Gammaherpesviruses (GHVs) have been identified in many species and are often associated with disease. Recently, we characterized three novel felid GHVs in domestic cats, bobcats, and pumas. In this study we sought to determine whether free ranging ocelots (Leopardus pardalis) and bobcats (Lynx rufus) are infected with additional GHVs. We screened DNA samples from ocelots on Barro Colorado Island, Panama and bobcats in western Colorado using a degenerate nested PCR that targets the GHV glycoprotein B gene. We identified a novel GHV glycoprotein B sequence in two ocelots and a second novel sequence in a bobcat which is distinct from the previously characterized bobcat GHV (Lynx rufus GHV1). Utilizing additional degenerate and virus-specific PCRs, we extended these sequences to include 3.4 kb of the GHV glycoprotein B and DNA polymerase genes. These sequences indicate the presence of the first GHV detected in ocelots and the second GHV detected in bobcats. These viruses were provisionally named Leopardus pardalis gammaherpesvirus 1 (LpaGHV1) and Lynx rufus gammaherpesvirus 2 (LruGHV2), respectively. Phylogenetic analysis indicates that these viruses are most closely related to recently identified GHVs of the Percavirus genus found in domestic cats (Felis catus GHV1) and bobcats (Lynx rufus GHV1), suggesting that a cluster of multiple felid GHVs exists within the Percavirus genus
