Source Jouranl of CSCD
Source Journal of Chinese Scientific and Technical Papers
Included as T2 Level in the High-Quality Science and Technology Journals in the Field of Environmental Science
Core Journal of RCCSE
Included in the CAS Content Collection
Included in the JST China
Indexed in World Journal Clout Index (WJCI) Report
Volume 44 Issue 3
Mar.  2026
Turn off MathJax
Article Contents
TIAN Tian, CUI Jinle. Distinct microbial assembly and structures of iron-dependent autotrophic denitrifying communities enriched by different ferrous salts[J]. ENVIRONMENTAL ENGINEERING , 2026, 44(3): 73-83. doi: 10.13205/j.hjgc.202603006
Citation: TIAN Tian, CUI Jinle. Distinct microbial assembly and structures of iron-dependent autotrophic denitrifying communities enriched by different ferrous salts[J]. ENVIRONMENTAL ENGINEERING , 2026, 44(3): 73-83. doi: 10.13205/j.hjgc.202603006

Distinct microbial assembly and structures of iron-dependent autotrophic denitrifying communities enriched by different ferrous salts

doi: 10.13205/j.hjgc.202603006
  • Received Date: 2025-11-28
    Available Online: 2026-04-11
  • Publish Date: 2026-03-01
  • Iron-dependent autotrophic denitrification (IDAD) is a promising biological technology for nitrogen removal from wastewater with a low C/N ratio. While significant differences in community structure among IDAD consortia from different studies have been reported, the underlying reasons remain unclear. Hypothesizing that the anion type of ferrous salts could be a key contributing factor, this study enriched two IDAD consortia (R1 and R2) using FeCl2 and FeSO4 as respective electron donors under identical inoculum sludge conditions. Their nitrogen removal performance, extracellular polymeric substance (EPS) composition, iron oxidation product properties, microbial community structure, and functional gene distribution characteristics during long-term operation were systematically compared. The results showed that R1 exhibited higher denitrification efficiency and Fe(Ⅱ) oxidation activity. Key iron-oxidizing bacterial genera such as Gallionella showed a significantly higher relative abundance in R1 than in R2, and R1 also maintained higher community diversity. Metagenomic analysis further revealed a higher abundance of functional genes related to iron oxidation and denitrification in R1. In contrast, R2 was enriched with more genera associated with sulfate metabolism and complex organic matter degradation. Distinct differences were also observed in EPS composition and iron mineral surface properties between the two consortia. This study confirms that the type of ferrous salt significantly regulates the structural assembly and metabolic functions of IDAD consortia, providing a theoretical basis for optimizing their application in practical wastewater treatment.
  • loading
  • [1]
    KUYPERS M M M,MARCHANT H K,KARTAL B. The microbial nitrogen-cycling network[J]. Nature Reviews Microbiology,2018,16(5):263-276.
    [2]
    PAN Y,SUN R Z,YU H Q. Research advances in biological denitrification technology driven by exogenous electron donors[J]. Environmental Engineering,2024,42(9):1-12. 潘元,孙睿哲,俞汉青. 外源电子供体驱动生物反硝化技术研究进展[J]. 环境工程,2024,42(9):1-12.
    [3]
    TIAN T,YU H Q. Denitrification with non-organic electron donor for treating low C/N ratio wastewaters[J]. Bioresource Technology,2020,299:122686.
    [4]
    TIAN T,ZHOU K,XUAN L,et al. Exclusive microbially driven autotrophic iron-dependent denitrification in a reactor inoculated with activated sludge[J]. Water Research,2020,170:115300.
    [5]
    STRAUB K L,BENZ M,SCHINK B,et al. Anaerobic,nitrate-dependent microbial oxidation of ferrous iron[J]. Applied and Environmental Microbiology,1996,62(4):1458-1460.
    [6]
    HE S,TOMINSKI C,KAPPLER A,et al. Metagenomic analyses of the autotrophic Fe(Ⅱ)-oxidizing,nitrate-reducing enrichment culture KS[J]. Applied and Environmental Microbiology,2016,82(9):2656-2668.
    [7]
    BRYCE C,BLACKWELL N,SCHMIDT C,et al. Microbial anaerobic Fe(Ⅱ)oxidation-ecology,mechanisms and environmental implications[J]. Environmental Microbiology,2018,20:3462-3483.
    [8]
    KLUEGLEIN N,ZEITVOGEL F,STIERHOF Y,et al. Potential role of nitrite for abiotic Fe(Ⅱ)oxidation and cell encrustation during nitrate reduction by denitrifying bacteria[J]. Applied and Environmental Microbiology,2014,80(3):1051-1061.
    [9]
    BLOTHE M,RODEN E E. Composition and activity of an autotrophic Fe(Ⅱ)-oxidizing,nitrate-reducing enrichment culture[J]. Applied and Environmental Microbiology,2009,75:6937-6940.
    [10]
    ETIQUE M,JORAND F P A,ZEGEYE A,et al. Abiotic process for Fe(Ⅱ)oxidation and green rust mineralization driven by a heterotrophic nitrate reducing bacteria(Klebsiella mobilis)[J]. Environmental Science& Technology,2014,48(7):3742-3751.
    [11]
    NORDHOFF M,TOMINSKI C,HALAMA M,et al. Insights into nitrate-reducing Fe(Ⅱ)oxidation mechanisms through analysis of cell-mineral associations,cell encrustation,and mineralogy in the chemolithoautotrophic enrichment culture KS[J]. Applied and Environmental Microbiology,2017,83(13):e00752-17.
    [12]
    JAKUS N,BLACKWELL N,OSENBRÜCK K,et al. Nitrate removal by a novel lithoautotrophic nitrate-reducing iron(Ⅱ)-oxidizing culture enriched from a pyrite-rich limestone aquifer[J]. Applied and Environmental Microbiology,2021,87(16):e0046021.
    [13]
    JAKUS N,MELLAGE A,HÖSCHEN C,et al. Anaerobic neutrophilic pyrite oxidation by a chemolithoautotrophic nitrate-reducing iron(Ⅱ)-oxidizing culture enriched from a fractured aquifer[J]. Environmental Science& Technology,2021,55(14):9876-9884.
    [14]
    DING P,REN Z Q,MA J Y,et al. Research status and prospect on autotrophic denitrification technology utilizing pyrite as the electron donor[J]. Environmental Engineering,2024,42(12):97-107. 丁沛,任之琪,马佳莹,等. 以黄铁矿为电子供体的自养反硝化技术研究现状与展望[J]. 环境工程,2024,42(12):97-107.
    [15]
    WANG R,YANG C,ZHANG M,et al. Chemoautotrophic denitrification based on ferrous iron oxidation:reactor performance and sludge characteristics[J]. Chemical Engineering Journal,2017,313:693-701.
    [16]
    ZHANG M,ZHANGZHU G,WEN S,et al. Chemolithotrophic denitrification by nitrate-dependent anaerobic iron oxidizing(NAIO)process:Insights into the evaluation of seeding sludge[J]. Chemical Engineering Journal,2018,345:345-352.
    [17]
    牟宏霖,刘绪振,熊新竹,等. 铁自养反硝化滤池强化高速公路服务区污水生物处理尾水的应用研究[J/OL]. 环境工程,2025,1-13[ 2026-01-20]. https://link.cnki.net/urlid/11.2097.X.20251107.0935.004.

    MU H L,LIU XZ,XIONG X Z,et al. Research on the application of iron autotrophic denitrification filter to enhance the biological treatment of tail water in expressway service areas[J/OL]. Environmental Engineering,2025,1-13[ 2026-01-20]. https://link.cnki.net/urlid/11.2097.X.20251107.0935.004..
    [18]
    TIAN T,ZHOU K,LI Y S,et al. Phosphate recovery from wastewater prominently through a Fe(Ⅱ)-P oxidizing pathway in the autotrophic iron-dependent denitrification process[J]. Environmental Science& Technology,2020,54(18):11576-11583.
    [19]
    LI R,MORRISON L,COLLINS G,et al. Simultaneous nitrate and phosphate removal from wastewater lacking organic matter through microbial oxidation of pyrrhotite coupled to nitrate reduction[J]. Water Research,2016,96:32-41.
    [20]
    LIU J,ZHANG H,WANG H,et al. Remediation of arsenic-and nitrate-contaminated groundwater through iron-dependent autotrophic denitrifying culture[J]. Environmental Research,2024,257:119239.
    [21]
    ZHOU K,PAN Y,TIAN T,et al. Deciphering the evolution of biological and chemical process in acclimatization of autotrophic iron-dependent denitrifying sludge[J]. Chinese Journal of Environmental Engineering,2021,15(8):2789-2800. 周可,潘元,田天,等. 铁自养反硝化污泥富集培养过程中化学与生物作用的变化规律[J]. 环境工程学报,2021,15(8):2789-2800.
    [22]
    WANG R,ZHENG P,ZHANG M,et al. Bioaugmentation of nitrate-dependent anaerobic ferrous oxidation by heterotrophic denitrifying sludge addition:a promising way for promotion of chemoautotrophic denitrification[J]. Bioresource Technology,2015,197:410-415.
    [23]
    ZHANG H B,WANG H F,LIU J B,et al. Anaerobic ammonium oxidation coupled to iron(Ⅲ)reduction catalyzed by a lithoautotrophic nitrate-reducing iron(Ⅱ)oxidizing enrichment culture[J]. The ISME Journal,2024,18(1):wrae149.
    [24]
    WANG R,ZHENG P,YANG C,et al. Stress of Cl-,SO42- and PO43- on the heterotrophic denitrifying sludge[J]. China Environmental Science,2016,36(4):1039-1044. 王茹,郑平,杨程,等. Cl-、SO42- 和 PO43- 对异养反硝化污泥的胁迫效应[J]. 中国环境科学,2016,36(4):1039-1044.
    [25]
    XU J,SHENG G,MA Y,et al. Roles of extracellular polymeric substances(EPS)in the migration and removal of sulfamethazine in activated sludge system[J]. Water Research,2018,47:5298-5306.
    [26]
    DUBOIS M,GILL K A,HAMILTON J K,et al. Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry,1956,2:350-356.
    [27]
    FRØLUND B,PALMGREN R,KEIDING K,et al. Extraction of extracellular polymers from activated sludge using a cation exchange resin[J]. Water Research,1996,30(8):1749-1758.
    [28]
    SCHAEDLER F,KAPPLER A,SCHMIDT C. A revised iron extraction protocol for environmental samples rich in nitrite and carbonate[J]. Geomicrobiology Journal,2018,35:23-30.
    [29]
    BI Z,QIAO S,ZHOU J,et al. Fast start-up of Anammox process with appropriate ferrous iron concentration[J]. Bioresource Technology,2014,170:506-512.
    [30]
    TIAN T,ZHOU K,LI Y,et al. Recovery of iron-dependent autotrophic denitrification activity from cell-iron mineral aggregation-induced reversible inhibition by low-intensity ultrasonication[J]. Environmental Science& Technology,2022,56(1):595-604.
    [31]
    LUO X K,WEI Z P,LÜ F J,et al. De-chlorination performance by bio-stimulants and characteristics of extracellular polymeric substances[J]. Environmental Science& Technology,2025,48(9):96-107. 罗啸康,魏展鹏,吕芳杰,等. 生物刺激微生物脱氯性能及其胞外聚合物特性分析[J]. 环境科学与技术,2025,48(9):96-107.
    [32]
    ZHANG X,SUN Y,MA F,et al. In-situ utilization of soluble microbial product(SMP)cooperated with enhancing SMP-dependent denitrification in aerobic-anoxic sequencing batch reactor[J]. Science of the Total Environment,2019,693:133558.
    [33]
    WU Z X,HUANG X W,SONG W F,et al. The EPS components variation and adsorption characteristics of Pseudomonas aeruginosa under different Cu(Ⅱ)stress/induction[J]. Acta Scientiae Circumstantiae,2025,45(7):202-214. 吴芷昕,黄祥武,宋卫锋,等. 不同Cu(Ⅱ)胁迫/诱导下P. aeruginosa EPS组分变化及其吸附特性[J]. 环境科学学报,2015,45(7):202-214.
    [34]
    DIPPON U,PANTKE C,PORSCH K,et al. Potential function of added minerals as nucleation sites and effect of humic substances on mineral formation by the nitrate-reducing Fe(Ⅱ)-oxidizer Acidovorax sp. BoFeN1[J]. Environmental Science& Technology,2012,46(12):6556-6565.
    [35]
    LIU X,EUSTERHUES K,THIEME J,et al. STXM and nanoSIMS investigations on EPS fractions before and after adsorption to Goethite[J]. Environmental Science& Technology,2013,47(7):3158-3166.
    [36]
    SU C,LI Y F,YAN X,et al. Diversity of iron minerals and their adsorption to Cd in ferrous oxidation and denitrification biofilm reactor[J]. Environmental Engineering,2020,38(5):76-83. 粟畅,李颖芬,严兴,等. 亚铁氧化反硝化生物膜反应器中铁矿物的多样性及其对重金属Cd的吸附[J]. 环境工程,2020,38(5):76-83.
    [37]
    XUE Y F,WANG S F,Research on Fe(Ⅱ)-mediated nitrite reduction coupled with arsenic oxidation[J]. Journal of Dalian University of Technology,2025,65(2):118-124. 薛杨峰,王少锋. Fe(Ⅱ)介导的亚硝酸盐还原耦合砷氧化研究[J]. 大连理工大学学报,2025,65(2):118-124.
    [38]
    HUANG Y,STRAUB D,BLACKWELL N,et al. Meta-omics reveal Gallionellaceae and Rhodanobacter species as interdependent key players for Fe(Ⅱ)oxidation and nitrate reduction in the autotrophic enrichment culture KS[J]. Applied and Environmental Microbiology,2021,87(15):e00496-21.
    [39]
    PANG Y,XIA L,LI S,et al. Metagenomic insights into a solid phase heterotrophic-autotrophic denitrification reactor driven by biodegradable polymers and pyrite[J]. Journal of Cleaner Production,2025,523:146411.
    [40]
    WANG X. Manganese redox regulates microbial nitrogen conversion metabolic pathways and community interactions[D]. Harbin:Harbin Institute of Technology,2025. 王暄. 锰氧化还原调控微生物氮转化代谢途径与群落互作机制[D]. 哈尔滨:哈尔滨工业大学,2025.
    [41]
    OKUBO T,TOYODA A,FUKUHARA K,et al. The physiological potential of anammox bacteria as revealed by their core genome structure[J]. DNA Research,2020,28(1):1-12.
    [42]
    EO J,PARK K. Long-term effects of imbalanced fertilization on the composition and diversity of soil bacterial community[J]. Agriculture,Ecosystems& Environment,2016,231:176-182.
    [43]
    ZHENG Y,ZHOU Z,YE X,et al. Identifying microbial community evolution in membrane bioreactors coupled with anaerobic side-stream reactor,packing carriers and ultrasonication for sludge reduction by linear discriminant analysis[J]. Bioresource Technology,2019,291:121920.
    [44]
    XU Q,WU Z,XU Z,et al. Soil moisture-dependent tire wear particles aging processes shift soil microbial communities and elevated nitrous oxide emission on drylands[J]. Science of the Total Environment,2024,952:175948.
    [45]
    FENG X,ZHAO J Q,DAI W,et al. Accumulation characteristics of NO and N2O in nitrite denitrifying phosphorus accumulation process[J]. Environmental Engineering,2019,37(12):1-5,54. 冯鑫,赵剑强,代伟,等. 亚硝酸盐反硝化聚磷过程中NO和N2O的累积特征[J]. 环境工程,2019,37(12):1-5,54.
    [46]
    CASTELLE C,GUIRAL M,MALARTE G,et al. A new iron-oxidizing/O2-reducing supercomplex spanning both inner and outer membranes,isolated from the extreme acidophile Acidithiobacillus ferrooxidans[J]. Journal of Biological Chemistry,2008,283:25803-25811.
    [47]
    YANG G,LI S,NIU R,et al. Insights into nitrate-reducing Fe(Ⅱ)oxidation by Diaphorobacter caeni LI3T through kinetic,nitrogen isotope fractionation,and genome analyses[J]. Science of the Total Environment,2024,912:168720.
    [48]
    DENG Q,SU C,CHEN Z,et al. Effect of hydraulic retention time on the denitrification performance and metabolic mechanism of a multi-chambered bio-electrochemical system[J]. Journal of Environmental Management,2021,299:113575.
    [49]
    SUN H,ZHAO X,DING J,et al. Unveiling dynamics of microbial communities,species interactions,and ecological assembly during low-temperature-induced sludge bulking in full-scale wastewater treatment systems[J]. Bioresource Technology,2025,435:132950.
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Article Metrics

    Article views (33) PDF downloads(0) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return