Surveys for disease agents were conducted in introduced free-ranging elk (Cervus elaphus nelsoni) in Arkansas and Kentucky. Elk had been captured in Colorado and Nebraska and released in Arkansas during 1981–1985. From 1997 through 2002 elk were captured in Arizona, Kansas, North Dakota, New Mexico, Oregon, and Utah and released in southeastern Kentucky. Specimens were collected from 170 hunter-killed elk in Arkansas during 1998–2006, and 44 elk in Kentucky during 2001–2004. Significant findings included isolation of Mycobacterium avium paratuberculosis from one elk in Kentucky and evidence of previous or current infections by Parelaphostrongylus tenuis in several animals in Arkansas. Serological tests provided evidence of previous infection by epizootic hemorrhagic disease virus, bluetongue virus, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza-3 virus, and multiple serovars of Leptospira interrogans. Mycobacterium bovis, Brucella abortus, chronic wasting disease (CWD), and hemoparasites such as Anaplasma spp. were not detected. Results from elk obtained through these surveys were consistent with exposure to disease agents endemic in livestock and wildlife in Arkansas and Kentucky.