不同水分管理下稻田土壤CH4和N2O排放与微生物菌群的关系

采用MPN计数法对黑土(海伦)和草甸棕壤(沈阳)稻田生长季4种微生物菌群数量进行了测定,同时采用封闭式箱法对CH4和N2O通量进行观测,以深入了解稻田生物源温室气体排放的微生物学过程,两地试验田均采用长期淹灌与间歇灌溉两种不同水分管理,对实验结果多元回归分析,结果表明,海伦与沈阳两地稻田两种水分管理条件下CH4通量季节变化与产甲烷菌数季节变化存在极显著正相关关系沈阳稻田生长季CH4通量季节变化与甲烷氧化菌数季节变化具有显著正相关性,间歇灌溉条件下黑土稻田N2O通量与反硝化菌数呈显著性正相关关系,两种水分管理条件下沈阳稻田N2O通量与硝化菌数具有显著正相关关系,间歇灌溉条件下沈阳稻田N2O通量与反硝化菌数呈显著性正相关关系。     

Inhibition of Acetoclastic Methanogenesis in Crude Oil- and Creosote-Contaminated Groundwater

Results from a series of studies of methanogenic processes in crude oil- and creosote-contaminated aquifers indicate that acetoclastic methanogenesis is inhibited near non-aqueous sources. At a crude oil-contaminated site, numbers of acetoclastic methanogens found close to crude oil were one hundred times fewer than those of hydrogen- and formate-utilizing methanogens. In laboratory toxicity assays, crude oil collected from the site inhibited methane production from acetate but not from formate or hydrogen. Toxicity assays with aqueous creosote extract completely inhibited acetate utilization over the range of tested dilutions but only mildly affected formate and hydrogen utilization. The combined results from the laboratory and field studies suggest that in methanogenic contaminated aquifers, inhibition of acetoclastic methanogenesis may lead to a buildup of acetate relative to dissolved organic carbon.     

Molecular characterization of mesophilic and thermophilic sulfate reducing microbial communities in expanded granular sludge bed (EGSB) reactors

The microbial communities established in mesophilic and thermophilic expanded granular sludge bed reactors operated with sulfate as the electron acceptor were analyzed using 16S rRNA targeted molecular methods, including denaturing gradient gel electrophoresis, cloning, and phylogenetic analysis. Bacterial and archaeal communities were examined over 450 days of operation treating ethanol (thermophilic reactor) or ethanol and later a simulated semiconductor manufacturing wastewater containing citrate, isopropanol, and polyethylene glycol 300 (mesophilic reactor), with and without the addition of copper(II). Analysis, of PCR-amplified 16S rRNA gene fragments using denaturing gradient gel electrophoresis revealed a defined shift in microbial diversity in both reactors following a change in substrate composition (mesophilic reactor) and in temperature of operation from 30°C to 55°C (thermophilic reactor). The addition of copper(II) to the influent of both reactors did not noticeably affect the composition of the bacterial or archaeal communities, which is in agreement with the very low soluble copper concentrations (3–310 μg l⁻¹) present in the reactor contents as a consequence of extensive precipitation of copper with biogenic sulfides. Furthermore, clone library analysis confirmed the phylogenetic diversity of sulfate-reducing consortia in mesophilic and thermophilic sulfidogenic reactors operated with simple substrates.     

Effects of influent DO/COD ratio on the performance of an anaerobic fluidized bed reactor fed low-strength synthetic wastewater

The effect of influent DO/COD (dissolved oxygen/chemical oxygen demand) ratio on the performance of an anaerobic fluidized bed reactor (AFBR) containing GAC was studied. A high influent DO concentration was found to have adverse impacts on organic removal efficiency, methane production, and effluent suspended solids (SS) concentration. These problems resulted with a DO/COD ratio of 0.12, but not at a lower ratio of 0.05. At first organic removal appeared satisfactory at the higher DO/COD ratio at a hydraulic retention time of 0.30 h, but soon a rapid growth of oxygen-consuming zoogloeal-like organisms resulted, eventually causing high effluent SS concentrations. The influent DO also had an inhibitory effect, resulting in a long recovery time for adequate methanogenic activity to return after influent DO removal began. With the growing interest in anaerobic treatment of low COD wastewaters, the increased possibility of similar adverse DO effects occurring needs consideration.     

Butachlor inhibits production and oxidation of methane in tropical rice soils under flooded condition

In laboratory incubation experiments, application of a commercial formulation of the herbicide butachlor (N-butoxymethyl-2-chloro-2',6'-diethyl acetanilide) to three tropical rice soils, widely differing in their physicochemical characteristics, under flooded condition inhibited methane (CH4) production. The inhibitory effect was concentration dependent and most remarkable in the alluvial soil. Thus, following application of butachlor at 5, 10, 50 and 100 microg g(-1) soil, respectively, cumulative CH4 production in the alluvial soil was inhibited by 15%, 31%, 91% and 98% over unamended control. Since CH4 production was less pronounced in the sandy loam and acid sulfate soil, the impact of amendment with butchalor, albeit inhibitory, was less extensive than the alluvial soil. Inhibition of CH4 production in butachlor-amended alluvial soil was related to the prevention in the drop in redox potential as well as low methanogenic bacterial population especially at high concentrations of butachlor. CH4 oxidation was also inhibited in butachlor-amended alluvial soil with the inhibitory effect being more prevalent under flooded condition. Inhibition in CH4 oxidation was related to a reduction in the population of soluble methane monooxygenase producing methanotrophs. Results demonstrate that butachlor, a commonly used herbicide in rice cultivation, even at very low concentrations can affect CH4 production and its oxidation, thereby influencing the biogeochemical cycle of CH4 in flooded rice soils.     

The archaeal flagellum: a different kind of prokaryotic motility structure

The archaeal flagellum is a unique motility apparatus distinct in composition and likely in assembly from the bacterial flagellum. Gene families comprised of multiple flagellin genes co-transcribed with a number of conserved, archaeal-specific accessory genes have been identified in several archaea. However, no homologues of any bacterial genes involved in flagella structure have yet been identified in any archaeon, including those archaea in which the complete genome sequence has been published. Archaeal flagellins possess a highly conserved hydrophobic N-terminal sequence that is similar to that of type IV pilins and clearly unlike that of bacterial flagellins. Also unlike bacterial flagellins but similar to type IV pilins, archaeal flagellins are initially synthesized with a short leader peptide that is cleaved by a membrane-located peptidase. With recent advances in genetic transfer systems in archaea, knockouts have been reported in several genes involved in flagellation in different archaea. In addition, techniques to isolate flagella with attached hook and anchoring structures have been developed. Analysis of these preparations is under way to identify minor structural components of archaeal flagella. This and the continued isolation and characterization of flagella mutants should lead to significant advances in our knowledge of the composition and assembly of archaeal flagella.     

A novel pathway of peptide biosynthesis found in methanogenic Archaea

The peptide subunits of the pseudomurein, the cell-wall peptidoglycan of some methanogens, are usually composed of glutamic acid, alanine and lysine. In order to get a more detailed picture of the biosynthetic pathway of the peptide subunit, we performed in vitro assays. Starting from glutamic acid a pentapeptide was obtained in seven steps:

Effect of methanogenic substrates on coenzyme F420-dependent N5,N10-methylene-H4MPT dehydrogenase,N5,N10-methenyl-H4MPT cyclohydrolase and F420-reducing hydrogenase activities in Methanosarcina barkeri

We measured F₄₂₀-dependent N⁵,N¹⁰-methylenetetrahydro-methanopterin dehydrogenase, N⁵, N¹⁰-methenyltetrahydro-methanopterin cyclohydrolase, and F₄₂₀-reducing hydrogenase levels in Methanosarcina barkeri grown on various substrates. Variation in dehydrogenase levels during growth on a specific substrate was usually <3-fold, and much less for cyclohydrolase. H₂–CO₂-, methanol-, and H₂–CO₂+ methanol-grown cells had roughly equivalent levels of dehydrogenase and cyclohydrolase. In acetate-grown cells cyclohydrolase level was lowered 2 to 3-fold and dehydrogenase 10 to 80-fold; this was not due to repression by acetate, since, if cultures growing on acetate were supplemented with methanol or H₂–CO₂, dehydrogenase levels increased 14 to 19-fold, and cyclohydrolase levels by 3 to 4-fold. Compared to H₂–CO₂- or methanol-grown cells, acetate-or H₂–CO₂ + methanol-grown cells had lower levels of and less growth phase-dependent variation in hydrogenase activity. Our data are consistent with the following hypotheses: 1. M. barkeri oxidizes methanol via a portion of the CO₂-reduction pathway operated in the reverse direction. 2. When steps from CO₂ to CH₃-S-CoM in the CO₂-reduction pathway (in either direction) are not used for methanogenesis, hydrogenase activity is lowered.Abbreviations MF methanofuran - H₄MPT 5,6,7,8-tetrahydromethanopterin - HS-HTP 7-mercaptoheptanoylthreonine phosphate - CoM-S-S-HTP heterodisulfide of HS-CoM and HS-HTP - F₄₂₀ coenzyme F₄₂₀ (a 7,8-didemethyl-8-hydroxy-5-deaza-riboflavin derivative) - H₂F₄₂₀ reduced coenzyme F₄₂₀ - HC⁺=H₄MPT N⁵,N¹⁰-methenyl-H₄MPT - H₂C=H₄MPT N⁵,N¹⁰-methylene-H₄MPT - H₃C=H₄MPT N⁵-methyl-H₄MPT - BES 2-bromoethanesulfonic acid     

Effect of various anionic species on net methane production in flooded rice soils

In a laboratory incubation study, effect of various anions on net methane production in two rice soils (alluvial and acid sulphate) under flooded conditions was examined. Methane production was considerable in alluvial soil and almost negligible in acid sulphate soil, albeit with a higher density of viable methanogens, during 30-day incubation without salts. Sodium salts of hydroxide and phosphate further stimulated methane production in alluvial soil and marginally in acid sulphate soil. But, addition of sodium molybdate, a selective inhibitor of sulphate-reducing bacteria, increased the production of methane in acid sulphate soil. In contrast, nitrite, nitrate, sulphite and sulphate suppressed the production of methane in both soils. Acetate served as an excellent substrate for methanogenesis in alluvial soil, but not in acid sulphate soil. Succinate and citrate also stimulated methane production especially in alluvial soil, but after a longer lag. In acid sulphate soil, most of the added carbon in the form of sodium salts of carboxylic acids was converted to CO2 and not methane, which is consistent with their preferential use by the sulphate-reducing bacteria. In general, none of the amendments could increase production of methane in acid sulphate soil to the same level as in alluvial soil.