Abstract

The health and environmental risks associated with antibiotic use in aquaculture have promoted bacterial probiotics as an alternative approach to control fish infections in vulnerable larval and juvenile stages. However, evidence-based identification of probiotics is often hindered by the complexity of bacteria-host interactions and host variability in microbiologically uncontrolled conditions. While these difficulties can be partially resolved using gnotobiotic models harboring no or reduced microbiota, most host-microbe interaction studies are carried out in animal models with little relevance for fish farming. Here we studied host-microbiota-pathogen interactions in a germ-free and gnotobiotic model of rainbow trout (Oncorhynchus mykiss), one of the most widely cultured salmonids. We demonstrated that germ-free larvae raised in sterile conditions displayed no significant difference in growth after 35 days compared to conventionally-raised larvae, but were extremely sensitive to infection by Flavobacterium columnare, a common freshwater fish pathogen causing major economic losses worldwide. Furthermore, re-conventionalization with 11 culturable species from the conventional trout microbiota conferred resistance to F. columnare infection. Using mono-re-conventionalized germ-free trout, we identified that this protection is determined by a commensal Flavobacterium strain displaying antibacterial activity against F. columnare. Finally, we demonstrated that use of gnotobiotic trout is a suitable approach for the identification of both endogenous and exogenous probiotic bacterial strains protecting teleostean hosts against F. columnare. This study therefore establishes an ecologically-relevant gnotobiotic model for the study of host-pathogen interactions and colonization resistance in farmed fish.

Highlights

  • As wild fish stock harvests have reached biologically unsustainable limits, aquaculture has grown to provide over half of all fish consumed worldwide [1]

  • We developed procedures to raise germ-free rainbow trout larvae to study the determinants of microbiota-associated resistance to infection

  • We showed that the fish pathogen Flavobacterium columnare rapidly kills infected germ-free but not conventional larvae

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Summary

Introduction

As wild fish stock harvests have reached biologically unsustainable limits, aquaculture has grown to provide over half of all fish consumed worldwide [1]. Intensive aquaculture facilities are prone to disease outbreaks and the high mortality rate in immunologically immature juveniles, in which vaccination is unpractical, constitutes a primary bottleneck for fish production [2,3,4] These recurrent complications prompt the prophylactic or therapeutic use of antibiotics and chemical disinfectants to prevent fish diseases [5,6] but may lead to final consumer safety risks, environmental pollution and spread of antibiotic resistance [7]. In this context, the use of bacterial probiotics to improve fish health and protect disease-susceptible juveniles is an economic and ecological sensible alternative to antibiotic treatments [8,9]. Selection of probiotic bacteria is often empirical or hampered by the poor reproducibility of in vivo challenges, frequently performed in relatively uncontrolled conditions with high inter-individual microbial compositions [15,19]

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