Long-term effect of dissolved oxygen on partial nitrification performance and microbial community structure

In this study, the performance of partial nitrification via nitrite and microbial community structure were investigated and compared in two sequencing batch reactors (SBR) with different dissolved oxygen (DO) levels. Both reactors achieved stable partial nitrification with nitrite accumulation ratio of above 95% by using real-time aeration duration control. Compared with high DO (above 3 mg/l on average) SBR, simultaneous nitrification and denitrification (SND) via nitrite was carried out in low DO (0.4–0.8 mg/l) SBR. The average efficiencies of SND in high DO and low DO reactor were 7.7% and 44.9%, and the specific SND rates were 0.20 and 0.83 mg N/(mg MLSS h), respectively. Low DO did not produce sludge with poorer settling properties but attained lower turbidities of the effluent than high DO. Fluorescence in situ hybridization (FISH) analysis in both the reactors showed that ammonia-oxidizing bacteria (AOB) were the dominant nitrifying bacteria and nitrite-oxidizing bacteria (NOB) did not be recovered in spite of exposing nitrifying sludge to high DO. The morphology of the sludge from both two reactors according to scanning electron microscope indicated that small rod-shaped and spherical clusters were dominant, although filamentous bacteria and few long rod-shaped coexisted in the low DO reactor. By selecting properly DO level and adopting process control method is not only of benefit to the achievement of novel biological nitrogen removal technology, but also favorable to sludge population optimization.     

Microbial community developments and biomass characteristics in membrane bioreactors under different organic loadings

Microbial community developments and biomass characteristics (concentration, particle size, extracellular polymeric substances (EPS), and membrane fouling propensity) were compared when three MBRs were fed with the synthetic wastewater at different organic loadings. Results showed that the bacterial communities dynamically shifted in different ways and the EPS displayed dissimilar profiles under various organic loadings, which were associated with the ratios of food to microorganism and dissolved oxygen levels in the MBRs. The membrane fouling tendency of biomass in the low-loading MBR (0.57 g COD/L day) was insignificantly different from that in the medium-loading MBR (1.14 g COD/L day), which was apparently lower than that in the high-loading MBR (2.28 g COD/L day). The membrane fouling propensity of biomass was strongly correlated with their bound EPS contents, indicating cake layer fouling (i.e., deposition of microbial flocs) was predominant in membrane fouling at a high flux of 30 L/m² h.     

Control of the production of individual fusicoccins at different dissolved oxygen concentrations

The regulatory effect of different concentrations of dissolved oxygen on the production of fusicoccins by the fungus Fusicoccum amygdali Del. was studied. The maximum output of total fusicoccins was obtained by using a profiled dissolved oxygen tension (DOT) regime, in which the DOT was maintained at 15–20% during the biomass growth phase and at 5–8% during the fusicoccins production phase. In comparison with the profiled regime, the maintenance of DOT at 15–20% during the whole fermentation shortened the fusicoccins production phase. The fermentation performance at a low DOT (5–8%) inhibited both the accumulation of biomass and the production of fusicoccins. At high DOT (40–50%), an accelerated accumulation of the biomass with an expressed autolysis of mycelia took place, and the production of fusicoccins was lowered. The qualitative composition of individual fusicoccins varied substantially at different DOTs. Fusicoccins, A, C, D, J, H, 16-O-demethyl-J, detretpentenylfusicoccin and some minor fusicoccin metabolites were found in the fermentation broth using the method of liquid secondary ion mass spectrometry. It was established that the profiled DOT regime (15–20% to 5–8%) provided both the maximum concentration of fusicoccins and an enhanced accumulation of the main metabolite – fusicoccin A (FC A). The performance of the fermentation at a DOT of 15–20% decreased the content of FC A by 2–6% in comparison with the profiled DOT regime, and increased the content of fusicoccin C to 14–20% of the total fusicoccins. Fermentation at DOT of 5–8% was characterized by the highest content of the precursors of FC A, the less oxidized fusicoccins H and J, the contents of which were in range 7–12% and 16–17% of total fusicoccins, respectively.     

不同溶氧对谷氨酸棒杆菌代谢的影响

【目的】以谷氨酸棒杆菌为研究对象,分别控制在0、30%、50%3种溶氧水平下进行发酵,分析不同溶氧水平下代谢的变化。【方法】通过检测发酵代谢物中有机酸、氨基酸的含量,以及测定代谢途径中关键酶活性及其编码基因的表达情况来考察不同溶氧水平下物质代谢发生的变化。通过检测胞内还原力和ATP的含量来分析不同溶氧水平对能量代谢产生的影响。【结果】谷氨酸棒杆菌代谢支路受溶氧的影响而发生改变,氨基酸、有机酸的产量也随之改变。特别是在低溶氧(0)情况下,细胞内氧化磷酸化减弱,导致维持生命活动所必需的ATP供应减少,因此细胞通过增强底物水平磷酸化来产生ATP以满足生命活动的需求。在此情况下,胞内NADH得到较多积累,TCA循环代谢流量减小,而转向糖酵解、乙醛酸循环等,并且这个过程伴随多种杂酸包括乳酸、缬氨酸、亮氨酸等的产生,必将影响目的产物的产量。【结论】研究结果对于进一步采取措施优化溶氧的控制策略,提高目的产物的产量具有指导意义。     

Linking microbial community composition and soil processes in a California annual grassland and mixed-conifer forest

To investigate the potential role of microbial community composition in soil carbon and nitrogen cycling, we transplanted soil cores between a grassland and a conifer ecosystem in the Sierra Nevada California and measured soil process rates (N-mineralization, nitrous oxide and carbondioxide flux, nitrification potential), soil water and temperature, and microbial community parameters (PLFA and substrate utilization profiles) over a 2 year period. Our goal was to assess whether microbial community composition could be related to soil process rates independent of soil temperature and water content. We performed multiple regression analyses using microbial community parameters and soil water and temperature as X-variables and soil process rates and inorganic N concentrations as Y-variables. We found that field soil temperature had the strongest relationship with CO₂ production and soil NH₄⁺ concentration, while microbial community characteristics correlated with N₂O production, nitrification potential, gross N-mineralization, and soil NO₃^– concentration, independent of environmentalcontrollers. We observed a relationship between specific components of the microbial community (as determined by PLFA) and soil processes,particularly processes tightly linked to microbial phylogeny (e.g. nitrification). The most apparent change in microbial community composition in response to the 2 year transplant was a change in relative abundance of fungi (there was only one significant change in PLFA biomarkers for bacteria during 2 years). The relationship between microbial community composition and soil processes suggests that prediction of ecosystem response to environmental change may be improved by recognizing and accounting for changes in microbial community composition and physiological ecology.     

Dynamic variations of microbial community structure in Myriophyllum aquaticum constructed wetlands in response to different NH4+-N concentrations

Constructed wetlands (CWs) have received increasing attentions for their N removal performances, especially regarding NH₄⁺-N. Different influent NH₄⁺-N concentration may influence N removal efficiency in practice, while the effects of different NH₄⁺-N concentrations on microorganisms removing N in CWs are poorly understood. In this study, surface flow CWs planted with Myriophyllum (M). aquaticum were established to investigate the influences of different NH₄⁺-N concentrations on the composition, structure, and interactions of microbial community. Our findings suggested 105 mg/L NH₄⁺-N CWs achieved highest N removal rate, removing 89.30 % NH₄⁺-N and 92.34 % TN from the influent. The results of real-time quantitative polymerase chain reactions (qPCR) indicated abundances of nitrifying genes (nxrA) and denitrifying genes (narG, nirS, nirK, and nosZ) were increased by increasing NH₄⁺-N concentrations, and the strongest effects were observed in narG (8-fold) and nosZ genes (11-fold). Different NH₄⁺-N concentrations was proved to alter composition and structure of microbial communities via high-throughput sequencing, e.g. denitrifiers including Brevendomonas.sp, Dokdonella.sp and Rhodococcus.sp were enriched obviously with increasing NH₄⁺-N concentrations. In addition, network showed interactions among microbial populations and positive interactions were dramatically shifted and enhanced by increasing NH₄⁺-N concentrations.     

Effects of dilution on dissolved oxygen depletion and microbial populations in the biochemical oxygen demand determination

The biochemical oxygen demand (BOD) value is still a key parameter that can determine the level of organics, particularly the content of biodegradable organics in water. In this work, the effects of sample dilution, which should be done inevitably to get appropriate dissolved oxygen (DO) depletion, on the measurement of 5-day BOD (BOD₅), was investigated with and without seeding using natural and synthetic water. The dilution effects were also evaluated for water samples taken in different seasons such as summer and winter because water temperature can cause a change in the types of microbial species, thus leading to different oxygen depletion profiles during BOD testing. The predation phenomenon between microbial cells was found to be dependent on the inorganic nutrients and carbon sources, showing a change in cell populations according to cell size after 5-day incubation. The dilution of water samples for BOD determination was linked to changes in the environment for microbial growth such as nutrition. The predation phenomenon between microbial cells was more important with less dilution. BOD₅ increased with the specific amount of inorganic nutrient per microbial mass when the natural water was diluted. When seeding was done for synthetic water samples, the seed volume also affected BOD due to the rate of organic uptake by microbes. BOD₅ increased with the specific bacterial population per organic source supplied at the beginning of BOD measurement. For more accurate BOD measurements, specific guidelines on dilution should be established.     

从微生物组到合成功能菌群

微生物组是指特定微环境中的微生物群落及其组学,自然界中的生物过程几乎都是通过微生物组完成的。随着测序技术的发展和成本降低,微生物组已经成为微生物生态学研究的热点领域。继合成生物学之后,基于微生物组的合成功能菌群研究正在兴起,在人类健康、农业、工业和生态等领域都有广泛的应用前景。本文从微生物组到合成功能菌群的角度系统论述了其在不同领域的研究现状与发展趋势,为微生物组从理论研究到应用提供思路。     

Soil microbial community responses to altered lignin biosynthesis in Populus tremuloides vary among three distinct soils

The development and use of transgenic plants has steadily increased, but there are still little data about the responses of soil microorganisms to these genetic modifications. We utilized a greenhouse trial approach to evaluate the effects of altered stem lignin in trembling aspen (Populus tremuloides) on soil microbial communities in three soils which differed in their chemical and physical properties; they included a sandy loam (CO-Colorado), a silt loam (KS-Kansas), and a clay loam (TX-Texas). Three transgenic aspen lines were developed from a natural clone common to the Great Lakes region of North America. The concentrations of stem lignin concentrations were reduced by 35% (Line 23), 40% (Line 141) and 50% (Line 72). Line 72 and Line 141 also had a 40 and 20% increase in syringyl-type stem lignin, respectively. Indirectly, these modifications resulted in increased (5–13%) and decreased (−5 to −57%) levels of root production across the lines and soil types. Responses of the soil microbial communities were investigated using: phospholipid fatty acids (PLFA), neutral lipid fatty acids (NLFA), and 3) extracellular enzyme assays. PLFA analyses indicated that there were large differences in microbial community composition between the three soils. Similarly, there were large differences in total NLFA between soils, with the KS soils having the highest amount and CO the lowest. Enzyme activities did not differ between soils, except for cellubiohydrolase, which was highest in CO soil. Across all three soils, responses to the four genetic lines were not consistent. Interactions between soil type and genetic line make it difficult to assess the potential ecological impacts of transgenic aspen on soil microbial communities and their associated functions. Given these interactions, field trials with transgenic aspen should encompass the wide range of soils targeted for commercial planting in order to determine their effect(s) on the resident soil microbial community. Responsible Editor: Barbara Wick     

Membrane biofouling characterization: effects of sample preparation procedures on biofilm structure and the microbial community

Ensuring the quality and reproducibility of results from biofilm structure and microbial community analysis is essential to membrane biofouling studies. This study evaluated the impacts of three sample preparation factors (ie number of buffer rinses, storage time at 4°C, and DNA extraction method) on the downstream analysis of nitrifying biofilms grown on ultrafiltration membranes. Both rinse and storage affected biofilm structure, as suggested by their strong correlation with total biovolume, biofilm thickness, roughness and the spatial distribution of EPS. Significant variations in DNA yields and microbial community diversity were also observed among samples treated by different rinses, storage and DNA extraction methods. For the tested biofilms, two rinses, no storage and DNA extraction with both mechanical and chemical cell lysis from attached biofilm were the optimal sample preparation procedures for obtaining accurate information about biofilm structure, EPS distribution and the microbial community.     

Impacts of 2,4-D application on soil microbial community structure and on populations associated with 2,4-D degradation

The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) application rate on microbial community structure and on the diversity of dominant 2,4-D degrading bacteria in an agricultural soil was examined using cultivation-independent molecular techniques coupled with traditional isolation and enumeration methods. Fingerprints of microbial communities established under increasing concentrations of 2,4-D (0-500 mg kg-1) in batch soil microcosms were obtained using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene segments. While a 2,4-D concentration of at least 100 mg kg-1 was required to obtain an apparent change in the community structure as visualized by DGGE, the greatest impact of 2,4-D concentration occurred in the 500 mg kg-1 treatment, resulting in significantly reduced diversity of the dominant populations and enrichment by Burkholderia-like populations. The greatest diversity of 2,4-D degrading isolates was cultivated from the 10 mg kg-1 treatment, indicating that under these conditions, cultivation was more sensitive than DGGE for detecting changes in community structure. Most of these isolates harbored homologs of Ralstonia eutrophus JMP134 and Burkholderia cepacia tfdA catabolic genes. Results from this study revealed that agriculturally relevant application rates of 2,4-D may provide a temporary selective advantage for organisms capable of utilizing 2,4-D as a carbon and energy source.     

Electricity generation and microbial community changes in microbial fuel cells packed with different anodic materials

Four materials, carbon felt cube (CFC), granular graphite (GG), granular activated carbon (GAC) and granular semicoke (GS) were tested as packed anodic materials to seek a potentially practical material for microbial fuel cells (MFCs). The microbial community and its correlation with the electricity generation performance of MFCs were explored. The maximum power density was found in GAC, followed by CFC, GG and GS. In GAC and CFC packed MFCs, Geobacter was the dominating genus, while Azospira was the most populous group in GG. Results further indicated that GAC was the most favorable for Geobacter adherence and growth, and the maximum power densities had positive correlation with the total biomass and the relative abundance of Geobacter, but without apparent correlation with the microbial diversity. Due to the low content of Geobacter in GS, power generated in this system may be attributed to other microorganisms such as Synergistes, Bacteroidetes and Castellaniella.     

Analysis of microbial communities developed on the fouling layers of a membrane-coupled anaerobic bioreactor applied to wastewater treatment

The structure of the biofouling layers formed on a pilot-scale membrane-coupled upflow anaerobic sludge blanket bioreactor (UASB) used to treat urban wastewater was analyzed by scanning electron microscopy and electron-dispersive X-ray microanalysis. For comparison, control samples of the membranes were fed either UASB effluent or raw wastewater in a laboratory-scale experiment. Microbial diversity in the fouling materials was analyzed by temperature gradient gel electrophoresis (TGGE) combined with sequence analysis of partial 16S rRNA. Significant differences in structure of the Bacteria communities were observed amongst the different fouling layers analyzed in the UASB membranes, particularly following a chemical cleaning step (NaClO), while the Archaea communities retained more similarity in all samples. The main Bacteria populations identified were evolutively close to Firmicutes (42.3%) and Alphaproteobacteria (30.8%), while Archaea were mostly affiliated to the Methanosarcinales and Methanospirillaceae. Sphingomonadaceae-related bacteria and methanogenic Archaea were persistently found as components of biofouling, regardless of chemical cleaning.     

Investigation of the microbial community structure and activity as indicators of compost stability and composting process evolution

In a bid to identify suitable microbial indicators of compost stability, the process evolution during windrow composting of poultry manure (PM), green waste (GW) and biowaste was studied. Treatments were monitored with regard to abiotic factors, respiration activity (determined using the SOUR test) and functional microflora. The composting process went through typical changes in temperature, moisture content and microbial properties, despite the inherent feedstock differences. Nitrobacter and pathogen indicators varied as a monotonous function of processing time. Some microbial groups have shown a potential to serve as fingerprints of the different process stages, but still they should be examined in context with respirometric tests and abiotic parameters. Respiration activity reflected well the process stage, verifying the value of respirometric tests to access compost stability. SOUR values below 1 mg O₂/g VS/h were achieved for the PM and the GW compost.     

Effect of dilution rate on the microbial structure of a mesophilic butyrate-degrading methanogenic community during continuous cultivation

We constructed two mesophilic anaerobic chemostats that were continuously fed with synthetic wastewater containing butyrate as the sole source of carbon and energy. Steady-state conditions were achieved at dilution rates between 0.025 and 0.7 day⁻¹. Butyrate, fed into the chemostat, was almost completely mineralized to CH₄ and CO₂ at dilution rates below 0.5 day⁻¹. The butyrate-degrading methanogenic communities in the chemostats at dilution rates between 0.025 and 0.7 day⁻¹ were monitored based on the 16S rRNA gene, using molecular biological techniques including clone library analysis, denaturing gradient gel electrophoresis, and quantitative real-time polymerase chain reaction. The aceticlastic methanogen Methanosaeta and the hydrogenotrophic methanogen Methanoculleus dominated in methanogens at low dilution rates, whereas the aceticlastic methanogen Methanosaeta, Methanosarcina, the hydrogenotrophic methanogen Methanoculleus, and Methanospirillum dominated at high dilution rates. Bacteria affiliated with the family Syntrophaceae in the phylum Proteobacteria predominated at the low dilution rate of 0.025 day⁻¹, whereas bacteria affiliated with the phylum Firmicutes and Candidate division OP3 predominated at high dilution rates. A significant quantity of bacteria closely related to the genus Syntrophomonas was detected at high dilution rates. Dilution rate showed an apparent effect on archaeal and bacterial communities in the butyrate-fed chemostats.     

Increases in soil respiration following labile carbon additions linked to rapid shifts in soil microbial community composition

Organic matter decomposition and soil CO₂ efflux are both mediated by soil microorganisms, but the potential effects of temporal variations in microbial community composition are not considered in most analytical models of these two important processes. However, inconsistent relationships between rates of heterotrophic soil respiration and abiotic factors, including temperature and moisture, suggest that microbial community composition may be an important regulator of soil organic matter (SOM) decomposition and CO₂ efflux. We performed a short-term (12-h) laboratory incubation experiment using tropical rain forest soil amended with either water (as a control) or dissolved organic matter (DOM) leached from native plant litter, and analyzed the effects of the treatments on soil respiration and microbial community composition. The latter was determined by constructing clone libraries of small-subunit ribosomal RNA genes (SSU rRNA) extracted from the soil at the end of the incubation experiment. In contrast to the subtle effects of adding water alone, additions of DOM caused a rapid and large increase in soil CO₂ flux. DOM-stimulated CO₂ fluxes also coincided with profound shifts in the abundance of certain members of the soil microbial community. Our results suggest that natural DOM inputs may drive high rates of soil respiration by stimulating an opportunistic subset of the soil bacterial community, particularly members of the Gammaproteobacteria and Firmicutes groups. Our experiment indicates that variations in microbial community composition may influence SOM decomposition and soil respiration rates, and emphasizes the need for in situ studies of how natural variations in microbial community composition regulate soil biogeochemical processes.     

Stability and microbial community structure of a partial nitrifying fixed-film bioreactor in long run

A partial nitrification system was investigated for 471 days under DO varying concentrations for assessing its stability and population dynamics. Within 130 days of operation at feed DO concentration of 1.0 ± 0.1 mg/L, more than 85% of nitrite was accumulated. Efficiency deteriorated when the feed DO concentration was increased to 4.2 ± 0.3 mg/L. Nitrite accumulation could not be re-established on decreasing feed DO to 1.0 ± 0.1 mg/L. Even at DO concentration of <0.05 mg/L, nitrate production was observed; a condition termed as anoxic nitrification. NOB was detected in the biomass even under this condition by Fluorescence in-situ hybridization (FISH) analysis. Through 16S rRNA gene sequencing a major fraction of unknown bacterial sequences closely resembling haloalkalophilic bacteria of marine origin were detected. The study indicated that these bacterial species might play a role in anoxic nitrification and that NOB could survive extreme low DO condition.     

Effects of temperature and temperature shock on the performance and microbial community structure of a submerged anaerobic membrane bioreactor

Effects of temperature and temperature shock on the performance and microbial community structure of a submerged anaerobic membrane bioreactor (SAnMBR) treating thermomechanical pulping pressate were studied for 416 days. The results showed that the SAnMBR system were highly resilient to temperature variations in terms of chemical oxygen demand (COD) removal. The residual COD in treated effluent was slightly higher at 55 °C than that at 37 and 45 °C. There were no significant changes in biogas production rate and biogas composition. However, temperature shocks resulted in an increase in biogas production temporarily. The SAnMBR could tolerate the 5 and 10 °C temperature shocks at 37 °C and the temperature variations from 37 to 45 °C. The temperature shock of 5 and 10 °C at 45 °C led to slight and significant disturbance of the performance, respectively. Temperature affected the richness and diversity of microbial populations.