Key gut microbiota components and functions in an aquatic keystone species across diets assessed by metaproteomics

Abstract The gut microbiota plays a crucial role in maintaining host fitness and modulating contaminant toxicity-related responses. However, information on how the gut microbiota of sentinel species responds to environmental factors is limited. In this study, we characterized the gut microbial community and its functions under normal, contaminant-free conditions by examining the effects of different diets over a 10-day period (alder leaf, carrot, spinach, and protein-rich granules) on the amphipod Gammarus fossarum , commonly used in bioassays for ecotoxicity assessment of contaminated rivers. Metaproteomic analysis of intestine samples enabled taxonomic characterization of the gut microbiota from this millimetric animal, assignment of biological functions to each microbial entity, and functional analysis of host proteins. The most abundant microbes detected in the gut belong to 37 bacterial and 5 fungal genera. Functional analyses of host and microbial proteins revealed complementary metabolic activities, allowing the degradation of complex polysaccharides such as cellulose and chitin. Diet was found to shape microbial community structure, with foodborne microorganisms strongly influencing structural changes during short-term feeding in amphipods. These microorganisms remained viable post-ingestion and contributed to food digestion. Functional stability was maintained across different diets, although the protein-rich granules diet induced functional shifts in both the host and its microbiota, reflecting their adaptation to a novel nutrient source. Finally, we identified a core microbiota driving key gut functions, less affected by dietary variations. These findings are significant for future ecotoxicological and biomonitoring investigations, leveraging the microbiomes of these sentinel animals as pivotal targets.