Journal of Oceanology and Limnology

, Volume 36, Issue 3, pp 913–925 | Cite as

Denitrification potential evaluation of a newly indigenous aerobic denitrifier isolated from largemouth bass Micropterus salmoides culture pond

  • Cuicui Wang (王翠翠)
  • Kai Zhang (张凯)
  • Jun Xie (谢骏)Email author
  • Qigen Liu (刘其根)
  • Deguang Yu (余德光)
  • Guangjun Wang (王广军)
  • Ermeng Yu (郁二蒙)
  • Wangbao Gong (龚望宝)
  • Zhifei Li (李志斐)


This work evaluates the application potential of a new indigenous aerobic denitrifier, strain Pseudomonas CW-2, isolated from a largemouth bass culture pond. The rate of ammonium-N removal by strain CW-2 was approximately 97% at a DO concentration of 5.2 mg/L. Furthermore, when nitrate and ammonia coexisted, the strain gave priority to assimilating ammonia, and thereafter to denitrification. Under optimal cultivation conditions, citrate and acetate were the carbon resources, C/N was 8, dissolved oxygen was 5.2 mg/L, and pH was 7; the removal rate of ammonium reached nearly 90%. The changing patterns of different bacteria in strain CW-2-treated and the control pond water were also compared. Lower levels of ammonia, nitrite, and phosphates were observed in the treated water as compared with the controls. Meanwhile, phylum-level distributions of the bacterial OTUs revealed that Proteobacteria, Bacteroidetes, Planctomycetes, and Nitrospirae continuously changed their relative abundances in relation to carbon and the addition of strain CW-2; this finding implies that the conventional denitrification process was weakened under the effects of carbon or the presence of strain CW-2. We propose that strain CW-2 is a promising organism for the removal of ammonium in intensive fish culture systems, according to our evaluations of its denitrification performance.


aerobic denitrification ammonium removal Micropterus salmoides Pseudomonas CW-2 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abufayed A A, Schroeder E D. 1986. Kinetics and stoichiometry of SBR/denitrification with a primary sludge carbon source. J. Water Pollut. Control Fed., 58: 58–398.Google Scholar
  2. Carpenter J H. 1965. The Chesapeake Bay institute technique for the Winkler dissolved oxygen method. Limnology and Oceanography, 10 (1): 141–143.CrossRefGoogle Scholar
  3. Chen P Z, Li J, Li Q X, Wang Y C, Li S P, Ren T Z, Wang L G. 2012. Simultaneous heterotrophic nitrification and aerobic denitrification by bacterium Rhodococcus sp. CPZ24. Bioresource Technology, 116: 116–266.Google Scholar
  4. Garbeva P, Baggs E M, Prosser J I. 2007. Phylogeny of nitrite reductase (nir K) and nitric oxide reductase (nor B) genes from Nitrosospira species isolated from soil. FEMS Microbiology Letters, 266 (1): 83–89.CrossRefGoogle Scholar
  5. GBT 7493–198. Water quality-determination of nitrogen (nitrite)-spectrophotometric method. 1987. The State Environmental Protection Administration of China, p.144-148.Google Scholar
  6. Grguric G, Wetmore S S, Fournier R W. 2000. Biological denitrification in a closed seawater system. Chemosphere, 40 (5): 549–555.CrossRefGoogle Scholar
  7. Guo L Y, Chen Q K, Fang F, Hu Z X, Wu J, Miao A J, Xiao L, Chen X F, Yang L Y. 2013. Application potential of a newly isolated indigenous aerobic denitrifier for nitrate and ammonium removal of eutrophic lake water. Bioresource Technology, 142: 142–45.CrossRefGoogle Scholar
  8. Huang H K, Tseng S K. 2001. Nitrate reduction by Citrobacter diversus under aerobic environment. Applied Microbiology & Biotechnology, 55 (1): 90–94.CrossRefGoogle Scholar
  9. Joo H S, Hirai M, Shoda M. 2005. Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis No. 4. Journal of Bioscience and Bioengineering, 100 (2): 184–191.CrossRefGoogle Scholar
  10. Kim M, Jeong S Y, Yoon S J, Cho S J, Kim Y H, Kim M J, Ryu E Y, Lee S J. 2008. Aerobic denitrification of Pseudomonas putida AD-21 at different C/N ratios. Journal of Bioscience and Bioengineering, 106 (5): 498–502.CrossRefGoogle Scholar
  11. Kolndadacha O D, Adikwu I A, Okaeme A N, Atiribom R Y, Mohammed A, Musa Y M. 2011. The role of probiotics in aquaculture in Nigeria-a review. Continental Journal of Fisheries and Aquatic Sciences, 5 (1): 8–15.Google Scholar
  12. Kumar G S, Ramakrishnan A, Hung Y T. 2010. Simultaneous removal of carbon and nitrogen from domestic wastewater in an aerobic RBC. In: Wang L K, Tay J H, Tay S T L, Hung Y T eds. Environmental Bioengineering: Volume 11. Humana Press, Totowa, NJ, USA. p.403-443.Google Scholar
  13. Liang Y H, Li D, Zhang X J, Zeng H P, Yang Z, Zhang J. 2014. Microbial characteristics and nitrogen removal of simultaneous partial nitrification, anammox and denitrification (SNAD) process treating low C/N ratio sewage. Bioresource Technology, 169: 103–109.CrossRefGoogle Scholar
  14. Lukow T, Diekmann H. 1997. Aerobic denitrification by a newly isolated heterotrophic bacterium strain TLl. Biotechnology Letters, 19 (11): 1 157–1 159.CrossRefGoogle Scholar
  15. Michael E T, Amos S O, Hussaini L T. 2014. A review on probiotics application in aquaculture. Fisheries and Aquaculture Journal, 5 (4): 111.CrossRefGoogle Scholar
  16. Miyahara M, Kim S W, Fushinobu S, Takaki K, Yamada T, Watanabe A, Miyauchi K, Endo G, Wakagi T, Shoun H. 2010. Potential of aerobic denitrification by Pseudomonas stutzeri TR2 to reduce nitrous oxide emissions from wastewater treatment plants. Applied and Environmental Microbiology, 76 (14): 4 619–4 625.CrossRefGoogle Scholar
  17. Obaja D, Macé S, Mata-Alvarez J. 2005. Biological nutrient removal by a sequencing batch reactor (SBR) using an internal organic carbon source in digested piggery wastewater. Bioresource Technology, 96 (1): 7–14.CrossRefGoogle Scholar
  18. Pai S L, Chong N M, Chen C H. 1999. Potential applications of aerobic denitrifying bacteria as bioagents in wastewater treatment. Bioresource Technology, 68 (2): 179–185.CrossRefGoogle Scholar
  19. Patureau D, Bernet N, Delgenès J P, Moletta R. 2000. Effect of dissolved oxygen and carbon-nitrogen loads on denitrification by an aerobic consortium. Applied Microbiology and Biotechnology, 54 (4): 535–542.CrossRefGoogle Scholar
  20. Qiu X F, Wang T W, Zhong X M, Du G C, Chen J. 2012. Screening and characterization of an aerobic nitrifyingdenitrifying bacterium from activated sludge. Biotechnology and Bioprocess Engineering, 17 (2): 353–360.CrossRefGoogle Scholar
  21. Robertson L A, van Niel E W J, Torremans R A M, Kuenen J G. 1988. Simultaneous nitrification and denitrification in aerobic chemostat cultures of Thiosphaera pantotropha. Applied and Environmental Microbiology, 54 (11): 2 812–2 818.Google Scholar
  22. Robertson L A, Kuenen J G. 1984. Aerobic denitrification: a controversy revived. Archives of Microbiology, 139 (4): 351–354.CrossRefGoogle Scholar
  23. Shao Q, Yu X B. 2008. Isolation and characterization of a strain denitrobacteria. Biotechnology, 18 (3): 63–65. (in Chinese with English abstract)Google Scholar
  24. Šiljeg M, Foglar L, Kukučka M. 2010. The ground water ammonium sorption onto Croatian and Serbian clinoptilolite. Journal of Hazardous Materials, 178 (1–3): 572–577.Google Scholar
  25. Skjermo J, Bakke I, Dahle S W, Vadstein O. 2015. Probiotic strains introduced through live feed and rearing water have low colonizing success in developing Atlantic cod larvae. Aquaculture, 438: 17–23.CrossRefGoogle Scholar
  26. Song Z F, An J, Fu G H, Yang X L. 2011. Isolation and characterization of an aerobic denitrifying Bacillus sp. YX-6 from shrimp culture ponds. Aquaculture, 319 (1–2): 188–193.Google Scholar
  27. Su J J, Yeh K S, Tseng P W. 2006. A strain of Pseudomonas sp. isolated from piggery wastewater treatment systems with heterotrophic nitrification capability in Taiwan. Current Microbiology, 53 (1): 77–81.CrossRefGoogle Scholar
  28. Takaya N, Catalan-Sakairi M A B, Sakaguchi Y, Kato I, Zhou Z M, Shoun H. 2003. Aerobic denitrifying bacteria that produce low levels of nitrous oxide. Applied and Environmental Microbiology, 6 9 (6): 3 152–3 157.CrossRefGoogle Scholar
  29. Tang J Y, Dai Y X, Li Y M, Qin J G, Wang Y. 2016. Can application of commercial microbial products improve fish growth and water quality in freshwater polyculture? North American Journal of Aquaculture, 78 (2): 154–160.CrossRefGoogle Scholar
  30. Taylor S M, He Y L, Zhao B, Huang J. 2009. Heterotrophic ammonium removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Providencia rettgeri YL. Journal of Environmental Sciences, 21 (10): 1 336–1 341.CrossRefGoogle Scholar
  31. Throbäck I N, Enwall K, Jarvis A, Hallin S. 2004. Reassessing PCR primers targeting nir S, nir K and nos Z genes for community surveys of denitrifying bacteria with DGGE. FEMS Microbiology Ecology, 49 (3): 401–417.CrossRefGoogle Scholar
  32. van Niel E W J, Braber K J, Robertson L A, Kuene J G. 1992. Heterotrophic nitrification and aerobic denitrification in Alcaligenes faecalis strain TUD. Antonie van Leeuwenhoek, 62 (3): 231–237.CrossRefGoogle Scholar
  33. Verschuere L, Rombaut G, Sorgeloos P, Verstraete W. 2000. Probiotic bacteria as biological control agents in aquaculture. Microbiology and Molecular Biology Reviews, 64 (4): 655–671.CrossRefGoogle Scholar
  34. Wan C L, Yang X, Lee D J, Du M A, Wan F, Chen C. 2011. Aerobic denitrification by novel isolated strain using NO ˉ 2-N as nitrogen source. B ioresource Technology, 102 (15): 7 244–7 248.CrossRefGoogle Scholar
  35. Wang P, Yuan Y Z, Li Q, Yang J Z, Zheng Y L, He M Q, Geng H, Xiong L, Liu D L. 2013. Isolation and immobilization of new aerobic denitrifying bacteria. International Biodeterioration & Biodegradation, 76: 12–17.CrossRefGoogle Scholar
  36. Xiang M F, Wang P, Liu X T, Zhai Q. 2006. Selection and identification of aerobic denitrifiers in denitrification process of wastewater. Food Science and Technology, 31 (7): 153–156. (in Chinese with English abstract)Google Scholar
  37. Xu Y T. 1994. The effect of different carbon sources on biological denitrification. Chinese Journal of Environmental Science, 15 (2): 28–32, 44. (in Chinese with English abstract)Google Scholar
  38. Xu Z N, Lin X T, Lin Q, Yang Y F, Wang Y X. 2007. Nitrogen, phosphorus, and energy waste outputs of four marine cage-cultured fish fed with trash fish. Aquaculture, 263 (1–4): 130–141.CrossRefGoogle Scholar
  39. Yao M J, Rui J P, Li J B, Dai Y M, Bai Y F, Heděnec P, Wang J M, Zhang S H, Pei K Q, Liu C, Wang Y F, He Z L, Frouz J, Li X Z. 2014. Rate-specific responses of prokaryotic diversity and structure to nitrogen deposition in the Leymus chinensis steppe. Soil Biology and Biochemistry, 79: 79–81.CrossRefGoogle Scholar
  40. Ye L, Shao M F, Zhang T, Tong A H Y, Lok S. 2011. Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing. Water Research, 45 (15): 4 390–4 398.CrossRefGoogle Scholar
  41. Ye L, Zhang T. 2010. Estimation of nitrifier abundances in a partial nitrification reactor treating ammonium-rich saline wastewater using DGGE, T-RFLP and mathematical modeling. Applied Microbiology and Biotechnology, 88 (6): 1 403–1 412.CrossRefGoogle Scholar
  42. Zhang J B, Wu P X, Hao B, Yu Z N. 2011. Heterotrophic nitrification and aerobic denitrification by the bacterium Pseudomonas stutzeri YZN-001. Bioresource Technology, 102 (21): 9 866–9 869.CrossRefGoogle Scholar
  43. Zhang Q L, Liu Y, Ai G M, Miao L L, Zheng H Y, Liu Z P. 2012. The characteristics of a novel heterotrophic nitrification-aerobic denitrification bacterium, Bacillus methylotrophicus strain L7. Bioresource Technology, 108: 108–35.CrossRefGoogle Scholar
  44. Zhao Y G, Fang Y, Jin Y L, Huang J, Ma X R, He K Z, He Z M, Wang F, Zhao H. 2015. Microbial community and removal of nitrogen via the addition of a carrier in a pilotscale duckweed-based wastewater treatment system. Bioresource Technology, 179: 179–549.Google Scholar
  45. Zheng H Y, Liu Y, Gao X Y, Ai G M, Miao L L, Liu Z P. 2012. Characterization of a marine origin aerobic nitrifyingdenitrifying bacterium. Journal of Bioscience and Bioengineering, 114 (1): 33–37.CrossRefGoogle Scholar
  46. Zhu L, Ding W, Feng L J, Dai X, Xu X Y. 2012a. Characteristics of an aerobic denitrifier that utilizes ammonium and nitrate simultaneously under the oligotrophic niche. Environmental Science and Pollution Research, 19 (8): 3 185–3 191.CrossRefGoogle Scholar
  47. Zhu L, Ding W, Feng L J, Kong Y, Xu J, Xu X Y. 2012b. Isolation of aerobic denitrifiers and characterization for their potential application in the bioremediation of oligotrophic ecosystem. Bioresource Technology, 108: 1–7.CrossRefGoogle Scholar

Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Cuicui Wang (王翠翠)
    • 1
    • 2
  • Kai Zhang (张凯)
    • 1
  • Jun Xie (谢骏)
    • 1
    Email author
  • Qigen Liu (刘其根)
    • 2
  • Deguang Yu (余德光)
    • 1
  • Guangjun Wang (王广军)
    • 1
  • Ermeng Yu (郁二蒙)
    • 1
  • Wangbao Gong (龚望宝)
    • 1
  • Zhifei Li (李志斐)
    • 1
  1. 1.Key Laboratory of Tropical and Subtropical Fishery Resource Application and Cultivation, Ministry of Agriculture, Pearl River Fisheries Research InstituteChinese Academy of Fishery SciencesGuangzhouChina
  2. 2.College of Fisheries and Life SciencesShanghai Ocean UniversityShanghaiChina

Personalised recommendations