Effect of Zinc Oxide Nanoparticles on Denitrification and Denitrifying Bacteria Communities in Typical Estuarine Sediments

For revealing the effects of increasing of zinc oxide nanoparticles(ZnO NPs)on denitrification and denitrifying bacteria communities in estuarine sediments,the surface sediments of two typical estuaries(the Yangtze River Estuary and the Yellow River Estuary)were added with medium concentration(170mgL−1)and high concentration(1700mgL−1)of ZnO NPs for anaerobic cul-ture in laboratory.The concentration of NO_(3)^(−)and NO_(2)^(−),the reductase activity and denitrification rate were measured by physico-chemical analysis,nirS gene abundance and denitrifying bacteria communities by molecular biological methods.The results showed that ZnO NPs inhibited NO_(3)^(−), NO_(2)^(−)reduction process and NO_(3)^(−), NO_(2)^(−)reductase activity,and a stronger inhibition effect resulting from the higher ZnO NPs concentration.ZnO NPs decreased nirS gene abundance and community diversity of denitrifying bacteria.In addition,the inhibition degree of ZnO NPs on the denitrification process of sediments in different estuaries was different.These results were of great significance for evaluating the potential ecological toxicity and risks of nanomaterials in estuaries.

Temporal characteristics of algae-denitrifying bacteria co-occurrence patterns and denitrifier assembly in epiphytic biofilms on submerged macrophytes in Caohai Lake,SW China

Denitrifying bacteria in epiphytic biofilms play a crucial role in nitrogen cycle in aquatic habitats.However,little is known about the connection between algae and denitrifying bacteria and their assembly processes in epiphytic biofilms.Epiphytic biofilms were collected from submerged macrophytes(Patamogeton lucens and Najas marina L.)in the Caohai Lake,Guizhou,SW China,from July to November 2020 to:(1)investigate the impact of abiotic and biotic variables on denitrifying bacterial communities;(2)investigate the temporal variation of the algae-denitrifying bacteria co-occurrence networks;and(3)determine the contribution of deterministic and stochastic processes to the formation of denitrifying bacterial communities.Abiotic and biotic factors influenced the variation in the denitrifying bacterial community,as shown in the Mantel test.The co-occurrence network analysis unveiled intricate interactions among algae to denitrifying bacteria.Denitrifying bacterial community co-occurrence network complexity(larger average degrees representing stronger network complexity)increased continuously from July to September and decreased in October before increasing in November.The co-occurrence network complexity of the algae and nirS-encoding denitrifying bacteria tended to increase from July to November.The co-occurrence network complexity of the algal and denitrifying bacterial communities was modified by ammonia nitrogen(NH_(4)^(+)-N)and total phosphorus(TP),pH,and water temperature(WT),according to the ordinary least-squares(OLS)model.The modified stochasticity ratio(MST)results reveal that deterministic selection dominated the assembly of denitrifying bacterial communities.The influence of environmental variables to denitrifying bacterial communities,as well as characteristics of algal-bacterial co-occurrence networks and the assembly process of denitrifying bacterial communities,were discovered in epiphytic biofilms in this study.The findings could aid in the appropriate understanding and use of epiphytic biofilms denitrification function,as well as the enhancement of water quality.

Isolation of Denitrifying Bacteria in Sea Sediment and Simulated Experiment of Removing Nitrate-N from Seawater

The objective of this research was to isolate denitrifying bacteria from sea sediment and simulate the removal efficiency of nitrate-N by denitrifying bacteria from seawater. The result showed that the isolated denitrifying bacteria could effectively remove nitrate-N from seawater. About 90 % of nitrate-N was removed by denitrifying bacteria from seawater within a week in the simulated experiment I （the initial concentration of nitrate-N was 100 mg/L）. The removal efficiency of nitrate-N reached about 70 % within one day in the simulated experiment Ⅱ （initial concentration of nitrate-N was 1 mg/L）. The final removal efficiency was about 98 % and 85 % in the simulated experiments Ⅰ and Ⅱ, respectively. It was found that there was positive correlation between the concentration of nitrate-N and the number of denitrifying bacteria in seawater. Lots of denitrifying bacteria would disappear and the seawater would become transparent once the process of bioremediation was completed.

Impact of Long-Term Fertilization on Community Structure of Ammonia Oxidizing and Denitrifying Bacteria Based on amoA and nirK Genes in a Rice Paddy from Tai Lake Region,China

Ammonia oxidizing （AOB） and denitrifying bacteria （DNB） play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community composition of AOB and DNB were studied with targeting ammonia monooxygenase （amoA） and nitrite reductase （nirK） genes using polymerase chain reaction- denaturing gradient gel electrophoresis （PCR-DGGE） and real-time PCR, respectively. A field trial with different fertilization treatments in a rice paddy from Tai Lake region, centre East China was used in this study, including no fertilizer application （NF）, balanced chemical fertilizers （CF）, combined organic/inorganic fertilizer of balanced chemical fertilizers plus pig manure （CFM）, and plus rice straw return （CFS）. The abundances and riehnesses of amoA and nirK were increased in CF, CFM and CFS compared to NF. Principle component analysis of DGGE profiles showed significant difference in nirK and amoA genes composition between organic amended （CFS and CFM） and the non-organic amended （CF and NF） plots. Number of amoA copies was significantly positively correlated with normalized soil nutrient richness （NSNR） of soil organic carbon （SOC） and total nitrogen （T-N）, and that of nirK copies was with NSNR of SOC, T-N plus total phosphorus. Moreover, nitrification potential showed a positive correlation with SOC content, while a significantly lower denitrification potential was found under CFM compared to under CFS. Therefore, SOC accumulation accompanied with soil nutrient richness under long-term balanced and organic/inorganic combined fertilization promoted abundance and diversity of AOB and DNB in the rice paddy.

Operation of three parallel AN/AO processes to enrich denitrifying phosphorus removing bacteria for low strength wastewater treatment

Three parallel anaerobic-anoxic/anaerobic-aerobic （AN/AO） processes were developed to enrich denitrifying phosphorus removal bacteria （DPB） for low strength wastewater treatment. The main body of the parallel AN/AO process consists of an AN （anaerobic-anoxic） process and an AO （anaerobic-aerobic） process. In the AO process, the common phosphorus accumulating organisms （PAOs） was dominate, while in the AN process, DPB was dominate, The volume of anaerobic zone（Vana）：anoxie zone（Vano） ： aerobic zone （Vaer） for the parallel AN/AO process is 1：1：1 in contrast with a Vana：Vaer and Vano：Vaer of 1：2 and 1：4 for a traditional biological nutrient removal process （BNR）. Process 3 excels in the 3 processes on the basis of COD, TN and TP removal. For 4 month operation, the effluent COD concentration of process 3 did not exceed 60 mg/L; the effluent TN concentration of process 3 was lower than 15 mg/L; and the effluent TP concentration of process 3 was lower than 1 mg/L.

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

To investigate the characteristics and metabolic mechanism of short-cut denitrifying phospho- rus-removing bacteria （SDPB） that are capable of enhanced biological phosphorus removal （EBPR） using nitrite as an electron acceptor, an aerobic/anoxic sequencing batch reactor was operated under three phases. An SDPB-strain YC was screened after the sludge enrichment and was identified by morphological, physiological, biochemical properties and 16S rDNA gene sequence analysis. Denitrifying phosphorus-removing experiments were conducted to study anaerobic and anoxic metabolic mechanisms by analyzing the changes of chemical oxygen demand （COD）, phosphate, nitrite, poly-fl-hydroxybutyrate （PHB）, and glycogen. The results show that strain YC is a non-fermentative SDPB similar to Paracoccus denitrificans. As a kind of non-fermentative bacteria, the energy of strain YC was mainly generated from phosphorus release （96.2%） under anaerobic conditions with 0.32 mg P per mg synthesized PHB. Under anoxic conditions, strain YC accumulated 0.45 mg P per mg degraded PHB, which produced most of energy for phosphate accumulation （91.3%） and a little for glycogen synthesis （8.7%）. This metabolic mechanism of strain YC is different from that of traditional phosphorus-accumulating organisms （PAOs）. It is also found that PHB, a kind of intracellular polymer, plays a very important role in denitrifying and accumulating phosphorus by supplying sufficient energy for phosphorous accumulation and carbon sources for denitrification. Therefore, monitoring AP/APHB and ANO2 -N/APHB is more necessary than monitoring AP/ACOD, ANO2 -N/ACOD, or AP / ANO2 -N.

Denitrifying Bacteria in Paddy Soils of the Taihu Lake Basin,China

This study attempted to determine the characteristics of the communities, the ecological factors, and the denitrifying enzyme activity for denitrifying bacteria found in the paddy soils of the Taihu Lake Basin, China. Samples of the six main soil types of the basin were taken from paddy fields with different fertilities. The total numbers of bacteria and denitrifying bacteria in the high fertility soils were much more than those in low fertility soils, and the number of denitrifying bacteria accounted for 49% to 80% of the total number of bacteria. The O2 content was an important ecological factor that affected denitrification. Of test the strains isolated from the paddy soils in the Taihu Lake Basin, some (e.g., Pseudomonas spp.) grew well under low oxygen partial pressure, while others (e.g., Bacillus spp.) had no strict predilection with O2 content. Another critical ecological factor was the nitrogen concentration. Three selected denitrifying bacteria grew better in aculture medium with 135 instead of 276mg L^-1 nitrogen. At the same time 67% of the test strains were able to reduce NO3^- to NO2^- and 56% had N2O reductase.

Community structure and assembly of denitrifying bacteria in epiphytic biofilms in a freshwater lake ecosystem

Denitrifying bacteria are a crucial component of aquatic ecosystem in nitrogen cycle.However,the denitrifying bacterial community dynamics and structure in epiphytic biofilms remain unexplored.The abundance of denitrification gene(nir)and structure of nirS-denitrifying bacterial community in the epiphytic biofilms collected in July and November of 2018 from a typical plateau lake(Caohai Wetland,Guizhou,China)were studied by Real-time Quantitative Polymerase Chain Reaction(qPCR)and highthroughput sequencing.Results show that the gene abundance of nirK was higher than that of nirS(P<0.05),and it was significantly different during the growth period(July)than the decline period(November).The denitrifying bacterial species was similar in the two months and shared 76.18%of OTUs.Proteobacteria(56.55%±22.15%)was the dominant phylum in all the samples.Epiphytic biofilms between growth period and decline period displayed significantly different microbial community structures due to differences in species abundance.Water temperature was the crucial factor that affected the denitrifying microbial community structure in our study.Environmental factors explain only partially the dynamic characteristics of denitrifying microbial communities,implying that the stochastic processes affected the construction of denitrifying microbial communities.As the null model analysis results show,dispersal limitation(stochastic)and undominated processes significantly influenced the assembly of denitrifying microbial communities.This study broadened our understanding of the denitrifying bacterial community structure and its function on epiphytic biofilms in freshwater ecosystems with new information provided.

Effect of carbon source and nitrate concentration on denitrifying phosphorus removal by DPB sludge

Effect of added carbon source and nitrate concentration on the denitrifying phosphorus removal by DPB sludge was systematically studied using batch experiments, at the same time the variation of ORP was investigated. Results showed that the denitrifying and phosphorus uptake rate in anoxic phase increased with the high initial anaerobic carbon source addition. However once the initial COD concentration reached a certain level, which was in excess to the PHB saturation of poly-P bacteria, residual COD carried over to anoxic phase inhibited the subsequent denitrifying phosphorus uptake. Simultaneously, phosphate uptake continued until all nitrate was removed, following a slow endogenous release of phosphate. High nitrate concentration in anoxic phase increased the initial denitrifying phosphorus rate. Once the nitrate was exhausted, phosphate uptake changed to release. Moreover, the time of this turning point occurred later with the higher nitrate addition. On the other hand, through on-line monitoring the variation of the ORP with different initial COD concentration, it was found ORP could be used as a control parameter for phosphorus release, but it is impossible to utilize ORP for controlling the denitrificaion and anoxic phosphorus uptake operations.

Denitrifying phosphorous removal in anaerobic/anoxic SBR system with different startup operation mode

To achieve stable and efficient nitrogen and phosphorus removal and to investigate the characteristics of the A/A SBR enriched with denitrifying phosphorus removal bacteria(DPB),the whole course of startup was studied with two reactors operated in different mode.The reactor I was operated under anaerobic/settling/anoxic/settling mode,and the reactor II was operated under anaerobic/anoxic/settling mode.Differences between the two reactors in removal efficiency of COD,nitrogen and phosphorus were examined.The results indicated that efficient performance could be achieved in both reactors with different startup operation mode,while the phosphorus removal efficiency was improved sooner in reactor I than in reactor II,which suggested that reactor I would supply a more favorable condition for DPB proliferation.Meanwhile,it was observed that the amount of organic substrates consumption had a linear correlation to that of phosphorus release in anaerobic phase when DPB was accumulated in the A/A SBR denitrifying phosphorus removal system.

Denitrification and Dephosphatation by Anaerobic/Anoxic Sequencing Batch Reactor

Removal of denitrifying phosphorus was verified in a laboratory anaerobic/anoxic sequencing batch reactor (A/A SBR). The results obtained demonstrated that the anaerobic/anoxic strategy can enrich the growth of denitrifying phosphorus removing bacteria (DPB) and take up phosphate under anoxic condition by using nitrate as the electron acceptor. The phosphorus removal efficiency was higher than 90% and the effluent phosphate concentration was lower than 1 mg·L-1 after the A/A SBR was operated in a steady-state. When the chemical oxygen demand(COD) of influent was lower than 180mg· L-1, the more COD in the influent was, the higher efficiency of phosphorus removal could be attained under anoxic condition. However, simultaneous presence of carbon and nitrate would be detrimental to denitrifying phosphorus removal. Result of influence of sludge retention time (SRT) on denitrifying phosphorus removal suggested that the decrease of SRT caused a washout of DPB and consequently the enhanced biological phosphorus removal decreased with 8 days SRT. When the SRT was restored to 16 days, however, the efficiency of phosphorus removal was higher than 90%.

减氮对麦玉轮作农田土壤反硝化细菌群落结构和多样性的影响

探究减氮对华北地区麦玉轮作农田土壤反硝化细菌群落结构和多样性影响,为华北地区麦玉轮作农田氮肥管理提供技术支持,本研究以不施氮为对照(CK),设置2个施氮量,分别为常规施氮量(纯氮300 kg·hm^(-2),N2)、减氮20%(纯氮240 kg·hm^(-2),N1),提取土壤DNA,用nirS(细胞色素cd1-亚硝酸还原酶)、nirK(Cu-亚硝酸还原酶)引物扩增后采用MiSeq PE300测序技术,研究施氮量对麦玉轮作农田土壤性状及nirS、nirK反硝化细菌群落结构及多样性的影响。结果表明,3个处理nirS、nirK反硝化细菌α-多样性指数无显著差异,nirS反硝化细菌α-多样性高于nirK。减氮显著影响nirS、nirK反硝化细菌物种组成。减氮对nirS、nirK反硝化细菌门水平及nirK纲水平物种组成无显著影响,但显著降低了nirS反硝化细菌Deltaproteobacteria(δ-变形菌纲)相对丰度;减氮显著影响nirS、nirK属水平物种组成。硝态氮、速效磷、pH值是影响土壤nirS反硝化细菌属水平群落结构主要环境因子;pH值是影响土壤nirK反硝化细菌属水平群落结构的主要环境因子。研究表明,适量减氮不影响反硝化细菌α-多样性,但显著影响反硝化细菌属水平群落组成和群落结构,减氮主要是通过影响土壤性状及微生物群落结构进而影响农田土壤N2O排放。

污水厂尾水对河流水质和底泥细菌群落的影响

污水厂的出水即尾水,其排放关系到受纳水体的水质和生态状况。分析了武汉市某污水厂尾水排放指标,采用水槽试验探究了污水厂尾水对河流水质指标和底泥细菌群落的影响。结果表明:尾水排放促进了河水中硝酸盐氮的降解,总磷去除更加稳定;尾水使河水pH值提高了0.3、电导率上升幅度55%、溶解氧降低了0.6 mg/L。尾水排放之后,底泥细菌群落的丰富度升高、多样性降低,抑制厌氧菌Anaerolineaceae和SBR1031,促进好氧菌Pseudomonas;反硝化菌Azospira、Dechloromonas、Pseudomonas、Sphingomonas增多,促进了NO_(3)^(-)-N的降解。底泥中致病菌奈瑟菌属、军团菌属、消毒残生细菌相对丰度上升。

接种两株有益菌的生物絮团净水性能及饲料化潜力

为强化新的絮团形成菌伪杜擀氏菌YN12(Pseudoduganella eburnea YN12,YN)利用养殖尾水生成生物絮团效果,并将絮团制成饲料,探讨在胡子鲶(Claris fuscus)饲养上的可行性。在生物絮团系统中,接种活性污泥(Activated sludge,AS)、伪杜擀氏菌YN12和枯草芽孢杆菌(Bacillus subtilis,BS)并配制成7组(AS、YN、BS、AS+YN、AS+BS、YN+BS和AS+YN+BS),分析絮团形态结构差异,评估出对模拟养殖尾水氮去除效果及微生物群落结构优势的最佳组合,最终收集最佳组合絮团烘干研磨成粉末添加到商业饲料制粒,分成对照组和絮团组两组,探讨对胡子鲶的生长性能、饲料利用率、肌肉营养成分及肝脏的消化酶和免疫酶活性的影响。结果表明:YN+BS组水质净化效果稳定,总氮去除效果(89.6%)良好,氨氮、亚硝酸盐和硝酸盐积累低,具有良好的絮团生成量(9.0 ml/L)及占据优势(47.5%)的反硝化菌群(包括Hydrogenophaga、Flavobacterium、Pseudoxanthomonas、Burkholderiaceae、Comamonas、Acinetobacter等)。该组絮团粉以5%比例与商业饲料混合、研磨加工制粒后用于胡子鲶养殖,发现胡子鲶生长性能[包括存活率:(100±0.00)%>(95±0.00)%;增重:(2.81±0.35)g>(2.52±0.52)g;体长增加(1.68±0.36)cm>(1.51±0.34)cm]和饲料系数55±0.03>46±0.12均优于纯商业饲料的对照组,然而在淀粉酶等消化酶、免疫酶包括过氧化氢酶、总超氧化物歧化物、谷胱氨酸过氧化物酶等及肌肉营养成分包括谷氨酸、天冬氨酸、丙氨酸和甘氨酸等鲜味氨基酸上均低于对照组。YN+BS组絮团饲料在生长性能占据优势,然而在消化能力、免疫活性及肌肉营养成分上并未表现出明显优势,需要进一步调整水平絮团添加水平以优化胡子鲶饲养效果。

葡萄籽纳米铁介导微氧污泥床生物脱氮性能研究

针对微氧活性污泥技术面临的启动周期长、氮去除效率低等问题,通过投加采用葡萄籽提取物为原材料制备的纳米铁(Grape Seed-nano iron,GS-nFe)构建了GS-nFe介导的微氧活性污泥新体系,从而实现了运行效能的提高。研究结果表明,GS-nFe的单个粒径范围在50~300 nm;反应器在投加GS-nFe后,平均出水NH_(4)^(+)-N、TIN和COD去除率分别维持在89.48%、76.24%和79.39%,GS-nFe的投加使得厌氧氨氧化菌属的相对丰度由1.98%增至2.87%。此外,norank_f_norank_o_C10-SB1A的相对丰度占比由1.59%增至3.02%,其可与好氧反硝化菌、铁自养反硝化菌、厌氧氨氧化等菌属共同协作,提高生物脱氮效能。

一株适用于短程反硝化的细菌筛选及其脱氮特性

从水稻田土壤中筛选出一株短程反硝化作用明显的菌株D5,结合细菌的形态结构以及16S rDNA序列分析,鉴定为肺炎克雷伯氏杆菌(Klebsiella pneumoniae),并进一步采用单因素实验方法探究了环境因子(碳源种类、C/N和温度)对该菌株脱氮性能的影响。结果表明,多种环境因子均对菌株D5的短程反硝化效果具有较大影响,最佳碳源为柠檬酸钠,最佳C/N为3,最佳温度为25℃和30℃。结果显示,菌株D5具有优良的短程反硝化性能,通过调控环境条件可以取得最佳的短程反硝化效果。

1株反硝化细菌的筛选鉴定及其基于响应面法的脱氮条件优化

从污水处理厂采集的污泥样品中分离、筛选出1株具有较高好氧反硝化性能的菌株,命名为X_(3)。通过菌落形态特征、染色特征观察以及16S rDNA分子鉴定,初步确定该菌株为赤红球菌(Rhodococcus ruber)。为了明确该菌株的脱氮特性,通过单因素试验,确定了菌株的最佳碳源为葡萄糖,最佳C/N比为10。利用响应面分析方法考察温度、pH以及转速三个因素相互作用对菌株脱氮能力的影响。研究结果表明,菌株X_(3)在32.24℃,pH为6.65,180.79 r/min为最佳培养条件。最佳条件下培养48 h后,NO_(3)^(-)-N去除率可达96.99%,同时没有NO_(2)^(-)-N的产生。筛选获得的好氧反硝化菌株X_(3)可以作为生物脱氮菌剂开发和相关脱氮机制研究的候选菌株。

对虾养殖环境脱氮脱硫复合菌的构建与效能分析

随着对虾集约化养殖的快速发展,由于追求高产量和过量投饲等原因导致养殖水体中氨氮(NH_(4)^(+)-N)、亚硝酸盐氮(NO_(2)^(-)-N)和硫化物污染加重,严重危害对虾生长和生理功能。本研究利用自对虾养殖环境中筛选出的异养硝化-好氧反硝化菌(HHVEN1和SDVEA2)和硫氧化菌(GHWS3和GHWS5),成功构建了两种高效脱氮脱硫复合菌NS1-1和NS2-2,并研究其脱氮、脱硫能力。结果显示:复合菌NS1-1对NH_(4)^(+)-N、NO_(2)^(-)-N和硫化物去除率均达到95%以上,NS2-2的去除率分别为90.27%、97.38%和89.62%,且均优于单一菌株;复合菌NS1-1和NS2-2在温度20℃~35℃、碳氮比(C/N) 5~20、盐度20~30和以葡萄糖或琥珀酸钠为碳源的培养条件下均能发挥最大脱氮脱硫效能,两者最适pH范围不同,分别为7.5~8.0和7.0~8.5。复合菌NS1-1和NS2-2在水产养殖尾水中各菌株之间具有良好的共存性,在实际养殖尾水中NH_(4)^(+)-N、NO_(2)^(-)-N、硫化物去除率分别为78.75%、75.09%、79.61%和81.44%、62.68%、72.64%。研究表明,复合菌NS1-1和NS2-2具有高效脱氮脱硫的去除效能和优异的环境适应能力,研究成果为水产养殖环境消除氮、硫污染提供了科学支持。

基于污水同步脱氮除磷的反硝化聚磷菌的研究与展望

与传统的好氧除磷技术相比,反硝化除磷具有节约碳源、一碳两用、减少污泥产量等优点。综述了反硝化脱氮除磷过程中的主要功能微生物反硝化聚磷菌的培养、分离筛选、电子受体利用等研究的相关进展,总结了近年来研究中常见的反硝化聚磷菌,并对其未来的发展前景做出展望。

异养硝化-好氧反硝化细菌的研究进展及在水产养殖中的应用

异养硝化-好氧反硝化细菌可以同时实现硝化和反硝化,具有高效的脱氮性能,是目前废水生物脱氮处理的研究热点之一。文章从异养硝化-好氧反硝化细菌的研究现状、作用机制、脱氮影响因素、在水产养殖中的应用等4个方面进行了综述,可为异养硝化-好氧反硝化细菌的研究提供理论依据,为异养硝化-好氧反硝化细菌在水产养殖中的应用提供新的思路。