Aim We use an integrative biogeographical approach to further understand the evolution of an important Southern Ocean marine benthic element, the limpet genus Nacella (Mollusca: Patellogastropoda). Location Southern Ocean.Methods We used multi-locus time-calibrated phylogeny of Nacella at the scale of the whole Southern Ocean to elucidate the underlying processes involved in the origin and diversification of the genus.Results Divergence-time estimates suggest that soon after its origin during the mid-Miocene (c. 12.5 Ma), Nacella separated into two main lineages currently distributed in (1) South America and (2) Antarctica and the sub-Antarctic islands. We identified two pulses of diversification, during the late Miocene (8 to 5.5 Ma) and the Pleistocene (< 1 Ma).Main conclusions Major periods of climatic and oceanographical change strongly affected the biogeography of Nacella and demonstrate both the longand short-term influence of the Antarctic Circumpolar Current across the Southern Ocean. Our analyses support the validity of all currently recognized Nacella species and reveal a new South-American lineage. This work constitutes the most detailed molecular-based study of an ecologically important, nearshore invertebrate Southern Ocean group and in so doing contributes to the improved understanding of the underlying patterns and processes in the origin and diversification of marine benthic fauna across this globally important region.

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Human driven changes such as increases in oceanic CO 2 , global warming, petroleum hydrocarbons and heavy metals may negatively affect the ability of marine calcifiers to build their skeletons/shells, especially in polar regions. We examine spatio-temporal variability of skeletal Mg-calcite in shallow water Antarctic marine invertebrates using bryozoan and spirorbids as models in a recruitment experiment of settlement tiles in East Antarctica. Mineralogies were determined for 754 specimens belonging to six bryozoan species (four cheilostome and two cyclostome species) and two spirorbid species from around Casey Station. Intra- and interspecific variability in wt% MgCO 3 in calcite among most species was the largest source of variation overall. Therefore, the skeletal Mg-calcite in these taxa seem to be mainly biologically controlled. However, significant spatial variability was also found in wt% MgCO 3 in calcite, possibly reflecting local environment variation from sources such as freshwater input and contaminated sediments. Species with high-Mg calcite skeletons (e.g. Beania erecta ) could be particularly sensitive to multiple stressors under predictions for near-future global ocean chemistry changes such as increasing temperature, ocean acidification and pollution.

Free-ocean CO2 enrichment (FOCE) experiments have been deployed in marine ecosystems to manipulate carbonate system conditions to those predicted in future oceans. We investigated whether the pH/carbonate chemistry of extremely cold polar waters can be manipulated in an ecologically relevant way, to represent conditions under future atmospheric CO2 levels, in an in-situ FOCE experiment in Antarctica. We examined spatial and temporal variation in local ambient carbonate chemistry at hourly intervals at two sites between December and February and compared these with experimental conditions. We successfully maintained a mean pH offset in acidified benthic chambers of −0.38 (±0.07) from ambient for approximately 8 weeks. Local diel and seasonal fluctuations in ambient pH were duplicated in the FOCE system. Large temporal variability in acidified chambers resulted from system stoppages. The mean pH, Ωarag and fCO2 values in the acidified chambers were 7.688 ± 0.079, 0.62 ± 0.13 and 912 ± 150 µatm, respectively. Variation in ambient pH appeared to be mainly driven by salinity and biological production and ranged from 8.019 to 8.192 with significant spatio-temporal variation. This experiment demonstrates the utility of FOCE systems to create conditions expected in future oceans that represent ecologically relevant variation, even under polar conditions.

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