Effect of sulfide on the denitrification potential of sediment in black-odor rivers
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摘要: 通过探究不同浓度硫化物对黑臭河道底泥反硝化过程的影响,同时分析底泥细菌、反硝化菌和硫酸盐还原菌的响应变化,为强化底泥反硝化脱氮提供理论依据与技术支撑.研究结果表明:较低浓度的硫化物(8 mg-1)对底泥反硝化潜势无明显影响;适宜浓度的硫化物(40和64 mg-1)对底泥反硝化有明显的促进作用,且浓度越高促进作用越明显;当硫化物浓度升高到96 mg-1及以上时,还原态硫对反硝化过程起抑制作用,浓度越高抑制作用越明显.底泥经过一段时间的反硝化培养,细菌多样性以变形菌门、绿弯菌门、拟杆菌门为主;同时,底泥细菌总数明显增加,代谢菌群的nirS丰度比、dsrB丰度比分别为1.42%和0.05%,相较原始底泥(0.15%,0.19%),反硝化细菌增值明显,但硫酸盐还原菌数量有所下降.Abstract: This study aimed to explore the effect of sulfide on sediment denitrification in different concentrations, analyze the response of bacteria, including denitrifying and sulfate-reducing bacteria, and provide a theoretical basis and technical support for improving the denitrification process of sediment in black-odor rivers. The results showed that a low concentration of sulfides (8 mg·L-1) didn't have a significant effect. In constrast, a moderate concentration of sulfides (40 mg·L-1, 64 mg·L-1) promoted the denitrification process; in this range the higher the concentration of sulfides, the faster the rate of nitrate degradation. When the sulfide concentration rises to 96 mg·L-1 and above, reductive sulfur inhibits denitrification, and the higher the concentration, the more obvious the inhibition. After a period of denitrification of the culture, the bacterial diversity is mainly composed of Proteobacteria, Chlofloflexi, and Bacillus. As the same time, the total number of bacteria in the sediment increased; the abundance ratio of nirS to 16S rRNA gene and dsrB to 16S rRNA gene were 1.42% and 0.05%, respectively. Compared with the original sediment (0.15%, 0.19%), the denitrifying bacteria increased significantly, but the number of sulfate-reducing bacteria decreased.
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Key words:
- black-odor rivers /
- reductive sulfur /
- denitrification /
- sediment
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表 1 不同培养实验组所添加基质
Tab. 1 ddition of substrates in each sediment incubation assays
mg·L-1 组号 1 2 3 4 5 硫化物浓度 0 8 40 64 96 注:实验组2在第66天将硫化物浓度提高至160 mg$\cdot $L$^{-1}$ 表 2 高通量测序及荧光定量所用引物
Tab. 2 Primers used in high-throughput sequencing and real-time PCR analysis
表 3 培养周期内添加不同浓度硫化物的各底泥培养组中硝酸盐还原速率分析
Tab. 3 Assessment of nitrate reduction rate for sediment incubations with the addition of different sulfide concentrations over different time periods
d 培养周期 $D_{1}$ $D_{2}$ $D_{3}$ $D_{4}$ $D_{5}$ 1 14 9 9 5 $-$ 2 14 10 10 6 $-$ 3 11 10 6 5 $-$ 4 10 10 6 6 $-$ 5 9 9 6 6 $-$ 6 10 $-$ 6 3 $-$ 7 9 $-$ 6 3 $-$ 8 9 $-$ 6 3 $-$ 注: $D_{1}$—$D_{5}$分别表示一个培养周期内实验组1—5硝酸盐还原率达到98%以上所需天数; "$-$"表示硝酸盐还原受到抑制; 从第6个培养周期开始实验组2的硫化物浓度增加到160 mg$\cdot $L$^{-1}$ 表 4 反硝化培养组底泥样品细菌多样性信息统计
Tab. 4 Statistics for bacterial diversity in sediment from denitrification incubations
样品 有效序列数量/条 OTUs Chao1指数 Shannon指数 Simpson指数 实验组3 22 036 872 1 285.7 6.247 4 0.956 32 对照组 22 418 1 130 1 655.1 6.711 9 0.971 12 注:实验组3添加硫化物浓度为40 mg$\cdot $L$^{-1}$, 对照组不添加硝酸盐, 只添加硫化物40 mg$\cdot $L$^{-1}$ 表 5 反硝化培养组底泥细菌门水平组成
Tab. 5 Composition of bacteria at the phylum level in sediment from denitrification-assessed
Actinobacteria Bacteroidetes Caldiserica Chloroflexi Firmicutes Proteobacteria 实验组3 1.13% 13.21% 0.49% 22.96% 3.01% 53.75% 对照组 1.33% 21.61% 2.13% 37.45% 1.73% 28.31% 注:实验组3添加硫化物浓度为40 mg$\cdot $L$^{-1}$, 对照组不添加硝酸盐, 只添加硫化物40 mg$\cdot $L$^{-1}$ 表 6 反硝化底泥中氮硫代谢关键菌群丰度响应
Tab. 6 Statistics for bacterial diversity in sediment from denitrification incbuation
样品 16S基因丰度 nirS基因丰度 dsrB基因丰度 nirS丰度比 dsrB丰度比 原始底泥 1.06$\times$10$^{9}$ 1.58$\times$10$^{6}$ 1.99$\times $10$^{6}$ 0.15% 0.19% 实验组3 2.61$\times $10$^{9}$ 3.71$\times $10$^{7}$ 1.32$\times $10$^{6}$ 1.42% 0.05% 注: nirS丰度比和dsrB丰度比均为对应功能基因丰度与16S基因丰度的比值 -
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