左氧氟沙星暴露对斑马鱼肠道微生物的影响

doi:10.16258/j.cnki.1674-5906.2025.04.009

生态环境学报 ›› 2025, Vol. 34 ›› Issue (4): 593-607.DOI: 10.16258/j.cnki.1674-5906.2025.04.009

• 研究论文【环境科学】 • 上一篇下一篇

左氧氟沙星暴露对斑马鱼肠道微生物的影响

张亚平¹^,^*(), 莫璀玲¹, 李迎鹏², 徐棵¹

1.集美大学港口与海岸工程学院，福建厦门 361021
2.集美大学水产学院，福建厦门 361021

收稿日期:2024-10-18 出版日期:2025-04-18 发布日期:2025-04-24
通讯作者: ^*
作者简介:张亚平（1978年生），教授，博士，主要从事环境科学与工程技术研究。E-mail: ypzhang@jmu.edu.cn
基金资助:
国家自然科学基金项目(21978246);国家自然科学基金项目(22478119);福建省自然科学基金项目(2014J01050)

Effects of Levofloxacin Exposure on Intestinal Microbes of Zebrafish

ZHANG Yaping¹^,^*(), MO Cuiling¹, LI Yingpeng², XU Ke¹

1. College of Harbour and Coastal Engineering, Jimei University, Xiamen 361021, P. R. China
2. Fisheries College, Jimei University, Xiamen 361021, P. R. China

Received:2024-10-18 Online:2025-04-18 Published:2025-04-24

摘要/Abstract

摘要： 左氧氟沙星作为一种抗生素类药物，在水产养殖、畜牧兽医、人类健康等方面应用广泛，属于药物与个人护理用品（PPCPs）的一种典型代表物。随着肠道微生物相关研究的开展与不断深入，越来越多的证据表明，PPCPs可以通过干扰肠道微生物稳态来造成各种疾病。为了探索左氧氟沙星对斑马鱼肠道微生物的影响及由此产生的毒性作用，采用环境浓度下的左氧氟沙星（0、0.01、0.1、1、10、100 mg·L⁻¹）浸泡成年斑马鱼10 d，然后采用石蜡切片与HE染色、16S rDNA高通量测序等方法分析斑马鱼肠道组织形态，以及肠道菌群组成的变化。结果表明，斑马鱼肠道组织切片显示肠腺体结构紊乱、黏膜层糜烂，绒毛脱落，炎症细胞数量增多，随着左氧氟沙星浓度的升高，损伤作用越强。16S rDNA高通量测序显示斑马鱼肠道细菌中放线菌门（Actinomycetota）、厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）丰度下降至对照组的70%，微生物多样性指数相较于对照组有明显下降。综上，左氧氟沙星会对斑马鱼造成显著的肠道损伤，导致肠道菌群丰度和多样性下降，进而使其免疫和代谢功能弱化。该研究为深入了解左氧氟沙星引起的肠道毒性相关机制提供了线索，并将为左氧氟沙星的合理使用提供理论参考。

关键词: 左氧氟沙星, 斑马鱼, 肠道菌群, 肠道稳态, 环境毒理, 抗生素

Abstract:

Levofloxacin is widely used in aquaculture, animal husbandry, veterinary medicine, and in human health as an antibiotic drug. This is a typical representation of a PPCPs. With the development and deepening of research on intestinal microbes, increasing evidence has shown that PPCPs can cause various diseases by disrupting gut microbiota homeostasis. Therefore, there is an urgent need to explore the relationship between levofloxacin and intestinal microbial homeostasis as well as the impact of levofloxacin on intestinal microbes. This study aimed to explore the effects of levofloxacin on the intestinal microbes of zebrafish and the resulting toxicity. Adult zebrafish were exposed to environmentally relevant concentrations of levofloxacin (0, 10 μg·L⁻¹, 100 μg·L⁻¹, 1 mg·L⁻¹, 10 mg·L⁻¹, and 100 mg·L⁻¹) for 10 days, after which the morphological changes in zebrafish intestinal tissue were analyzed. The composition of the intestinal flora was analyzed using paraffin sectioning, HE staining, and 16S rDNA high-throughput sequencing. The results showed that zebrafish intestinal tissue sections revealed disordered intestinal gland structure, erosion of the mucosal layer, shedding of villi, and an increased number of inflammatory cells. These damaging effects increased with increasing concentrations of levofloxacin. 16S rDNA high-throughput sequencing revealed that the abundances of Actinobacteria, Firmicutes, Bacteroidetes, and Proteobacteria in the intestinal bacteria of zebrafish decreased to 70% of that in the control group, and the microbial diversity index decreased significantly compared to that in the control group. In summary, levofloxacin can cause significant intestinal damage, immune and metabolic dysfunction, and other toxic effects in zebrafish, thereby reducing the abundance and diversity of the intestinal flora. This study provides a deeper understanding of the mechanisms underlying levofloxacin-induced gut toxicity, and offers a theoretical reference for the rational use of levofloxacin.

Key words: levofloxacin, zebrafish, intestinal flora, intestinal homeostasis, environmental toxicology, antibiotic

中图分类号:

X171.5

张亚平, 莫璀玲, 李迎鹏, 徐棵. 左氧氟沙星暴露对斑马鱼肠道微生物的影响[J]. 生态环境学报, 2025, 34(4): 593-607.

ZHANG Yaping, MO Cuiling, LI Yingpeng, XU Ke. Effects of Levofloxacin Exposure on Intestinal Microbes of Zebrafish[J]. Ecology and Environment, 2025, 34(4): 593-607.

图/表 10

图1 左氧氟沙星暴露对斑马鱼肠道显微结构影响

Figure 1 Effects of levofloxacin exposure on Intestinal microstructure in zebrafish

图2 OTUs的upset图 A代表对照组，B为10 μg·L?1组，C为100 μg·L?1组，D为1 mg·L?1组，E为10 mg·L?1组，F为100 mg·L?1组。下同

Figure 2 Upset diagram of OTUs

图3 菌门分布堆叠图和菌种分布堆叠图

Figure 3 Stacked diagrams of phylum and bacterial species distribution

图4 属和种水平上的物种丰度热图（a）图为各组别变化显著的前20菌属；（b）图为各组别变化显著的前10菌种。数值越大，表示丰度越高，数值越小，表示丰度越低

Figure 4 Species abundance heatmap at the genus and species levels

图5 斑马鱼肠道菌群α多样性指数

Figure 5 Alpha diversity index of zebrafish gut microbiota

图6 OTU聚类分析

Figure 6 OTU clustering analysis

图7 PCoA分析

Figure 7 PCoA analysis

图8 基于LEFSe分析的不同组肠道微生物LDA和多级物种差异判别表由内至外辐射的圆圈代表了由界（单个圆圈）至属（或种）的分类级别，不同分类级别上的每一个小圆圈代表该水平下的一个分类，小圆圈直径大小与物种相对丰度大小成正比；无显著差异的物种统一着色为黄色，差异显著的物种跟随组别进行着色，不同颜色表示在各自组别中起到重要作用的微生物类群。生物标志物对应的物种名展示在右侧，字母编号与图中对应

Figure 8 LDA and multi-level species differentiation table for different groups of gut microbiota based on LEFSe analysis

图9 通路Level2功能丰度热图

Figure 9 Channel Level2 functional abundance heatmap

图10 KW秩和检验

Figure 10 Kruskal-Wallis rank sum test

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左氧氟沙星暴露对斑马鱼肠道微生物的影响

Effects of Levofloxacin Exposure on Intestinal Microbes of Zebrafish

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