In wine, one method of limiting the addition of sulphites, a harmful and allergenic agent, is bio-protection. This practice consists of the early addition of microorganisms on grape must before fermentation. Non-Saccharomycesyeasts have been proposed as an interesting alternative to sulphite addition. However, scientific data proving the effectiveness of bio-protection remains sparse. This study provides the first analysis of the chemical and microbiological effects of aMetschnikowia pulcherrimastrain inoculated at the beginning of the red winemaking process in three wineries as an alternative to sulphiting. Like sulphiting, bio-protection effectively limited the growth of spoilage microbiota and had no influence on the phenolic compounds protecting musts and wine from oxidation. The bio-protection had no effect on the volatile compounds and the sensory differences were dependent on the experimental sites. However, a non-targeted metabolomic analysis by FTICR-MS highlighted a bio-protection signature.