Immobilization of nitrifying bacteria by polyethylene glycol prepolymer

The effects of immobilizing materials on the activity of nitrifying bacteria were investigated by using 11 kinds of prepolymers of polyethylene glycol. Relative respiratory activity of immobilized nitrifying bacteria with polyethylene glycol metacrylate prepolymer was higher than that of polyethylene glycol acrylate prepolymer, and there was a tendency for relative respiratory activity to be higher with a prepolymer of greater molecular weight. With the polyethylene glycol prepolymer, there was a drastic improvement over the conventional method of immobilization by acrylamide in the relative respiratory activity of the pellet. Inorganic synthetic wastewater was treated under a high loading rate of 1.14 kg-N/m³·d. Influent NH₄-N could be removed to 2 mg/l or less and the nitrogen removal was 90%.     

两株异养硝化细菌的分离鉴定及其脱氮特性

【目的】利用异养硝化培养基,从华中农业大学实验猪场污水中筛选得到2株具有较高脱氮效率的细菌。【方法】通过形态学特征及16S rDNA序列的系统发育分析,对分离菌株进行了鉴定。且对菌株P2和P9降解氨氮的相关特性也作了研究。此外,将菌株单独或混合接种于猪场污水,检测其处理实际污水的脱氮效果。【结果】初步判断菌株P2为副球菌属(Paracoccus sp.),P9为申氏杆菌属(Shinella sp.)。2株细菌能在有机物存在下进行异养硝化作用,经24h培养,菌株P2和P9对氨氮的去除率可达80%左右,同时未发现亚硝酸盐、硝酸盐积累;但菌株P2,P9不能以NO 3-或NO 2-为唯一氮源发生好氧反硝化作用。菌株P2和P9异养硝化的最适碳源为丁二酸钠,最适C/N比为9,且脱氮过程中pH值从6.8到8.9一直呈上升趋势。菌株对小分子碳源具有较强的依赖性,在加入小分子碳源的情况下,其对污水具有较强的脱氮能力,且这两个菌株混合施用较单独作用氨氮去除效果更好。【结论】菌株P2和P9脱氮能力较强,其在污水处理行业具有重要的应用前景。     

Ammonia removal from prawn aquaculture water using immobilized nitrifying bacteria

Intensive prawn aquaculture in tropical regions is associated with high concentrations of total ammoniacal nitrogen (TAN) as a result of high rates of prawn excretion and feed loading. Excessive TAN can adversely effect productivity and result in adverse impacts on coastal waters. Cultures of indigenous nitrifying bacteria were enriched from intensive prawn aquaculture pond water using continuous and batch enrichment techniques. Cultures were capable of TAN removal over a wide range of initial TAN concentrations - up to 200 mg/l. Cultures were immobilized onto porous clay pellets to enhance cell density and applied to culture medium and TAN-augmented pond water under aerobic conditions to determine TAN removal proficiency. Immobilized cultures were able to achieve a high TAN removal proficiency in pond water--even at a low density of 0.1 pellet per liter. A concentration of less than 0.5 mg TAN/l could be maintained under a fed-batch condition of 3.2 mg TAN/l per day, after an initial 2-day lag phase. A simplified and effective culture enrichment process was developed for culture immobilization onto pellets using TAN-augmented pond water. Overall, pellet immobilization of indigenous nitrifying bacteria represents a potentially effective TAN control system for prawn aquaculture in low-cost, but intensive tropical prawn farms.     

Ammonia removal from freshwater using nitrifying bacteria enriched from a seawater aquaculture pond

Intensively cultured fish stock when fed protein-rich feeds typically excrete high concentrations of total ammoniacal-nitrogen (TAN) into the water column which can have adverse effects on productivity, and upon the environment when aquaculture water is discharged. An immobilized culture of nitrifying bacteria isolated from prawn pond water and known to effectively remove TAN from saline water was tested for its ability to remove TAN from freshwater. The culture was readily adaptable to non-saline conditions and maintained TAN at less than 0.25 mg l^–1, even with a daily addition of 3.2 to 4.2 mg TAN l^–1 per d. The use of the immobilized culture of nitrifying bacteria represents an innovative and economical in situ treatment technology for removal of TAN in both saline and freshwater.     

厌氧氨氧化生物脱氮技术的研究进展

厌氧氨氧化是指在厌氧条件下,厌氧氨氧化混合菌直接以NH4 为电子供体,以NO3-或NO2-为电子受体,将NH4^ 、NO3-或NO2-转变成N2的过程。厌氧氨氧化作为一种新型的污水处理工艺具有较高的理论意义和良好的应用前景。本文主要阐述了厌氧氨氧化生物脱氮技术原理、厌氧氨氧化的可能途径、方法及其应用现状,并且讨论了厌氧氨氧化反应的微生物学机理和厌氧氨氧化工艺的开发,提出了今后研究的主要方向。     

Role of anammox bacteria in removal of nitrogen compounds from wastewater

The review deals with the unique microbial group responsible for anaerobic ammonium oxidation with nitrite (anammox), and with the role of this process in development of the biotechnology for removal of nitrogen compounds from wastewater. The history of the study of this process is briefly related. Up-to date knowledge on the intracellular organization, energy metabolism, growth stoichiometry, and physiology of anammox bacteria is described, and the main methods for cultivation of these microorganisms are characterized. Special attention is paid to the problems associated with practical application of anammox bacteria, which result from their extremely slow growth, the absence of pure cultures, and the interaction with other microbial groups.     

Complete and simultaneous removal of ammonium and m-cresol in a nitrifying sequencing batch reactor

The kinetic behavior, oxidizing ability and tolerance to m-cresol of a nitrifying sludge exposed to different initial concentrations of m-cresol (0–150 mg C L^?1) were evaluated in a sequencing batch reactor fed with 50 mg NH₄ ⁺-N L^?1 and operated during 4 months. Complete removal of ammonium and m-cresol was achieved independently of the initial concentration of aromatic compound in all the assays. Up to 25 mg m-cresol-C L^?1 (C/N ratio of 0.5), the nitrifying yield (Y-NO₃ ^?) was 0.86 ± 0.05, indicating that the nitrate was the main product of the process; no biomass growth was detected. From 50 to 150 mg m-cresol-C L^?1 (1.0 ≤ C/N ≤ 3.0), simultaneous microbial growth and partial ammonium-to-nitrate conversion were obtained, reaching a maximum microbial total protein concentration of 0.763 g L^?1 (247 % of its initial value) and the lowest Y-NO₃ ^? 0.53 ± 0.01 at 150 mg m-cresol-C L^?1. m-Cresol induced a significant decrease in the values of both specific rates of ammonium and nitrite oxidation, being the ammonium oxidation pathway the mainly inhibited. The nitrifying sludge was able to completely oxidize up to 150 mg m-cresol-C L^?1 by SBR cycle, reaching a maximum specific removal rate of 6.45 g m-cresol g^?1 microbial protein-N h^?1. The number of SBR cycles allowed a metabolic adaptation of the nitrifying consortium since nitrification inhibition decreased and faster oxidation of m-cresol took place throughout the cycles.     

Immobilization of Saccharomyces uvarum cells in porous beads of polyacrylamide gel for ethanolic fermentation

Summary A procedure which does not involve the use of an immiscible organic solvent phase is described for the entrapment of yeast cells in porous beads of polyacrylamide gel. The cells are rapidly dispersed at 4° C in an aqueous solution containing sodium alginate and acrylamide-N,Nmethylene-bis-acrylamide monomer, and the suspension is immediately dropped into a solution of calcium formate to give calcium alginate coated beads. Polyacrylamide gel forms within the bead. The calcium alginate is subsequently leached out of the composite bead with either sodium citrate or potassium phosphate buffer solution. Cells of Saccharomyces uvarum ATCC 26 602 entrapped in such polyacrylamide beads ferment cane molasses in batch mode at higher specific ethanol productivity than a free cell suspension. Their volumetric productivity in continuous fermentation is higher than that of Ca²⁺-alginate immobilized cells.NCL Communication No. 4383     

Fate of 14C-labeled microbial products derived from nitrifying bacteria in autotrophic nitrifying biofilms

The cross-feeding of microbial products derived from 14C-labeled nitrifying bacteria to heterotrophic bacteria coexisting in an autotrophic nitrifying biofilm was quantitatively analyzed by using microautoradiography combined with fluorescence in situ hybridization (MAR-FISH). After only nitrifying bacteria were labeled with [14C]bicarbonate, biofilm samples were incubated with and without NH4+ as a sole energy source for 10 days. The transfer of 14C originally incorporated into nitrifying bacterial cells to heterotrophic bacteria was monitored with time by using MAR-FISH. The MAR-FISH analysis revealed that most phylogenetic groups of heterotrophic bacteria except the beta-Proteobacteria showed significant uptake of 14C-labeled microbial products. In particular, the members of the Chloroflexi were strongly MAR positive in the culture without NH4+ addition, in which nitrifying bacteria tended to decay. This indicated that the members of the Chloroflexi preferentially utilized microbial products derived from mainly biomass decay. On the other hand, the members of the Cytophaga-Flavobacterium cluster gradually utilized 14C-labeled products in the culture with NH4+ addition in which nitrifying bacteria grew. This result suggested that these bacteria preferentially utilized substrate utilization-associated products of nitrifying bacteria and/or secondary metabolites of 14C-labeled structural cell components. Our results clearly demonstrated that the coexisting heterotrophic bacteria efficiently degraded and utilized dead biomass and metabolites of nitrifying bacteria, which consequently prevented accumulation of organic waste products in the biofilm.     

异养硝化复合菌强化处理含氮废水脱氮性能研究

针对传统污水处理脱氮工艺过程中工艺流程复杂、处理高氨氮废水效率低等问题,利用三株不同种属的高效异养硝化-好氧反硝化细菌构建异养硝化复合菌YM,探讨其异养硝化-好氧反硝化特性及其生物强化脱氮效能研究,从而为异养硝化菌强化处理高氨氮废水工程应用提供理论依据。结果表明:异养硝化复合菌YM的增殖速率、异养氨氧化、好氧反硝化能力均优于单一菌种,YM强化后的污泥系统氨氧化速率较未强化系统从7.04 mg/L/h提高到12.2 mg/L/h,并且生物强化作用可有效提高污泥系统的抗冲击负荷能力,一定程度上提高了系统的处理能力。研究表明异养硝化菌强化污水脱氮处理具有显著的应用潜能,尤其对于目前尚缺少经济高效处理技术的高污染物浓度废水处理而言,无疑是一条具有高潜在应用价值的新途径。     

Combined anaerobic ammonium and methane oxidation for nitrogen and methane removal

Anammox (anaerobic ammonium oxidation) is an environment-friendly and cost-efficient nitrogen-removal process currently applied to high-ammonium-loaded wastewaters such as anaerobic digester effluents. In these wastewaters, dissolved methane is also present and should be removed to prevent greenhouse gas emissions into the environment. Potentially, another recently discovered microbial pathway, n-damo (nitrite-dependent anaerobic methane oxidation) could be used for this purpose. In the present paper, we explore the feasibility of simultaneously removing methane and ammonium anaerobically, starting with granules from a full-scale anammox bioreactor. We describe the development of a co-culture of anammox and n-damo bacteria using a medium containing methane, ammonium and nitrite. The results are discussed in the context of other recent studies on the application of anaerobic methane- and ammonia-oxidizing bacteria for wastewater treatment.     

Competition for ammonium between plant roots and nitrifying and heterotrophic bacteria and the effects of protozoan grazing

The competition for limiting amounts of ammonium between the chemolithotrophic ammonium-oxidizing species Nitrosomonas europaea, the heterotrophic species Arthrobacter globiformis and roots of Plantago lanceolata (Ribwort plantain) was studied in a series of model systems of increasing complexity, i.e. energy-limited continuous cultures, non-water-saturated continuously percolated soil columns and pots with γ-sterilized soil planted with axetic P. lanceolata seedlings. The effects of bacterial grazing by the flagellate species Adriamonas peritocrescens on the competition for ammonium were also investigated in the three model systems. It was found that N. europaea was a weaker competitor for ammonium than either A. globiformis or plant roots of P. lanceolata. It is assumed that the heterotrophic bacteria have a higher affinity for ammonium than the nitrifying bacteria, whereas growing plant roots have a greater capacity to exploit the soil for ammonium than the immobile nitrifying bacteria. It is not very likely that allelochemicals were involved in suppressing the nitrification process. Four reasons are given for this assumption. Presence of the flagellates strongly stimulated the potential nitrification rate in all model systems. It is assumed that there is a more even distribution over the soil of either nitrifying bacteria or their substrate ammonium in the presence of flagellates. In addition to the distribution effect, there is a stimulation of the potential ammonium oxidation rate. The results are discussed in the light of the function of nitrate as nitrogen sink in the biogeochemical nitrogen cycle.     

混合培养真菌和细菌对废水的生物去氮作用

Denitrificationis a biological processin which nitrateand/or nitrite is reduced to gaseous nitrogen,dinitrogen(N2)or nitrous oxide(N2O)while carbon dioxide is thesecond gaseous product of the process.This is one of themain mechanisms of the global nitrogen cycle,and playsanimportant role as the reverse reaction of nitrogen fixa-tion in maintaining global environmental homeostasis[1].Denitrification has beenlongthought to be a unique char-acteristic of prokaryotes[2,3].Anumber of bacteria(such…     

Effects of aeration cycles on nitrifying bacterial populations and nitrogen removal in intermittently aerated reactors

The effects of the lengths of aeration and nonaeration periods on nitrogen removal and the nitrifying bacterial community structure were assessed in intermittently aerated (IA) reactors treating digested swine wastewater. Five IA reactors were operated in parallel with different aeration-to-nonaeration time ratios (ANA). Populations of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were monitored using 16S rRNA slot blot hybridizations. AOB species diversity was assessed using amoA gene denaturant gradient gel electrophoresis. Nitrosomonas and Nitrosococcus mobilis were the dominant AOB and Nitrospira spp. were the dominant NOB in all reactors, although Nitrosospira and Nitrobacter were also detected at lower levels. Reactors operated with the shortest aeration time (30 min) showed the highest Nitrosospira rRNA levels, and reactors operated with the longest anoxic periods (3 and 4 h) showed the lowest levels of Nitrobacter, compared to the other reactors. Nitrosomonas sp. strain Nm107 was detected in all reactors, regardless of the reactor's performance. Close relatives of Nitrosomonas europaea, Nitrosomonas sp. strain ENI-11, and Nitrosospira multiformis were occasionally detected in all reactors. Biomass fractions of AOB and effluent ammonia concentrations were not significantly different among the reactors. NOB were more sensitive than AOB to long nonaeration periods, as nitrite accumulation and lower total NOB rRNA levels were observed for an ANA of 1 h:4 h. The reactor with the longest nonaeration time of 4 h performed partial nitrification, followed by denitrification via nitrite, whereas the other reactors removed nitrogen through traditional nitrification and denitrification via nitrate. Superior ammonia removal efficiencies were not associated with levels of specific AOB species or with higher AOB species diversity.     

Effect of high concentrations of nitrogen on mixed populations of nitrifying bacteria isolated from industrial nitrogenous wastewaters.

The effect of high concentrations of different forms of nitrogen (NH4+, NO2-, NO3- and urea N) on nitrification by mixed populations of nitrosobacters isolated from nitrogen fertilizer plant wastewaters and nitrobacters isolated from effluents from a biological bed treating these wastewaters was determined. The inhibitory activity (within the concentration values for industrial wastewaters) of only the reaction products was observed, i.e. nitrites for nitrification phase I and nitrates for nitrification phase II. A mixed population of phase II nitrifying bacteria is highly resistant to high concentrations of ammonia nitrogen (50% inhibition at 3,000 mg N/l).     

极端条件下异养硝化-好氧反硝化菌脱氮的研究进展

异养硝化-好氧反硝化(HN-AD)是对传统自养硝化异养反硝化理论的丰富与突破。HN-AD菌在好氧条件下可快速实现氨氮、硝态氮(NO_3~–-N)、亚硝态氮(NO_2~–-N)三氮同步脱除。它们不仅具有分布范围广、适应能力强、代谢通路特殊等特点,而且还具有世代时间短、脱氮速率快、高活性持久等独特优势,在高盐、低温、高氨氮等极端条件表现出了巨大的脱氮潜力,因此在废水生物脱氮领域受到广泛关注。文中在介绍HN-AD菌属类别及代谢机理的基础上,重点总结了在高盐、低温、高氨氮等极端条件下进行氨氮脱除的HN-AD种属,系统分析了它们在极端条件下的脱氮特性及潜力,并简述了HN-AD菌在极端条件下的工艺应用研究进展,最后展望了HN-AD脱氮技术的应用前景和研究方向。     

异养硝化细菌的生物脱氮

正氮类污染物是造成水体污染和富营养化的主要原因,水体中氮的去除对于清洁水体有着重要的意义~([1-2])。因此,高效与便捷地去除污水中的氮是污水处理中的关键问题,而生物脱氮又被认为是目前废水脱氮中最经济有效的方法之一~([3])。最初研究者们认为只有自养硝化细菌才能进行高效的硝化作用,近年来研究表明,异养硝化微生物同样在生物硝化过程中起着不可忽视的作用~([4])。异养硝化微生物相比传统的自养微生物在废水生物脱氮中占有明显优势,具有良好的应用前景。