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Perfluoroalkyl substances (PFAS), such as perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), are persistent, globally distributed, anthropogenic compounds. The primary source(s) for human exposure are not well understood although within home exposure is likely important since many consumer products have been treated with different PFAS, and people spend much of their lives indoors. Herein, domestic cats were used as sentinels to investigate potential exposure and health linkages. PFAS in serum samples of 72 pet and feral cats, including 11 healthy and 61 with one or more primary disease diagnoses, were quantitated using high-resolution time-of-flight mass spectroscopy. All but one sample had detectable PFAS, with PFOS and perfluorohexane sulfonate (PFHxS) ranging from < LOQ to 121 and < LOQ to 235 ng/mL, respectively. PFAS prevalence and geometric means in cats were very similar to contemporary NHANES reports of human sera in the U. S. population. The highest PFAS serum concentrations detected were in indoor cats due to disproportionately elevated PFHxS levels. Ranked by quartile, contingency testing indicated that total PFAS levels were positively associated with living indoors and with higher body weight and body condition scores. Individual PFAS quartile rankings suggested positive associations with respiratory effusion, thyroid, liver, and possibly chronic kidney disease. Domestic cats appear to be useful sentinels for assessing primary PFAS exposure routes, especially indoor sources of relevance to children. Additional case-control studies in pet cats are warranted to better define the potential health associations observed herein. A "One Health" approach assessing humans, pets, and their common environment may improve our understanding of chronic, low-level, largely indoor, PFAS exposure and effects in humans and animals alike. Published by Elsevier Inc.