洛美沙星和诺氟沙星对水中生物反硝化过程的影响模拟试验

doi:10.13745/j.esf.sf.2022.1.22

摘要/Abstract

摘要：

近年来水环境中硝酸盐污染与抗生素污染的现象引起了人们的广泛关注,但目前复合抗生素污染对反硝化过程产生的影响并不明确。本研究以水环境中检出率较高的诺氟沙星和洛美沙星为代表进行模拟实验,探究ng级的两种抗生素复合对反硝化过程的影响。硝氮和亚硝氮的降解情况表明,实验浓度条件下,洛美沙星、诺氟沙星单用与二者联用对水环境中的反硝化过程存在不同的抑制作用。洛美沙星单用前期轻微促进反硝化,后期表现抑制作用,而诺氟沙星单用始终表现出抑制作用;洛美沙星和诺氟沙星联用抑制作用小于诺氟沙星单用,联用表现为拮抗作用。各体系的抑制作用大小为诺氟沙星>诺氟沙星+洛美沙星>洛美沙星。该模拟试验条件虽与实际条件有所差异,但在一定程度上表明抗生素联用表现出拮抗作用与反硝化体系内微生物数量、活性,反硝化酶活性,反硝化菌优势物种Achromobacter xylosoxidans、Acinetobacter baumannii、Pseudomonas sp.KY及功能基因nosZ和aac的丰度的变化有关。抗生素加入反硝化体系后会产生持续的影响,随反应时间的增加,反硝化菌逐渐适应有低浓度抗生素存在的环境,喹诺酮类抗生素耐药基因数量增加,微生物的耐药性增强,反硝化菌在数量和活性、反硝化酶活性及微生物群落层面均有回升趋势。

关键词: 硝酸盐, 抗生素复合, 反硝化作用, 拮抗作用

Abstract:

Nitrate and antibiotic pollutions in water have attracted much attention in recent years, but the effects of combination antibiotic pollution on the biological water denitrification process is not clear. To address this issue, we tested norfloxacin and lomefloxacin under simulated experimental conditions. The $\text{NO}_{3}^{-}$-N and $\text{NO}_{2}^{-}$-N degradation results showed that lomefloxacin and norfloxacin had different inhibitory effects on water denitrification: Lomefloxacin promoted denitrification slightly in the early stage but inhibited it later on, whereas norfloxacin only inhibited it. Lomefloxacin combined with norfloxacin were less inhibitory than norfloxacin alone, and the combination showed antagonistic activity. The inhibitory activities of antibiotics ranked as norfloxacin > norfloxacin + lomefloxacin > lomefloxacin. Although the simulated experimental conditions did not fully match the actual water condition, the above results implied that combination antibiotics have antagonistic effects on the denitrification process. Such effects were related largely to the microbial numbers and activities, activities of denitrification enzymes, as well as abundance variations of dominant denitrifying bacteria Achromobacter xylosoxidans, Acinetobacter baumannii, and Pseudomonas sp. KY and the nosZ and aac genes. As reaction time increased, the denitrifying bacteria gradually adapted to the environment of low-level antibiotics and the number of quinolone resistance genes increased; also on the rise were the amount and activities of denitrifying bacteria, activities of denitrification enzymes, and microbial communities.

Key words: nitrate, antibiotic compound, denitrification, antagonism

中图分类号:

X523

张玉叶, 何江涛, 邓璐, 邹华, 张金刚, 杨美萍. 洛美沙星和诺氟沙星对水中生物反硝化过程的影响模拟试验[J]. 地学前缘, 2022, 29(5): 497-507.

ZHANG Yuye, HE Jiangtao, DENG Lu, ZOU Hua, ZHANG Jingang, YANG Meiping. Effects of lomefloxacin and norfloxacin on the biological water denitrification process—an experimental study[J]. Earth Science Frontiers, 2022, 29(5): 497-507.

图/表 11

图1 抗生素的化学结构式

Fig.1 Chemical structural formula of antibiotics

图2 实验装置图

Fig.2 Experimental device diagram

图3 反应体系中硝氮、亚硝氮和氨氮变化情况

Fig.3 Changes of Nitrate, Nitrite and Ammonia in the reaction system

表1 试验周期各反应体系硝氮、亚硝氮降解情况统计表

Table 1 Statistical table of Nitrate and Nitrite degradation in each reaction system during the test period

组别		硝氮		亚硝氮
组别		降解时间/d	作用效果	降解时间/d	作用效果
空白周期V	各组	4	—	0	—
投加抗生素VI	Blank	4	—	0	—
	159 ng/L LOM	6	前期轻微促进,后期抑制	0	—
	509 ng/L NOR	10	明显抑制	10	抑制
	LOM+NOR	8	抑制作用减弱	7	轻微抑制
恢复期VII	Blank	4	—	0	—
	159 ng/L LOM	5	抑制作用	0	—
	509 ng/L NOR	7	抑制作用	0	—
	LOM+NOR	7	抑制作用	0	—

表2 投加抗生素期(0天和8天)时体系内气体含量变化

Table 2 Changes of gas content in the system during antibiotic administration period (0 days) and (8 days)

组别	N₂含量/%
组别	0d	8d	变化量	0d		8d		变化量		0d		8d		变化量
Blank	69.43	77.63	8.20		778.0		840.2		62.2		44.1		127.0		82.9
LOM	72.20	80.96	8.76		761.1		817.9		56.8		66.6		136.5		69.9
NOR	71.46	73.56	2.10		774.0		817.3		43.3		72.6		123.1		50.5
LOM+NOR	68.83	75.44	6.61		763.9		812.7		48.8		65.1		136.6		71.5

图4 正式试验各体系内反硝化菌OD490、OD600的变化(误差棒为3个平行样测试结果的标准差)

Fig.4 Changes of Denitrifying bacteria OD490 and OD600 in formal tests of each system (Error bar is the standard deviation of test results of three parallel samples)

图5 第VI周期各体系内反硝化菌活性与数量的抑制率(误差棒为3个平行样测试结果的标准差)

Fig.5 Inhibition rate of denitrifying bacteria activity and quantity in each system in the VI cycle (Error bar is the standard deviation of test results of three parallel samples)

图6 正式试验各体系内OTU数量变化注:空白周期(V)、投加抗生素期(VI)、恢复期(VII)中Blank、LOM(L)、NOR(N)、LOM+NOR(LN)体系内的OTU数量,如V-L表示空白周期LOM体系的OTU数量,下同。

Fig.6 Variation of OTU quantity in each system in formal test

图7 正式试验各样本物种群落结构聚类热图

Fig.7 Cluster heat map of community structure of each sample species in formal test

图8 正式试验各反应体系内Nar、Nir活性变化

Fig.8 Nar and Nir activity changes of each reaction system in formal test

图9 正式试验各样本的功能基因丰度

Fig.9 Functional gene abundance of each sample in formal test

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