An in situ transplantation experiment followed by metagenomic analysis revealed shifts in gut bacterial communities of deep-sea cold seep mussels (Gigantidas haimaensis) when deprived of their natural methane-rich environment. Gammaproteobacteria were enriched and Bacteroidetes were slightly depleted, with functional changes linked to carbon source acquisition, ammonia and sulphide utilisation, and self-protection mechanisms.