Fe(III) mineral formation and cell encrustation by the nitrate-dependent Fe(II)-oxidizer strain BoFeN1

Understanding the mechanisms of anaerobic microbial iron cycling is necessary for a full appreciation of present‐day biogeochemical cycling of iron and carbon and for drawing conclusions about these cycles on the ancient Earth. Towards that end, we isolated and characterized an anaerobic nitrate‐dependent Fe(II)‐oxidizing bacterium from a freshwater sediment. The 16SrRNA gene sequence of the isolated bacterium (strain BoFeN1) places it within the β‐Proteobacteria, with Acidovorax sp. strain G8B1 as the closest known relative. During mixotrophic growth with acetate plus Fe(II) and nitrate as electron acceptor, strain BoFeN1 forms Fe(III) mineral crusts around the cells. The amount of the organic cosubstrate acetate present seems to control the rate and extent of Fe(II) oxidation and the viability of the cells. The crystallinity of the mineral products is influenced by nucleation by Fe minerals that are already present in the inoculum.     

Nutrient removal and sludge age in a sequencing batch reactor

The aim of this work was to establish a relation between the mean cellular retention time and the ability of activated sludge to remove phosphate and ammonium. A sequencing batch reactor (SBR) with a total volume of 1.94 m³ was fed with municipal wastewater and was operated under four different organic loading rates to obtain sludge ages of 23, 16, 6, and 3 days. The operational strategy included fill, anaerobic, aerobic, settling and draw phases. The experimental work lasted 445 days. Biological phosphate removal was achieved with sludge ages from 6 to 23 days. The highest PO₄-P removal rate observed was of 98% and corresponds to a 16-day sludge age; phosphate removal increased with the sludge age. A sludge age of 3 days resulted in a chemical oxygen demand (COD) removal rate of 81% and a sludge age of 23 days in a removal rate of 99%. Full nitrification was observed with a sludge age of 16 days. Nitrification increased with the sludge age. The 3-day sludge age did not allow nitrification. The phosphate concentrations in the biomass were inversely proportional to the sludge age.     

Aerobic treatment of a concentrated urea wastewater with simultaneous stripping of ammonia

An industrial wastewater containing a total Kjeldahl nitrogen (TKN) of 12.80 g l(-1) was treated in a continuously fed activated sludge reactor. The main contaminant was urea (21.52 g l(-1)), together with minor amounts of the nitrification inhibitor dicyandiamide (0.46 g l(-1)) and free ammonia (0.56 g l(-1)). The wastewater was diluted 1:1 with water and treated under alkaline conditions (pH 9.4), enabling the simultaneous hydrolysis of urea and stripping of free ammonia in one aerobic reactor. Experiments were conducted to eliminate the remaining ammonia in a separate treatment unit by nitrification/denitrification. An adapted nitrifying bacterial population was isolated which was able to nitrify at a rate of 0.1 g nitrogen l(-1) day(-1) at a dicyandiamide concentration of 0.22 g l(-1). However, this was found to be too slow for an industrial-scale operation. Therefore, separate stripping with air or steam after pH adjustment to > or =10.5 is proposed. The diluted wastewater was treated with a hydraulic retention time of 6 days, corresponding to a volumetric nitrogen loading rate of 1.1 g nitrogen l(-1) day(-1) with an overall TKN reduction of 78.0%.     

Denitrification in a nitrogen-limited stream ecosystem

Denitrification was measured in hyporheic, parafluvial, and bank sediments of Sycamore Creek, Arizona, a nitrogen-limited Sonoran Desert stream. We used three variations of the acetylene block technique to estimate denitrification rates, and compared these estimates to rates of nitrate production through nitrification. Subsurface sediments of Sycamore Creek are typically well-oxygenated, relatively low in nitrate, and low in organic carbon, and therefore are seemingly unlikely sites of denitrification. However, we found that denitrification potential (C & N amended, anaerobic incubations) was substantial, and even by our conservative estimates (unamended, oxic incubations and field chamber nitrous oxide accumulation), denitrification consumed 5–40% of nitrate produced by nitrification. We expected that denitrification would increase along hyporheic and parafluvial flowpaths as dissolved oxygen declined and nitrate increased. To the contrary, we found that denitrification was generally highest at the upstream ends of subsurface flowpaths where surface water had just entered the subsurface zone. This suggests that denitrifiers may be dependent on the import of surface-derived organic matter, resulting in highest denitrification rate at locations of surface-subsurface hydrologic exchange. Laboratory experiments showed that denitrification in Sycamore Creek sediments was primarily nitrogen limited and secondarily carbon limited, and was temperature dependent. Overall, the quantity of nitrate removed from the Sycamore Creek ecosystem via denitrification is significant given the nitrogen-limited status of this stream.     

Nitrification and ammonia volatilisation losses from urea and dicyandiamide-treated urea in a sandy loam soil

Summary Nitrification and ammonia volatilisation losses from urea and dicyandiamide (DCD)-treated urea were studied in a sandy loam soil. Laboratory experiments indicated that 20 ppm (of soil) DCD effectively inhibited nitrification of urea over sixty days. If the urea was treated with DCD (20 ppm), ammonia emission from the soil was extended over 105 days; with urea alone, it was negligible after 15 days. A field study indicated that DCD treatment increased volatilisation losses of ammonia tremondously if urea was applied to the soil surface; these losses were minimised if the urea was placed at 5 cm depth. It would seem that nitrification inhibitors must be combined with a placement technique.     

Assessment of a bioaugmentation strategy with polyphosphate accumulating organisms in a nitrification/denitrification sequencing batch reactor

Different alternative configurations and strategies for the simultaneous biological removal of organic matter and nutrients (N and P) in wastewater have been proposed in the literature. This work demonstrates a new successful strategy to bring in enhanced biological phosphorus removal (EBPR) to a conventional nitrification/denitrification system by means of bioaugmentation with an enriched culture of phosphorus accumulating organisms (PAO). This strategy was tested in a sequencing batch reactor (SBR), where an 8 h configuration with 3 h anoxic, 4.5 h aerobic and 25 min of settling confirmed that nitrification, denitrification and PAO activity could be maintained for a minimum of 60 days of operation after the bioaugmentation step. The successful bioaugmentation strategy opens new possibilities for retrofitting full-scale WWTP originally designed for only nitrification/denitrification. These systems could remove P simultaneously to COD and N if they were bioaugmented with waste purge of an anaerobic/aerobic SBR operated in parallel treating part of the influent wastewater.     

Transformation of urea and ammonium sulphate in different soils

Summary The transformation of urea and ammonium sulphate in Ladwa sandy loam and Balsamand sand was studied in laboratory. Urea took at least one week in sandy loam and 2 weeks in sandy soils to hydrolyse completely. The process of hydrolysis was faster in finer soil with high organic matter than in coarse soil having low organic matter. There was no nitrification upto 3 days in sandy loam and upto 7 days in sandy soils, respectively, but there was immobilization of NO₃-N during these initial periods. The NO₃-N content at the end of incubation period (35 days) was more in case of urea than in case of ammonium sulphate treated samples in sandy loam soil and reverse was true in sandy soil. The hydrolysis of urea did not follow zero or first order kinetics as proposed in previous studies.     

Interpreting the effect of increasing COD loading rates on the performance of a pre-anoxic MBBR system: implications on the attached and suspended biomass dynamics and nitrification–denitrification activity

A pre-anoxic MBBR system was subjected to increasing organic loading rates up to 18 gCOD/(m² day). At 3 gCOD/(m² day), most of the incoming organic matter was removed via denitrification. However, at higher loads, anoxic COD removal became limited by the nitrite/nitrate supply from the aerobic reactor, which assumed an important role in this conversion. Despite the application of low dissolved oxygen (DO) levels (<2 mg/L) in this tank, nitrification was observed to be nearly complete until 8 gCOD/(m² day). As the organic input was increased, the maximum specific nitrifying activity gradually declined. Activity tests suggested that an oxygen-limited environment was established in the biofilm. At lower loads [3–8 gCOD/(m² day)], the nitrification product obtained was affected by the DO concentration, whereas from 16 to 21 gCOD/(m² day), nitrite/nitrate profiles were likely associated with microbial stratification in the biofilm. The results also indicated that the role of the suspended biomass in the overall nitrification and denitrification can be very significant in high loaded MBBRs and should not be neglected, even at low HRTs.     

Quantification of ammonia-oxidizing bacteria and factors controlling nitrification in salt marsh sediments

To elucidate the geomicrobiological factors controlling nitrification in salt marsh sediments, a comprehensive approach involving sediment geochemistry, process rate measurements, and quantification of the genetic potential for nitrification was applied to three contrasting salt marsh habitats: areas colonized by the tall (TS) or short (SS) form of Spartina alterniflora and unvegetated creek banks (CBs). Nitrification and denitrification potential rates were strongly correlated with one another and with macrofaunal burrow abundance, indicating that coupled nitrification-denitrification was enhanced by macrofaunal burrowing activity. Ammonia monooxygenase (amoA) gene copy numbers were used to estimate the ammonia-oxidizing bacterial population size (5.6 x 10(4) to 1.3 x 10(6) g of wet sediment(-1)), which correlated with nitrification potentials and was 1 order of magnitude higher for TS and CB than for SS. TS and CB sediments also had higher Fe(III) content, higher Fe(III)-to-total reduced sulfur ratios, higher Fe(III) reduction rates, and lower dissolved sulfides than SS sediments. Iron(III) content and reduction rates were positively correlated with nitrification and denitrification potential and amoA gene copy number. Laboratory slurry incubations supported field data, confirming that increased amounts of Fe(III) relieved sulfide inhibition of nitrification. We propose that macrofaunal burrowing and high concentrations of Fe(III) stimulate nitrifying bacterial populations, and thus may increase nitrogen removal through coupled nitrification-denitrification in salt marsh sediments.     

Control of urea hydrolysis and nitrification in soil by chemicals—Prospects and problems

Summary A review is made of the recent work to assess the prospects of regulating urea hydrolysis and nitrification processes in soils by employing chemicals that can retard urea hydrolysis and nitrification. The possible benefits from control of nitrogen transformations in terms of conserving and enhancing fertilizer nitrogen efficiency for crop production and the problems associated with their use with regard to N metabolism of plants have also been discussed with examples. Prospects of using cheap and effective indigenous materials and chemicals for control of urea hydrolysis and nitrification under specific soil situations appear eminent in improving the fertilizer nitrogen efficiency. Urease inhibitors may be helpful in reducing problems associated with ammonia volatilization if this is not offset by leaching of urea. On the other hand retardation of nitrification appears useful in reducing losses that accompany nitrification due to leaching and denitrification, and with the plants that metabolize equally well with relatively higher amounts of NH₄–N may be more effective in improving the utilization of fertilizer N under these situations.     

Phosphate removal from the returned liquor of municipal wastewater treatment plant using iron-reducing bacteria

AIM: The application of iron-reducing bacteria (IRB) to phosphate removal from returned liquor (liquid fraction after activated sludge digestion and anaerobic sludge dewatering) of municipal wastewater treatment plant (WWTP) was studied. METHODS AND RESULTS: An enrichment culture and two pure cultures of IRB, Stenotrophomonas maltophilia BK and Brachymonas denitrificans MK identified by 16S rRNA gene sequencing, were produced using returned liquor from a municipal WWTP as carbon and energy source, and iron hydroxide as oxidant. The final concentration of phosphate increased from 70 to 90 mg l(-1) in the control and decreased from 70 to 1 mg l(-1) in the experiment. The mass ratio of removed P to produced Fe(II) was 0.17 g P g(-1) Fe(II). The strain S. maltophilia BK showed the ability to reduce Fe(III) using such xenobiotics as diphenylamine, m-cresol, 2,4-dichlorphenol and p-phenylphenol as sole sources of carbon under anaerobic conditions. CONCLUSIONS: Bacterial reduction of ferric hydroxide enhanced the phosphate removal from the returned liquor. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability of the facultative anaerobes S. maltophilia BK and B. denitrificans MK to reduce Fe(III) was shown. These micro-organisms can be used for anaerobic removal of phosphate and xenobiotics by bacterial reduction of ferric ions.     

脲酶/硝化抑制剂在黑土和褐土中对尿素氮转化的调控效果

本试验研究脲酶/硝化抑制剂不同组合在黑土和褐土中对尿素水解和硝化作用的调控效果,旨在筛选出适合东北黑土、褐土的高效抑制剂组合。采用室内恒温、恒湿培养试验,以不施氮肥(CK)和施用普通尿素肥料(U)为对照,研究分别添加脲酶抑制剂N-丁基硫代磷酰三胺(NBPT)及其与硝化抑制剂双氰胺(DCD)、3,4-二甲基吡唑磷酸盐(DMPP)、2-氯-6(三氯甲基)-吡啶(CP)、2-氨基-4-氯-6-甲基嘧啶(AM)、3-甲基吡唑(MP)组合制成的6种高效稳定性尿素在黑土和褐土中的尿素水解和氨氧化特征。在培养125 d内分别取土壤样品15次,通过测定2种土壤中尿素态氮、铵态氮和硝态氮含量,及氨氧化作用强度,计算硝化抑制率,确定最适合2种土壤的抑制剂或组合。结果表明: 尿素在黑土和褐土中水解时间约7 d,添加NBPT以及其与不同硝化抑制剂组合均能将尿素水解时间延长21 d以上。与U处理相比,添加抑制剂可显著增加土壤NH₄⁺-N含量,降低NO₃^--N生成量,维持土壤中高NH₄⁺-N含量的时间更久。黑土中,添加硝化抑制剂的处理均能显著抑制土壤硝化作用,有效硝化抑制时间超过125 d;DMPP、CP与NBPT配施使黑土NH₄⁺-N含量提高1.6～1.8倍,培养125 d时其硝化抑制率分别为47.9%和24.1%。褐土中,U处理培养80 d左右基本完成硝化过程,而添加硝化抑制剂使硝化过程延长至少30 d;DCD、DMPP与NBPT配施使土壤NH₄⁺-N含量提高2.1～3.4倍,培养125 d时其硝化抑制率分别为25.3%和23.2%。因此,尿素与NBPT+DMPP和NBPT+DCD制成的高效稳定性尿素分别在黑土和褐土中施用效果最好,其次分别是NBPT+CP和NBPT+DMPP。     

Enzymes of carbohydrate metabolism in the developing endosperm of maize

A number of enzymes presumably implicated in starch synthesis were assayed at various stages of endosperm development ranging from 8 days to 28 days after pollination. Activity for invertase, hexokinase, the glucose phosphate isomerases, the phosphoglucomutases, phosphorylase I, uridine diphosphate glucose pyrophosphorylase, and the starch granule-bound nucleoside diphosphate glucose-starch glucosyltransferase was present at the earliest stage of development (8 days) studied. Activity was detectable for phosphorylase III, the soluble adenosine diphosphate glucose-starch glucosyltransferase, adenosine diphosphate glucose pyrophosphorylase, and sucrose-uridine diphosphate glucosyltransferase at 12 days. For phosphorylase II and cytidine diphosphate glucose pyrophosphorylase, activity was first detectable at the 14- and 16-day stages, respectively. Rapid increases in starch content are observed prior to detectable activity for adenosine diphosphate glucose pyrophosphorylase, the soluble adenosine diphosphate glucose-starch glucosyltransferase and phosphorylases II and III. For all enzymes, except invertase, activity per endosperm rises to a peak at 22 or 28 days. Greatest activity for invertase is found at 12 days with a steady decline thereafter. The pattern of invertase activity in comparison with that of sucrose-uridine diphosphate glucosyltransferase supports previous suggestions, that the latter plays a key role in the conversion of sucrose to starch. In addition to phosphorylases I, II, and III, multiple forms of glucosephosphate isomerase and phosphoglucomutase were detected.     

Enhancing nitrogen removal in an Orbal oxidation ditch by optimization of oxygen supply: practice in a full-scale municipal wastewater treatment plant

Seven different aeration modes, in which oxygen supply was changed by adjusting the number of aerators, were designed and applied in a full-scale municipal wastewater treatment plant with Orbal oxidation ditch to investigate the influence of dissolved oxygen (DO) on nitrogen removal performance. The full-scale experiment results of 574 days showed that nitrogen removal efficiency depended on the degree of nitrification and denitrification in the outer channel, which was the largest contributor for TN removal in the Orbal oxidation ditch. Appropriate aeration control in the outer channel was essential to balance nitrification and denitrification in the Orbal oxidation ditch. When DO was as low as about 0.2 mg/L in the outer channel, the highest TN removal efficiency of 75% was obtained. Microbial analysis confirmed that aerobic and anaerobic bacteria coexisted in the outer channel. The greater species diversity and more intensive activities of these bacteria in aeration Mode V may be responsible for the higher TN removal efficiency compared with Mode III. These results suggest that different aerated conditions in the Orbal oxidation ditch might have a significant effect on microbial community characteristics and nitrogen removal efficiencies.     

Availability of Ferric Iron for Microbial Reduction in Bottom Sediments of the Freshwater Tidal Potomac River

The distribution of Fe(III), its availability for microbial reduction, and factors controlling Fe(III) availability were investigated in sediments from a freshwater site in the Potomac River Estuary. Fe(III) reduction in sediments incubated under anaerobic conditions and depth profiles of oxalate-extractable Fe(III) indicated that Fe(III) reduction was limited to depths of 4 cm or less, with the most intense Fe(III) reduction in the top 1 cm. In incubations of the upper 4 cm of the sediments, Fe(III) reduction was as important as methane production as a pathway for anaerobic electron flow because of the high rates of Fe(III) reduction in the 0- to 0.5-cm interval. Most of the oxalate-extractable Fe(III) in the sediments was not reduced and persisted to a depth of at least 20 cm. The incomplete reduction was not the result of a lack of suitable electron donors. The oxalate-extractable Fe(III) that was preserved in the sediments was considered to be in a form other than amorphous Fe(III) oxyhydroxide, since synthetic amorphous Fe(III) oxyhydroxide, amorphous Fe(III) oxyhydroxide adsorbed onto clay, and amorphous Fe(III) oxyhydroxide saturated with adsorbed phosphate or fulvic acids were all readily reduced. Fe₃O₄ and the mixed Fe(III)-Fe(II) compound(s) that were produced during the reduction of amorphous Fe(III) oxyhydroxide in an enrichment culture were oxalate extractable but were not reduced, suggesting that mixed Fe(III)-Fe(II) compounds might account for the persistence of oxalate-extractable Fe(III) in the sediments. The availability of microbially reducible Fe(III) in surficial sediments demonstrates that microbial Fe(III) reduction can be important to organic matter decomposition and iron geochemistry. However, the overall extent of microbial Fe(III) reduction is governed by the inability of microorganisms to reduce most of the Fe(III) in the sediment.     

微生物驱动硝酸盐还原耦合亚铁氧化成矿过程的锌胁迫

【目的】探究中性厌氧条件下,金属锌影响下硝酸盐依赖型铁氧化菌Pseudomonas stutzeri LS-2驱动的硝酸盐还原耦合亚铁氧化成矿过程机制,对深入理解中性厌氧环境中微生物亚铁氧化驱动的反硝化作用及重金属固定机制具有重要意义。【方法】以不同Zn(Ⅱ)浓度构建LS-2驱动的亚铁氧化成矿体系,分析不同体系中亚铁氧化速率、硝酸盐还原速率以及形成矿物的结构变化规律。【结果】LS-2驱动的硝酸盐还原耦合亚铁氧化成矿过程中,共存Zn(Ⅱ)降低该过程中硝酸盐的还原速率和亚铁氧化速率。同时,随着Zn(Ⅱ)浓度提高,抑制作用增强。微生物亚铁氧化形成的矿物通过吸附、共沉淀和离子置换等过程固定Zn(Ⅱ),降低Zn(Ⅱ)活性。Zn(Ⅱ)浓度对形成的矿物结构有较大的影响:低浓度Zn(Ⅱ)体系中,形成的矿物为纤铁矿;随着Zn(Ⅱ)浓度的提高,矿物结构与结晶度都有一定程度的变化,当Zn(Ⅱ)达到4 mmol/L时,形成的矿物主要为铁锌尖晶石。【结论】明确了重金属锌对LS-2菌株反硝化及亚铁氧化过程的抑制规律,同时阐明了Zn(Ⅱ)浓度对形成矿物结构的影响。研究结果有助于深入认识中性厌氧环境中重金属与微生物驱动的铁循环和反硝化过程的耦合作用,为土壤重金属污染防治提供理论支撑。     

The fate of labelled15N urea and ammonium nitrate applied to a winter wheat crop

Labelled urea or ammonium nitrate was applied to winter wheat growing on a loamy soil in Northern France. Two applications of fertilizer were given: 50 kg N ha^–1 at tillering (early March) and 110 kg N ha^–1 at the beginning of stem elongation (mid-April). The kinetics of urea hydrolysis, nitrification of ammonium and the disappearance of inorganic nitrogen were followed at frequent intervals. Inorganic nitrogen soon disappeared, mainly immobilized by soil microflora and absorbed by the crop. Net immobilization of fertilizer N occured at a very similar rate for urea and ammonium nitrate. Maximum immobilization (16 kg N ha¹) was found at harvest for the first dressing and at anthesis for the second dressing (23 kg N ha¹). During the nitrification period, the labelled ammonium pool was immobilized two to three times faster than the labelled nitrate pool. No significant net¹⁵N remineralization was found during the growth cycle.The actual denitrification and volatilization losses were probably more important than indicated from calculations made by extrapolation of fluxes measured over short intervals. However microbial immobilization was the most important of the processes which compete with plant uptake for nitrogen.     

湖泊微生物反硝化过程及速率研究进展

湖泊中微生物介导的反硝化过程对于区域乃至全球的气候环境变化有着深远的影响。因此,研究湖泊微生物反硝化过程及速率有助于我们深刻理解湖泊氮元素生物地球化学循环规律,全面认识湖泊生境对全球氮循环的贡献。本文综述了湖泊生境中反硝化过程(包括典型的反硝化过程及与其他物质循环耦合的反硝化过程,如与有机氮耦合的共反硝化作用、与碳循环耦合的硝酸盐/亚硝酸盐依赖型厌氧甲烷氧化、与铁循环耦合的硝酸盐依赖型铁氧化、与硫循环耦合的硝酸盐还原硫氧化)的速率、驱动微生物及其影响因素。最后对湖泊反硝化过程研究现状和未来发展方向提出总结与展望。     

新型脱氮微生物与水体脱氮新工艺研究进展

氨氮是河流等淡水资源有机污染的主要污染指标之一。生物脱氮具有低成本、高效、无二次污染和易操作等优点,极具发展前景。重点概述了水体净化系统中新型脱氮微生物的种类及研究进展,介绍了厌氧氨氧化、短程硝化-反硝化和分段进水生物脱氮等高效节能新工艺的工艺原理。     

Who contributes more to N2O emission during sludge bio-drying with two different aeration strategies,nitrifiers or denitrifiers?

Global warming effects have drawn more and more attention to studying all sources and sinks of nitrous oxide (N₂O). Sludge bio-drying, as an effective sludge treatment technology, is being adopted worldwide. In this study, two aeration strategies (piles I and II) were compared to investigate the primary contributors to N₂O emission during sludge bio-drying through studying the evolution of functional genes involved in nitrification (amoA, hao, and nxrA) and denitrification (narG, nirS, nirK, norB, and nosZ) by quantitative PCR (qPCR). Results showed that the profile of N₂O emission can be divided into three stages, traditional denitrification contributed largely to N₂O emission at stage I (days 1–5), but N₂O emission mainly happened at stage II (days 5–14) due to nitrifier denitrification and NH₂OH accumulation by ammonia-oxidizing bacteria (AOB), accounting for 51.4% and 58.2% of total N₂O emission for piles I and II, respectively. At stage III (days 14–21), nitrifier denitrification was inhibited because sludge bio-drying proceeded mainly by the physical aeration, thus N₂O emission decreased and changed little. The improved aeration strategy availed pile I to reduce N₂O emission much especially at stages II and III, respectively. These results indicated that nitrifier denitrification by AOB and biological NH₂OH oxidation due to AOB made more contribution to N₂O emission, and aeration strategy was crucial to mitigate N₂O emission during sludge bio-drying.