(Table 1) Prey species identified from the scats of Antarctic fur seals (Arctocephalus gazella) on Bouvet Island

Intraspecific differences in the diets of many species of pinnipeds are to be expected in view of the great differences in morphology, life history and foraging behaviour between the sexes of many species. We examined the diet of the Antarctic fur seal population at Bouvetøya, Southern Ocean, to assess intersexual differences. This was made possible by the analysis of prey remains extracted from scats and regurgitations collected in areas used primarily by one or the other sex. The results indicate that both males and females feed primarily on Antarctic krill Euphausia superba with several species of fish and squid being taken, likely opportunistically given their prevalence. Significant differences were identified in the frequency of occurrence of otoliths in scats and the percentage numerical abundance of the major fish prey species in the diet. Adult males ate a smaller quantity of fish overall, but ate significantly more of the larger fish species. The greater diving capabilities of males and the fact that they are not limited in the extent of their foraging area by having to return regularly to feed dependant offspring may play a role in the differences found between the diets of males and females. Additionally, females might be more selective, favouring myctophids because they are richer in energy than krill. The absence of major differences in the diet between the sexes at this location is likely due to the high overall abundance of prey at Bouvetøya.

DNA alignment and Bayesian trees of partial sequences of COI, COIII and the haemocyanin functional unit f-g of 28 octopod species

Background: Octopods have successfully colonised the world's oceans from the tropics to the poles. Yet, successful persistence in these habitats has required adaptations of their advanced physiological apparatus to compensate impaired oxygen supply. Their oxygen transporter haemocyanin plays a major role in cold tolerance and accordingly has undergone functional modifications to sustain oxygen release at sub-zero temperatures. However, it remains unknown how molecular properties evolved to explain the observed functional adaptations. We thus aimed to assess whether natural selection affected molecular and structural properties of haemocyanin that explains temperature adaptation in octopods. Results: Analysis of 239 partial sequences of the haemocyanin functional units (FU) f and g of 28 octopod species of polar, temperate, subtropical and tropical origin revealed natural selection was acting primarily on charge properties of surface residues. Polar octopods contained haemocyanins with higher net surface charge due to decreased glutamic acid content and higher numbers of basic amino acids. Within the analysed partial sequences, positive selection was present at site 2545, positioned between the active copper binding centre and the FU g surface. At this site, methionine was the dominant amino acid in polar octopods and leucine was dominant in tropical octopods. Sites directly involved in oxygen binding or quaternary interactions were highly conserved within the analysed sequence. Conclusions: This study has provided the first insight into molecular and structural mechanisms that have enabled octopods to sustain oxygen supply from polar to tropical conditions. Our findings imply modulation of oxygen binding via charge-charge interaction at the protein surface, which stabilize quaternary interactions among functional units to reduce detrimental effects of high pH on venous oxygen release. Of the observed partial haemocyanin sequence, residue 2545 formed a close link between the FU g surface and the active centre, suggesting a role as allosteric binding site. The prevalence of methionine at this site in polar octopods, implies regulation of oxygen affinity via increased sensitivity to allosteric metal binding. High sequence conservation of sites directly involved in oxygen binding indicates that functional modifications of octopod haemocyanin rather occur via more subtle mechanisms, as observed in this study.