碳源对反硝化细菌的反硝化速率和群落结构的影响

周梦娟; 缪恒锋; 陆震明; 阮文权

doi:10.13198/j.issn.1001-6929.2018.10.03

碳源对反硝化细菌的反硝化速率和群落结构的影响

doi: 10.13198/j.issn.1001-6929.2018.10.03

周梦娟^1,,
缪恒锋^{1, 2, 3, ,},
陆震明⁴,
阮文权^{1, 2, 3}

1.
江南大学环境与土木工程学院, 江苏无锡 214122
2.
江苏省厌氧生物技术重点实验室, 江苏无锡 214122
3.
江苏省水处理技术与材料协同创新中心, 江苏苏州 215009
4.
粮食发酵工艺与技术国家工程实验室, 江苏无锡 214122

基金项目:

国家水体污染控制与治理科技重大专项 2017ZX07203-003

详细信息

作者简介:
周梦娟(1994-), 女, 安徽合肥人, 2387334376@qq.com

通讯作者:
缪恒锋(1980-), 男, 江苏无锡人, 教授, 博士, 主要从事环境生物技术、环境化学研究, hfmiao@jiangnan.edu.cn

中图分类号: X703
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- 被引次数: 0
出版历程
- 收稿日期: 2018-04-05
- 修回日期: 2018-09-04
- 刊出日期: 2018-12-25

The Influence of Different Carbon Sources on Denitrification Rate and Community Structure of Denitrifying Bacteria

ZHOU Mengjuan^1
,,
MIAO Hengfeng^{1, 2, 3
, ,},
LU Zhenming⁴,
RUAN Wenquan^{1, 2, 3}

1.
School of Environmental & Civil Engineering, Jiangnan University, Wuxi 214122, China
2.
Jiangsu Key Laboratory of Anaerobic Biotechnology, Wuxi 214122, China
3.
Jiangsu Collaborative Innovation Center of Technology and Material of Water Treatment, Suzhou 215009, China
4.
National Engineering Laboratory for Grain Fermentation Technology and Technology, Wuxi 214122, China

Funds:

National Major Science and Technology Program for Water Pollution Control and Treatment, China 2017ZX07203-003

摘要

摘要: 为探究碳源类型在反硝化过程中对氮素转化和微生物群落组成的影响，分别建立R1（以C₆H₁₂O₆为碳源）和R2（以CH₃COONa为碳源）反应器，通过分析R1和R2反应器中反硝化过程的氮素转化情况，评价C₆H₁₂O₆和CH₃COONa对脱氮效果的影响，并运用动力学模型对R1和R2反应器中反硝化能力进行评价；同时，采用高通量测序技术表征2种碳源对反应器中微生物群落结构和多样性的影响.结果表明：①运行后期的R1、R2反应器中单位生物量的反硝化速率（以NO₃^--N计，下同）分别为8.56、11.26 mg/（g·h），R1反应器中NO₂^--N累积平均值为11.34 mg/L，显著高于R2反应器（0.20 mg/L），且R1反应器中NH₄⁺-N累积平均值为6.58 mg/L，是R2反应器（0.65 mg/L）的10.11倍.②反应器中NO₃^--N还原过程均符合Haldane模型，其中R1、R2反应器中单位生物量的r_max（最大降解速率）分别为35.61、47.79 mg/（g·h），表明R2反应器中的反硝化能力强于R1反应器.③微生物经过富集后，其细菌多样性和物种丰度下降，但发挥反硝化作用的微生物相对丰度逐渐增加.R1和R2反应器中共同的优势菌门有Proteobacterias、Bacteroidetes、Firmicutes和Gracilibacters，其在R1反应器中的相对丰度依次为96.14%、2.06%、0.66%和0.47%，在R2反应器中依次为79.75%、6.88%、9.47%和2.13%，优势菌门在不同运行时间的丰度表达上存在消长变化状态.研究显示，C₆H₁₂O₆和CH₃COONa在反硝化过程的氮素转化上存在明显差异，对各类优势菌群的相对丰度有明显影响.
- 连续流培养 /
- 反硝化细菌 /
- 外加碳源 /
- 氮素变化 /
- 生物群落结构
Abstract: In order to investigate the influence of carbon sources on nitrogen change and microorganism community structure in the denitrification process, R1 (C₆H₁₂O₆) and R2 (CH₃COONa) continuous-flowing reactors were set up for comparison of nitrogen conversion, which can be used to evaluate the removal effect of C₆H₁₂O₆ and CH₃COONa. The NO₃^--N removal kinetics for the denitrification capacity of R1 and R2 were conducted, and the high-throughput sequencing was utilized to characterize and compare the community structure and diversity. The results showed that: (1) The denitrification rate per unit biomass of R1 was 8.56 mg/(g·h) when the denitrification rate per unit biomass of R2 was 11.26 mg/(g·h). The average accumulation value of NO₂^--N in R1 was 11.34 mg/L, which was significantly higher than that in R2 (0.20 mg/L). The average value of NH₄⁺-N accumulation in R1 was 6.58 mg/L, which was 10.11 times of the cumulative amount of NH₄⁺-N in R2. (2) The Haldane model fit the experimental data in the NO₃^--N reduction process well, in which the r_max (the maximum degradation rate) value per unit biomass of R1 was 35.61 mg/(g·h), and the r_max value per unit biomass of R2 was 47.79 mg/(g·h), indicating that the denitrification ability of R2 is stronger than the denitrification ability of R1. (3) High-throughput sequencing analysis showed that the bacterial diversity and the species abundance were decreased after a period, but the relative abundance of denitrifying bacteria was raised. Proteobacterias, Bacteroidetes, Firmicutes and Gracilibacters were the dominant groups on the phylum level in the R1 and R2. And the relative proportions of the groups in R1 were 96.14%, 2.06%, 0.66% and 0.47%, respectively, and the relative abundance of the groups in R2 were 79.75%. 6.88%, 9.47% and 2.13%. The abundance of the dominant bacteria experienced changing state of growth and decline during the operation time. It showed that C₆H₁₂O₆ and CH₃COONa have significant differences in nitrogen conversion in the process of denitrification, and have a significant impact on the relative abundance of various prevailing species.
- continuous-flowing culture /
- denitrifying bacteria /
- external carbon source /
- nitrogen change /
- microbial community structure

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图 1 反硝化细菌富集装置

注：1—进水箱; 2—计量泵; 3—水浴循环泵; 4—气体流量计; 5—出水箱; 6—布水板.

Figure 1. The device of culturing denitrifying bacteria

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图 2 R1和R2反应器中单位生物量反硝化速率的变化

Figure 2. Changes of the denitrification rate for per unit biomass of R1 and R2 reactors

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图 3 不同碳源反应器ρ(NO₂^--N)的变化

Figure 3. The concentrations of NO₂^--N in the different carbon source reactors

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图 4 不同碳源反应器ρ(NH₄⁺-N)的变化

Figure 4. The concentrations of NH₄⁺-N in the different carbon source reactors

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图 5 单位生物量反硝化速率变化及反应动力学模型拟合结果

Figure 5. The denitrification rate value for per unit biomass and reaction kinetics fitting

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图 6 不同分类水平上细菌群落结构及分布

Figure 6. Bacterial community structure and distribution at different taxonomic levels

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表 1 微生物样本多样性指数

Table 1. Diversity index of microbial samples

样品编号	序列数/条	OTUs/个	Chao 1指数	ACE指数	Shannon-Wiener指数	Simpson指数	覆盖值/%
GS0	36 690	1 321	1 423.64	1 408.78	5.97	0.01	99.60
G5	47 870	562	970.86	1 135.95	3.79	0.04	99.50
G10	55 157	506	886.91	1 071.84	2.74	0.14	99.50
S5	46 768	549	1 064.68	965.55	2.53	0.16	99.40
S10	41 748	399	698.52	881.13	2.85	0.20	99.50

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