The whale shark ( Smith 1828) is the largest of the filter-feeding sharks and inhabits tropical and sub-tropical oceans worldwide. Evidence from anecdotal observations of feeding events and stomach content analysis have identified a wide range of planktonic and nektonic organisms including copepods, krill and small fish as whale shark prey. However, recent studies based on biochemical analysis (e.g. signature fatty acids (FA)) indicated that whale sharks in the western Indian Ocean had a wider foraging range than previous studies suggested, with important contributions from meso- and bathypelagic sources. However, it remains unknown if these results characterize the diet of whale sharks over the wider Indian Ocean. Here, we investigate the feeding ecology of whale sharks in the eastern Indian Ocean at Ningaloo Reef, Western Australia, by identifying differences in whale shark diet according to time of collection, sex and size-class and by examining likely food web linkages. To examine whale shark feeding ecology we used signature FA analysis of both whale shark subdermal tissue and an extensive set of potential prey collected at Ningaloo Reef (Western Australia) in 2013 and 2014. Compared to other methods such as stomach content analysis, signature FA analysis provides both longer-term (up to months) dietary information and an assessment of spatial and temporal changes in the diet of predators. This is possible because some FA in animal tissues (e.g. long-chain (≥C20) polyunsaturated FA, LC-PUFA) can be used as biomarkers as they pass relatively unchanged from the low trophic levels where they are biosynthesized up the food chain. Whale shark sub-dermal tissue was low in lipid content (4 mg g–1 dry mass) which was dominated by phospholipids (72% of total lipid) with an energy density of 18.66 kJ g–1 dry mass. A significant intraspecific variability in whale shark FA profiles was observed resulting in four distinct groups of sharks in 2013 and five in 2014. As this variability was not related to sex or size-class, we suggest that it may be attributed to differences in the feeding habitats and thus different prey consumed by these groups of whale sharks. Variation in dietary patterns was also observed between years likely due to changes in the primary and secondary producers. Overall, examination of food web interactions showed that fatty acid profiles of whale sharks and their presumed collected prey were significantly different, suggesting that sharks fed over a wider range of habitats, including deeper waters, than we were able to access. A significant component of whale shark diet may originate from benthic and deeper water habitats. High intraspecific variation in diet indicates that whale sharks are likely to forage over a range of distances and depths due to the challenge of inhabiting the patchy prey habitats of tropical and open ocean waters. Future studies should seek to combine signature FA analysis with other techniques such as stable isotopes, genetic and longterm tagging data to help better elucidate the feeding ecology of this iconic species.


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