Brettanomyces yeasts play a relevant role in the fermentation industry, showing controversial behavior. There is growing interest in these yeasts in the fermentation industry as beer and bioethanol production, while in winemaking, they are considered spoilage microorganisms mainly used to produce ethyl phenols. These compounds may alter wine’s organoleptic characteristics, leading to significant economic loss. In this work, 45 Brettanomyces strains from seven different environments were genotyped and assayed for some oenological characters to investigate the possible relationship among sources of isolation, genotype characterization, and oenological characters. The results of biotyping showed four main clusters which were also distinguished according to the oenological characters. The oenological characters also distinguished the strains based on the isolation source, suggesting an overall relation between origin and biotypes. The negative correlation between fermentation rate and ethyl phenols production in the Brettanomyces population may indicate the adaptation to hostile environments differently from crop strains that showed the opposite behavior. The overall results contribute to clarifying some features of Brettanomyces yeasts, even if further investigations into the ability of these yeasts to colonize winemaking environments are needed.