Multisystemic enterococcosis, caused by the bacterium Enterococcus lacertideformus, is a potentially fatal infection affecting multiple lizard species. Initially described on Christmas Island in 2014, E. lacertideformus has since infected Anolis sagrei (Brown Anole) populations in Pinellas County, FL. In 2023, we found a Brown Anole exhibiting clinical signs consistent with multisystemic enterococcosis. Molecular and histopathological analyses confirmed the presence of E. lacertideformus infection. Thus, we report the first documented case of multisystemic enterococcosis in Hillsborough County, FL, a previously undocumented locale for this pathogen. This range expansion underscores the potential risk E. lacertideformus poses to Florida's lizard populations, highlighting the necessity for increased surveillance and further research to better understand the transmission dynamics and ecological impacts of this emerging pathogen.