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DNA barcoding – the sequencing of a c. 650 base pair region of the mitochondrial cytochrome c oxidase I gene – strongly suggests that barramundi (Lates calcarifer) from Australia and from Myanmar are different species (Kimura 2 parameter distance of c. 9·5%). Cytochrome b sequence data support this conclusion (distance c. 11·3%). Further examination, both genetic and morphological, of L. calcarifer throughout its range is recommended.