A survey of more than 200 trees has documented the widespread occurrence of Xylella fastidiosa in Florida oak populations. The pathogen was detected readily via enzyme-linked immunosorbent assay in oaks exhibiting decline or leaf scorch symptoms and was infrequently detected in asymptomatic trees. It was also associated with reduced growth in Quercus laevis as measured by current-year shoot length. The occurrence of X. fastidiosa in Q. laevis and the evidence for its occurrence in Q. incana represent first reports for these oak hosts. The role of X. fastidiosa in oak decline scenarios deserves further attention.