Invasive rats are among the most damaging animals to native species on many island ecosystems including those in Hawaii. On Oahu Island, U.S. Army Garrison Natural Resources Program currently manages invasive rat populations to protect natural resources by using grids of A24 automated traps, and previously snap-trap grids and rodenticide bait stations. Despite these control efforts generally suppressing rats, some lands with natural resources that are at risk to rat predation are not easily accessible for implementing these traditional rat control methods. In a 430-ha mesic forest on Oahu where ungulates are excluded and site access is limited due to military training and presence of live ordnance, we tested the efficacy of aerial application of anticoagulant rodenticide bait pellets (Diphacinone-50 Conservation), applied in two applications at a rate of 12.82 kg/ha per application. We measured the effectiveness of the rodenticide bait application by deploying tracking tunnels (inked and baited cards to identify rat presence) before, during, and after applications within treated and nearby untreated areas. Due to restricted access, we failed to estimate nest success of an endangered bird; yet previous research showed rat control increases this bird’s population. We also measured diphacinone residues in stream water at the treatment site to determine this method’s risk level to the aquatic ecosystem. The aerial application resulted in immediate and sustained reduction in the rat population, as evidenced by rat activity decreasing from ~44% to 3.8% during the first three months after bait application and maintained <20% rat activity for 10 months. Trail cameras and recovered rat carcasses also highlighted effectiveness. One of 34 stream samples analyzed had detectable diphacinone residues and this single sample was taken one week after application and it had very low levels of diphacinone (below levels quantifiable). Aerial application of diphacinone appears to be an efficient and effective rat suppression technique for natural resource protection in complex landscapes.