# Exogenous indole promotes florfenicol tolerance in Edwardsiella tarda

## TL;DR

Exogenous indole increases resistance to the antibiotic florfenicol in the fish pathogen Edwardsiella tarda by altering its metabolism and reducing antibiotic effectiveness.

## Contribution

This study reveals a novel mechanism by which indole promotes florfenicol tolerance in E. tarda through metabolic reprogramming and oxidative stress.

## Key findings

- Exogenous indole promotes florfenicol tolerance in Edwardsiella tarda by reprogramming its metabolome.
- Indole disrupts the TCA cycle and activates oxidative stress pathways, increasing intracellular Fe2+ and reactive oxygen species.
- In vivo, indole reduces the efficacy of florfenicol in protecting fish and eliminating the pathogen.

## Abstract

Bacterial metabolism is important for antibiotic resistance and tolerance. However, the impact of indole on bacterial metabolism and antibiotic efficacy has not been fully elucidated. In this study, we investigated the effect and specific mechanism of exogenous indole on the antibiotic susceptibility of Edwardsiella tarda, a common pathogen in freshwater and marine fish farming. We found that exogenous indole promoted E. tarda tolerance to the antibiotic florfenicol, and reprogrammed the E. tarda metabolome. A total of 108 metabolites were detected, including 66 differential metabolites that regulate various metabolic pathways, such as the tricarboxylic acid (TCA) cycle and nucleotide metabolism. Exogenous indole disrupted the TCA cycle in E. tarda by increasing the intracellular NADH contents and activating the respiratory chain to increase the reactive oxygen species levels, thereby increasing the intracellular Fe2+ content to activate the Fenton reaction, which in turn promotes the oxidative stress response. Furthermore, indole inhibited antibiotic entry into the cell and activated efflux pumps to reduce the intracellular antibiotic content, ultimately promoting antibiotic tolerance. In vivo, exogenous indole compromised the ability of florfenicol to protect fish survival and eliminate pathogenic bacteria. These results shed light on the metabolic changes induced by indole and suggest future directions for addressing antibiotic tolerance and clinical infections of E. tarda in aquaculture. This study serves as a reminder of the adverse effects of combining antibiotics with metabolites in aquaculture.