Metabolomics approach for predicting stomach and colon contents in dead Arctocephalus pusillus pusillus, Arctocephalus tropicalis, Lobodon carcinophaga and Ommatophoca rossii from sub-Antarctic region.

The dietary habits of seals play a pivotal role in shaping management and administration policies, especially in regions with potential interactions with fisheries. Previous studies have utilized various methods, including traditional approaches, to predict seal diets by retrieving indigestible prey parts, such as calcified structures, from intestines, feces, and stomach contents. Additionally, methods evaluating nitrogen and stable isotopes of carbon have been employed. The metabolomics approach, capable of quantifying small-scale molecules in biofluids, holds promise for specifying dietary exposures and estimating disease risk. This study aimed to assess the diet composition of five seal species-Arctocephalus pusillus pusillus, Lobodon carcinophaga, Ommatophoca rossii, and Arctocephalus tropicalis 1 and 2-by analyzing stomach and colon contents collected from stranded dead seals at various locations. Metabolite concentrations in the seal stomach and colon contents were determined using Nuclear Magnetic Resonance Spectroscopy. Among the colon and stomach contents, 29 known and 8 unknown metabolites were identified. Four metabolites (alanine, fumarate, lactate, and proline) from stomach contents and one metabolite (alanine) from colon contents showed no significant differences between seal species (p>0.05). This suggests that traces of these metabolites in the stomach and colon contents may be produced by the seals' gut microbiome or derived from other animals, possibly indicating reliance on fish caught at sea. Despite this insight, the cause of death for stranded seals remains unclear. The study highlights the need for specific and reliable biomarkers to precisely indicate dietary exposures across seal populations. Additionally, there is a call for the development of relevant metabolite and disease interaction networks to explore disease-related metabolites in seals. Ultimately, the metabolomic method employed in this study reveals potential metabolites in the stomach and colon contents of these seal species.

Individual variation in life-history timing: synchronous presence, asynchronous events and phenological compensation in a wild mammal.

Many animals and plants have species-typical annual cycles, but individuals vary in their timing of life-history events. Individual variation in fur replacement (moult) timing is poorly understood in mammals due to the challenge of repeated observations and longitudinal sampling. We examined factors that influence variation in moult duration and timing among elephant seals (Mirounga angustirostris). We quantified the onset and progression of fur loss in 1178 individuals. We found that an exceptionally rapid visible moult (7 days, the shortest of any mammals or birds), and a wide range of moult start dates (spanning 6-10× the event duration) facilitated high asynchrony across individuals (only 20% of individuals in the population moulting at the same time). Some of the variation was due to reproductive state, as reproductively mature females that skipped a breeding season moulted a week earlier than reproductive females. Moreover, individual variation in timing and duration within age-sex categories far outweighed (76-80%) variation among age-sex categories. Individuals arriving at the end of the moult season spent 50% less time on the beach, which allowed them to catch up in their annual cycles and reduce population-level variance during breeding. These findings underscore the importance of individual variation in annual cycles.

Foraging habits of Northwest Atlantic hooded seals over the past 30 years: Future habitat suitability under global warming.

The Arctic is a global warming 'hot-spot' that is experiencing rapid increases in air and ocean temperatures and concomitant decreases in sea ice cover. These environmental changes are having major consequences on Arctic ecosystems. All Arctic endemic marine mammals are highly dependent on ice-associated ecosystems for at least part of their life cycle and thus are sensitive to the changes occurring in their habitats. Understanding the biological consequences of changes in these environments is essential for ecosystem management and conservation. However, our ability to study climate change impacts on Arctic marine mammals is generally limited by the lack of sufficiently long data time series. In this study, we took advantage of a unique dataset on hooded seal (Cystophora cristata) movements (and serum samples) that spans more than 30 years in the Northwest Atlantic to (i) investigate foraging (distribution and habitat use) and dietary (trophic level of prey and location) habits over the last three decades and (ii) predict future locations of suitable habitat given a projected global warming scenario. We found that, despite a change in isotopic signatures that might suggest prey changes over the 30-year period, hooded seals from the Northwest Atlantic appeared to target similar oceanographic characteristics throughout the study period. However, over decades, they have moved northward to find food. Somewhat surprisingly, foraging habits differed between seals breeding in the Gulf of St Lawrence vs those breeding at the "Front" (off Newfoundland). Seals from the Gulf favoured colder waters while Front seals favoured warmer waters. We predict that foraging habitats for hooded seals will continue to shift northwards and that Front seals are likely to have the greatest resilience. This study shows how hooded seals are responding to rapid environmental change and provides an indication of future trends for the species-information essential for effective ecosystem management and conservation.

From land to ocean: One month for southern elephant seal pups to acquire aquatic skills prior to their first departure to sea.

Weaned southern elephant seals (SES) quickly transition from terrestrial to aquatic life after a 5- to 6-week post-weaning period. At sea, juveniles and adult elephant seals present extreme, continuous diving behaviour. Previous studies have highlighted the importance of the post-weaning period for weanlings to prepare for the physiological challenges of their future sea life. However, very little is known about how their body condition during this period may influence the development of their behaviour and brain activities. To characterise changes in the behavioural and brain activity of weanlings prior to ocean departure, we implemented a multi-logger approach combining measurements of movements (related to behaviour), pressure (related to diving), and brain electrical activity. As pups age, the amount of time allocated to resting decreases in favour of physical activity. Most resting (9.6 ± 1.2 h/day) takes place during daytime, with periods of slow-wave sleep representing 4.9 ± 0.9 h/day during the first 2 weeks. Furthermore, an increasing proportion of physical activity transitions from land to shore. Additionally, pups in poorer condition (lean group) are more active earlier than those in better condition (corpulent group). Finally, at weaning, clear circadian activity with two peaks at dawn and dusk is observed, and this pattern remains unchanged during the 4 weeks on land. This circadian pattern matches the one observed in adults at sea, with more prey catches at dawn and dusk, raising the question of whether it is endogenous or triggered by the mother during lactation.

Circadian coupling of mitochondria in a deep-diving mammal.

Regulation of mitochondrial oxidative phosphorylation is essential to match energy supply to changing cellular energy demands, and to cope with periods of hypoxia. Recent work implicates the circadian molecular clock in control of mitochondrial function and hypoxia sensing. Because diving mammals experience intermittent episodes of severe hypoxia, with diel patterning in dive depth and duration, it is interesting to consider circadian-mitochondrial interaction in this group. Here, we demonstrate that the hooded seal (Cystophora cristata), a deep-diving Arctic pinniped, shows strong daily patterning of diving behaviour in the wild. Cultures of hooded seal skin fibroblasts exhibit robust circadian oscillation of the core clock genes per2 and arntl. In liver tissue collected from captive hooded seals, expression of arntl was some 4-fold higher in the middle of the night than in the middle of the day. To explore the clock-mitochondria relationship, we measured the mitochondrial oxygen consumption in synchronized hooded seal skin fibroblasts and found a circadian variation in mitochondrial activity, with higher coupling efficiency of complex I coinciding with the trough of arntl expression. These results open the way for further studies of circadian-hypoxia interactions in pinnipeds during diving.

Diverse papillomaviruses identified from Antarctic fur seals, leopard seals and Weddell seals from the Antarctic.

Papillomaviruses (family Papillomaviridae) are non-enveloped, circular, double-stranded DNA viruses known to infect squamous and mucosal epithelial cells. In the family Papillomaviridae there are 53 genera and 133 viral species whose members infect a variety of mammalian, avian, reptilian, and fish species. Within the Antarctic context, papillomaviruses (PVs) have been identified in Adélie penguins (Pygoscelis adeliae, 2 PVs), Weddell seals (Leptonychotes weddellii, 7 PVs), and emerald notothen (Trematomus bernacchii, 1 PV) in McMurdo Sound and Ross Island in eastern Antarctica. Here we identified 13 diverse PVs from buccal swabs of Antarctic fur seals (Arctocephalus gazella, 2 PVs) and leopard seal (Hydrurga leptonyx, 3 PVs) in western Antarctica (Antarctic Peninsula), and vaginal and nasal swabs of Weddell seals (8 PVs) in McMurdo Sound. These PV genomes group into four genera representing 11 new papillomavirus types, of which five are from two Antarctic fur seals and a leopard seal and six from Weddell seals.

Genomics of post-bottleneck recovery in the northern elephant seal.

Populations and species are threatened by human pressure, but their fate is variable. Some depleted populations, such as that of the northern elephant seal (Mirounga angustirostris), recover rapidly even when the surviving population was small. The northern elephant seal was hunted extensively and taken by collectors between the early 1800s and 1892, suffering an extreme population bottleneck as a consequence. Recovery was rapid and now there are over 200,000 individuals. We sequenced 260 modern and 8 historical northern elephant seal nuclear genomes to assess the impact of the population bottleneck on individual northern elephant seals and to better understand their recovery. Here we show that inbreeding, an increase in the frequency of alleles compromised by lost function, and allele frequency distortion, reduced the fitness of breeding males and females, as well as the performance of adult females on foraging migrations. We provide a detailed investigation of the impact of a severe bottleneck on fitness at the genomic level and report on the role of specific gene systems.

Contrasting trophic transfer patterns of cadmium and mercury in the Arctic marine food web of east Hudson Bay, Canada.

We investigated trophic transfer of cadmium (Cd) through an Arctic marine food web in Hudson Bay and compared it with mercury (Hg), a metal known to strongly biomagnify. We evaluated blue mussel, sea urchin, common eider, sculpin, Arctic cod, and ringed seal for the influence of dietary and biological variables on variation in Cd and Hg concentrations. Age and size influenced metal concentrations among individuals within a vertebrate species. Consumer carbon and sulfur isotope values were correlated with their Cd and Hg concentrations, indicating habitat-specific feeding influenced metal bioaccumulation. Trophic transfer patterns for Cd depended on the vertebrate tissue, with food web biodilution observed for the muscle but not the liver. Liver Cd concentrations were higher in ringed seal and some common eider relative to prey. In contrast, we observed mercury biomagnification for both tissues. Tissue- and species-specific physiology can explain discrepancies of Cd trophic transfer in this Arctic marine food web.

Anthropogenic debris in three sympatric seal species of the Western Antarctic Peninsula.

Litter pollution is a growing concern, including for Antarctica and the species that inhabit this ecosystem. In this study, we investigated the microplastic contamination in three seal species that inhabit the Western Antarctic Peninsula: crabeater (Lobodon carcinophaga), leopard (Hydrurga leptonyx) and Weddell (Leptonychotes weddellii) seals. Given the worldwide ubiquity of this type of contaminant, including the Southern Ocean, we hypothesized that the three seal species would present anthropogenic debris in their feces. We examined 29 scat samples of crabeater (n = 5), leopard (n = 13) and Weddell (n = 11) seals. The chemical composition of the items found were identified using micro-Raman and micro-FTIR spectroscopies. All the samples of the three species presented anthropic particles (frequency of occurrence - %FO - 100 %). Fibers were the predominant debris, but fragments and filaments were also present. Particles smaller than 5 mm (micro debris) were predominant in all the samples. Leopard seals ingested significantly larger micro-debris in comparison with the other seal species. The dominant color was black followed by blue and white. Micro-Raman and micro-FTIR Spectroscopies revealed the presence of different anthropogenic pigments such as reactive blue 238, Indigo 3600 and copper phthalocyanine (blue and green). Carbon black was also detected in the samples, as well as plastic polymers such as polystyrene, polyester and polyethylene terephthalate (PET), polyamide, polypropylene and polyurethane These results confirm the presence of anthropogenic contamination in Antarctic seals and highlight the need for actions to mitigate the effects and reduce the contribution of debris in the Antarctic ecosystem.

Wavelength-induced shedding frequency modulation of seal whisker inspired cylinders.

The spanwise undulated cylinder geometry inspired by seal whiskers has been shown to alter shedding frequency and reduce fluid forces significantly compared to smooth cylindrical geometry. Prior research has parameterized the whisker-inspired geometry and demonstrated the relevance of geometric variations on force reduction properties. Among the geometric parameters, undulation wavelength was identified as a significant contributor to forcing changes. To analyze the effect of undulation wavelength, a thorough investigation isolating changes in wavelength is performed to expand upon previous research that parameterized whisker-inspired geometry and the relevance of geometric variations on the force reduction properties. A set of five whisker-inspired models of varying wavelength are computationally simulated at a Reynolds number of 250 and compared with an equivalent aspect ratio smooth elliptical cylinder. Above a critical non-dimensional value, the undulation wavelength reduces the amplitude and frequency of vortex shedding accompanied by a reduction in oscillating lift force. Frequency shedding is tied to the creation of wavelength-dependent vortex structures which vary across the whisker span. These vortices produce distinct shedding modes in which the frequency and phase of downstream structures interact to decrease the oscillating lift forces on the whisker model with particular effectiveness around the wavelength values typically found in nature. The culmination of these location-based modes produces a complex and spanwise-dependent lift frequency spectra at those wavelengths exhibiting maximum force reduction. Understanding the mechanisms of unsteady force reduction and the relationship between undulation wavelength and frequency spectra is critical for the application of this geometry to vibration tuning and passive flow control for vortex-induced vibration (VIV) reduction.

The decline of kilkas, sturgeons and seals in the Caspian Sea: The potential of qualitative loop analysis for the cumulative assessment of multiple drivers of stress.

Multiple stressors often act concomitantly on ecosystems but detection of species responses follows the "single species-single driver" strategy, and cumulative impacts are seldom considered. During 1990-2010, multiple perturbations in the Caspian Sea, led to the decline of kilka, sturgeon and Caspian seal populations. Specific causes for their collapse were identified but a cumulative assessment has never been carried out. Using loop analysis, a qualitative modelling technique suitable in poor-data contexts, we show how multiple drivers can be combined to assess their cumulative impact. We confirm that the decline of kilka, sturgeon and Caspian seal populations is compatible with a net effect of the concomitant perturbations. Kilkas collapse was certainly due to the outburst of M. leidyi and overfishing. In addition, the excess nutrient might have conspired to reduce these populations. The interplay between concurrent drivers produces trade-offs between opposite effects and ecosystem management must face this challenge.

Response of Eurasian otters (Lutra lutra) to underwater acoustic harassment device sounds.

Seal scarers (or acoustic harassment devices, AHDs) are designed to deter seals from fishing gear and aquaculture operations, as well as to prevent seals from entering rivers to avoid predation on valuable fish. Our study investigated the potential effects of AHDs on non-target species, specifically the Eurasian otters (Lutra lutra), by testing the reaction of two rehabilitated otters to simulated AHDs sounds at 1 and 14 kHz, with a received sound intensity of 105-145 dB re 1 µPa rms. The 1 kHz sounds were used to investigate alternative frequencies for scaring seals without scaring otters. The otters reacted to both 1 and 14 kHz tonal signals when retrieving fish from a feeding station 0.8 m below the surface. Their diving behaviour and time to extract food progressively increased as sound intensity increased for all tested sound levels. Notably, the sound levels used in our tests were significantly lower (40-80 dB) than the source levels from commercial AHDs. These findings highlight the importance of caution when using AHDs in river and sea habitats inhabited by otters, as AHDs can change their behaviour and potentially result in habitat exclusion.

Foraging behavior and age affect maternal transfer of mercury to northern elephant seal pups.

Deep ocean foraging northern elephant seals (Mirounga angustirostris) consume fish and squid in remote depths of the North Pacific Ocean. Contaminants bioaccumulated from prey are subsequently transferred by adult females to pups during gestation and lactation, linking pups to mercury contamination in mesopelagic food webs (200-1000 m depths). Maternal transfer of mercury to developing seal pups was related to maternal mercury contamination and was strongly correlated with maternal foraging behavior (biotelemetry and isotopes). Mercury concentrations in lanugo (hair grown in utero) were among the highest observed worldwide for young pinnipeds (geometric mean 23.01 µg/g dw, range 8.03-63.09 µg/g dw; n = 373); thus, some pups may be at an elevated risk of sub-lethal adverse health effects. Fetal mercury exposure was affected by maternal foraging geographic location and depth; mercury concentrations were highest in pups of the deepest diving, pelagic females. Moreover, pup lanugo mercury concentrations were strongly repeatable among successive pups of individual females, demonstrating relative consistency in pup mercury exposure based on maternal foraging strategies. Northern elephant seals are biosentinels of a remote deep-sea ecosystem. Our results suggest that mercury within North Pacific mesopelagic food webs may also pose an elevated risk to other mesopelagic-foraging predators and their offspring.

Predation and spatial connectivity interact to shape ecosystem resilience to an ongoing regime shift.

Ecosystem regime shifts can have severe ecological and economic consequences, making it a top priority to understand how to make systems more resilient. Theory predicts that spatial connectivity and the local environment interact to shape resilience, but empirical studies are scarce. Here, we use >7000 fish samplings from the Baltic Sea coast to test this prediction in an ongoing, spatially propagating shift in dominance from predatory fish to an opportunistic mesopredator, with cascading effects throughout the food web. After controlling for the influence of other drivers (including increasing mesopredator densities), we find that predatory fish habitat connectivity increases resilience to the shift, but only when densities of fish-eating top predators (seals, cormorants) are low. Resilience also increases with temperature, likely through boosted predatory fish growth and recruitment. These findings confirm theoretical predictions that spatial connectivity and the local environment can together shape resilience to regime shifts.

Endogenous lipid pneumonia in a ringed seal (Pusa hispida subsp. ochotensis).

An adult female ringed seal died suddenly and was subsequently examined for diagnostic purposes. The animal's lungs demonstrated mild non-collapsibility and multifocal white to yellow patches. Histopathological examination revealed multifocal pulmonary histiocytosis. Alveoli were filled with numerous foamy macrophages cytoplasm and scattered multinucleated giant cells containing cholesterol clefts. The foamy cytoplasm of the macrophages stained with oil red O stain. Further, lipid droplets within the cytoplasm were detected by electron microscopy. To the author's knowledge, this is the first case report describing the histochemical staining and electron microscopic findings associated with endogenous lipid pneumonia in ringed seal.

Optimization design of fiber rubber asphalt gravel sealing layer based on fatigue crack resistance test.

Crack is one of the main diseases of pavement structure. In order to improve the anti-reflective crack ability of pavement, fiber rubber gravel sealing layer is proposed as the stress absorbing layer. In view of the shortcoming that Mcleod design method can not be associated with road performance, a sealing layer optimization design method based on fatigue crack test is proposed. Firstly, the reinforcement effect of fiber on rubber asphalt was studied through force ductility testing. Secondly, the optimum dosage of fiber, asphalt and gravel was optimized through fatigue cracking resistance test. Finally, the cracking resistance of fiber rubber gravel seal was verified through fracture energy test. The results show that fibers can significantly increase the maximum tensile force and strain yield energy of rubber asphalt, and basalt fiber has the best reinforcement effect. The most obvious effect on cracking resistance performance in the sealing layer is the amount of fiber, followed by the amount of asphalt, and finally the amount of gravel. The optimized material combination with the best crack resistance is 120g/m2 fiber, 14kg/m2 gravel and 2.4kg/m2 rubber asphalt, and the fatigue resistance times can reach 19532 times. The fracture energy of the composite pavement treated by the optimized sealing layer is nearly double that of the non-treated pavement structure, and it has a good anti-crack effect.

Survival and adaptation of Streptococcus phocae in host environments.

Marine mammals are sentinel species representing the "health" of our oceans on which we are dependent. There are many threats to marine mammals including infectious diseases that increase with climate change and pollution of the marine environment. Streptococcus phocae has frequently been isolated from diseased or dead marine mammals. However, its pathogenicity and contribution to disease in marine mammals is still unknown. As bacteria including (potential) pathogens has to deal with different host environments during colonization or infection, we investigated the survival of S. phocae in fresh porcine and phocid blood, in seawater and in the presence of macrophages and (epithelial) cells from harbor seals and pigs. Furthermore, we tested adherence on and invasion of different (marine) mammalian cells by S. phocae. Our results showed that S. phocae can survive in seawater for at least 11 and 28 days at 16°C and 4°C, respectively. It is able to grow in blood of harbor and grey seals, but not in porcine blood. Furthermore, S. phocae is adherent and invasive to cells from seals and pigs, while the portion of invasive cells was higher in seal derived cells. Macrophages of harbor seals were more efficient in killing S. phocae than porcine macrophages. Our results indicate that S. phocae has strategies enabling it to adapt to the marine environment and seal hosts.

Attachment performance of the ectoparasitic seal louse Echinophthirius horridus.

Marine mammals host a great variety of parasites, which usually co-evolved in evolutionary arms races. However, little is known about the biology of marine mammal insect parasites, and even less about physical aspects of their life in such a challenging environment. One of 13 insect species that manage to endure long diving periods in the open sea is the seal louse, Echinophthirius horridus, parasitising true seals. Its survival depends on its specialised adaptations for enduring extreme conditions such as hypoxia, temperature changes, hydrostatic pressure, and strong drag forces during host dives. To maintain a grip on the seal fur, the louse's leg morphology is equipped with modified snap hook claws and soft pad-like structures that enhance friction. Through techniques including CLSM, SEM, and histological staining, we have examined the attachment system's detailed structure. Remarkably, the seal louse achieves exceptional attachment forces on seal fur, with safety factors (force per body weight) reaching 4500 in average measurements and up to 18000 in peak values, indicating superior attachment performance compared to other insect attachment systems. These findings underscore the louse's remarkable adaptations for life in a challenging marine environment, shedding light on the relationship between structure and function in extreme ecological niches.

Breaking the fast: first report of dives and ingestion events in molting southern elephant seals.

Southern elephant seals (SES) experience a 'catastrophic molt', a costly event characterized by the renewal of both hair and epidermis that requires high peripheral vascular circulation. Molting animals are therefore constrained by high metabolic heat loss and are thought to fast and remain on land. To examine the ability of individuals to balance the energetic constraints of molting on land we investigate the stomach temperature and movement patterns of molting female SES. We find that 79% of females swam and 61% ingested water or prey items, despite the cost of cold-water exposure while molting. This behavior was related to periods of warm and low wind conditions, and females that dived and ingested more often, lost less body mass. We conclude that the paradigm of fasting during the molt in this species, and the fitness consequences of this behavior should be reconsidered, especially in the context of a changing climate.

Monk seals (Monachus monachus) in the Mediterranean Sea: The threat of organochlorine contaminants and polycyclic aromatic hydrocarbons.

The Mediterranean monk seal (Monachus monachus) is an emblematic species of conservation concern. Anthropogenic pressures have led to severe population declines and significant fragmentation of the remaining populations. Because of their close relationship with coastal areas, Mediterranean monk seals may be potentially exposed to pollution from agricultural sources, as well as from oil tanker spills. Although monitoring of pollution has long been considered a priority for this species, data on monk seal contamination levels are scarce. In this study, 55 blubber samples of all genders and age classes collected during necropsies (1995-2013) from seals of the Eastern Mediterranean subpopulation were analyzed for organochlorine compounds (OCs), i.e., hexachlorobenzene (HCB), polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDTs), and polycyclic aromatic hydrocarbons (PAHs). Overall, PCBs > DDTs ≫ PAHs > HCB in all samples. Results showed a significant downward trend over the 19-year period for DDTs and HCB. No marked pattern was found for PAHs, even though relative abundance of cancerogenic PAH fraction rose in recent years. PCB levels in subadult specimens increased noticeably over time despite worldwide ban. Our findings did not suggest recent releases of DDT or new pesticides (e.g., Dicofol) in the environment, but may indicate an improper disposal of ancient storages of PCBs or a remobilization from reservoirs. OC levels exceeded thresholds that are commonly associated with immunosuppression and reproductive impairment in other pinnipeds. Hence, OCs could be responsible of potential toxicological effects in this subpopulation. This study represents the first report on PAH and one of the few reports on OC bioaccumulation in Mediterranean monk seals. Based on the levels found in the blubber of monk seals from the Eastern Mediterranean, and given the known harmful effects associated to the prolonged exposure to these substances and the reduced long-term expectations of species recovery, regular monitoring is advocated.