Species with biological traits favourable to long-distance marine larval dispersal might show a phylogeographic structuring over broad regions, even when they are genetically connected within smaller scales. Here, we evaluated this hypothesis by using the widespread amphidromous shrimp Atya scabra, predicting a genetic discontinuity across biogeographical barriers throughout the Western Gulf of Mexico (WGM), Caribbean Sea (CS), south-western Atlantic (SWA) and eastern Atlantic (EA). Using cytochrome oxidase subunit 1 (COI) and 16S ribosomal unit (16S) gene fragments, we did a phylogeographic assessment and genetic characterisation with Bayesian clustering, AMOVA, haplotype networks and demographic analyses. As predicted, three discrete genetic groups, corresponding to the regions WGM, CS and EA, were uncovered by COI, as well an unpredicted SWA+CS group. The 16S fragment detected a low genetic variation, probably owing to a recent lineage differentiation, which was estimated by the COI molecular clock. We evaluated the role of the biological traits of A. scabra, as well as the consequences of Panama Isthmus closure and Pleistocene glaciation cycles in the lineage isolation of WGM and EA, as well as the genetic connectivity shown within regions and between CS and SWA. Our results highlighted that amphidromous species genetically connected over large scales should be genetically characterised in their wide distribution to provide more comprehensive systematics and to assist decision-making in biological conservation.