We evaluated spatial and temporal variability in parasite communities from the commercially important tropical marine fish the black-spotted croaker (Protonibea diacanthus) (Teleostei: Sciaenidae) to examine its population structure off the coast of the Northern Territory, Australia. Differences in parasite assemblage between four locations, across three seasons of the year, were used to evaluate the degree of connectivity of the sciaenid across coastal study areas. Analysis of parasite prevalence and mean intensity in these fish suggested the four sampling sites are distinct host populations. Across time, parasite assemblages at the four sites were distinct during the mid-dry (April–August) and build-up (September–November) seasons. During the wet season (December–March) there was substantial overlap in the parasite assemblages at three of the four sites indicating that fish population mixing may be occurring. Parasite assemblages at one nearshore site remained distinct across spatial and temporal scales. Our findings support the utility of parasitic organisms for elucidating the population structure of host species and reiterate the need to account for both spatial and temporal variability when performing stock discrimination analyses.