Herpesviruses are important pathogens in zoologic chelonian collections and have been associated with fatal disease in turtles of the family Emydidae. In this report, three western pond turtles (Actinemys marmorata), living in a mixed-species freshwater turtle exhibit, presented with varying degrees of lethargy, pallor, generalized edema, and cloacal hemorrhage before death within a 2-wk period. Postmortem findings included necrohemorrhagic enterocolitis, necrotizing splenitis, hepatic necrosis, esophagitis, thymic necrosis, and pneumonia with epithelial necrosis and degeneration of the trachea and kidneys. Intraepithelial, intranuclear, amphophilic to eosinophilic, Cowdry type A viral inclusion bodies were identified in the intestinal tract, liver, spleen, kidney, trachea, lung, and thymus. PCR amplification and sequencing of liver tissue produced amplicons that were 100% homologous with emydid herpesvirus 1 (EmyHV-1). Molecular screening of cohoused emydid turtles revealed that a red-eared slider (Trachemys scripta elegans) and a western pond turtle, both asymptomatic, were PCR-positive for EmyHV-1 on combined oral–cloacal swabs. This report describes, for the first time, EmyHV-1–associated disease in western pond turtles and molecularly identifies EmyHV-1 in an asymptomatic red-eared slider.