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Lipids are biomolecules present in all living organisms that, apart from their physiological functions, can be involved in different pathologies. One of these pathologies is fat embolism, which has been described histologically in the lung of cetaceans in association with ship strikes and with gas and fat embolic syndrome. To assess pathological lung lipid composition, previous knowledge of healthy lung tissue lipid composition is essential; however, these studies are extremely scarce in cetaceans. In the present study we aimed first, to characterize the lipids ordinarily present in the lung tissue of seven cetacean species; and second, to better understand the etiopathogenesis of fat embolism by comparing the lipid composition of lungs positive for fat emboli, and those negative for emboli in Physeter macrocephalus and Ziphius cavirostris (two species in which fat emboli have been described). Results showed that lipid content and lipid classes did not differ among species or diving profiles. In contrast, fatty acid composition was significantly different between species, with C16:0 and C18:1ω9 explaining most of the differences. This baseline knowledge of healthy lung tissue lipid composition will be extremely useful in future studies assessing lung pathologies involving lipids. Concerning fat embolism, non-significant differences could be established between lipid content, lipid classes, and fatty acid composition. However, an unidentified peak was only found in the chromatogram for the two struck whales and merits further investigation.

Diving air-breathing vertebrates have long been considered protected against decompression sickness (DCS) through anatomical, physiological, and behavioural adaptations. However, an acute systemic gas and fat embolic syndrome similar to DCS in human divers was described in beaked whales that stranded in temporal and spatial association with military exercises involving high-powered sonar. More recently, DCS has been diagnosed in bycaught sea turtles. Both cases were linked to human activities. Two Risso’s dolphin (Grampus griseus) out of 493 necropsied cetaceans stranded in the Canary Islands in a 16-year period (2000–2015), had a severe acute decompression sickness supported by pathological findings and gas analysis. Deadly systemic, inflammatory, infectious, or neoplastic diseases, ship collision, military sonar, fisheries interaction or other type of lethal inducing associated trauma were ruled out. Struggling with a squid during hunting is discussed as the most likely cause of DCS.

Strikes between vessels and cetaceans have significantly increased worldwide in the last decades. The Canary Islands archipelago is a geographical area with an important overlap of high cetacean diversity and maritime traffic, including high-speed ferries. Sperm whales (Physeter macrocephalus), currently listed as a vulnerable species, are severely impacted by ship strikes. Nearly 60% of sperm whales’ deaths are due to ship strikes in the Canary Islands. In such cases, subcutaneous, muscular and visceral extensive hemorrhages and hematomas, indicate unequivocal antemortem trauma. However, when carcasses are highly autolyzed, it is challenging to distinguish whether the trauma occurred ante- or post-mortem. The presence of fat emboli within the lung microvasculature is used to determine a severe “in vivo” trauma in other species. We hypothesized fat emboli detection could be a feasible, reliable and accurate forensic tool to determine ante-mortem ship strikes in stranded sperm whales, even in decomposed carcasses. In this study, we evaluated the presence of fat emboli by using an osmium tetroxide (OsO4)-based histochemical technique in lung tissue of 24 sperm whales, 16 of them with evidence of ship strike, stranded and necropsied in the Canaries between 2000 and 2017. About 70% of them presented an advanced autolysis. Histological examination revealed the presence of OsO4-positive fat emboli in 13 out of the 16 sperm whales with signs of ship strike, and two out of eight of the “control” group, with varying degrees of abundance and distribution. A classification and regression tree was developed to assess the cut off of fat emboli area determining the high or low probability for diagnosing ship-strikes, with a sensitivity of 89% and a specificity of 100%. The results demonstrated: (1) the usefulness of fat detection as a diagnostic tool for “in vivo” trauma, even in decomposed tissues kept in formaldehyde for long periods of time; and (2) that, during this 18-year period, at least, 81% of the sperm whales with signs of ship strike were alive at the moment of the strike and died subsequently. This information is highly valuable in order to implement proper mitigation measures in this area.

Rationale: Isotope ratios from skin samples have been widely used to study cetacean trophic ecology. Usually, isotopic skin uniformity has been assumed, despite the heterogeneity of this tissue. This study aims to investigate (1) regional isotopic variation within the skin in cetaceans, and (2) isotopic variation among internal tissues.Methods: Stable carbon (delta C-13 values) and nitrogen (delta N-15 values) isotope ratios were measured in 11 skin positions in 10 common dolphins (Delphinus delphis) and 9 striped dolphins (Stenella coeruleoalba). In addition, the isotope ratios in the muscle, liver and kidney of both species were determined and compared with those from the skin and from all tissues combined. The signatures were determined by means of elemental analyser/isotope ratio mass spectrometry (EA/IRMS).Results: In both species, no differences between isotope ratios of the skin positions were found. Moreover, the isotope ratios of skin were similar to those of muscle. In contrast, liver and kidney showed higher isotope ratios than muscle and skin.Conclusions: Isotopic homogeneity within the skin suggests that the isotope ratios of a sample from a specific skin position can be considered representative of the ratios from the entire skin tissue in dolphins. This conclusion validates the results of previous stable isotope analyses in dolphins that used skin samples as representative of the whole skin tissue. Isotopic similarities or dissimilarities among tissues should be considered when analysing different tissues and comparing results from the same or different species.