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A three-year-old ferret presented with a three-month history of rapid clinical deterioration necessitating euthanasia shortly after initial veterinary assessment. Postmortem PCR testing confirmed Mycobacterium xenopi which is most commonly identified in amphibians, reptiles and aquatic life. Infection of a captive-bred domestic ferret is highly unusual. A collaborative effort involving medical doctors, clinical veterinarians and veterinary pathologists investigated the potential sources of human-animal, animal-animal and environmental-animal M xenopi transmission. No human-animal or animal-animal risks were identified. As the affected ferret was the only ferret to have regular exposure to the owner's aquarium, a postmortem study of a dead guppy and aquarium water analysis were performed which confirmed mycobacteriosis. Although M xenopi was not specifically cultured, as a slow-growing organism, M xenopi may have been outgrown by more rapidly growing mycobacteria or Gram-positive bacilli present in the water. Thus, transmission of M xenopi via aquarium exposure was certainly plausible. This is the second documented case of M xenopi in a ferret and the first to determine a source of infection. This report highlights the previously recognised risk of mycobacterial exposure from aquaria and that caution is required before allowing domestic ferrets to have contact with potentially infected water reservoirs due to its fatal nature in this vulnerable species.