A study was conducted to determine the release rates of piperonyl butoxide (PBO) and permethrin from synergized insecticidal cattle ear tags and their effects on mortality of the horn fly, Hematobia irritans irritans (L.) (Diptera: Muscidae). PBO was released from the ear tags at a higher rate than permethrin in both winter and summer trials. The cumulative release of PBO and permethrin from the ear tags at the end of 18 wk in the winter trial was 50.4 and 30.3%, respectively. The cumulative release of PBO and permethrin from the ear tags at the end of 18 wk in the summer trial was 66.7 and 44.7%, respectively. There was a significant correlation between the cumulative daily high ambient temperature (°C) and the cumulative release of both PBO and permethrin. Compared with the susceptible horn fly strain, the permethrin-resistant strain demonstrated 7.9- and 12.8-fold resistance to permethrin at the levels of LC50 and LC90, respectively. When exposed to filter paper wipes taken from the shoulders of cattle treated with the PBO-synergized permethrin tags from the summer trial, the resistant strain demonstrated reduced mortality compared with the susceptible strain. The mortality of the resistant strain at 2- and 3-h exposure exhibited a pattern of declining fly mortalities as a result of the decreased release of PBO and permethrin, as well as the decline in the ratio of PBO:permethrin released from the tags after 8 wk. A similar decline in horn fly mortalities was observed in the susceptible strain at 30-min exposure time that coincided with the pattern of reduced release of PBO and permethrin from the ear tags over the course of summer trial.