Aim Identification of taxonomically cryptic species is essential for the effective conservation of biodiversity. Freshwater-limited organisms tend to be genetically isolated by drainage boundaries, and thus may be expected to show substantial cryptic phylogenetic and taxonomic diversity. By comparison, populations of diadromous taxa, that migrate between freshwater and marine environments, are expected to show less genetic differentiation. Here we test for cryptic diversity in Australasian populations (both diadromous and non-diadromous) of two widespread Southern Hemisphere fish species. Location Throughout both their Australian ranges (including Lord Howe Island) and localities in New Zealand (including the Subantarctic Islands and Chatham Island). Taxon Galaxias brevipinnis and Galaxias maculatus. Methods mtDNA and nuclear markers were used to assess the presence of cryptic species and to determine if differences in species ecology could influence the degree of cryptic diversity. Results Both mtDNA and nuclear markers reveal putative cryptic species within these taxa. The substantial diversity detected within G. brevipinnis may be explained by its strong climbing ability which allows it to form isolated inland populations. In island populations, G. brevipinnis similarly show deeper genetic divergence than those of G. maculatus, which may be explained by the greater abundance of G. maculatus larvae in the sea allowing more ongoing dispersal. Main conclusions Our study highlights that even widespread, ‘high-dispersal’ species can harbour substantial cryptic diversity and therefore warrant increased taxonomic and conservation attention.

The Southern Ocean surrounding Antarctica harbours some of the most pristine marine environments remaining, but is increasingly vulnerable to anthropogenic pressures, climate change, and invasion by non-native species. Monitoring biotic responses to cumulative impacts requires spatiotemporal baselines and ongoing monitoring - traditionally, this has been obtained by continuous plankton recorder (CPR) surveys. Here, we conduct a 3000 nautical mile environmental DNA (eDNA) transect from Hobart (Australia) to Davis Station (Antarctica). We evaluate eDNA sampling strategies for long-term open ocean biomonitoring by comparing two water volume and filter pore size combinations: large (12 L with 20 μm) and small (2 L with 0.45 μm). Employing a broad COI metabarcoding assay, we found the large sample/pore combination was better suited to open-ocean monitoring, detecting more target DNA and rare or low abundance species. Comparisons with four simultaneously conducted CPR transects revealed that eDNA detections were more diverse than CPR, with 7 (4 unique) and 4 (1 unique) phyla detections respectively. While both methods effectively delineated biodiversity patterns across the Southern Ocean, eDNA enables surveys in the presence of sea-ice where CPR cannot be conducted. Accordingly, 16 species of concern were detected along the transect using eDNA, notably in the Antarctic region (south of 60°S). These were largely attributed to hull biofouling, a recognized pathway for marine introductions into Antarctica. In a warming Southern Ocean, continued biomonitoring is vital for conserving Antarctic ecosystems. We advocate for the long-term implementation of eDNA metabarcoding alongside CPR surveys to facilitate ecosystem-based management of these vulnerable environments.