Resuscitation of aerobic denitrifying bacteria in the VBNC state and their nitrogen conversion capacity

doi:10.7523/j.issn.2095-6134.2019.04.014

Abstract

Abstract: At the present,the biological treatment of high ammonium nitrogen wastewater has attracted much attention because of its environmental friendly characteristics and high efficiency. Most of strains can not be excavated due to entry into a viable but non-culturable (VBNC) state. The resuscitation of VBNC strains is important for biological treatment of wastewater with high ammonium nitrogen. In this study,VBNC strain was cultured in high ammonium nitrogen wastewater by resuscitation-promoting factors (Rpf) resuscitation,and its homology was analyzed by 16S rRNA gene sequencing. The isolated strains,which were originally in the VBNC state, were further conducted to investigate the aerobic denitrification capacities with the help of the analyses of nitrate, nitrite, total nitrogen, and ammonium nitrogen. The results showed that the strains belonged to Gordonia and Pseudomonas, among which the aerobic denitrification ability of strain ZYR51 was the highest. This study provides a new method for excavation of VBNC bacteria with aerobic denitrifying properties and a new way for screening efficient strains in the treatment of ammonium nitrogen wastewater.

Key words: high ammonia nitrogen wastewater, VBNC bacteria, Rpf protein, microbial resource, nitrogen removal

CLC Number:

X522

XU Luning, WANG Yuyang, XUE Binbing, MEI Rongwu, ZHANG Yu, DING Linxian, SU Xiaomei. Resuscitation of aerobic denitrifying bacteria in the VBNC state and their nitrogen conversion capacity[J]. , 2019, 36(4): 545-551.

References

[1] Frette L, Gejlsbjerg B, Westermann P. Aerobic denitrifiers isolated from an alternating active sludge system[J]. FEMS Microbiology Ecology, 1997, 24(4):363-370.
[2] Robertson L A, Kuenen J G. Aerobic denitrification:a controversy revived[J]. Archives of Microbiology, 1984, 139(4):351-354.
[3] 苏晓梅, 丁林贤, 沈超峰. 活的但非可培养功能菌群对多氯联苯降解的探索进展[J]. 微生物学报, 2013, 53(9):908-914.
[4] Mukamolova G V, Murzin A G, Salina E G, et al. Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation[J]. Molecular Microbiology, 2006, 59(1):84-98.
[5] Li Y, Chen J, Zhao M, et al. Promoting resuscitation of viable but nonculturable cells of Vibrio harveyi by a resuscitation-promoting factor-like protein YeaZ[J]. Journal of Applied Microbiology, 2017, 122(2):338-346.
[6] Mukamolova G V, Kaprelyants A S, Young D I, et al. A bacterial cytokine[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(15):8916-8921.
[7] 丁林贤, 张萍华, 洪华嫦. 藤黄微球菌Rpf活性蛋白的制取及对红球菌VBNC菌体的复苏作用[J]. 微生物学报, 2012, 52(1):77-89.
[8] Kumar S, Stecher G, Tamura K. MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolution, 2016, 33(7):1870-1874.
[9] 国家环境保护总局. 水和废水监测分析方法[M]. 4版. 北京:中国环境科学出版社, 2002:105-218.
[10] 王存政, 王兴春, 牛倩, 等. N-(1-萘基)-乙二胺光度法测定水中亚硝酸盐氮的测量不确定度评定[J]. 环境工程学报, 2013, 31(5):147-150.
[11] Jin Y, Gan G, Yu X, et al. Isolation of viable but non-culturable bacteria from printing and dyeing wastewater bioreactor based on resuscitation promoting factor[J]. Current Microbiology, 2017, 74(7):787-797.
[12] Lin C L, Shen F T, Tan C C, et al. Characterization of Gordonia sp. strain CC-NAPH129-6 capable of naphthalene degradation[J]. Microbiological Research, 2012, 167(7):395-404.
[13] 刘磊, 李习武, 刘双江, 等. 降解多环芳烃的菌株Gordonia sp. He4的分离鉴定及其在菲污染土壤修复过程中的动态变化[J]. 环境科学, 2007, 28(3):617-622.
[14] Chen J, Zhan P, Koopman B, et al. Bioaugmentation with Gordonia strain JW8 in treatment of pulp and paper wastewater[J]. Clean Technologies and Environmental Policy, 2012, 14(5):899-904.
[15] Wang B, Wang C, Liu W, et al. Biosurfactant-producing microorganism Pseudomonas sp. SB assists the phytoremediation of DDT-contaminated soil by two grass species[J]. Chemosphere, 2017, 182:137-142.
[16] Postnikova O A, Shao J, Mock N M, et al. Gene expression profiling in viable but nonculturable (VBNC) cells of Pseudomonas syringae pv. syringae[J]. Frontiers in Microbiology, 2015, 6:1419.
[17] He T, Li Z, Sun Q, et al. Heterotrophic nitrification and aerobic denitrification by Pseudomonas tolaasii Y-11 without nitrite accumulation during nitrogen conversion[J]. Bioresource Technology, 2016, 200:493-499.
[18] 胡朝松, 李春强, 刘志昕, 等. 海洋沉积物中反硝化细菌的分离鉴定及反硝化性能研究[J]. 环境科学研究, 2009, 22(1):114-118.
[19] Yang L, Ren Y X, Liang X, et al. Nitrogen removal characteristics of a heterotrophic nitrifier Acinetobacter junii YB and its potential application for the treatment of high-strength nitrogenous wastewater[J]. Bioresource Technology, 2015, 193:227-233.
[20] 孙庆花, 于德爽, 张培玉, 等. 1株海洋异养硝化-好氧反硝化菌的分离鉴定及其脱氮特性[J]. 环境科学, 2016, 37(2):647-654.
[21] 张峰峰, 谢凤行, 赵玉洁, 等. 一株反硝化细菌的分离鉴定及脱氮能力[J]. 环境工程学报, 2013, 7(11):4415-4420.