Applied Microbiology and Biotechnology

, Volume 102, Issue 19, pp 8551–8560 | Cite as

Comprehensive analyses of functional bacteria and genes in a denitrifying EGSB reactor under Cd(II) stress

  • Yu Miao
  • Xu-Xiang ZhangEmail author
  • Shuyu Jia
  • Runhua Liao
  • Aimin Li
Environmental biotechnology


A bench-scale expanded granular sludge bioreactor (EGSB) was continuously operated to treat synthesized high-nitrate industrial wastewater with increasing bivalent cadmium (Cd(II)) stress. The bioreactor showed nearly complete nitrate removal regardless of Cd(II) loadings, while nitrite accumulated in the effluent when influent Cd(II) loading was over 64 mg/L. Mi-seq sequencing of 16S rRNA gene amplicons elucidated that denitrifiers had decreasing abundances while biodiversity showed increasing trend as the Cd(II) loading increased. In the bioreactor, genera Halomonas, Thauera, Pseudomonas, and Zoogloea played major roles in the denitrification under lower Cd(II) loadings (< 32 mg/L), while Halomonas sp. KM-1 and Halomonas sp. BC04 acted as the crucial Cd-resistant denitrifiers under 128 mg/L Cd(II) loading. Metagenomic analyses and real-time quantitative PCR consistently indicated that napA encoding nitrate reductase was the predominant denitrifying gene, that could be mainly functioning on the efficient nitrate removal. Statistical analyses revealed the significantly positive correlation between Halomonas and nirS gene, both of which were functionally responsible for nitrite reduction. The obtained results may be practically useful for regulation and optimization of the biological processes to treat industrial wastewater containing high levels of nitrate and Cd(II).


Bacterial denitrifiers Bivalent cadmium Denitrifying genes High-throughput sequencing Microbial community 



We would like to thank the High Performance Computing Center (HPCC) of Nanjing University for the help of computation.

Funding information

This study was financially supported by the National Key Technology Support Program of China (2014BAC08B04), the Jiangsu Science Foundation for Distinguished Young Scholars (BK20150019), and the Six Talent Peaks Project in Jiangsu Province (2015-JNHB-001).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9228_MOESM1_ESM.pdf (3.5 mb)
ESM 1 (PDF 3588 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yu Miao
    • 1
  • Xu-Xiang Zhang
    • 1
    Email author
  • Shuyu Jia
    • 1
  • Runhua Liao
    • 1
    • 2
  • Aimin Li
    • 1
  1. 1.State Key Laboratory of Pollution Control and Resource Reuse, School of the EnvironmentNanjing UniversityNanjingChina
  2. 2.School of Materials Science and EngineeringJingdezhen Ceramic InstituteJingdezhenChina

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