the equilibrium aqueous concentration of TCE from 0 to 10mg/l led to shifts in community composition at 16, but not at 143mg/l of toluene. In closed system co-degradation studies, TCE at an aqueous concentration of 1mg/1 was effectively degraded by toluene-degrading soil enrichments once the aqueous toluene concentration dropped below 25mg/l. Little TCE degradation was observed at higher toluene concentrations (50–250mg/l). The results indicate that TCE changes microbial community composition under conditions where it is being actively metabolized.     

Association of microbial community composition and activity with lead,chromium, and hydrocarbon contamination

Microbial community composition and activity were characterized in soil contaminated with lead (Pb), chromium (Cr), and hydrocarbons. Contaminant levels were very heterogeneous and ranged from 50 to 16,700 mg of total petroleum hydrocarbons (TPH) kg of soil(-1), 3 to 3,300 mg of total Cr kg of soil(-1), and 1 to 17,100 mg of Pb kg of soil(-1). Microbial community compositions were estimated from the patterns of phospholipid fatty acids (PLFA); these were considerably different among the 14 soil samples. Statistical analyses suggested that the variation in PLFA was more correlated with soil hydrocarbons than with the levels of Cr and Pb. The metal sensitivity of the microbial community was determined by extracting bacteria from soil and measuring [(3)H]leucine incorporation as a function of metal concentration. Six soil samples collected in the spring of 1999 had IC(50) values (the heavy metal concentrations giving 50% reduction of microbial activity) of approximately 2.5 mM for CrO(4)2- and 0.01 mM for Pb2+. Much higher levels of Pb were required to inhibit [14C]glucose mineralization directly in soils. In microcosm experiments with these samples, microbial biomass and the ratio of microbial biomass to soil organic C were not correlated with the concentrations of hydrocarbons and heavy metals. However, microbial C respiration in samples with a higher level of hydrocarbons differed from the other soils no matter whether complex organic C (alfalfa) was added or not. The ratios of microbial C respiration to microbial biomass differed significantly among the soil samples (P < 0.05) and were relatively high in soils contaminated with hydrocarbons or heavy metals. Our results suggest that the soil microbial community was predominantly affected by hydrocarbons.     

Bioremediation of a polyaromatic hydrocarbon contaminated soil by native soil microbiota and bioaugmentation with isolated microbial consortia

Biodegradation of a mixture of PAHs was assessed in forest soil microcosms performed either without or with bioaugmentation using individual fungi and bacterial and a fungal consortia. Respiratory activity, metabolic intermediates and extent of PAH degradation were determined. In all microcosms the low molecular weight PAH’s naphthalene, phenanthrene and anthracene, showed a rapid initial rate of removal. However, bioaugmentation did not significantly affect the biodegradation efficiency for these compounds. Significantly slower degradation rates were demonstrated for the high molecular weight PAH’s pyrene, benz[a]anthracene and benz[a]pyrene. Bioaugmentation did not improve the rate or extent of PAH degradation, except in the case of Aspergillus sp. Respiratory activity was determined by CO₂ evolution and correlated roughly with the rate and timing of PAH removal. This indicated that the PAHs were being used as an energy source. The native microbiota responded rapidly to the addition of the PAHs and demonstrated the ability to degrade all of the PAHs added to the soil, indicating their ability to remediate PAH-contaminated soils.     

Impact of an indigenous microbial enhanced oil recovery field trial on microbial community structure in a high pour-point oil reservoir

Based on preliminary investigation of microbial populations in a high pour-point oil reservoir, an indigenous microbial enhanced oil recovery (MEOR) field trial was carried out. The purpose of the study is to reveal the impact of the indigenous MEOR process on microbial community structure in the oil reservoir using 16Sr DNA clone library technique. The detailed monitoring results showed significant response of microbial communities during the field trial and large discrepancies of stimulated microorganisms in the laboratory and in the natural oil reservoir. More specifically, after nutrients injection, the original dominant populations of Petrobacter and Alishewanella in the production wells almost disappeared. The expected desirable population of Pseudomonas aeruginosa, determined by enrichment experiments in laboratory, was stimulated successfully in two wells of the five monitored wells. Unexpectedly, another potential population of Pseudomonas pseudoalcaligenes which were not detected in the enrichment culture in laboratory was stimulated in the other three monitored production wells. In this study, monitoring of microbial community displayed a comprehensive alteration of microbial populations during the field trial to remedy the deficiency of culture-dependent monitoring methods. The results would help to develop and apply more MEOR processes.     

N-acyl-l-homoserine lactones (AHLs) affect microbial community composition and function in activated sludge

The role of intercellular signalling in the regulation of genes and phenotypes in a broad range of bacterial species is now firmly established. In contrast, the impact of intercellular signalling on microbial community parameters, such as species diversity and function, is less well understood. In this study the role of N-acyl-l-homoserine lactones (AHLs) in microbial community dynamics in an industrial wastewater treatment system is addressed. Seven proteobacterial strains producing compounds with AHL-like activity were isolated from the treatment plant. Three of these belong to genera with no previously identified AHL producing species. Addition of AHLs at 2 micro M to sludge samples generated changes in both community function (phenol degradation) and composition as determined by length heterogeneity PCR and denaturing gradient gel electrophoresis. Phenol degradation was more stable as a result of the AHL augmentation. A dominant functional member of the Thauera genus was transiently supplanted by a member of the Comomonas genus in response to AHL addition. This suggests that AHLs can play a role in mediating microbial community parameters and has implications for ecosystem function and industrial wastewater treatment.     

Changes in functional abilities of the microbial community during composting of manure

The objective of this study was (a) to detect changes of the functional abilities of the microflora during composting of manure as a result of windrow turning frequency and (b) to detect differences between distinct zones within the windrows. Biolog GN microtiter plates containing 95 different carbon sources were inoculated with diluted suspensions of compost material containing 15,000 microorganisms per well (120 l). We found a dramatic shift in functional microbial community structure during the 8-week composting process. The shift was more rapid when the compost windrows were turned. The substrate use pattern in the outer, well-aerated zone of the unturned windrow was similar to that of the turned windrows. Microbial biomass and respiration decreased more rapidly in the turned than in the unturned windrows, indicating a different pace of compost maturation. The data suggest that the Biolog assay may be a suitable approach to determine compost maturity. Correspondence to: H. Insam     

Short-term lime application impacts microbial community composition and potential function in an acid black soil

Plant and Soil - Soil acidification is a natural process that can be accelerated by intensive nitrogen fertilization. Lime application is a typical agricultural practice to enhance soil pH and...     

Link between microbial composition and carbon substrate-uptake preferences in a PHA-storing community

The microbial community of a fermented molasses-fed sequencing batch reactor (SBR) operated under feast and famine conditions for production of polyhydroxyalkanoates (PHAs) was identified and quantified through a 16 S rRNA gene clone library and fluorescence in situ hybridization (FISH). The microbial enrichment was found to be composed of PHA-storing populations (84% of the microbial community), comprising members of the genera Azoarcus, Thauera and Paracoccus. The dominant PHA-storing populations ensured the high functional stability of the system (characterized by high PHA-storage efficiency, up to 60% PHA content). The fermented molasses contained primarily acetate, propionate, butyrate and valerate. The substrate preferences were determined by microautoradiography-FISH and differences in the substrate-uptake capabilities for the various probe-defined populations were found. The results showed that in the presence of multiple substrates, microbial populations specialized in different substrates were selected, thereby co-existing in the SBR by adapting to different niches. Azoarcus and Thauera, primarily consumed acetate and butyrate, respectively. Paracoccus consumed a broader range of substrates and had a higher cell-specific substrate uptake. The relative species composition and their substrate specialization were reflected in the substrate removal rates of different volatile fatty acids in the SBR reactor.     

荒漠-绿洲土壤微生物群落组成与其活性对比

结合野外观测与实验室研究方法,对比研究了准葛尔盆地南缘盐生荒漠与绿洲农田土壤微生物活性与其群落组成的变化特征,并分析了土壤温度与湿度对荒漠-绿洲土壤微生物活性的影响。结果表明:荒漠开垦为绿洲后,土壤细菌明显增加,真菌无明显变化,放线菌显著减少。细菌在绿洲农田土壤矿化作用中占主导,真菌则在荒漠中占优势,绿洲农田土壤微生物活性(包括真菌与细菌活性)明显高于荒漠。温度对荒漠-绿洲土壤微生物活性的影响只在一定土壤湿度范围内作用显著,绿洲农田受其影响较大;荒漠有机质含量明显高于绿洲农田,但水分与盐分因素抑制了微生物对其的分解和矿化。不同土地利用方式导致了荒漠绿洲间土壤湿度及盐份的较大差异,加之与土壤温度极显著的交互作用,使得开垦后土壤有机碳的易得性增强,微生物群落结构发生显著改变,进而有机碳的矿化速率加快,土壤碳库随之消减。     

微生物菌剂对猪粪堆肥中细菌群落结构的影响

以猪粪和小麦秸秆做堆肥试验,处理组添加外源微生物菌剂,利用常规方法对堆肥样品进行理化性状测定,采用高通量测序技术分析堆肥过程中细菌群落特征。理化性状测定结果表明: 添加外源菌剂可延长堆肥高温时间,降低堆肥发酵末期的pH,增加全氮含量,加快C/N的下降。主成分分析表明: 外源菌剂影响堆肥样品细菌群落的稳定性。门分类水平上,厚壁菌门、变形菌门和绿弯菌门的相对丰度在处理组中较高;纲分类水平上,梭状芽孢杆菌纲、α-变形菌纲和γ-变形菌纲在处理组的升温期和高温期相对丰度增加;科分类水平上,小单孢菌科和梭状芽孢杆菌纲的消化链球菌科、梭菌科以及盐厌氧菌科的相对丰度在处理组的升温期和高温期均呈上升趋势。Pearson相关性分析表明,盐胞菌属与外源菌剂呈显著正相关,而氨苄芽孢杆菌属与外源菌剂呈显著负相关。研究表明,猪粪堆肥中添加外源菌剂可使堆肥的理化性质和细菌群落结构均发生显著变化。