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Azithromycin can induce SOS response and horizontal gene transfer of SXT element in Vibrio cholerae

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Abstract

Background

The emergence and spread of drug resistance in Vibrio cholerae are mainly attributed to horizontal gene transfer of mobile genetic elements, especially the SXT (sulfamethoxazole and trimethoprim) element, an integrative conjugative element carrying multiple drug resistance genes. SOS (save our souls) bacterial stress response in Vibrio cholerae plays a pivotal role in inducing the SXT element transfer and induction of the CTX prophage, carrying the important virulence factor cholera toxin encoded by the ctxAB gene.

Methods

This study investigated whether the subinhibitory concentration of antibiotics like ciprofloxacin, tetracycline, and azithromycin induce SOS response by detecting the expression of recA and lexA, the key genes of SOS response by reverse transcriptase real time PCR (RT-qPCR). We also studied the transfer of SXT element in response to these three antibiotics by bacterial conjugation. Transfer of SXT elements was confirmed by detecting the SXT element-specific conserved genes.

Results

The results of the real-time PCR showed that all three antibiotics induced SOS response with more robust induction by tetracycline and azithromycin relative to ciprofloxacin. We observed a higher frequency of transfer of SXT elements in cultures exposed to these antibiotics and the control mitomycin C compared to unexposed cultures.

Conclusion

Our study indicates that antibiotics including azithromycin, ciprofloxacin, and tetracycline activate SOS response in Vibrio cholerae and demonstrates a robust mechanism for wide dissemination of drug resistance.

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Data availability

All data generated or analysed during this study are included in this published article and its supplementary information files (ESF1).

Code availability

Not applicable.

Abbreviations

SXT:

Sulfamethoxazole and trimethoprim

SOS:

Save our souls/ship

MIC:

Minimum inhibitory concentration

HGT:

Horizontal gene transfer

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Acknowledgements

We thank Dr. Asish K Muhkopadhyay, Scientist F (Sr. Deputy Director), Division of Bacteriology, NICED, Kolkata, for kindly providing us the standard strains; classical (O395), El Tor (N16961), and variant (IDH 03709) and E coli J53 strain. We thank Dr. Sudhakar Baluchamy, Professor of Biotechnology Pondicherry University, Pondicherry, for his valuable comments and suggestion given throughout the study. We are also thankful to Mr. Mailan Natrajan, Ms. Ankita Das and Ms. Kamali Doss research scholars, for their time, help and support throughout the project.

Funding

The Council of Scientific and Industrial Research, (CSIR), New Delhi, supported this research, Grant No. Ref: 27(0336)/18/EMR-II.

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Authors and Affiliations

  1. Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education & Research, (JIPMER), Dhanvantri Nagar, Gorimedu, Puducherry, 605 006, India

    Regina Sumitha Mohanraj & Jharna Mandal

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  1. Regina Sumitha Mohanraj
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Contributions

JM conceived and designed the experiments. RM performed the experiments. RM and JM analyzed the data. JM & RM discussed the results and reviewed them, RM wrote the manuscript, and JM edited it.

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Correspondence to Jharna Mandal.

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The authors declare that they have no conflict of interest.

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This study was reviewed and approved by the Institute Ethics Committee at Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER), Pondicherry (Approval No. JIP/IEC/2017/0130) and by the Institute Bio-Safety Committee (Approval No. JIP/IBSC/2017/01) for performing the genetic transfer.

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Mohanraj, R.S., Mandal, J. Azithromycin can induce SOS response and horizontal gene transfer of SXT element in Vibrio cholerae. Mol Biol Rep 49, 4737–4748 (2022). https://doi.org/10.1007/s11033-022-07323-2

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