We report on the discovery of a wild, reproducing population of Channa aurolineata (Pisces: Channidae) in west-central Florida (USA), and first documented occurrence of snakeheads in the Gulf Coast region. Channa aurolineata is a large, predatory fish of the bullseye snakehead “Marulius group” species complex from Asia. Adult and juvenile specimens were captured in June 2020 in a 1.8-hectare pond that connects during high water to a small stream within the Manatee River-Tampa Bay Basin. The pond site is 250-km from the only other wild C. aurolineata population in the USA (present in southeast Florida since ca. 2000) and is considered a separate introduction and not the result of natural dispersal. Morphological and molecular comparisons revealed high overlap between the two Florida populations, evidence humans may have transported fish between sites. To verify identification, we compared Florida samples to C. aurolineata from Thailand and found mtDNA-COI barcode sequences to be identical or to differ by only a single base pair. Life body coloration of Florida samples matched their Asian counterparts, but Florida specimens averaged fewer dorsal fin rays (53.6 vs. 56.0), anal fin rays (34.2 vs 36.1), lateral line scales (65.3 vs. 67.4), and vertebrae (62.1 vs. 64.3), differences implying possible founder effect or sampling bias. Existence of this invasive predator is a concern because of the risk of spread and negative ecological effects, including an observation of terrestrial hunting behavior. In 2020–2021, several hundred C. aurolineata were removed from the pond by nets and electrofishing, and surveys suggested the population had not spread to nearby waters. In May 2021 the pond was treated with rotenone and 48 more specimens were recovered. No additional snakeheads have been sighted since the piscicide operation, although verification of eradication will require monitoring of the watershed.