• First record of the invasion and establishment of the invasive arbovirus vector Ae. vittatus in the Americas confirmed by integrated morphological and molecular approaches. • Median-joining network analysis of COI sequences supports multiple introductions into the Caribbean from the Indian subcontinent. • Heightened biosurveillance efforts to expose Ae. vittatus populations elsewhere in the Caribbean, southern USA, and in Central and South America is recommended. Routine biosurveillance efforts at the Naval Station Guantanamo Bay, Cuba, on 18 June 2019, detected two unusual mosquitos in a CO 2 -baited CDC light trap. Morphological and molecular analysis confirmed the presence of Aedes (Fredwardsius) vittatus (Bigot, 1861) – the first record of the Old World dengue, chikungunya, Zika and yellow fever virus vector into the Americas – and provides evidence for its establishment in Cuba. Newly submitted GenBank sequences from Dominican Republic further evidence its establishment in the Caribbean, and a median-joining network analysis using mitochondrial COI gene sequences clearly supports multiple introductions of Ae. vittatus into the Caribbean from the Indian subcontinent. It was determined that many Ae. vittatus COI barcode sequences in GenBank are currently misidentified as Aedes (Fredwardsius) cogilli Edwards, 1922. Image, graphical abstract [ABSTRACT FROM AUTHOR]