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Oxytetracycline and heavy metals promote the migration of resistance genes in the intestinal microbiome by plasmid transfer

The ISME Journal volume 17, pages 2003–2013 (2023)Cite this article

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Abstract

Horizontal gene transfer (HGT) has been considered the most important pathway to introduce antibiotic resistance genes (ARGs), which seriously threatens human health and biological security. The presence of ARGs in the aquatic environment and their effect on the intestinal micro-ecosystem of aquatic animals can occur easily. To investigate the HGT potential and rule of exogenous ARGs in the intestinal flora, a visual conjugative model was developed, including the donor of dual-fluorescent bacterium and the recipient of Xenopus tropicalis intestinal microbiome. Some common pollutants of oxytetracycline (OTC) and three heavy metals (Zn, Cu and Pb) were selected as the stressor. The multi-techniques of flow cytometry (FCM), scanning electron microscopy (SEM), atomic force microscopy (AFM), single-cell Raman spectroscopy with sorting (SCRSS) and indicator analysis were used in this study. The results showed that ARG transfer could occur more easily under stressors. Moreover, the conjugation efficiency mainly depended on the viability of the intestinal bacteria. The mechanisms of OTC and heavy metal stressing conjugation included the upregulation of ompC, traJ, traG and the downregulation of korA gene. Moreover, the enzymatic activities of SOD, CAT, GSH-PX increased and the bacterial surface appearance also changed. The predominant recipient was identified as Citrobacter freundi by SCRSS, in which the abundance and quantity of ARG after conjugation were higher than those before. Therefore, since the diversity of potential recipients in the intestine are very high, the migration of invasive ARGs in the microbiome should be given more attention to prevent its potential risks to public health.

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Fig. 1: Schematic depicting experimental design and conjugation results of ARGs induced by OTC and heavy metals.
Fig. 2: Effects of OTC and heavy metals on the cell viability of donors and recipients.
Fig. 3: Effects of mRNA expression in different exposures.
Fig. 4: Effects of antibiotics and heavy metals on reactive oxygen species in mating systems of each group.
Fig. 5: Effects of antibiotics and heavy metals on the characteristics of the cell membrane surface in conjugation solution.
Fig. 6: Transconjugant sorting and ARG analysis.
Fig. 7: A model depicting the mechanisms underlying the RP4 plasmid-mediated conjugation promoted by OTC and heavy metals.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (No. 42277260, 41977340) and the Guangdong Basic and Applied Basic Research Foundation (No. 2019A1515110937). The authors acknowledge the help of the Guangdong University of Technology analysis and test center.

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Authors and Affiliations

  1. School of Environmental Science and Engineering, Guangdong University of Technology, No. 100 Wai Huan Xi Road, Guangzhou Higher Education Mega Center, Panyu District, Guangzhou, 510006, PR China

    Xiaojun Lin, Chaonan Zhang, Ruiqi Han, Huishi Peng & Yanbin Xu

  2. Analysis and Test Center, Guangdong University of Technology, No. 100 Wai Huan Xi Road, Guangzhou Higher Education Mega Center, Panyu District, Guangzhou, 510006, PR China

    Shoupeng Li, Xiao Zhou & Lu Huang

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YX conceived and supervised the project. XL conceived and designed the experiments. XL, CZ, RH, SL, HP, XZ and LH performed the experiments. XL analyzed data and wrote the manuscript.

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Correspondence to Yanbin Xu.

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Lin, X., Zhang, C., Han, R. et al. Oxytetracycline and heavy metals promote the migration of resistance genes in the intestinal microbiome by plasmid transfer. ISME J 17, 2003–2013 (2023). https://doi.org/10.1038/s41396-023-01514-w

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