Heterotrophic nitrifying bacteria from activated sludge in DHS reactor for ammonium removal of natural rubber processing wastewater treatment

Tran Minh Duc, Phan Thi Thanh Thuy, Nguyen Thi Huyen, Nguyen Lan Huong
Author affiliations


  • Tran Minh Duc School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet, Hai Ba Trung, Hanoi, Viet Nam
  • Phan Thi Thanh Thuy School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet, Hai Ba Trung, Hanoi, Viet Nam
  • Nguyen Thi Huyen School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet, Hai Ba Trung, Hanoi, Viet Nam
  • Nguyen Lan Huong School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet, Hai Ba Trung, Hanoi, Viet Nam




heterotrophic nitrifying bacterium, downflow hanging sponge, ammonium removal, natural rubber processing wastewater


Two heterotrophic nitrifying bacterial strains, D2 and D7 were isolated from an activated sludge of sponges in a laboratory-scale downflow hanging sponge reactor. Both strains exhibited efficient ammonium removal ability over a wide range of ammonium loads. At the initial concentration of 100 mg/L, NH4+-N was completely degraded within 20 h by both strains. When the initial concentration was increased to 200 mg/L, the NH4+-N removal efficiency was 99.6 % within 20 h by the strain D2 and 61.3 % by the strain D7. In natural rubber processing wastewater, the ammonium removal efficiencies of strain D2 and D7 were 38 % and 99 % with the initial N-NH4+ concentration of 280 and 380 mg/L after 88 h, respectively. The 16S rRNA gene sequence of D2 and D7 showed the highest similarity to the Pseudomonas aeruginosa and Glutamicibacter nicotianae, respectively. This is the first report to demonstrate the ability to remove ammonium in NRPW by ​heterotrophic nitrifying bacteria isolated from activated sludge in DHS reactor.


Download data is not yet available.


Tanikawa D., Syutsubo K., Hatamoto M., Fukuda M., Takahashi M., Choeisai P. K., Yamaguchi, T. - Treatment of natural rubber processing wastewater using a combination system of a two-stage up-flow anaerobic sludge blanket and down-flow hanging sponge system. Water Sci. Technol. 73 (2016) 1777-1784. doi.org/10.2166/wst.2016.019.

Tanikawa D., Kataoka T., Hirakata Y., Hatamoto M., Yamaguchi, T. - Pre-treatment and post-treatment systems for enhancing natural rubber industrial wastewater treatment. Process Saf. Environ. Prot. 138 (2020) 256–262. doi.org/10.1016/j.psep.2020.03.030.

Tran P.T., Watari T., Hirakata Y., Nguyen T.T., Hatamoto M., Tanikawa D., Syutsubo K., Nguyen M.T., Fukuda M., Nguyen L.H., Yamaguchi, T. - Anaerobic baffled reactor in treatment of natural rubber processing wastewater: reactor performance and analysis of microbial community. J. Water Environ. Technol. 15 (2017) 241-251. doi.org/10.2965/jwet.17-038.

Watari T., Mai T. C., Tanikawa D., Hirakata Y., Hatamoto M., Syutsubo K., Fukuda M., Nguyen N. B., Yamaguchi T. - Development of downflow hanging sponge (DHS) reactor as post treatment of existing combined anaerobic tank treating natural rubber processing wastewater. Water Sci. Technol. 75 (2017) 57-68. doi.org/10.2166/wst.2016.487.

Watari T., Nguyen T.T., Natsumi T., Tanikawa D., Kuroda K., Nguyen L.H., Nguyen M.T., Huynh T.T., Hatamoto M., Syutsubo K., Fukuda M., Yamaguchi T. - Development of a BR-UASB-DHS system for natural rubber processing wastewater treatment. Environ. Technol. 37 (2016) 459-465. doi.org/10.1080/09593330.2015.1117042.

Tanikawa D., Kataoka T., Sonaka H., Hirakata Y., Hatamoto M., Yamaguchi T. - Evaluation of key factors for residual rubber coagulation in natural rubber processing wastewater, J. Water Process. Eng. 33 (2020) 101041. doi.org/10.1016/j.jwpe.2019.101041.

Yang M., Lu D., Yang J., Zhao Y., Zhao Q., Sun Y., Liu H., Ma, J. - Carbon and nitrogen metabolic pathways and interaction of cold-resistant heterotrophic nitrifying bacteria under aerobic and anaerobic conditions. Chemosphere 234 (2019) 162-170. doi.org/10.1016/j.chemosphere.2019.06.052.

Zhang Q.L., Liu Y., Ai G.M., Miao L.L., Zheng H.Y., Liu, Z.P. - The characteristics of a novel heterotrophic nitrification-aerobic denitrification bacterium, Bacillus methylotrophicus strain L7. Bioresour. Technol. 108 (2012) 35-44. doi.org/10.1016/j.biortech.2011.12.139.

Zhang J., Wu P., Hao B., and Yu Z. - Heterotrophic nitrification and aerobic denitrification by the bacterium Pseudomonas stutzeri YZN-001. Bioresour. Technol. 102 (2011) 9866-9869. doi.org/10.1016/j.biortech.2011.07.118.

Sun Y., Li A., Zhang X., Ma F. - Regulation of dissolved oxygen from accumulated nitrite during the heterotrophic nitrification and aerobic denitrification of Pseudomonas stutzeri T13. Appl. Microbiol. Biotechnol., 99 (2015) 3243-3248. doi.org/10.1007/s00253-014-6221-6.

Yu Y., An Q., Zhou Y., Deng S., Miao Y., Zhao B., Yang L. - Highly synergistic effects on ammonium removal by the co-system of Pseudomonas stutzeri XL-2 and modified walnut shell biochar. Bioresour. Technol. 280 (2019) 239-246. doi.org/10.1016/j.biortech.2019.02.037.

Gao J., Zhu T., Liu C., Zhang J., Gao J., Zhang J., Cai M.,

Li Y. - Ammonium removal characteristics of heterotrophic nitrifying bacterium Pseudomonas stutzeri GEP 01 with potential for treatment of ammonium rich wastewater. Bioprocess. Biosyst. Eng. 43 (2020) 959-969. doi.org/10.1007/s00449-020-02292-x.

Motamedi H., Jafari M. - Screening heterotrophic ammonia removal and aerobic denitrifying bacteria from wastewater of ammonia production units of a petrochemical industry. Curr. Microbiol. 77 (2020) 2207-2214. doi.org/10.1007/s00284-020-02065-5.

Yang L., Ren Y. X., Zhao S. Q., Liang X., Wang J. - Isolation and characterization of three heterotrophic nitrifying-aerobic denitrifying bacteria from a sequencing batch reactor. Ann. Microbiol. 66 (2016) 737-747. doi.org/10.1007/s13213-015-1161-7.

Zhao B., Tian M., An Q., Ye J., Guo J. S., - Characteristics of a heterotrophic nitrogen removal bacterium and its potential application on treatment of ammonium-rich wastewater. Bioresour. Technol. 226 (2017) 46-54. doi.org/10.1016/j.biortech.2016.11.120.

Cai X., Li K., He T., Wang Y., Zhang X., Xie E., Ding N.,

and Li Z. - Characteristics of Heterotrophic Nitrifying and Aerobic Denitrifying Arthrobacter nicotianae D51 Strain in the Presence of Copper. Water 11 (2019) 434. doi.org/10.3390/w11030434.

Zhang N., Zhang Y., Bohu T., Wu S., Bai Z., Zhuang.X. - Nitrogen removal characteristics and constraints of an Alphaproteobacteria with potential for high nitrogen content heterotrophic nitrification-aerobic denitrification. Microorganisms 10 (2022) 235. doi.org/10.3390/microorganisms10020235

Nguyen L.H., Itoh K., Suyama K. - Diversity of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T)-degrading bacteria in Vietnamese soils. Microbes Environ. 22 (2007) 243-256. doi.org/10.1264/jsme2.22.243

Lane D.J. - 16S/23S rRNA sequencing. In: Stackebrandt E., Goodfellow M. - Nucleic acid techniques in bacterial systematics, John Wiley & Sons, New York (1991), 115-175.

Tamura K., Stecher G., and Kumar S. - MEGA 11: Molecular Evolutionary Genetics Analysis Version 11. Mol. Biol. Evol. 38 (2021) 3022-3027. doi.org/10.1093/molbev/msab120.

Zhang, Q. - Research on key issues in determination of ammomia nitrogen in water and waswater by Nessler's reagent spectrophotometry. Environ. Eng. 27 (2009) 85-88.

Hach Company, - Nitrite Method 8153. Water Anal. Handb. 3 (2014) 4-11.

Huang X., Ni J., Yang C., Mi Feng M., Zhenlun Li Z., Xie D. - Efficient ammonium removal by bacteria Rhodopseudomonas isolated from natural landscape water: China case study. Water 10 (2018) 1107. doi.org/10.3390/w10081107.




How to Cite

T. Minh Duc, P. Thi Thanh Thuy, N. Thi Huyen, and N. L. Huong, “Heterotrophic nitrifying bacteria from activated sludge in DHS reactor for ammonium removal of natural rubber processing wastewater treatment”, Vietnam J. Sci. Technol., vol. 61, no. 5, pp. 865–874, Oct. 2023.




Most read articles by the same author(s)