Garvicin KS, a Broad-Spectrum Bacteriocin Protects Zebrafish Larvae against Lactococcus garvieae Infection

Saurabh Dubey,Øystein Evensen,Hetron M Munang’Andu,Dzung B Diep

doi:10.3390/ijms23052833

Saurabh Dubey, Øystein Evensen + Show 2 more

Open Access

https://doi.org/10.3390/ijms23052833

Copy DOI

Abstract

Bacteriocins are emerging as a viable alternative to antibiotics due to their ability to inhibit growth or kill antibiotic resistant pathogens. Herein, we evaluated the ability of the bacteriocin Garvicin KS (GarKS) produced by Lactococcus garvieae KS1546 isolated from cow milk to inhibit the growth of fish and foodborne bacterial pathogens. We found that GarKS inhibited the growth of five fish L. garvieae strains isolated from infected trout and eels. Among fish pathogens, GarKS inhibited the growth of Streptococcus agalactiae serotypes Ia and Ib, and Aeromonas hydrophila but did not inhibit the growth of Edwardsiella tarda. In addition, it inhibited the growth of A. salmonicida strain 6421 but not A. salmonicida strain 6422 and Yersinia ruckeri. There was no inhibition of three foodborne bacterial species, namely Salmonella enterica, Klebsiella pneumoniae, and Escherichia coli. In vitro cytotoxicity tests using different GarKS concentrations showed that the highest concentration of 33 µg/mL exhibited low cytotoxicity, while concentrations ≤3.3 µg/mL had no cytotoxicity on CHSE-214 and RTG-2 cells. In vivo tests showed that zebrafish larvae treated with 33 µg/mL and 3.3 µg/mL GarKS prior to challenge had 53% and 48% survival, respectively, while concentrations ≤0.33 µg/mL were nonprotective. Altogether, these data show that GarKS has a broad inhibitory spectrum against Gram positive and negative bacteria and that it has potential applications as a therapeutic agent for a wide range of bacterial pathogens. Thus, future studies should include clinical trials to test the efficacy of GarKS against various bacterial pathogens in farmed fish.

Highlights

Aquaculture is one of the fastest growing food producing sectors in the world [1] and bacterial diseases cause high economic losses in aquaculture
Our findings show that all foodborne bacterial pathogens, K. pneumoniae, E. coli, and S. enterica were not inhibited at 33 μg/mL Garvicin KS (GarKS) at all three temperatures of 25 ◦C, 30 ◦C and 37 ◦C after overnight culture (Figure 3A–C)
We have shown the inhibitory properties of GarKS on growth of different fish bacterial pathogens, foodborne bacteria, and its protective ability against lethal challenge with L. garvieae in zebrafish larvae

Summary

Introduction

Aquaculture is one of the fastest growing food producing sectors in the world [1] and bacterial diseases cause high economic losses in aquaculture. Vaccine development has contributed to reducing the occurrence of bacterial diseases in aquaculture, there are still many pathogens that have not been successfully controlled through vaccination [2]. The use of antibiotics for the treatment of bacterial diseases poses the danger of selecting for antibiotic resistant bacteria. There is need for novel antimicrobials with broad therapeutic spectrums. Bacteriocins which are widely used in food preservation [3], have great potential as a source for antimicrobials to help reduce bacterial infections in aquaculture. Most bacteriocins have a narrow antimicrobial inhibition spectrum, the inhibition of food-borne zoonoses, food spoilage, and antimicrobial resistant (AMR) bacteria has been reported [6]

Objectives

Methods

Results

Conclusion