有机碳对混合菌群亚硝酸盐氮氧化的影响

自养硝化过程在自然界氮素循环和污水处理系统脱氮过程中起着关键作用。因此，了解有机碳对硝化的影响和硝化菌与异养菌之间的竞争对微生物生态学和污水处理系统设计都很重要。目前对氨氧化到硝酸盐氮过程的研究文献很多，但对亚硝酸盐氧化过程在异养菌的存在下如何受到有机碳影响的研究甚少。本文从生理生化指标、基因组学、蛋白组学三方面考察了在实验室条件下有机碳（乙酸钠）对硝化细菌和异养菌组成的混合菌群的硝化性能、菌群结构及代谢功能的变化的影响。 全文分为两大部分： 第一部分为乙酸钠对游离态硝化混合菌群的硝化性能和菌群结构的短期影响。混合菌株先在自养条件下进行连续培养，两个月后硝化速率达到20 mg N/(L·d)；而后离心收集菌体进行批式实验。在批式反应器中，初始亚硝氮均为126mg N/ L，乙酸钠-C 与亚硝酸盐-N 的比分别为0，0.44，0.88，4.41，8.82。结果表明：在低C/N 比（0.44 和0.88）时，亚硝酸盐去除速率比C/N=0 下高，细菌呈现一次生长；而在高C/N 比（4.41 和8.82）时，出现连续的硝化反硝化，亚硝酸盐去除率仍比对照下高，细菌呈现二次生长。不同C/N 比下微生物群落明显不同，优势菌群从自养和寡营养细菌体系（包括亚硝酸盐氧化菌，拟杆菌门,α-变形菌纲,浮霉菌门和绿色非硫细菌下的一些菌株）过渡到异养和反硝化菌体系 （γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas stutzeri 和P. nitroreducens 占主导）。 第二部分为乙酸钠对硝化混合菌群生物膜的硝化性能和菌群结构的长期影响。接种富集的硝化混合菌群于装有组合式填料的三角瓶中，于摇床中自养培养；两个月后填料上形成生物膜的硝化速率达到20 mg N/ (L·d)；而后进行长期实验，每12 小时更换混合营养培养基（亚硝氮约200 mg N/ L，C/N 比同上）。结果显示：相较于C/N 比=0 时的亚硝酸盐氧化反应来说，低C/N 比出现了部分的反硝化，而高C/N 比则是几乎完全的反硝化。与对照比，C/N=0.44 时亚硝酸盐氧化速率并未受乙酸钠的影响，反而上升了，但C/N=0.88 时亚硝酸盐氧化速率有所下降。菌群结构分析表明自养对照与混合营养下微生物群落的不同；PCR-DGGE未检测出混合营养下硝化杆菌的存在，而显示异养菌尤其是反硝化菌的大量存 在。荧光定量PCR 结果表明随C/N 比上升，硝化杆菌数量从2.42 × 104 下降到1.34× 103 16S rRNA gene copies/ ng DNA，反硝化菌由0 增加至2.51 × 104 nosZgene copies/ ng DNA。SDS-PAGE 的结果表明不同C/N 比下的蛋白组较为复杂且呈现一定的差异性。 有机碳对亚硝氮氧化及微生物群落的影响很复杂，本文分别讨论了对游离态和生物膜固定态两种状态的混合菌群相应的短期和长期影响研究。研究发现，有机碳并非一定带来硝化的负影响，如果控制在适当的C/N 比范围，有机碳是有利于亚硝氮氧化的。这些发现阐明了有机碳和硝化反硝化的关系，填补了硝化微生物生态学上的空白，对污水处理系统中减少异养菌的影响并提高氮去除率有一定理论指导意义。 Nitrification plays a key role in the biological removal of nitrogen in both nature and wastewater treatment plant (WWTP). So, understanding of the effect of organic carbon on nitrification and the competition between nitrifying bacteria and heterotrophic bacteria is important for both microbial ecology and WWTP design and operation. Despite the fact that the nitrification process of ammonia to nitrate has been extensively investigated, it is not known how the process of nitrite oxidization is affected by organic carbon when heterotrophic bacteria are present. By measuring different physiological and biochemical parameters, as well as using genomic DNA and proteome analysis, we investigated the influence of organic (acetate) on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria under laboratory conditions. The dissertation involves two parts: Part one deals with the effect of organic matter on functional performance and bacterial community shift of nitrite-oxidizing and heterotrophic bacteria under suspended state. The bacteria were prepared in a continuous-flow stirred reactor under autotrophic condition; after two months, the nitrification rate of the culture reached about 20 mg N/ (L·d); then the bacteria were harvested for the next batch experiments. The initial concentrations of nitrite were 126 ± 6 mg N/ L in all flasks, and sodium acetate (C) to nitrite (N) ratios were 0, 0.44, 0.88, 4.41, and 8.82, respectively. The results showed that at low C/N ratios (0.44 or 0.88), the nitrite removal rate was higher than that obtained under autotrophic condition and the bacteria had single growth phase, while at high C/N ratios (4.41 or 8.82), continuous aerobic nitrification and denitrification occurred besides higher nitrite removal rates, and the bacteria had double growth phases. The community structure of total bacteria strikingly varied with the different C/N ratios; the dominant populations shifted from autotrophic and oligotrophic bacteria (NOB, and some strains of Bacteroidetes, Alphaproteobacteria, Actinobacteria, and green nonsulfur bacteria) to heterotrophic and denitrifying bacteria (strains of Gammaproteobacteria, especially Pseudomonas stutzeri and P. nitroreducens). Part two describes the influence of acetate on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria in biofilms. Bacterial enrichments was transferred into flasks with polypropylene carriers and cultured under agitated and autotrophic condition. After two month, the biofilms grown on the carriers had a nitrification rate of about 20 mg N/ (L·h); then the biofilms were refreshed with mixotrophic medium (nitrite were 200 mg N/ L in all flasks, and C/N ratios was the same as above) every 12 h. the results show: normal nitrite oxidization reactions were performed when C/N = 0, but nitrite oxidization and partial denitrification occurred with low C/N ratios (0.44 or 0.88). At high C/N ratios (4.41 or 8.82), we mainly observed denitrification. In contrast to C/N = 0, the nitrite oxidization rate was unaffected when C/N = 0.44, but decreased with C/N = 0.88. The structure of bacterial communities varied significantly between autotrophic and mixotrophic conditions. Nitrobacter was hard to detect by PCR-DGGE while heterotrophs and especially denitrifiers were in the majority under mixotrophic conditions. Real-time PCR indicated that the Nitrobacter population decreased from 2.42 × 104 to 1.34 × 103 16S rRNA gene copies/ ng DNA, while the quantity of denitrifiers obviously increased from 0 to 2.51×104 nosZ gene copies/ ng DNA with an increasing C/N ratio. SDS-PAGE indicated the complexity of and a certain difference between the proteome of nitrite-oxidizing and heterotrophic bacteria at different C/N ratios. We conclude that the influence of organic matter on nitrite oxidation and the community structure of NOB and heterotrophic bacteria is complex. In this dissertation, we focused on how sodium acetate influenced the system both under suspended state and in biofilms. We observed that acetate did not necessarily have a negative impact on nitrification. Instead, an appropriate amount of acetate benefited both nitrite oxidization and denitrification. These findings provide a greater understanding about the relationship between organics and nitrification; they fill the gaps in the field of microbial ecology of nitrifying bacteria; they also provide insight into how to minimize the negative impact of heterotrophic bacteria and maximize the benefit of nitrogen removal in biological treatment systems.

A 2-kW COIL with a square pipe-array JSOG and nitrogen buffer gas

A 2-kW-class chemical oxygen-iodine laser (COIL) using nitrogen buffer gas has been developed and tested since industrial applications of COIL devices will require the use of nitrogen as the buffer gas. The laser, with a gain length of 11.7 cm, is energized by a square pipe-array jet-type singlet oxygen generator (SPJSOG) and employs a nozzle bank with a designed Mach number of 2.5. The SPJSOG has advantages over the traditional plate-type JSOG in that it has less requirements on basic hydrogen peroxide (BHP) pump, and more important, it has much better operational stability. The SPJSOG without a cold trap and a gas-liquid separator could provide reliable operations for a total gas flow rate up to 450 mmol/s and with a low liquid driving pressure of around 0.7 atm or even lower. The nozzle bank was specially designed for a COIL using nitrogen as the buffer gas. The cavity was designed for a Mach number of 2.5, in order to provide a gas speed and static temperature in the cavity similar to that for a traditional COIL with helium buffer gas and a Mach 2 nozzle. An output power of 2.6 kW was obtained for a chlorine flow rate of 140 mmol/s, corresponding to a chemical efficiency of 20.4%. When the chlorine flow rate was reduced to 115 mmol/s, a higher chemical efficiency of 22.7% was attained. Measurements showed that the SPJSOG during normal operation could provide a singlet oxygen yield Y greater than or equal to 55%, a chlorine utilization U greater than or equal to 85%, and a relative water vapor concentration w = [H2O]/([O-2] + [Cl-2]) less than or equal to 0.1.

Ammonia synthesis on graphitic-nanofilament supported Ru catalysts

Graphitic-nanofilaments (GNFs) supported ruthenium catalysts were prepared and characterized by NZ physisorption, X-ray diffraction (XRD), transmission electron microscope (TEM) and temperature programmed reduction-mass spectroscopy (TPR-MS) and used for ammonia synthesis in a fixed bed microreactor. The TEMs of the Ru/GNFs and Ru-Ba/GNFs catalysts indicate that the Ru particles are in the range of 2-4 nm, which is the optimum size of Ru particles for the maximum number of B5 type sites. The activity of Ru-Ba/GNFs catalysts is higher than that of Ru-Ba/AC by about 25%. The methanation reaction on the Ru/GNFs catalyst is remarkably inhibited compared with a Ru/AC catalyst. High graphitization of GNFs is likely to be the reason for the high resistance to the methanation reaction. The power rate law for ammonia synthesis on Ru-Ba/GNFs catalysts can be expressed by r = Kp(NH3)(-0.4) P-N2(0.8) P-H2(-0.7), indicating that H-2 is an inhibitor for N-2 activation on the catalyst. Catalysts with the promoters Ba, K and Cs show large differences in activity for ammonia synthesis. The catalyst promoted with Ba (Ba/Ru = 0.2 molar ratio) was found to be the most active, whereas that with a K promoter was the least active. (C) 2003 Elsevier B.V. All rights reserved.

半湿润农田生态系统长期施肥对土壤团聚体中有机氮组分的影响

通过对黄土高原半湿润农田生态系统25年的田间肥料定位试验,研究了长期不同施肥模式对土壤有机氮组分及其在各级团聚体中分布的影响.结果表明:长期施肥对水解氨态氮、水解未知氮在土壤各级团聚体中分布的影响最大,对氨基酸态氮的分布有一定影响,而对氨基糖态氮分布的影响较小.长期施用化肥和有机肥能有效地影响水解氨态氮和水解未知氮与团聚体的结合作用,而氨基糖态氮在土壤氮循环转化过程中具有较强的稳定性.长期施肥条件下土壤水解全氮与有机碳、全氮以及团聚体分形维数均呈极显著正相关,其r分别为0.942,0.981,0.910(P<0.001),说明土壤有机氮组分对土壤团聚体的形成和性质具有显著影响.相关分析表明,土壤全氮或有机质对1～2mm和0.25～1mm土壤团聚体中各有机氮组分的影响较大

黄绵土不同形态有机氮径流流失规律

利用不同坡度径流小区 ,在自然降水条件下 ,研究侵蚀泥沙不同形态有机氮的流失规律 ,结果表明 :在 4次产流泥沙中 ,除氨基糖氮含量减少外 ,其它有机形态氮均存在增加趋势 ,以氨态氮含量增加最为显著。随坡度的下降 ,水解全氮、氨态氮和氨基酸氮含量呈增加趋势 ;泥沙中水解全氮、氨基酸氮、氨态氮、非酸水解氮均存在不同程度的富集 ,其富集率随坡度的下降而增大 ,以氨基酸氮富集程度高 ,泥沙中氨基糖态氮反而无富集现象存在 ;当坡度分别为 2 5°、2 0°、15°、10°和 5°时 ,水解全氮流失量分别为 1135 .0、76 3.7、485 .5、313.0和 15 4.4kg/ km2 ,而氨基酸氮流失量则为 2 34.8、182 .7、12 1.1、76 .8和 33.9kg/ km2 ,占水解全氮的 2 0 .6 %～ 2 4.9%。氨态氮流失量为 742 .5、5 0 3.3、32 2 .2、2 0 8.9、97.4kg/ km2 ,占水解全氮的 6 3.1%～ 6 6 .7%。氨基糖氮占酸水解氮 1.1%～ 2 .9%。非鉴别氮占水解全氮的 5 .8%～ 12 .6 %。而酸水解氮与非酸水解氮之比分别为 ...

Nitrogen-15 signals of leaf-litter-soil continuum as a possible indicator of ecosystem nitrogen saturation by forest succession and N loads

National Key Research and Development Program [2010CB833502]; National Natural Science Foundation of China [30600071, 40601097, 30590381]; Chinese Academy of Sciences [KZCX2-YW-432, O7V70080SZ, LENOM07LS-01]; GUCAS [O85101PM03]

Short-term effect of increasing nitrogen deposition on CO2, CH4 and N2O fluxes in an alpine meadow on the Qinghai-Tibetan Plateau, China

National Key Research and Development Program [2010CB833500]; Natural Science Foundation of China [30590381]; Knowledge Innovation Project of Chinese Academy of Sciences [KZCX2-YW-432]

Effects of temperature, glucose and inorganic nitrogen inputs on carbon mineralization in a Tibetan alpine meadow soil

High levels of available nitrogen (N) and carbon (C) have the potential to increase soil N and C mineralization We hypothesized that with an external labile C or N supply alpine meadow soil will have a significantly higher C mineralization potential and that temperature sensitivity of C mineralization will increase To test the hypotheses an incubation experiment was conducted with two doses of N or C supply at temperature of 5 15 and 25 C Results showed external N supply had no significant effect on CO2 emission However external C supply increased CO2 emission Temperature coefficient (Q(10)) ranged from 113 to 1 29 Significantly higher values were measured with C than with N addition and control treatment Temperature dependence of C mineralization was well-represented by exponential functions Under the control CO2 efflux rate was 425 g CO2-Cm-2 year(-1) comparable to the in situ measurement of 422 g CO2-Cm-2 year(-1) We demonstrated if N is disregarded microbial decomposition is primarily limited by lack of labile C It is predicted that labile C supply would further increase CO2 efflux from the alpine meadow soil (C) 2010 Elsevier Masson SAS All rights reserved