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.

Effects of long-term elevated CO_2 on N_2-fixing,denitrifying and nitrifying enzyme activities in forest soils under Pinus sylvestriformis in Changbai Mountain

A study was conducted to determine the effects of elevated CO2 on soil N process at Changbai Mountain in Jilin Province, northeastern China （42°24＂N, 128°06＂E, and 738 m elevation）. A randomized complete block design of ambient and elevated CO2 was established in an open-top chamber facility in the spring of 1999. Changpai Scotch pine （Pinus sylvestris var. sylvestriformis seeds were sowed in May, 1999 and CO2 fumigation treatments began after seeds germination. In each year, the exposure started at the end of April and stopped at the end of October. Soil samples were collected in June and August 2006 and in June 2007, and soil nitrifying, denitrifying and N2-fixing enzyme activities were measured. Results show that soil nitrifying enzyme activities （NEA） in the 5-10 cm soil layer were significantly increased at elevated CO2 by 30.3% in June 2006, by 30.9% in August 2006 and by 11.3% in June 2007. Soil denitrifying enzyme activities （DEA） were significantly decreased by elevated CO2 treatment in June 2006 （P 〈 0.012） and August 2006 （P 〈 0.005） samplings in our study; no significant difference was detected in June 2007, and no significant changes in N2-fixing enzyme activity were found. This study suggests that elevated CO2 can alter soil nitrifying enzyme and denitrifying enzyme activities.

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.

养殖池塘系统脱氮硫杆菌(Thiobacillus denitrificans)的分离、生长特性及脱氮特征研究

分别采集集约化养殖吉富罗非鱼(Oreochromis niloticus)的池塘底泥、表层水、塘边裸露土壤样品,采用脱氮硫杆菌选择培养基富集培养、分离得到3株细菌,通过菌落特征、形态学特征、生理生化检验和16S rDNA序列分析等手段,鉴定为脱氮硫杆菌(Thiobacillus denitrificans),命名为TD1、TD2和TD3。在30℃下,采用静置、厌氧培养实验研究这3株脱氮硫杆菌的生长繁殖特征、对硝酸盐氮的去除速率特征、硫酸根产生状况、亚硝酸盐产生情况、培养液pH值和氧化还原电位变化特征等方面的差异进行研究,探索养殖环境修复中应用的可能性。结果表明:在一定的时间内菌株生长速率、介质pH降低速率和氧化还原电位上升幅度均表现为TD1>TD2>TD3;菌株硝态氮脱除效率和硫酸根浓度升高速率均表现为TD1>TD2>TD3,亚硝酸盐产生情况则表现为TD1<TD2<TD3;TD1更适合异位修复,TD2更适合原位修复。

Effect of nitrification inhibitor DMPP on nitrogen leaching, nitrifying organisms, and enzyme activities in a rice-oilseed rape cropping system

DMPP （3,4-dimethylpyrazole phosphate） has been used to reduce nitrogen （N） loss from leaching or denitrification and to improve N supply in agricultural land. However, its impact on soil nitrifying organisms and enzyme activities involved in N cycling is largely unknown. Therefore, an on-farm experiment, for two years, has been conducted, to elucidate the effects of DMPP on mineral N （NH4^＋- N and NO3^--N） leaching, nitrifying organisms, and denitrifying enzymes in a rice-oilseed rape cropping system. Three treatments including urea alone （UA）, urea ＋ 1% DMPP （DP）, and no fertilizer （CK）, have been carded out. The results showed that DP enhanced the mean NH4^＋-N concentrations by 19.1%-24.3%, but reduced the mean NO3^--N concentrations by 44.9%-56.6% in the leachate, under a two-year rice-rape rotation, compared to the UA treatment. The population of ammonia oxidizing bacteria, the activity of nitrate reductase, and nitrite reductase in the DP treatment decreased about 24.5%-30.9%, 14.9%-43.5%, and 14.7%-31.6%, respectively, as compared to the UA treatment. However, nitrite oxidizing bacteria and hydroxylamine reductase remained almost unaffected by DMPP. It is proposed that DMPP has the potential to either reduce NO3^--N leaching by inhibiting ammonia oxidization or N losses from denitrification, which is in favor of the N conversations in the rice-oilseed rape cropping system.

Influence of pH on short-cut denitrifying phosphorus removal

Through a series of experiments using denitrifying phosphorus-accumulating sludge in sequencing batch reactors （SBRs）, the variations of the intracellular polymers during the anaerobic phosphorus release process at different pH values were compared, the probable reasons for different performances of phosphorus removal were examined, and system operations in a typical cycle were investigated. The results show that the phosphorus removal rate was positively correlated with pH values in a range of 6.5-8.5. When the pH value was 8.0, the anaerobic phosphorus release rate and anoxic phosphorus uptake rate of the activated sludge were 20.95 mg/（g, h） and 23.29 mg/（g, h）, respectively; the mass fraction of poly-13-hydroxybutyrate （PHB） increased to 62.87 mg/g under anaerobic conditions; the mass fraction of polyphosphate was 92.67 mg/g under anoxic conditions; and the effluent concentration of total phosphorus （TP） was 1.47 mg/L. With the increase of pH, the mass fraction of acetic acid and PHB also increased, and the absorption rate of acetic acid was equal to the disintegration rate of polyphosphate. When the pH value was above 8.0, biological phosphorus removal was achieved by chemical phosphorus precipitation, and the phosphorus removal rate decreased.

Denitrifying phosphorus removal in a step-feed CAST with alternating anoxic-oxic operational strategy

A bench-scale cyclic activated sludge technology （CAST） was operated to study the biological phosphorus removal performance and a series of batch tests was carried out to demonstrate the accumulation of denitrifying polyphosphate-accumulating organisms （DNPAOs） in CAST system. Under all operating conditions, step-feed CAST with enough carbon sources in influent had the highest nitrogen and phosphorus removal efficiency as well as good sludge settling performance. The average removal rate of COD, NH4^＋-N, PO4^3--P and total nitrogen （TN） was 88.2%, 98.7%, 97.5% and 92.1%, respectively. The average sludge volume index （SVI） was 133 mL/g. The optimum anaerobic/aerobic/anoxic （AOA） conditions for the cultivation of DNPAOs could be achieved by alternating anoxic/oxic operational strategy, thus a significant denitrifying phosphorus removal occurred in step-feed CAST. The denitrification of NO^x--N completed quickly due to step-feed operation and enough carbon sources, which could enhance phosphorus release and further phosphorus uptake capability of the system. Batch tests also proved that polyphosphate-accumulating organisms （PAOs） in the step-feed process had strong denitrifying phosphorus removal capacity. Both nitrate and nitrite could be used as electron acceptors in denitrifying phosphorus removal. Low COD supply with step-feed operation strategy would favor DNPAOs accumulation.

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.

Responses of N_2O reductase gene(nosZ)-denitrifier communities to long-term fertilization follow a depth pattern in calcareous purplish paddy soil

The effect of long-term fertilization on soil denitrifying communities was analysed by measuring the abundance and diversity of the nitrous oxide （N2O） reductase gene, nosZ. Soil samples were collected from plots of a long-term fertilization experiment established in 1982 in Suining City, China. The fertilizer treatments were no fertilizer （CK）, three chemical fertilizer （CF） treatments （N, NP, NPK）, manure （M） alone, and manure with chemical fertilizers （NM, NPM, NPKM）. The abundance and diversity of the denitrifying bacteria were assessed by real-time quantitative PCR, terminal restriction fragment length polymorphism （T-RFLP）, and cloning and sequencing of nosZ genes. The diversity and abundance of nosZ-denitrifiers was higher in soil amended with manure and chemical fertilizers （CFM） than in soil amended with CF alone, and the highest in topsoil （0-20 cm）. The nosZ-denitrifier community composition was more complex in CFM soil than in CF soil： Specific species were detected only in the CFM soil. The abundance of nosZ-denitrifier in the NPKM treatment was approximately two times higher than that in the CK, N, and NPK treatments. Most of the cloned nosZ sequences were closely related to nosZ sequences from Bradyrhizobiaceae and Rhodospirillaceae in Alphaproteobacteria. Of the measured abiotic factors, soil organic matter correlated significantly with the abundance （P〈0.01）; available phosphorus correlated significantly with the topsoil community composition （P〈0.01）, whereas soil organic matter correlated significantly with the subsoil （20-90 cm） community composition （P〈0.01）. This study demonstrated that long-term CFM fertilization affected both the abundance and composition of the nosZ-denitrifier community.

Effect of organic carbon on nitrification effciency and community composition of nitrifying biofilms

The effects of organic carbon/inorganic nitrogen （C/N） ratio on the nitrification processes and the community shifts of nitrifying biofilms were investigated by kinetic comparison and denaturing gradient gel electrophoresis （DGGE） analysis. The results showed that the nitrification rate decreased with an increasing organic concentration. However, the effect became weak when the carbon concentration reached a sufficiently high level. Denitrification was detected after organic carbon was added. The 12 h ammonium removal rate ranged from 85% to 30% at C/N = 0.5, 1, 2, 4, 8, and 16, as compared to the control （C/N = 0）. The loss of nitrogen after 24 h at C/N = 0.5, 1, 2, 4, 8, and 16 was 31%, 18%, 24%, 65%, 59%, and 62%, respectively. The sequence analysis of 16S rRNA gene fragments revealed that the dominant populations changed from nitrifying bacteria （Nitrosomonas europaea and Nitrobacter sp.） to denitrifying bacteria （Pseudomonas sp., Acidovorax sp. and Comamonas sp.） with an increasing C/N ratio. Although at high C/N ratio the denitrifying bacteria were the dominant populations, nitrifying bacteria grew simultaneously. Consequently, nitrification process coexisted with denitrification.

Experimental study on simultaneous desulfurization and denitrifi- cation based on highly active absorbent

Simultaneous removal of SO2 and NO from flue gas by the highly active absorbent prepared from fly ash, lime and a few oxidizing manganese compound additive was studied using a flue gas circulating fluidized bed （CFB） under different experimental conditions. The effects influencing the removal effiencies were discussed. The optimal flue gas temperature, flue gas humidity, gas velocity of CFB and Ca/（S＋N） molar ratio with this process were approximately 110℃, 6%, 1.8 m/s and 1.05, respectively. Removal efficiencies of 92.3% for SO2 and 60.88% for NO were obtained under the optimal experimental conditions. While the spent absorbent appeared in the form of dry powder, the mechanism of removal for SO2 and NO based on the highly active absorbent was investigated by a scanning electron microscope （SEM）, a X-ray energy spectrometer and the chemical analysis methods. The valuable references can be provided for industrial application by the process. The foreground of application will be vast in China and in the world.

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.

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.

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%.

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.

Simultaneous Nitrogen and Phosphorus Removal by Denitrifying Dephosphatation in a (AO)_2 Sequencing Batch Reactor

A 24 L working volume reactor was used for the research on simultaneous phosphorus (P) and nitrogen (N) removal by denitrifying dephosphatation in an anaerobic-oxid-anoxic-oxid sequencing batch reactor ((AO)_2SBR) system. The durations of each phase are: anaerobic 1.5 h, aerobic 2.5 h, anoxic 1.5 h, post-aerobic 0.5 h, settling 1.0 h, fill 0.5 h. The successful removal of nitrogen and phosphorus is achieved in a stable (AO)_2SBR. The effluent P concentrations is below 1 mg/L, and the COD,TN and P average removal efficiency is 88.9%, 77.5% and 88.7%, respectively. The batch experiment results show that the durations of aerobic and anoxic phase influence the P removal efficiency. Some feature points are found on the DO, ORP and pH curves to demonstrate the complete of phosphate release and phosphate uptake. These feature points can be used for the control of (AO)_2 SBR.

Microbial Characterization of Denitrifying Sulfide Removal Sludge Using High-Throughput Amplicon Sequencing Method

The denitrifying sulfide removal(DSR) process has recently been studied extensively from an engineering perspective. However, the importance of microbial communities of this process was generally underestimated. In this study, the microbial community structure of a lab-scale DSR reactor was characterized in order to provide a comprehensive insight into the key microbial groups in DSR system. Results from high-throughput sequencing analysis revealed that the fraction of autotrophic denitrifiers increased from 2.34 % to 10.93% and 44.51% in the DSR system when the influent Na Cl increased from 0 g/L, to 4 g/L and 30 g/L, respectively. On the contrary, the fraction of heterotrophic denitrifiers decreased from 61.74% to 39.57%, and 24.12%, respectively. Azoarcus and Thiobacillus were the main autotrophic denitrifiers, and Thauera was the main hetetrophic denitrifier during the whole process. This study could be useful for better understanding the interaction between autotrophs and heterotrophs in DSR system.