Disease is often considered a key threat to species of conservation significance. For some, it has resulted in localised extinctions and declines in range and abundance. However, for some species, the assertion that a disease poses a significant threat of extinction is based solely on correlative or anecdotal evidence, often inferred from individual clinical case reports. While a species’ susceptibility to a disease may be demonstrated in a number of individuals, investigations rarely extend to measuring the impact of disease at the population level and its contribution, if any, to population declines. The eastern quoll (<i>Dasyurus viverrinus</i>) is a medium-sized Australian marsupial carnivore that is undergoing severe and rapid decline in Tasmania, its last refuge. Reasons for the decline are currently not understood. Feral cats (<i>Felis catus</i>) may be undergoing competitive release following the ongoing decline of the Tasmanian devil (<i>Sarcophilus harrisii</i>), with cats suppressing eastern quolls through increased predation, competition, exclusion or exposure to diseases such as toxoplasmosis. To investigate the effects of <i>Toxoplasma gondii</i> infection, eastern quoll populations at four sites were regularly screened for the seroprevalence of <i>T. gondii</i>-specific IgG antibodies. Seroprevalence was approximately five times higher at sites with declining quoll populations, and there was a negative association between seroprevalence and quoll abundance. However, <i>T. gondii</i> infection did not reduce quoll survival or reproduction. Despite a high susceptibility to T. gondii infection, eastern quoll populations do not appear to be limited by the parasite or its resultant disease. Significantly higher seroprevalence is a signal of greater exposure to feral cats at sites where eastern quolls are declining, suggesting that increased predation, competition or exclusion by feral cats may be precipitating population declines.