The southern river otter (Lontra provocax) and the marine otter (Lontra felina) are endangered species that inhabit Chile. In southern Chile, both species cohabit with the American mink (Neogale vison), an invasive exotic species. The Chilean aquaculture industry has grown exponentially since the late 1980s, with salmon farming taking place from central Chile to the Patagonian fjords and channels. This study assessed co-occurrence between otters, mink, and aquaculture in Patagonia by (1) distributing a survey among workers, fisheries personnel, and aquaculture inspectors concerning observations of otters and mink inside or around aquaculture facilities and outcomes; and (2) a geographical assessment of distribution overlap between known otter territory and salmon farming-registered facilities. We recorded the first anecdotal evidence of interaction, described as co-occurrence, among native otters, American mink, and salmon aquaculture in Patagonia, which varied among seasons and seems to be increasing. We also recorded evidence of difficulty in recognition of the three mustelids among respondents. There is a geographically extended interaction between otters and salmon farms in Chile. The evidence of interaction among alien American mink, native endangered otters, and aquaculture is an early alarm for human–wildlife conflict, and further studies are recommended to ensure native otter conservation.