The striped eel catfish, Plotosus lineatus was first described by Thunberg in 1787 from the Indo-Pacific region in the East Indian Ocean. Since then, the species has been recorded in various marine and brackish habitats in Japan, southern Korea, the Ogasawara Islands, Australia, Lord Howe Island, Palau and Yap in Micronesia, East Africa to Samoa, Madagascar, Red Sea and the Persian Gulf. Occurrences of this species have also been registered in the Mediterranean Sea, a non-native location, indicating a possible biological invasion event. Despite its long history, the taxonomic status of the P. lineatus species complex remains puzzling and uncertain. Here, we analysed all the available mitochondrial cytochrome c oxidase subunit 1 sequences (NCBI and BOLD) from specimens covering the current known distribution range, in an attempt to clarify the evolutionary relationships of different lineages within this species. We deduce nine monophyletic Lineages - I–IX, of P. lineatus with Kimura-2P distances ranging from 2% to 16%, with a mean intraspecific distance of 6%. Strikingly, Lineage V is composed uniquely of Mediterranean-captured individuals, with an unknown evolutionary origin. These findings strongly suggest the need for a careful species taxonomic reassessment. Some Lineages are composed of individuals from specific geographic locations (e.g., Australia and Indonesia), while others include specimens from broader geographic areas (e.g., almost all Indo-Pacific coastline). Additionally, several deposited sequences are most likely the result of morphology-based misidentifications. Due to the biological invasive potential, as well as the use of the species as a valuable physiology model, the P. lineatus species complex requires further attention. Overall our study offers a clear framework for future comparative studies of striped eel catfish individuals captured from different ecosystems.