Abstract

There is an increasing interest from the aquafeed industry in functional feeds containing selected additives that improve fish growth performance and health status. Functional feed additives include probiotics, prebiotics, organic acids, and phytogenics (substances derived from plants and their extracts). This study evaluated the effects of dietary inclusion of a mucilage extract rich in galactomannan oligosaccharides (GMOS), a mixture of garlic and labiatae-plants oils (PHYTO), and a combination of them (GMOSPHYTO), on gut microbiota composition of European sea bass (Dicentrarchus labrax) fed with a low fishmeal (FM) and fish oil (FO) diet. Three experimental diets and a control diet (plant-based formulation with 10% FM and 6% FO) were tested in a 63-days feeding trial. To analyze the microbiota associated to feeds and the intestinal autochthonous (mucosa-adhered) and allochthonous (transient) microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and QIIME2 pipeline were used. Metabarcoding analysis of feed-associated bacteria showed that the microbial communities of control (CTRL) feed deeply differed from those of experimental diets. The number of reads was significantly lower in CTRL feed than in other feeds. The OTU (operational taxonomic unit) number was instead similar between the feeds, ranging from 42 to 50 OTUs. The variation of resident gut microbiota induced by diet was lower than the variation of transient intestinal microbiota, because feedstuffs are a major source of allochthonous bacteria, which can temporarily integrate into the gut transient microbiome. However, the composition of transient bacterial communities was not simply a mirror of feed-borne bacteria. Indeed, the microbial profile of feeds was different from both faecal and mucosa profiles. Our findings suggest that the dietary inclusion of GMOS (0.5%) and PHYTO (0.02%) in a low FM and FO diet induces changes in gut microbiota composition of European sea bass. However, if on allochthonous microbiota the combined inclusion of GMOS and PHYTO showed an antagonistic effect on bactericidal activity against Vibrionales, at mucosa level, only GMOSPHYTO diet increased the relative abundance of Bacteroidales, Lactobacillales, and Clostridiales resident bacterial orders. The main beneficial effects of GMOS and PHYTO on gut microbiota are the reduction of coliforms and Vibrionales bacteria, which include several potentially pathogenic species for fish, and the enrichment of gut microbiota composition with butyrate producer taxa. Therefore, these functional ingredients have a great potential to be used as health-promoting agents in the farming of European sea bass and other marine fish.

Highlights

  • Farming of carnivorous fish species still relies on fishmeal (FM) and fish oil (FO) that represent the optimal sources of protein and lipids in aquafeeds

  • Based on the aforementioned properties of galactomannans and phytogenics, the aim of this study was to investigate the effects of dietary inclusion of galactomannan oligosaccharides (GMOS), a mixture of garlic and labiatae-plants oils (PHYTO), and a combination of them (GMOSPHYTO), on gut microbiota composition of European sea bass fed with a plant-based diet

  • Four isoenergetic and isonitrogenous experimental diets were manufactured by an extrusion process in the BioMar Tech-Centre (Brande, Denmark) starting from a plant-based formulation (10% FM, 6% FO) and containing different additives: 0.5% of galactomannan oligosaccharides from mucilage (GMOS; Delacon, Austria), 0.02% of a mixture of garlic and labiate-plants oils (PHYTO; Delacon, Austria) and a combination of both additives (GMOSPHYTO, 0.52%)

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Summary

Introduction

Farming of carnivorous fish species still relies on fishmeal (FM) and fish oil (FO) that represent the optimal sources of protein and lipids in aquafeeds. The shortening of the primary and secondary intestinal mucosal folds, higher level of infiltrated leucocytes in the lamina propria and submucosa, damaged microvilli, altered gut microbiota profiles, and increased gut permeability are the most typical signs of such inflammatory response [5, 6, 7] These negative-side effects reduce the capacity of enterocytes to absorb nutrients and can promote the translocation of indigenous or opportunistic bacteria, potentially pathogenic for the host [8, 7]. Functional feed additives include probiotics, prebiotics, immune-stimulants, organic acids, nucleotides, exogenous enzymes, and phytogenics (substances derived from herbs, spices, other plants and their extracts) Their application into diet formulations targets a specific purpose: probiotics, prebiotics, phytogenics, and immune-stimulants target the improvement of intestinal health, stress, and disease resistance, whereas acidifiers, exogenous enzymes and indirectly probiotics are used to improve fish performance by enhancing feed digestibility, or counteracting the negative effects of antinutrients [14]

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