一株好氧反硝化菌的反硝化性能研究

从长期运行的生物滤塔中筛选出一株好氧反硝化菌株A1,经鉴定为恶臭假单胞菌Pseudomonas putida。文章目的是对A1的反硝化特性进行研究,结果表明A1菌株在好氧条件下能有效去除培养液中的硝酸盐氮,24h脱氮率可达到94.84%。C/N对菌株A1的好氧反硝化能力有很大影响,当C/N>5时,基本能够进行完全的反硝化。和其他已报道的好氧反硝化菌相比,A1菌株有着更高的氧耐受浓度。菌株A1能够以硝酸盐或亚硝酸盐和氧气为电子受体进行协同呼吸,硝酸盐呼吸和亚硝酸盐呼吸都具有较高的脱氮效率,并且亚硝酸盐呼吸要较硝酸盐呼吸更容易进行。以丁二酸盐、葡萄糖和乙酸盐作为碳源时,其脱氮效果均要明显好于乙醇作为碳源。     

Effect of dissolved oxygen on methane production from bottom sediment in a eutrophic stratified lake

Clarifying the role of sulfate and dissolved oxygen (DO) in methane production may allow for precise and accurate modeling of methane emissions in eutrophic lakes. We conducted field observations of sulfate, methane, and DO concentrations in Lake Abashiri, a typical brackish and eutrophic lake in a cold region, to develop a DO-based method for quantitively estimating methane production in a eutrophic lake and analyzed the results. We found that sulfate concentrations decreased rapidly from 900.0 mg/L in water overlying the sediments to nearly 0.0 mg/L in the bottom sediment. Methane production was almost uniform across sediment depths of 0.05 to 0.25 m, ranging from 1400 to 1800 µmol/m²/day. Also, methane production was found to be a function of DO concentrations in water overlying the bottom and could be modeled by a logistic function: constant production at 1,400 µmol/m²/day for DO concentrations of 0.0 to 3.0 mg/L, rapidly decreasing to 0 µmol/m²/day for DO concentrations of 3.0 to 6.0 mg/L. This methane model was verified using a simple one-dimensional numerical model that showed good agreement with field observations. Our results thus suggest that the proposed methane model reduces uncertainty in estimating methane production in a eutrophic lake.     

温度和DO对MBBR系统硝化和反硝化的影响

在缺氧/好氧/好氧串联运行的移动床生物膜反应器（MBBR）系统中考察了温度和好氧反应器中溶解氧（DO）水平对生物膜硝化和反硝化过程氮素去除的影响,并通过高通量测序技术探究温度和DO的变化造成的MBBR系统中脱氮功能菌群结构的差异,从而在微观水平解释硝化和反硝化受温度和DO影响的生物学机理.结果表明,系统温度的升高可以同时强化生物膜硝化和反硝化过程,且好氧反应器中DO水平的提高对硝化过程有利,从而提高系统的脱氮效果.本研究中,在系统连续运行阶段,当系统温度和好氧O₁反应器的DO浓度为本研究范围内的最高水平时（即温度=20~22℃、DO=5~8mg O₂/L）,比硝化负荷可达1.60g NH₄⁺-N/（m²·d）以上,而相同温度范围内比反硝化负荷可高达2.84g NO₃^--N/（m²·d）,从而使MBBR系统在该工况条件下获得了最佳的NH₄⁺-N和TN去除率（分别达到了98.7%和85.7%）.温度和DO影响硝化和反硝化的根本原因是温度和DO变化引起了脱氮功能菌群数量和群落结构的改变：当好氧反应器的DO水平下降时,硝化功能细菌的OTUs比例显著降低,尤其是异养硝化细菌的生长受到了严重的抑制;而温度的变化对反硝化细菌的影响主要体现在群落结构的变化.     

Elevated CO2 facilitates C and N accumulation in a rice paddy ecosystem

Elevated CO₂ can stimulate wetland carbon (C) and nitrogen (N) exports through gaseous and dissolved pathways, however, the consequent influences on the C and N pools are still not fully known. Therefore, we set up a free-air CO₂ enrichment experiment in a paddy field in Eastern China. After five year fumigation, we studied C and N in the plant–water–soil system. The results showed: (1) elevated CO₂ stimulated rice aboveground biomass and N accumulations by 19.1% and 12.5%, respectively. (2) Elevated CO₂ significantly increased paddy soil TOC and TN contents by 12.5% and 15.5%, respectively in the 0–15 cm layer, and 22.7% and 26.0% in the 15–30 cm soil layer. (3) Averaged across the rice growing period, elevated CO₂ greatly increased TOC and TN contents in the surface water by 7.6% and 11.4%, respectively. (4) The TOC/TN ratio and natural δ¹⁵N value in the surface soil showed a decreasing trend under elevated CO₂. The above results indicate that elevated CO₂ can benefit C and N accumulation in paddy fields. Given the similarity between the paddies and natural wetlands, our results also suggest a great potential for long-term C and N accumulation in natural wetlands under future climate patterns.     

从不同反应器筛选、鉴别好氧反硝化菌

采用2个序批式反应器A和B,以硝态氮为唯一氮源,采用间歇曝气,以驯化、富集耐氧脱氮污泥.反应器A,其pH约为6.3,ρ(DO)为2.2～6.1 mg/L,碳氮比(ρ(C)/ρ(N),ρ(C)以ρ(COD_Cr)计)约为9;反应器B,其pH约为6.8～7.8,ρ(DO)为 2.2～3.0 mg/L,ρ(C)/ρ(N)约为15.2个反应器的ρ(NO₃－-N)均保持为80 mg/L.当2个反应器的总氮去除率达到60%以上,则认为完成好氧反硝化菌的富集.从2个反应器中共筛选得到20株BTB阳性菌,其中8株菌株的DNA样品经PCR成功扩增,进行16S rRNA测序.测序结果提交GenBank进行Blast同源性检索,并分析比对鉴别,判断8株菌株分属于假单胞菌(Pseudomonas),戴尔福特菌(Delftia),草螺菌(Herbaspirillum)和丛毛单胞菌(Comamonas)菌属.反硝化性能测定证实8株菌株均为好氧反硝化菌.      

Temporal and spatial variation of nitrogen and phosphorus and eutrophication assessment for a typical arid river—Fuyang River in northern China

Based on water quality surveys over 2 years(July to December,in 2014 and 2015) in a typical arid river in northern China the Xingtai segment of the Fuyang River basin — the variation of nitrogen(N) and phosphorus(P) was analyzed.The extent of water eutrophication of this segment was also assessed using a universal index formula for eutrophic evaluation and a logarithmic power function.The results showed that the average concentration of total N(TN) was 27.2 mg/L(NH_3-N,63.5%),total P(TP) was 2.0 mg/L(solution reactive phosphorus,68.8%).Temporal and spatial variations of N and P in this segment were observed.Concentrations of N and P in the arid season were higher than those in the rainy season.Spatially,the N and P concentrations followed the same trend;i.e.,higher in the city segment than in the suburbs,and decreasing along the river.The water eutrophication in the studied segment reached extremely high levels at all times(eutrophication index ≥76.3).Spatially,its trend was clearly linked with N and P.Water shortage,pollution accumulation and a weak self-purification function are the main reasons for the prominent eutrophication in this segment.     

Effect of dissolved oxygen on nitrate removal using polycaprolactone as an organic carbon source and biofilm carrier in fixed-film denitrifying reactors

Nitrate-nitrogen(NO_3~--N) always accumulates in commercial recirculating aquaculture systems(RASs) with aerobic nitrification units. The ability to reduce NO_3~--N consistently and confidently could help RASs to become more sustainable. The rich dissolved oxygen(DO)content and sensitive organisms stocked in RASs increase the difficulty of denitrifying technology. A denitrifying process using biologically degradable polymers as an organic carbon source and biofilm carrier was proposed because of its space-efficient nature and strong ability to remove NO_3~--N from RASs. The effect of dissolved oxygen(DO) levels on heterotrophic denitrification in fixed-film reactors filled with polycaprolactone(PCL) was explored in the current experiment. DO conditions in the influent of the denitrifying reactors were set up as follows: the anoxic treatment group(Group A, average DO concentration of 0.28 ± 0.05 mg/L), the low-oxygen treatment DO group(Group B, average DO concentration of 2.50 ± 0.24 mg/L) and the aerated treatment group(Group C, average DO concentration of 5.63 ± 0.57 mg/L). Feeding with 200 mg/L of NO_3~--N, the NO_3~--N removal rates were 1.53, 1.60 and 1.42 kg/m3PCL/day in Groups A, B and C, respectively. No significant difference in NO_3~--N removal rates was observed among the three treatments. It was concluded that the inhibitory effects of DO concentrations lower than 6 mg/L on heterotrophic denitrification in the fixed-film reactors filled with PCL can be mitigated.     

湖库沉积物好氧反硝化菌群脱氮特性及种群结构

好氧反硝化菌是水环境生物修复的热点研究领域,本研究从西安市金盆水库、兴庆公园和长乐公园采集沉积物,在运用传统的间歇曝气富集和选择培养基筛选好氧反硝化菌的基础上,引入超声波预处理沉积物-水悬液,成功筛选出3组高效好氧反硝化菌群H-30、X-10和C-30,并对其反硝化能力进行研究.结果表明,该3组混合好氧反硝化菌群具有高效的脱氮能力,在溶解氧为(7.2±0.6)mg·L-1的培养基中,好氧反硝化菌群H-30、X-10和C-30总氮(TN)去除率分别达到83.04%,83.40%和82.68%,仅有少量亚硝氮(NO-2-N)积累.Illumina高通量DNA测序结果表明,3组nir S型好氧反硝化优势混合菌群种群结构差异显著,H-30,X-10和C-30菌群的优势菌种分别是枯草芽孢杆菌(Bacillus subtilis),Paracoccus pantotrophus和斯氏假单胞菌(Pseudomonas stutzeri),其中斯氏假单胞菌(P.stutzeri)在H-30、X-10菌群中所占比例差异不显著,Pseudomonas xiamenensis仅存在于H-30.本研究筛选的3组高效好氧反硝化菌群将为城市内湖污染水体的微生物修复工程提供菌源保障.     

Effects of sulfate-reducing bacteria on methylmercury at the sediment–water interface

Sediment cores(containing sediment and overlying water) from Baihua Reservoir(SW China)were cultured under different redox conditions with different microbial activities, to understand the effects of sulfate-reducing bacteria(SRB) on mercury(Hg) methylation at sediment–water interfaces. Concentrations of dissolved methyl mercury(DMe Hg) in the overlying water of the control cores with bioactivity maintained(BAC) and cores with only sulfate-reducing bacteria inhibited(SRBI) and bacteria fully inhibited(BACI) were measured at the anaerobic stage followed by the aerobic stage. For the BAC and SRBI cores, DMe Hg concentrations in waters were much higher at the anaerobic stage than those at the aerobic stage, and they were negatively correlated to the dissolved oxygen concentrations(r =- 0.5311 and r =- 0.4977 for BAC and SRBI, respectively). The water DMe Hg concentrations of the SRBI cores were 50% lower than those of the BAC cores, indicating that the SRB is of great importance in Hg methylation in sediment–water systems, but there should be other microbes such as iron-reducing bacteria and those containing specific gene cluster(hgc AB), besides SRB,causing Hg methylation in the sediment–water system.     

活性污泥中好氧反硝化菌的富集筛选及鉴别

采用SBR反应器,以硝基氮为底物,通过间歇曝气方式,DO保持5mg/L以上,对活性污泥进行强化驯化,实现好氧反硝化细菌的富集培养。利用琥珀酸钠作为碳源,溴百里酚蓝(BTB)作为pH指示剂,共筛选得到20株BTB琼脂平板阳性菌。通过反硝化性能测定,复筛得到4株好氧反硝化细菌。实验结果表明,琥珀酸盐为碳源、硝酸盐为惟一氮源、C/N<10的条件下,4株菌在4d内的TIN去除率均达到60%以上。通过16SrRNA序列同源性比较成功鉴定出3株菌,初步判断2株属于Pseudomonas菌属、1株属于Delftia菌属。     

Enhancing anoxic denitrification of low C/N ratio wastewater with novel ZVI composite carriers

External organic carbon sources are needed to provide electron donors for the denitrification of wastewater with a low COD/NO₃^--N（C/N） ratio, increasing the treatment cost. The economic strategy is to enhance the bioactivity and/or biodiversity of denitrifiers to efficiently utilize organic substances in wastewater. In this study, novel zero-valent iron（ZVI） composite carriers were prepared and implemented in a suspended carrier biofilm reactor to enhance the bioactivity an...     

溶解氧对碳氮硫共脱除工艺中微生物群落影响解析

为了研究溶解氧对碳氮硫共脱除工艺中微生物群落的影响,探索供氧条件下提高单质硫转化率的机制,实验采用基因芯片GeoChip技术来解析不同曝气量下采集的污泥样品中的微生物群落结构特征.研究结果表明,微量的溶解氧能显著影响工艺中的微生物群落结构(P<0.001),相对较高浓度的溶解氧可以增加微生物的多样性但是却抑制了对溶解氧比较敏感的反硝化微生物的活性,导致工艺硝酸盐去除率下降.与反硝化微生物相比硫酸盐还原菌(sulfate-reducing bacteria,SRB)能抵抗更高浓度的溶解氧,因此工艺的硫酸盐还原过程未受到影响,一直保持较高的硫酸盐去除率.当曝气量为20 mL·min-1时单质硫的转化率最高,通过对sox(sulfur-oxidation)基因丰度分析和聚类分析发现,在该曝气量条件下sox基因的丰度最高,与厌氧条件相比有极其显著的差异.此外,GeoChip还检测到一些硫氧化细菌(sulfur-oxidizing bacteria,SOB)的丰度明显提高,证明适量的溶解氧能够刺激一部分SOB的生长与代谢.     

异养硝化-好氧反硝化菌的筛选及脱氮性能的实验研究

通过极限稀释和显色培养基相结合筛选的方法,从畜禽养殖废水样品中筛选到1株同时具有异养硝化-好氧反硝化双重功能的细菌CPZ24.该菌株革兰氏染色呈阳性,杆状,菌落颜色为橙红色.经形态、生理生化特性,16S rDNA序列分析,初步鉴定该菌为嗜吡啶红球菌(Rhodococuus pyridinivorans).对该菌进行异养硝化功能和好氧反硝化功能进行研究,结果表明,在异养硝化过程中,该菌可将培养基中的氨氮全部去除,其中对总氮的去除率可达98.70%;在好氧反硝化过程中,该菌对硝酸盐氮的去除率可达到66.74%,总氮的去除率达到64.27%.此高效脱氮降解菌可实现自身同步硝化-反硝化脱氮功能,能够独立完成生物脱氮的全过程.     

Nitrous oxide reductase gene (nosZ) and N2O reduction along the littoral gradient of a eutrophic freshwater lake

Lake littoral zones are characterized by heterogeneity in the biogeochemistry of nutrient elements. This study aimed to explore the relationship between the nitrous oxide reductase gene (nosZ)-encoding denitrifier community composition/abundance and N2O reduction. Five samples (deep sediment, near-transition sediment, transition site, near-transition land and land soil) were collected along a littoral gradient of eutrophic Baiyangdian Lake, North China. To investigate the relationship between the nosZ-encoding denitrifier community structure and N2O reduction, the nosZ-encoding denitrifier community composition/abundance, potential denitrification rate (DNR) and potential N2O production rate (pN2O) were investigated using molecular biological technologies and laboratory incubation experiments. The results showed that the average DNR of sediments was about 25 times higher than that of land soils, reaching 282.5 nmol N/(g dry weight (dw)·hr) and that the average pN2O of sediments was about 3.5 times higher than that of land soils, reaching 15.7 nmol N/(g dw·hr). In the land area, the nosZ gene abundance showed a negative correlation with the N2O/(N2O+N2) ratio, indicating that nosZ gene abundance dominated N2O reduction both in the surface soils of the land area and in the soil core of the transition site.Phylogenetic analysis showed that all the nosZ sequences recovered from sediment clustered closely with the isolates Azospirillum largimobile and Azospirillum irakense affiliated to Rhodospirillaceae in alpha-Proteobacteria, while about 92.3% (12/13) of the nosZ sequences recovered from land soil affiliated to Rhizobiaceae and Bradyrhizobiaceae in α-Proteobacteria. The community composition of nosZ gene-encoding denitrifiers appeared to be coupled with N2O reduction along the littoral gradient.     

厌氧/好氧SNEDPR系统处理低C/N污水的优化运行

为实现同步硝化内源反硝化除磷（SNEDPR）系统的优化运行,以实际生活污水为处理对象,采用厌氧（180min）/好氧运行的SBR反应器,并通过联合调控好氧段溶解氧（DO）浓度（0.3～1.0mg/L）和好氧时间（150～240min）,考察了该系统脱氮除磷特性.并结合荧光原位杂交（FISH）技术对系统优化过程中各功能菌群的结构变化情况进行了分析.试验结果表明,当系统好氧段DO浓度由约1.0mg/L逐渐降至0.3mg/L,且好氧时间由150min逐渐延长至240min后,出水PO₄^3--P浓度稳定在0.4mg/L左右,但出水TN浓度由14.3mg/L降至8.7mg/L,TN去除率由75%提高至84%.此外,随着好氧段DO浓度的降低,SNED现象愈加明显,SNED率由34.7%逐渐升高至63.8%.SNED的加强,降低了出水NO₃^--N浓度,并提高了系统的脱氮性能和厌氧段的内碳源储存量.FISH结果表明：经127d的优化运行,系统内PAOs,GAOs和AOB（氨氧化菌）仍保持在较高水平（分别全菌的29%±3%,20%±3%和13%±3%）,其保证了系统除磷、硝化和反硝化脱氮性能;但NOB（亚硝酸盐氧化菌）含量减少了50%,为系统内实现短程硝化内源反硝化提供了可能.     

微气泡曝气生物膜反应器同步硝化反硝化研究

同步硝化反硝化(SND)是废水处理中的新型生物脱氮工艺,和传统生物脱氮工艺相比具有显著的应用优势.本研究采用微气泡曝气固定床生物膜反应器,研究了SND过程中污染物去除效果并检测了生物膜功能菌群的变化情况.结果表明,在微气泡曝气固定床生物膜反应器内可以实现同步硝化反硝化,通过提高进水COD负荷和C∶N比,降低溶解氧(DO)浓度,同时增加填料床层孔隙率,可以改善SND效果.当进水COD负荷和总氮(TN)负荷为0.86 kg·(m3·d)-1和0.10 kg·(m3·d)-1,且填料床层孔隙率为81%时,COD和TN的去除率分别为97.6%和70.2%,实现了COD和TN的同步高效去除;同时,微气泡曝气对氧传质的强化作用使得氧利用率高达91.8%.此外,生物膜活性和硝化及反硝化功能菌群的变化,与反应器COD、氨氮和TN去除能力的变化基本一致.     

三峡蓄水期间汉丰湖消落区营养状态时间变化

为探明三峡蓄水后汉丰湖消落区水质营养状态的变化特征,于2013年10月至2014年2月对水质进行连续观察,测定了水质物理参数、营养盐与叶绿素(Chl-a)的质量浓度.结果表明,水体中营养盐与Chl-a质量浓度的增加,在淹水后营养程度有升高现象,2014年2月与2013年10月相比,TN、TP、高锰酸盐指数与Chl-a质量浓度分别增加了4.7、1.0、0.2、3.27倍,TN、TP质量浓度均超过藻类生长限值,随滞留时间延长易造成水体富营养化,应引起重视.Chl-a单因子评价反映出水质由贫营养向富营养演变.TN/TP结果表明,TN、TP分别在不同时间内制约着藻类的生长;2013年10~12月与2014年2月,藻类生长受TN限制;2014年1月,藻类生长受TP限制.Chl-a与p H、DO、NH+4-N、NO-3-N、TN、高锰酸盐指数及TP呈显著正相关,而与SD、水温呈显著负相关;蓄水期间,水质受到了同一污染源的影响.因子分析结果表明,汉丰湖消落区水质主要受p H、DO、NO-3-N、TN的影响,同时Chl-a、TP、NH+4-N与好氧性有机物的污染不可忽视;在蓄水稳定初期水体具有自净能力,随蓄水滞留时间的延长,水质污染程度整体上呈现逐步恶化的趋势,应加以控制;三峡蓄水期间,南河、东河营养程度相对较高,应加强治理.     

不同品种黑麦草对富营养化水体净化能力的比较

利用浮床植物系统研究了不同品种黑麦草在富营养化水体中的生长特性和净化效果差异.结果表明,12个黑麦草品种在冬春季富营养化水体中均能良好生长,净化效果非常明显,对TN、TP、CODMn和BOD5的去除率分别高达73.68%、84.69%、87.87%和75.85%;同时,还能使富营养化水体中pH值降低,溶解氧浓度明显升高.室内试验植物地上部N、P积累量分别在46.86～145.25 mg·m-2·d-1之间和11.39～42.74mg·m-2·d-1之间;室外试验植物地上部N、P积累量分别为61.5～168.2 mg·m-2·d-1和11.9～47.2mg·m-2·d-1;且植物体内N、P积累量与其生物量之间存在着明显的相关性(r≥0.90).品种同差异显著(P<0.05),其中,以邦德、安格斯2号和牧杰对富营养化水体净化的综合能力表现最好.     

味精废水处理系统中高效细菌的分离鉴定及其

利用BTB培养基快速高效地从某味精厂废水处理系统的活性污泥中筛选出一株高效异养硝化-好氧反硝化细菌LH2-3.通过形态观察,生理生化特征测定,Biolog鉴定以及16S rDNA分子鉴定,认定该菌株是巨大芽孢杆菌(Bacillus megaterium).菌株脱氮性能研究表明:在异养硝化过程中,该菌株可将培养液中的NH+₄-N　100%去除,对TN的去除率可达98.16%;在好氧反硝化过程中,其对NO-₂-N的去除率达到97.14%,对TN的去除率达到94.97%.      

河道底泥中异养硝化-好氧反硝化菌群富集培养及其脱氮性能

采用选择性富集培养基,从河道底泥中成功筛选出具有氮素去除效果的优势微生物菌群:异养硝化-好氧反硝化菌群(H)、异养硝化菌群(Y)和好氧反硝化菌群(F).高通量测序分析结果显示,各混合菌群中脱氮功能菌的相对丰度分别为54.72%、46.26%和91.57%.在此基础上,考察了微量元素对3组混合菌群脱氮性能的影响,结果表明,混合菌群H和F在添加微量元素浓缩液0.5 mL·L~(-1)时脱氮性能最佳,在72 h时混合菌群H对氨氮、硝酸盐氮的去除率分别为69.11%和99.06%,混合菌群F对硝酸盐氮的去除率达99.62%;而混合菌群Y在不添加微量元素时氨氮的去除率最高,为82.60%.采用优化培养的3组混合菌群及其复合菌群(H+Y、Y+F、H+F、H+Y+F)分别对模拟河水进行处理,模拟河水的COD和氮素指标可从地表水V类水质净化至Ⅳ类水质,其中,复合菌群H+Y+F的氮素去除效果最佳,在36 h时氨氮、硝酸盐氮和总氮的浓度分别为0.182、0.000和0.501 mg·L~(-1),说明本研究富集筛选出的混合脱氮菌群在实际河水净化中具有潜在应用价值.