Wildfires are particularly prevalent in the Western United States, home to more than 2 million dairy cows that produce more than 25% of the nation's milk. Wildfires emit fine particulate matter (PM 2.5 ) in smoke, which is a known air toxin and is thought to contribute to morbidity in humans by inducing inflammation. The physiological responses of dairy cows to wildfire PM 2.5 are unknown. Herein we assessed the immune, metabolic, and production responses of lactating Holstein cows to wildfire PM 2.5 inhalation. Cows (primiparous, n = 7; multiparous, n = 6) were monitored across the wildfire season from July to September 2020. Cows were housed in freestall pens and thus were exposed to ambient air quality. Air temperature, relative humidity, and PM 2.5 were obtained from a monitoring station 5.7 km from the farm. Animals were considered to be exposed to wildfire PM 2.5 if daily average PM 2.5 exceeded 35 µg/m 3 and wildfire and wind trajectory mapping showed that the PM 2.5 derived from active wildfires. Based on these conditions, cows were exposed to wildfire PM 2.5 for 7 consecutive days in mid-September. Milk yield was recorded daily and milk components analysis conducted before, during, and after exposure. Blood was taken from the jugular vein before, during, and after exposure and assayed for hematology, blood chemistry, and blood metabolites. Statistical analysis was conducted using mixed models including PM 2.5 , temperature-humidity index (THI), parity (primiparous or multiparous), and their interactions as fixed effects and cow as a random effect. Separate models included lags up to 7 d to identify delayed and persistent effects from wildfire PM 2.5 exposure. Exposure to elevated PM 2.5 from wildfire smoke resulted in lower milk yield during exposure and for 7 d after last exposure and higher blood CO 2 concentration, which persisted for 1 d following exposure. We observed a positive PM 2.5 by THI interaction for eosinophil and basophil count and a negative PM 2.5 by THI interaction for red blood cell count and hemoglobin concentration after a 3-d lag.Neutrophil count was also lower with a combination of higher THI and PM 2.5 . We found no discernable effect of PM 2.5 on haptoglobin concentration. Effects of PM 2.5 and THI on metabolism were contingent on day of exposure. On lag d 0, blood urea nitrogen (BUN) was reduced with higher combined THI and PM 2.5 , but on subsequent lag days, THI and PM 2.5 had a positive interaction on BUN. Conversely, THI and PM 2.5 had a positive interacting effect on nonesterified fatty acids (NEFA) on lag d 0 but subsequently caused a reduction in circulating NEFA concentration. Our results suggest that exposure to high wildfire-derived PM 2.5 , alone or in concert with elevated THI, alters systemic metabolism, milk production, and the innate immune system.