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Field studies suggest that changes in the stable isotope ratios of phytoplankton communities can be used to track changes in the utilization of different nitrogen sources, i.e., to detect shifts from dissolved inorganic nitrogen (DIN) uptake to atmospheric nitrogen (N2) fixation by diazotrophic cyanobacteria as an indication of nitrogen limitation. We explored changes in the stable isotope signature of the diazotrophic cyanobacterium Trichormus variabilis in response to increasing nitrate (NO3-) concentrations (0 to 170 mg L-1) under controlled laboratory conditions. In addition, we explored the influence of nitrogen utilization at the primary producer level on trophic fractionation by studying potential changes in isotope ratios in the freshwater model Daphnia magna feeding on the differently grown cyanobacteria. We show that δ 15N values of the cyanobacterium increase asymptotically with DIN availability, from -0.7 ‰ in the absence of DIN (suggesting N2 fixation) to 2.9 ‰ at the highest DIN concentration (exclusive DIN uptake). In contrast, δ 13C values of the cyanobacterium did not show a clear relationship with DIN availability. The stable isotope ratios of the consumer reflected those of the differently grown cyanobacteria but also revealed significant trophic fractionation in response to nitrogen utilization at the primary producer level. Nitrogen isotope turnover rates of Daphnia were highest in the absence of DIN as a consequence of N2 fixation and resulting depletion in 15N at the primary producer level. Our results highlight the potential of stable isotopes to assess nitrogen limitation and to explore diazotrophy in aquatic food webs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Helmer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Top carnivores can influence the structure of ecological communities, primarily through competition and predation; however, communities are also influenced by bottom-up forces such as anthropogenic habitat disturbance. Top carnivore declines will likely alter competitive dynamics within and amongst sympatric carnivore species. Increasing intraspecific competition is generally predicted to drive niche expansion and/or individual specialisation, while interspecific competition tends to constrain niches. Using stable isotope analysis of whiskers, we studied the effects of Tasmanian devil Sarcophilus harrisii declines upon the population- and individual-level isotopic niches of Tasmanian devils and sympatric spotted-tailed quolls Dasyurus maculatus subsp. maculatus. We investigated whether time since the onset of devil decline (a proxy for severity of decline) and landscape characteristics affected the isotopic niche breadth and overlap of devil and quoll populations. We quantified individual isotopic niche breadth for a subset of Tasmanian devils and spotted-tailed quolls and assessed whether between-site population niche variation was driven by individual-level specialisation. Tasmanian devils and spotted-tailed quolls demonstrated smaller population-level isotopic niche breadths with increasing human-modified habitat, while time since the onset of devil decline had no effect on population-level niche breadth or interspecific niche overlap. Individual isotopic niche breadths of Tasmanian devils and spotted-tailed quolls were narrower in human-modified landscapes, likely driving population isotopic niche contraction, however, the degree of individuals' specialisation relative to one another remained constant. Our results suggest that across varied landscapes, mammalian carnivore niches can be more sensitive to the bottom-up forces of anthropogenic habitat disturbance than to the top-down effects of top carnivore decline., (© 2024. The Author(s).)

Rationale: Metabolism and diet quality play an important role in determining delay mechanisms between an animal ingesting an element and depositing the associated isotope signal in tissue. While many isotope mixing models assume instantaneous reflection of diet in an animal- tissue, this is rarely the case. Here we use data from wildebeest to measure the lag time between ingestion of 34 S and its detection in tail hair., Methods: We use time-lagged regression analysis of δ 34 S data from GPS-collared blue wildebeest from the Serengeti ecosystem in combination with δ 34 S isoscape data to estimate the lag time between an animal ingesting and depositing 34 S in tail hair., Results: The best fitting regression model of δ 34 S in tail hair and an individual- position on the δ 34 S isoscape is generated assuming an average time delay of 78 days between ingestion and detection in tail hair. This suggests that sulfur may undergo multiple metabolic transitions before being deposited in tissue., Conclusion: Our findings help to unravel the underlying complexities associated with sulfur metabolism and are broadly consistent with results from other species. These findings will help to inform research aiming to apply the variation of δ 34 S in inert biological material for geolocation or understanding dietary changes, especially for fast moving migratory ungulates such as wildebeest., (© 2023 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Rationale: By combining precision satellite-tracking with blood sampling, seabirds can be used to validate marine carbon and nitrogen isoscapes, but it is unclear whether a comparable approach using low-precision light-level geolocators (GLS) and feather sampling can be similarly effective., Methods: Here we used GLS to identify wintering areas of northern gannets (Morus bassanus) and sampled winter grown feathers (confirmed from image analysis of non-breeding birds) to test for spatial gradients in δ 13 C and δ 15 N in the NE Atlantic., Results: By matching winter-grown feathers with the non-breeding location of tracked birds we found latitudinal gradients in δ 13 C and δ 15 N in neritic waters. Moreover, isotopic patterns were best explained by sea surface temperature. Similar isotope gradients were found in fish muscle sampled at local ports., Conclusions: Our study reveals the potential of using seabird GLS and feathers to reconstruct large-scale isotopic patterns., (© 2023 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Air pollution and poor air quality is impacting human health globally and is a major cause of respiratory and cardiovascular disease and damage to human organ systems. Automated air quality monitoring stations continuously record airborne pollutant concentrations, but are restricted in number, costly to maintain and cannot document all spatial variability of airborne pollutants. Biomonitors, such as lichens, are commonly used as an inexpensive alternative to assess the degree of pollution and monitor air quality. However, only a few studies combined lichen carbon, nitrogen and sulfur contents, with their stable-isotope-ratio signatures (δ 13 C, δ 15 N and δ 34 S values) to assess spatial variability of air quality and to 'fingerprint' potential pollution sources. In this study, a high-spatial resolution lichen biomonitoring approach (using Xanthoria parietina and Physcia spp.) was applied to the City of Manchester (UK), the centre of the urban conurbation Greater Manchester, including considerations of its urban characteristics (e.g., building heights and traffic statistics), to investigate finer spatial detail urban air quality. Lichen wt% N and δ 15 N signatures, combined with lichen nitrate (NO 3 - ) and ammonium (NH 4 + ) concentrations, suggest a complex mixture of airborne NO x and NH x compounds across Manchester. In contrast, lichen S wt%, combined with δ 34 S strongly suggest anthropogenic sulfur sources, whereas C wt% and δ 13 C signatures were not considered reliable indicators of atmospheric carbon emissions. Manchester's urban attributes were found to influence lichen pollutant loadings, suggesting deteriorated air quality in proximity to highly trafficked roads and densely built-up areas. Lichen elemental contents and stable-isotope-ratio signatures can be used to identify areas of poor air quality, particularly at locations not covered by automated air quality measurement stations. Therefore, lichen biomonitoring approaches provide a beneficial method to supplement automated monitoring stations and also to assess finer spatial variability of urban air quality., (© 2023. The Author(s).)

Between the sixteenth and nineteenth century, British agriculture underwent a 'revolutionary' transformation. Yet despite over a century of research and the recognised centrality of agricultural developments to industrialisation and population growth, the character or chronology of any 'revolution' during this period remains contentious. Enquiry has been hampered by the fragmented and locally specific nature of historic accounts and the broad dating of early-modern zooarchaeological assemblages. To address this, we conducted stable isotope analysis on 658 legal documents written on sheepskin parchment; a unique biological resource that records the day, month and year of use (AD 1499 to 1969). We find these provide a high temporal resolution analysis of changing agricultural practices and episodes of disease. Most significantly, they suggest that if an 'Agricultural Revolution' occurred in livestock management, it did so from the mid-nineteenth century, in the aftermath of the Napoleonic Wars., (© 2023. The Author(s).)

Classic evolutionary theory suggests that sexual dimorphism evolves primarily via sexual and fecundity selection. However, theory and evidence are beginning to accumulate suggesting that resource competition can drive the evolution of sexual dimorphism, via ecological character displacement between sexes. A key prediction of this hypothesis is that the extent of ecological divergence between sexes will be associated with the extent of sexual dimorphism. As the stable isotope ratios of animal tissues provide a quantitative measure of various aspects of ecology, we carried out a meta-analysis examining associations between the extent of isotopic divergence between sexes and the extent of body size dimorphism. Our models demonstrate that large amounts of between-study variation in isotopic (ecological) divergence between sexes is nonrandom and may be associated with the traits of study subjects. We, therefore, completed meta-regressions to examine whether the extent of isotopic divergence between sexes is associated with the extent of sexual size dimorphism. We found modest but significantly positive associations across species between size dimorphism and ecological differences between sexes, that increased in strength when the ecological opportunity for dietary divergence between sexes was greatest. Our results, therefore, provide further evidence that ecologically mediated selection, not directly related to reproduction, can contribute to the evolution of sexual dimorphism., Competing Interests: The authors declare that they have no competing interests., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Knowledge of the uptake and fate of mercury (Hg) compounds in biota is important in understanding the global cycling of Hg and its transfer pathways through food chains. In this study, we analysed total mercury (T-Hg) and methylmercury (MeHg) concentrations in 117 livers of Scottish birds of prey that were found across Scotland and submitted for post-mortem examination through the Raptor Health Scotland project between 2009 and 2019. Statistical comparisons focussed on six species (barn owl, Tyto alba; Eurasian common buzzard, Buteo buteo; golden eagle, Aquila chrysaetos; hen harrier, Circus cyaneus; Eurasian sparrowhawk, Accipiter nisus; and tawny owl, Strix aluco) and showed that golden eagles had a statistically lower fraction of MeHg compared to other raptor species. Further investigation using stable carbon and stable nitrogen isotope ratio measurements carried out for the golden eagles (n = 15) indicated that the increased presence of inorganic mercury (iHg) correlated with a marine influence on the primarily terrestrial diet. Additional bioimaging (n = 1) with laser ablation - inductively coupled plasma - mass spectrometry indicated the co-location of Hg and selenium (Se) within the liver tissue and transmission electron microscopy showed evidence of nanoparticles within the range of 10-20 nm. Further analysis using single particle - inductively coupled plasma - mass spectrometry (n = 4) confirmed the presence of Hg nanoparticles. Together, the evidence suggests the presence of mercury selenide (HgSe) nanoparticles in the liver of some golden eagles that, to our knowledge, has never been directly observed in terrestrial birds of prey. This study points to two alternative hypotheses: these golden eagles may be efficient at breaking down MeHg and form HgSe nanoparticles as a detoxification mechanism (as previously observed in cetaceans), or some golden eagles with elevated iHg may have accumulated these nanoparticles by foraging on stranded cetaceans or seabirds., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Stable isotopes from archaic Falkland Islands wolves ( Dusicyon australis ) indicate a high trophic, marine diet. Hamley et al. argue that this is consistent with mutualism with Yaghan people. However, most D. australis had similar isotopic signatures in the European era, despite human persecution. These data therefore neither support nor refute human-mediated introduction of D. australis to the Falklands.

Dietary studies in birds of prey involve direct observation and examination of food remains at resting and nesting sites. Although these methods accurately identify diet in raptors, they are time-consuming, resource-intensive, and associated with biases from the feeding ecology of raptors like Gyps vultures. Our study set out to estimate diet composition in Gyps vultures informed by stable isotopes that provide a good representation of assimilated diet from local systems.We hypothesized that differences in Gyps vulture diet composition is a function of sampling location and that these vultures move between Serengeti National Park and Selous Game Reserve to forage. We also theorized that grazing ungulates are the principal items in Gyps vulture diet.Through combined linear and Bayesian modeling, diet derived from δ 13 C in Gyps vultures consisted of grazing herbivores across sites, with those in Serengeti National Park consuming higher proportions of grazing herbivores (>87%). δ 13 C differences in vulture feather subsets did not indicate shifts in vulture diet and combined with blood δ 13 C, vultures fed largely on grazers for ~159 days before they were sampled. Similarly, δ 15 N values indicated Gyps vultures fed largely on herbivores. δ 34 S ratios separated where vultures fed when the two sites were compared. δ 34 S variation in vultures across sites resulted from baseline differences in plant δ 34 S values, though it is not possible to match δ 34 S to specific locations.Our findings highlight the relevance of repeated sampling that considers tissues with varying isotopic turnover and emerging Bayesian techniques for dietary studies using stable isotopes. Findings also suggested limited vulture movement between the two local systems. However, more sampling coupled with environmental data is required to fully comprehend this observation and its implications to Gyps vulture ecology and conservation., Competing Interests: The authors declare that they have no conflict of interest and that the views expressed herein are those of the authors., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

We present the isotopic discrimination between paired skin and bone collagen from animals of known life history, providing a modern baseline for the interpretation of archaeological isotopic data. At present, the interpretation of inter-tissue variation (Δ (skin-bone) ) in mummified remains is based on comparisons with other archaeological material, which have attributed divergence to their contrasting turnover rates, with rapidly remodelling skin collagen incorporating alterations in environmental, cultural and physiological conditions in the months prior to death. While plausible, the lack of baseline data from individuals with known life histories has hindered evaluation of the explanations presented. Our analysis of a range of animals raised under a variety of management practices showed a population-wide trend for skin collagen to be depleted in 13 C by -0.7‰ and enriched in 15 N by +1.0‰ relative to bone collagen, even in stillborn animals. These results are intriguing and difficult to explain using current knowledge; however, on the basis of the findings reported here, we caution any results which interpret simply on differing turnover rates. We hypothesize that there may be a consistent difference in the routing of dietary protein and lipids between skin and bone, with potentially on-site synthesis of non-essential amino acids using carbon and nitrogen that have been sourced via different biochemical pathways., (© 2022 The Authors.)

Background: The endogenous cannabinoid system modulates inflammatory signaling in a variety of pathological states, including traumatic brain injury (TBI). The selective expression of diacylglycerol lipase-β (DAGL-β), the 2-arachidonylglycerol biosynthetic enzyme, on resident immune cells of the brain (microglia) and the role of this pathway in neuroinflammation, suggest that this enzyme may contribute to TBI-induced neuroinflammation. Accordingly, we tested whether DAGL-β -/- mice would show a protective phenotype from the deleterious consequences of TBI on cognitive and neurological motor functions. Materials and Methods: DAGL-β -/- and -β +/+ mice were subjected to the lateral fluid percussion model of TBI and assessed for learning and memory in the Morris water maze (MWM) Fixed Platform (reference memory) and Reversal (cognitive flexibility) tasks, as well as in a cued MWM task to infer potential sensorimotor/motivational deficits. In addition, subjects were assessed for motor behavior (Rotarod and the Neurological Severity Score assays) and in the light/dark box and the elevated plus maze to infer whether these manipulations affected anxiety-like behavior. Finally, we also examined whether brain injury disrupts the ceramide/sphingolipid lipid signaling system and if DAGL-β deletion offers protection. Results: TBI disrupted all measures of neurological motor function and reduced body weight, but did not affect body temperature or performance in common assays used to infer anxiety. TBI also impaired performance in MWM Fixed Platform and Reversal tasks, but did not affect cued MWM performance. Although no differences were found between DAGL-β -/- and -β +/+ mice in any of these measures, male DAGL-β -/- mice displayed an unexpected survival-protective phenotype, which persisted at increased injury severities. In contrast, TBI did not elicit mortality in female mice regardless of genotype. TBI also produced significant changes in sphingolipid profiles (a family of lipids, members of which have been linked to both apoptotic and antiapoptotic pathways), in which DAGL-β deletion modestly altered levels of select species. Conclusions: These findings indicate that although DAGL-β does not play a necessary role in TBI-induced cognitive and neurological function, it appears to contribute to the increased vulnerability of male mice to TBI-induced mortality, whereas female mice show high survival rates irrespective of DAGL-β expression.

Rationale: The study of insect migration is problematic due to the small size of insects. Stable isotope analysis can be used to elucidate movement, either by geographic assignment of location of a species, or by simply distinguishing migrant from resident populations. There are few isoscapes of any kind in the UK/Ireland available for interrogation. Thus, I have measured stable isotope ratios (of H, C, N and S) of 299 individuals of the non-migratory Brimstone moth (Opisthograptis luteolata) collected from 93 locations around the UK and Ireland by citizen scientists., Methods: After removing lipids, stable isotope ratios were measured by continuous flow isotope ratio mass spectrometry, using either a conventional elemental analyser (C, N and S) or a high-temperature, thermal conversion elemental analyser in reductive mode., Results: Maps (isoscapes) were constructed that illustrate the stable isotope spatial distribution of this insect. These are the first isoscapes of H, C, N and S of biological samples covering both UK and Ireland., Conclusions: The insect isoscape patterns can be explained from what we know of moth diet, climate and geology. Sulfur isotopes may be of particular use for distinguishing individuals from areas of unique geology. Isoscape patterns may (with care) predict isotope compositions of other, herbivorous, non-aquatic, chitinous taxa. Such isoscapes, when extended beyond the UK and Ireland, would provide a useful tool to elucidate insect migration., (© 2021 The Author. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Background: Nothing is known about the mechanisms by which increased ceramide levels in the lung contribute to allergic responses and asthma severity., Objective: We sought to investigate the functional role of ceramide in mouse models of allergic airway disease that recapitulate the cardinal clinical features of human allergic asthma., Methods: Allergic airway disease was induced in mice by repeated intranasal administration of house dust mite or the fungal allergen Alternaria alternata. Processes that can be regulated by ceramide and are important for severity of allergic asthma were correlated with ceramide levels measured by mass spectrometry., Results: Both allergens induced massive pulmonary apoptosis and also significantly increased reactive oxygen species in the lung. Prevention of increases in lung ceramide levels mitigated allergen-induced apoptosis, reactive oxygen species, and neutrophil infiltration. In contrast, dietary supplementation of the antioxidant α-tocopherol decreased reactive oxygen species but had no significant effects on elevation of ceramide level or apoptosis, indicating that the increases in lung ceramide levels in allergen-challenged mice are not mediated by oxidative stress. Moreover, specific ceramide species were altered in bronchoalveolar lavage fluid from patients with severe asthma compared with in bronchoalveolar lavage fluid from individuals without asthma., Conclusion: Our data suggest that elevation of ceramide level after allergen challenge contributes to the apoptosis, reactive oxygen species generation, and neutrophilic infiltrate that characterize the severe asthmatic phenotype. Ceramide might be the trigger of formation of Creola bodies found in the sputum of patients with severe asthma and could be a biomarker to optimize diagnosis and to monitor and improve clinical outcomes in this disease., (Copyright © 2020 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

The serine palmitoyltransferase (SPT) complex catalyzes the rate-limiting step in the de novo biosynthesis of ceramides, the precursors of sphingolipids. The mammalian ORMDL isoforms (ORMDL1-3) are negative regulators of SPT. However, the roles of individual ORMDL isoforms are unclear. Using siRNA against individual ORMDLs, only single siORMDL3 had modest effects on dihydroceramide and ceramide levels, whereas downregulation of all three ORMDLs induced more pronounced increases. With the CRISPR/Cas9-based genome-editing strategy, we established stable single ORMDL3 KO (ORMDL3-KO) and ORMDL1/2/3 triple-KO (ORMDL-TKO) cell lines to further understand the roles of ORMDL proteins in sphingolipid biosynthesis. While ORMDL3-KO modestly increased dihydroceramide and ceramide levels, ORMDL-TKO cells had dramatic increases in the accumulation of these sphingolipid precursors. SPT activity was increased only in ORMDL-TKO cells. In addition, ORMDL-TKO but not ORMDL3-KO dramatically increased levels of galactosylceramides, glucosylceramides, and lactosylceramides, the elevated N-acyl chain distributions of which broadly correlated with the increases in ceramide species. Surprisingly, although C16:0 is the major sphingomyelin species, it was only increased in ORMDL3-KO, whereas all other N-acyl chain sphingomyelin species were significantly increased in ORMDL-TKO cells. Analysis of sphingoid bases revealed that although sphingosine was only increased 2-fold in ORMDL-TKO cells, levels of dihydrosphingosine, dihydrosphingosine-1-phosphate, and sphingosine-1-phosphate were hugely increased in ORMDL-TKO cells and not in ORMDL3-KO cells. Thus, ORMDL proteins may have a complex, multifaceted role in the biosynthesis and regulation of cellular sphingolipids., Competing Interests: Conflict of interest The authors declare no conflicts of interest., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: The Schreiber's bat, Miniopterus schreibersii, is adapted to long-distance flight, yet long distance movements have only been recorded sporadically using capture-mark-recapture. In this study, we used the hydrogen isotopic composition of 208 wing and 335 fur specimens from across the species' European range to test the hypothesis that the species migrates over long distances., Results: After obtaining the hydrogen isotopic composition (δ 2 H) of each sample, we performed geographic assignment tests by comparing the δ 2 H of samples with the δ 2 H of sampling sites. We found that 95 bats out of 325 showed evidence of long-distance movement, based on the analysis of either fur or wing samples. The eastern European part of the species range (Greece, Bulgaria and Serbia) had the highest numbers of bats that had moved. The assignment tests also helped identify possible migratory routes, such as movement between the Alps and the Balkans., Conclusions: This is the first continental-scale study to provide evidence of migratory behaviour of M. schreibersii throughout its European range. The work highlights the need for further investigation of this behaviour to provide appropriate conservation strategies.

Background: Current animal tracking studies are most often based on the application of external geolocators such as GPS and radio transmitters. While these technologies provide detailed movement data, they are costly to acquire and maintain, which often restricts sample sizes. Furthermore, deploying external geolocators requires physically capturing and recapturing of animals, which poses an additional welfare concern. Natural biomarkers provide an alternative, non-invasive approach for addressing a range of geolocation questions and can, because of relatively low cost, be collected from many individuals thereby broadening the scope for population-wide inference., Methods: We developed a low-cost, minimally invasive method for distinguishing between local versus non-local movements of cattle using sulfur isotope ratios (δ 34 S) in cattle tail hair collected in the Greater Serengeti Ecosystem, Tanzania., Results: We used a Generalized Additive Model to generate a predicted δ 34 S isoscape across the study area. This isoscape was constructed using spatial smoothers and underpinned by the positive relationship between δ 34 S values and lithology. We then established a strong relationship between δ 34 S from recent sections of cattle tail hair and the δ 34 S from grasses sampled in the immediate vicinity of an individual's location, suggesting δ 34 S in the hair reflects the δ 34 S in the environment. By combining uncertainty in estimation of the isoscape, with predictions of tail hair δ 34 S given an animal's position in the isoscape we estimated the anisotropic distribution of travel distances across the Serengeti ecosystem sufficient to detect movement using sulfur stable isotopes., Conclusions: While the focus of our study was on cattle, this approach can be modified to understand movements in other mobile organisms where the sulfur isoscape is sufficiently heterogeneous relative to the spatial scale of animal movements and where tracking with traditional methods is difficult., Competing Interests: Competing interestsAuthors Declare that no competing interest exist., (© The Author(s) 2020.)

Niemann-Pick type C (NPC) disease is a lysosomal storage disorder arising from mutations in the cholesterol-trafficking protein NPC1 (95%) or NPC2 (5%). These mutations result in accumulation of low-density lipoprotein-derived cholesterol in late endosomes/lysosomes, disruption of endocytic trafficking, and stalled autophagic flux. Additionally, NPC disease results in sphingolipid accumulation, yet it is unique among the sphingolipidoses because of the absence of mutations in the enzymes responsible for sphingolipid degradation. In this work, we examined the cause for sphingosine and sphingolipid accumulation in multiple cellular models of NPC disease and observed that the activity of sphingosine kinase 1 (SphK1), one of the two isoenzymes that phosphorylate sphingoid bases, was markedly reduced in both NPC1 mutant and NPC1 knockout cells. Conversely, SphK1 inhibition with the isotype-specific inhibitor SK1-I in WT cells induced accumulation of cholesterol and reduced cholesterol esterification. Of note, a novel SphK1 activator (SK1-A) that we have characterized decreased sphingoid base and complex sphingolipid accumulation and ameliorated autophagic defects in both NPC1 mutant and NPC1 knockout cells. Remarkably, in these cells, SK1-A also reduced cholesterol accumulation and increased cholesterol ester formation. Our results indicate that a SphK1 activator rescues aberrant cholesterol and sphingolipid storage and trafficking in NPC1 mutant cells. These observations highlight a previously unknown link between SphK1 activity, NPC1, and cholesterol trafficking and metabolism., Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article., (© 2020 Newton et al.)

Introduced mammals have devastated island nesting seabird populations worldwide. Declines in breeding seabirds on St Kilda, UK, have been linked to climate change and predation from great skuas Stercorarius skuas, but the introduced St Kilda field mouse Apodemus sylvaticus hirtensis may also play a role by feeding on adults, chicks or eggs. Here, we use stable isotopes in St Kilda mouse blood and potential dietary items to investigate their foraging ecology, specifically focussing on the importance of seabirds and marine foods in their diet. Mice were seasonally sampled at three sites on Hirta, St Kilda over three consecutive years (2010-2012). The δ 13 C and δ 15 N ratios were used in analyses, including isotope niche and dietary source mixing models, to examine foraging behaviour among locations and between seabird breeding seasons. Mice sampled in Carn Mor - where the majority of the island's seabirds nest - had consistently higher δ 13 C than other locations throughout the year, with δ 15 N also being significantly higher for all but one comparison. The isotopic niche width (SEAs) of Carn Mor mice in each season were distinct from the other locations, and became smaller during the seabird breeding season. Dietary mixing models revealed that seabirds made up a large proportion of the diet for mice from Carn Mor, particularly during the seabird breeding season. In conclusion, our work reveals that seabird-derived foods are likely to form a significant part of the diet of St Kilda mice populations located in and around breeding colonies. It is unclear however, whether this is from scavenging or predation of seabirds, or through their discarded food items. Given that mice have had significant effects on seabird populations elsewhere, it is important to carry out further work to determine whether mice are a significant cause of seabird mortality in this island ecosystem.

Carbon sequestration by sediments and vegetated marine systems contributes to atmospheric carbon drawdown, but little empirical evidence is available to help separate the effects of climate change and other anthropogenic activities on carbon burial over centennial timescales. We used marine sediment organic carbon to determine the role of historic climate variability and human habitation in carbon burial over the past 5,071 years. There was centennial-scale sensitivity of carbon supply and burial to climatic variability, with Little Ice Age cooling causing an abrupt ecosystem shift and an increase in marine carbon contributions compared to terrestrial carbon. Although land use changes during the late 1800s did not cause marked alteration in average carbon burial, they did lead to marked increases in the spatial variability of carbon burial. Thus, while carbon burial by vegetated systems is expected to increase with projected climate warming over the coming century, ecosystem restructuring caused by abrupt climate change may produce unexpected change in carbon burial whose variability is also modulated by land use change., (© 2020 John Wiley & Sons Ltd.)

Competition for resources within a population can lead to niche partitioning between sexes, throughout ontogeny and among individuals, allowing con-specifics to co-exist. We aimed to quantify such partitioning in Antarctic fur seals, Arctocephalus gazella, breeding at South Georgia, which hosts ~95% of the world's population. Whiskers were collected from 20 adult males and 20 adult females and stable isotope ratios were quantified every 5 mm along the length of each whisker. Nitrogen isotope ratios (δ 15 N) were used as proxies for trophic position and carbon isotope ratios (δ 13 C) indicated foraging habitat. Sexual segregation was evident: δ 13 C values were significantly lower in males than females, indicating males spent more time foraging south of the Polar Front in maritime Antarctica. In males δ 13 C values declined with age, suggesting males spent more time foraging south throughout ontogeny. In females δ 13 C values revealed two main foraging strategies: 70% of females spent most time foraging south of the Polar Front and had similar δ 15 N values to males, while 30% of females spent most time foraging north of the Polar Front and had significantly higher δ 15 N values. This niche partitioning may relax competition and ultimately elevate population carrying capacity with implications for ecology, evolution and conservation.

Sympatric species may partition resources to reduce competition and facilitate co-existence. While spatial variation and specialization in feeding strategies may be prevalent among large marine predators, studies have focussed on sharks, birds, and marine mammals. We consider for the first time the isotopic niche partitioning of co-occurring, teleost reef predators spanning multiple families. Using a novel tri-isotope ellipsoid approach, we investigate the feeding strategies of seven of these species across an atoll seascape in the Maldives. We demonstrate substantial spatial variation in resource use of all predator populations. Furthermore, within each area, there was evidence of intraspecific variation in feeding behaviors that could not wholly be attributed to individual body size. Assessing species at the population level will mask these intraspecific differences in resource use. Knowledge of resource use is important for predicting how species will respond to environmental change and spatial variation should be considered when investigating trophic diversity., Competing Interests: None declared., (© 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Patterns of behavioural variation and migratory connectivity are important characteristics of populations, particularly at the edges of species distributions, where processes involved in influencing evolutionary trajectories, such as divergence, mutual persistence, and natural hybridization, can occur. Here, we focused on two closely related seabird species that breed in the Mediterranean: Balearic shearwaters (Puffinus mauretanicus) and Yelkouan shearwaters (Puffinus yelkouan). Genetic and phenotypic evidence of hybridization between the two species on Menorca (the eastern and westernmost island in the breeding ranges of the two shearwaters, respectively) has provided important insights into relationships between these recently diverged species. Nevertheless, levels of behavioural and ecological differentiation amongst these populations remain largely unknown. Using geolocation and stable isotopes, we compared the at-sea movement behaviour of birds from the Menorcan 'hybrid' population with the nearest neighbouring populations of Balearic and Yelkouan shearwaters. The Menorcan population displayed a suite of behavioural features intermediate to those seen in the two species (including migration strategies, breeding season movements and limited data on phenology). Our findings provide new evidence to support suggestions that the Menorcan population is admixed, and indicate a role of non-breeding behaviours in the evolutionary trajectories of Puffinus shearwaters in the Mediterranean.

Coral reef food webs are complex, vary spatially and remain poorly understood. Certain large predators, notably sharks, are subsidized by pelagic production on outer reef slopes, but how widespread this dependence is across all teleost fishery target species and within atolls is unclear. North Malé Atoll (Maldives) includes oceanic barrier as well as lagoonal reefs. Nine fishery target predators constituting ca. 55% of the local fishery target species biomass at assumed trophic levels 3-5 were selected for analysis. Data were derived from carbon (δ 13 C), nitrogen (δ 15 N) and sulphur (δ 34 S) stable isotopes from predator white dorsal muscle samples, and primary consumer species representing production source end-members. Three-source Bayesian stable isotope mixing models showed that uptake of pelagic production extends throughout the atoll, with predatory fishes showing equal planktonic reliance between inner and outer edge reefs. Median plankton contribution was 65%-80% for all groupers and 68%-88% for an emperor, a jack and snappers. Lagoonal and atoll edge predators are equally at risk from anthropogenic and climate-induced changes, which may impact the linkages they construct, highlighting the need for management plans that transcend the boundaries of this threatened ecosystem., (© 2019 The Authors. Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.)

Southeast-Asian peat swamp forests have been significantly logged and converted to plantation. Recently, to mitigate land degradation and C losses, some areas have been left to regenerate. Understanding how such complex land use change affects greenhouse gas emissions is essential for modelling climate feedbacks and supporting land management decisions. We carried out field research in a Malaysian swamp forest and an oil palm plantation to understand how clear-felling, drainage, and illegal and authorized conversion to oil palm impacted the C cycle, and how the C cycle may change if such logging and conversion stopped. We found that both the swamp forest and the plantation emit centuries-old CO 2 from their drainage systems in the managed areas, releasing sequestered C to the atmosphere. Oil palm plantations are an iconic symbol of tropical peatland degradation, but CO 2 efflux from the recently-burnt, cleared swamp forest was as old as from the oil palm plantation. However, in the swamp forest site, where logging had ceased approximately 30 years ago, the age of the CO 2 efflux was modern, indicating recovery of the system can occur. 14 C dating of the C pool acted as a tracer of recovery as well as degradation and offers a new tool to assess efficacy of restoration management. Methane was present in many sites, and in higher concentrations in slow-flowing anoxic systems as degassing mechanisms are not strong. Methane loading in freshwaters is rarely considered, but this may be an important C pool in restored drainage channels and should be considered in C budgets and losses.

Migration can influence host-parasite dynamics in animals by increasing exposure to parasites, by reducing the energy available for immune defense, or by culling of infected individuals. These mechanisms have been demonstrated in several comparative analyses; however, few studies have investigated whether conspecific variation in migration distance may also be related to infection risk. Here, we ask whether autumn migration distance, inferred from stable hydrogen isotope analysis of summer-grown feathers ( δ 2 H f ) in Europe, correlates with blood parasite prevalence and intensity of infection for willow warblers ( Phylloscopus trochilus ) wintering in Zambia. We also investigated whether infection was correlated with individual condition (assessed via corticosterone, scaled mass index, and feather quality). We found that 43% of birds were infected with Haemoproteus palloris (lineage WW1). Using generalized linear models, we found no relationship between migration distance and either Haemoproteus infection prevalence or intensity. There was spatial variation in breeding ground origins of infected versus noninfected birds, with infected birds originating from more northern sites than noninfected birds, but this difference translated into only slightly longer estimated migration distances (~214 km) for infected birds. We found no relationship between body condition indices and Haemoproteus infection prevalence or intensity. Our results do not support any of the proposed mechanisms for migration effects on host-parasite dynamics and cautiously suggest that other factors may be more important for determining individual susceptibility to disease in migratory bird species., Competing Interests: The authors declare no competing interests.

Background: Novel crystalloid solutions containing polyethylene glycol polymers (PEG-20k) produce dramatic resuscitation effects but dose-dependently produce a hypocoagulative state. The objective of this study was to examine possible mechanisms of this effect. Based on previous thromboelastography data, we hypothesize the effect is largely due to platelet interactions with the polymers., Methods: Whole citrated blood from healthy volunteers was diluted ex-vivo 10% with crystalloids and tested for coagulation and platelet function. The specific tests included prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen and von Willebrand factor (vWf) activity, thrombin generation, thromboelastography with and without platelet mapping, platelet flow cytometry, and erythrocyte sedimentation rate., Findings: Fibrinogen and vWF activities, PT, and aPTT were not affected by PEG-20k dilutions. Thrombin activity was mildly suppressed with PEG-20k (TTP- 20%). Platelet mapping demonstrated significantly greater % inhibition of both ADP and arachidonic acid-induced platelet aggregation with PEG-20k, but direct ADP-activated gpIIa/IIIb (PAC1) and P-selectin (CD62P) binding site expression was not altered. Mild dose-dependent suppression of TEG-MA was seen with PEG-20k using platelet poor plasma. Erythrocyte Sedimentation Rates (ESR) were dramatically accelerated after dilution with 10% PEG-20k, which was competitively blocked by smaller PEG polymers, suggesting nonspecific PEG-20k cell binding effects., Conclusions: PEG-20k creates a mild hypocoagulative state in whole blood at concentrations ≥10%, which may be due to platelet-PEG interactions at the IIb/IIIa interface with lesser effects on fibrin polymerization. This interaction may cause a functional thrombasthenia induced by nonspecific platelet surface passivation by the PEG polymer., Competing Interests: The authors have declared that no competing interests exist.

Essentials Platelets in trauma-induced coagulopathy (TIC) are impaired, but the mechanism is not known. We performed comprehensive longitudinal platelet function testing in trauma patient samples. Platelets in TIC are widely impaired early after injury, but platelet activatability is intact. This suggests a mechanism of transient platelet cytoskeletal/integrin dysfunction during TIC. SUMMARY: Background Trauma-induced coagulopathy (TIC) is a common and deadly bleeding disorder. Platelet dysfunction is present during TIC, but its mechanisms remain unclear. Platelets are currently thought to become "exhausted," a state in which they have released their granule contents and can no longer aggregate or contract. Methods This prospective observational cohort study tested the hypothesis that platelet exhaustion is present during TIC and characterized the early time course of platelet dysfunction. Blood was collected from 95 adult trauma patients at a Level I trauma center at time of Emergency Department arrival and several time points over 72 h. Platelet activation state and function were characterized using CD62P (P-selectin) and PAC-1 surface membrane staining, platelet function analyzer (PFA-100), aggregometry, viscoelastic platelet mapping, and, to test for exhaustion, their ability to express CD62P after ex vivo adenosine diphosphate (ADP) agonism. Platelet function was compared between patients with and without TIC, defined by prothrombin time ≥18 s. Results Platelets in TIC showed no initial increase in their level of surface activation markers or impairment of their capacity to express CD62P in response to ADP stimulation. However, TIC platelets were impaired in nearly all functional assays, spanning adhesion, aggregation, and contraction. These effects largely remained after controlling for platelet count and fibrinogen concentration and resolved after 8 h. Conclusion The TIC platelets exhibit early impairment of adhesion, aggregation, and contraction with retained alpha granule secretion ability, suggesting a specific mechanism of cytoskeletal or integrin dysfunction that is not a result of more general platelet exhaustion., (© 2019 International Society on Thrombosis and Haemostasis.)

Niemann-Pick type C (NPC) disease is a lysosomal storage disorder resulting from mutations in either the NPC1 (95%) or NPC2 (5%) genes. NPC typically presents in childhood with visceral lipid accumulation and complex progressive neurodegeneration characterized by cerebellar ataxia, dysphagia, and dementia, resulting in a shortened lifespan. While cholesterol is widely acknowledged as the principal storage lipid in NPC, multiple species of sphingolipids accumulate as well. This accumulation of sphingolipids led to the initial assumption that NPC disease was caused by a deficiency in a sphingolipid catabolism enzyme, similar to sphingomyelinase deficiencies with which it shares a family name. It took about half a century to determine that NPC was in fact caused by a cholesterol trafficking defect, and still as we approach a century after the initial identification of the disease, the mechanisms by which sphingolipids accumulate remain poorly understood. Here we focus on the defects of sphingolipid catabolism in the endolysosomal compartment and how they contribute to the biology and pathology observed in NPC disease. This review highlights the need for further work on understanding and possibly developing treatments to correct the accumulation of sphingolipids in addition to cholesterol in this currently untreatable disease., (Copyright © 2018. Published by Elsevier Ltd.)

In breast cancer, 17β-estradiol (E2) plays critical roles mainly by binding to its canonical receptor, estrogen receptor (ER) α66, and eliciting genomic effects. E2 also triggers rapid, nongenomic responses. E2 activates sphingosine kinase 1 (SphK1), increasing sphingosine-1-phosphate (S1P) that binds to its receptors, leading to important breast cancer signaling. However, the E2 receptor responsible for SphK1 activation has not yet been identified. Here, we demonstrate in triple-negative breast cancer cells, which lack the canonical ERα66 but express the novel splice variant ERα36, that ERα36 is the receptor responsible for E2-induced activation of SphK1 and formation and secretion of S1P and dihydro-S1P, the ligands for S1PRs. Tamoxifen, the first-line endocrine therapy for breast cancer, is an antagonist of ERα66, but an agonist of ERα36, and, like E2, activates SphK1 and markedly increases secretion of S1P. A major problem with tamoxifen therapy is development of acquired resistance. We found that tamoxifen resistance correlated with increased SphK1 and ERα36 expression in tamoxifen-resistant breast cancer cells, in patient-derived xenografts, and in endocrine-resistant breast cancer patients. Our data also indicate that targeting this ERα36 and SphK1 axis may be a therapeutic option to circumvent endocrine resistance and improve patient outcome., (Copyright © 2018 Maczis et al.)

Background: Low volume resuscitation (LVR) in shock prevents deleterious effects of crystalloid loading in pre-hospital settings. Polyethylene glycol 20,000 (PEG-20k) based LVR solutions are 20-fold more effective at maintaining perfusion and survival in shock compared to conventional crystalloids. The aim of this study was to determine coagulation and platelet function of whole blood treated with 10% PEG-20k., Methods: Citrated blood from volunteers (n = 25) or early admission severely injured trauma patients (n = 9) were diluted 10% with various LVR solutions in a matched design with a paired volume control (saline), and studied using thromboelastography (TEG)., Findings: In healthy volunteers and patients, 10% PEG-20k significantly increased clot amplification time (k), decreased propagation (angle), maximal clot size and strength (MA), and the overall coagulation index (CI), but not clot initiation (R) or fibrinolysis (Ly30), relative to paired saline dilutional controls. Clinically, K, angle, and MA were just outside of the normal limits in volunteers but not in patients. No statistical differences existed between PEG-20k and Hextend (HES) in either patient population. In a dose response series using volunteer blood, all effects of 10% PEG-20k on TEG were reversed and normalized by lower concentrations (7.5% and 5%). Furthermore, 7.5% PEG-20k produced similar resuscitation effects as 10% PEG in rodent hemorrhagic shock models (n = 5)., Conclusions: In conclusion, PEG-20k based LVR solutions produced a dose-dependent minor hypocoagulative state, possibly associated with changes in clot propagation and platelet function, which can be reversed by dose reduction in concentration while providing superior LVR, microvascular rescue, and lactate clearance compared to saline or starch., Competing Interests: The authors have declared that no competing interests exist.

Incrementally grown, metabolically inert tissues such as fish otoliths provide biochemical records that can used to infer behavior and physiology throughout the lifetime of the individual. Organic tissues are particularly useful as the stable isotope composition of the organic component can provide information about diet, trophic level and location. Unfortunately, inert, incrementally grown organic tissues are relatively uncommon. The vertebrate eye lens, however, is formed via sequential deposition of protein-filled fiber cells, which are subsequently metabolically inert. Lenses therefore have the potential to serve as biochemical data recorders capturing life-long variations in dietary and spatial ecology. Here we review the state of knowledge regarding the structure and formation of fish eye lenses in the context of using lens tissue for retrospective isotopic analysis. We discuss the relationship between eye lens diameter and body size, describe the successful recovery of expected isotopic gradients throughout ontogeny and between species, and quantify the isotopic offset between lens protein and white muscle tissue . We show that fish eye lens protein is an attractive host for recovery of stable isotope life histories, particularly for juvenile life stages, and especially in elasmobranchs lacking otoliths, but interpretation of lens-based records is complicated by species-specific uncertainties associated with lens growth rates., Competing Interests: The authors declare there are no competing interests.

The bioactive sphingolipid metabolite sphingosine-1-phosphate (S1P) and the enzyme that produces it, SPHK1 (sphingosine kinase 1), regulate many processes important for the etiology of cancer. It has been suggested that SPHK1 levels are regulated by the tumor suppressor protein TP53, a key regulator of cell cycle arrest, apoptosis, and macroautophagy/autophagy. However, little is still known of the relationship between TP53 and SPHK1 activity in the regulation of these processes. To explore this link, we examined the effects of inhibiting SPHK1 in wild-type and TP53 null cancer cell lines. SK1-I, an analog of sphingosine and isozyme-specific SPHK1 inhibitor, suppressed cancer cell growth and clonogenic survival in a TP53-dependent manner. It also more strongly enhanced intrinsic apoptosis in wild-type TP53 cells than in isogenic TP53 null cells. Intriguingly, SK1-I induced phosphorylation of TP53 on Ser15, which increases its transcriptional activity. Consequently, levels of TP53 downstream targets such as pro-apoptotic members of the BCL2 family, including BAX, BAK1, and BID were increased in wild-type but not in TP53 null cells. Inhibition of SPHK1 also increased the formation of autophagic and multivesicular bodies, and increased processing of LC3 and its localization within acidic compartments in a TP53-dependent manner. SK1-I also induced massive accumulation of vacuoles, enhanced autophagy, and increased cell death in an SPHK1-dependent manner that also required TP53 expression. Importantly, downregulation of the key regulators of autophagic flux, BECN1 and ATG5, dramatically decreased the cytotoxicity of SK1-I only in cells with TP53 expression. Hence, our results reveal that TP53 plays an important role in vacuole-associated cell death induced by SPHK1 inhibition in cancer cells.

Horse mussel reefs (Modiolus modiolus) are biodiversity hotspots afforded protection by Marine Protected Areas (MPAs) in the NE Atlantic. In this study, horse mussel reefs, cobble habitats and sandy habitats were assessed using underwater visual census and drop-down video techniques in three UK regions. Megafauna were enumerated, differences in community composition and individual species abundances were analysed. Samples of conspicuous megafauna were also collected from horse mussel reefs in Orkney for stable isotope analysis. Communities of conspicuous megafauna were different between horse mussel habitats and other habitats throughout their range. Three commercially important species: whelks (Buccinum undatum), queen scallops (Aequipecten opercularis) and spider crabs (Maja brachydactyla) were significantly more abundant (by as much as 20 times) on horse mussel reefs than elsewhere. Isotopic analysis provided insights into their trophic relationship with the horse mussel reef. Protection of M. modiolus habitat can achieve biodiversity conservation objectives whilst benefiting fisheries also., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Niemann-Pick type C (NPC) disease is a fatal neurodegenerative disorder caused by mutations in NPC1 or NPC2 with decreased functions leading to lysosomal accumulation of cholesterol and sphingolipids. FTY720/fingolimod, used for treatment of multiple sclerosis, is phosphorylated by nuclear sphingosine kinase 2, and its active phosphorylated form (FTY720-P) is an inhibitor of class I histone deacetylases. In this study, administration of clinically relevant doses of FTY720 to mice increased expression of NPC1 and -2 in brain and liver and decreased cholesterol in an SphK2-dependent manner. FTY720 greatly increased expression of NPC1 and -2 in human NPC1 mutant fibroblasts that correlated with formation of FTY720-P and significantly reduced the accumulation of cholesterol and glycosphingolipids. In agreement with this finding, FTY720 pretreatment of human NPC1 mutant fibroblasts restored transport of the cholera toxin B subunit, which binds ganglioside GM1, to the Golgi apparatus. Together, these findings suggest that FTY720 administration can ameliorate cholesterol and sphingolipid storage and trafficking defects in NPC1 mutant fibroblasts. Because neurodegeneration is the main clinical feature of NPC disease, and FTY720 accumulates in the CNS and has several advantages over available histone deacetylase inhibitors now in clinical trials, our work provides a potential opportunity for treatment of this incurable disease.-Newton, J., Hait, N. C., Maceyka, M., Colaco, A., Maczis, M., Wassif, C. A., Cougnoux, A., Porter, F. D., Milstien, S., Platt, N., Platt, F. M., Spiegel, S. FTY720/fingolimod increases NPC1 and NPC2 expression and reduces cholesterol and sphingolipid accumulation in Niemann-Pick type C mutant fibroblasts., (© FASEB.)

The Caribbean spiny lobster, Panulirus argus, is one of the most valuable fisheries commodities in the Central American region, directly employing 50,000 people and generating >US$450 million per year [1]. This industry is particularly important to small island states such as The Bahamas, which exports more lobster than any other country in the region [1]. Several factors contribute to this disproportionally high productivity, principally the extensive shallow-water banks covered in seagrass meadows [2], where fishermen deploy artificial shelters for the lobsters to supplement scarce reef habitat [3]. The surrounding seabed communities are dominated by lucinid bivalve mollusks that live among the seagrass root system [4, 5]. These clams host chemoautotrophic bacterial symbionts in their gills that synthesize organic matter using reduced sulfur compounds, providing nutrition to their hosts [6]. Recent studies have highlighted the important role of the lucinid clam symbiosis in maintaining the health and productivity of seagrass ecosystems [7, 8], but their biomass also represents a potentially abundant, but as yet unquantified, food source to benthic predators [9]. Here we undertake the first analysis of Caribbean spiny lobster diet using a stable isotope approach (carbon, nitrogen, and sulfur) and show that a significant portion of their food (∼20% on average) is obtained from chemosynthetic primary production in the form of lucinid clams. This nutritional pathway was previously unrecognized in the spiny lobster's diet, and these results are the first empirical evidence that chemosynthetic primary production contributes to the productivity of commercial fisheries stocks., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Amongst migratory species, it is common to find individuals from different populations or geographical origins sharing staging or wintering areas. Given their differing life histories, ecological theory would predict that the different groups of individuals should exhibit some level of niche segregation. This has rarely been investigated because of the difficulty in assigning migrating individuals to breeding areas. Here, we start by documenting a broad geographical gradient of hydrogen isotopes (δ 2 H) in robin Erithacus rubecula feathers across Europe. We then use δ 2 H, as well as wing-tip shape, as surrogates for broad migratory origin of birds wintering in Iberia, to investigate the ecological segregation of populations. Wintering robins of different sexes, ages and body sizes are known to segregate between habitats in Iberia. This has been attributed to the despotic exclusion of inferior competitors from the best patches by dominant individuals. We find no segregation between habitats in relation to δ 2 H in feathers, or to wing-tip shape, which suggests that no major asymmetries in competitive ability exist between migrant robins of different origins. Trophic level (inferred from nitrogen isotopes in blood) correlated both with δ 2 H in feathers and with wing-tip shape, showing that individuals from different geographic origins display a degree of ecological segregation in shared winter quarters. Isotopic mixing models indicate that wintering birds originating from more northerly populations consume more invertebrates. Our multi-scale study suggests that trophic-niche segregation may result from specializations (arising in the population-specific breeding areas) that are transported by the migrants into the shared wintering grounds.

Ecomorphological differentiation is a key feature of adaptive radiations, with a general trend for specialization and niche expansion following divergence. Ecological opportunity afforded by invasion of a new habitat is thought to act as an ecological release, facilitating divergence, and speciation. Here, we investigate trophic adaptive morphology and ecology of an endemic clade of oreochromine cichlid fishes (Alcolapia) that radiated along a herbivorous trophic axis following colonization of an isolated lacustrine environment, and demonstrate phenotype-environment correlation. Ecological and morphological divergence of the Alcolapia species flock are examined in a phylogenomic context, to infer ecological niche occupation within the radiation. Species divergence is observed in both ecology and morphology, supporting the importance of ecological speciation within the radiation. Comparison with an outgroup taxon reveals large-scale ecomorphological divergence but shallow genomic differentiation within the Alcolapia adaptive radiation. Ancestral morphological reconstruction suggests lake colonization by a generalist oreochromine phenotype that diverged in Lake Natron to varied herbivorous morphologies akin to specialist herbivores in Lakes Tanganyika and Malawi., (© 2016 The Author(s). Evolution published by Wiley Periodicals, Inc. on behalf of The Society for the Study of Evolution.)

We investigate the origin of archaeological wool textiles preserved by anoxic waterlogging from seven medieval archaeological deposits in north-western Europe (c. 700-1600 AD), using geospatial patterning in carbon (δ13C), nitrogen (δ15N) and non-exchangeable hydrogen (δ2H) composition of modern and ancient sheep proteins. δ13C, δ15N and δ2H values from archaeological wool keratin (n = 83) and bone collagen (n = 59) from four sites were interpreted with reference to the composition of modern sheep wool from the same regions. The isotopic composition of wool and bone collagen samples clustered strongly by settlement; inter-regional relationships were largely parallel in modern and ancient samples, though landscape change was also significant. Degradation in archaeological wool samples, examined by elemental and amino acid composition, was greater in samples from Iceland (Reykholt) than in samples from north-east England (York, Newcastle) or northern Germany (Hessens). A nominal assignment approach was used to classify textiles into local/non-local at each site, based on maximal estimates of isotopic variability in modern sheep wool. Light element stable isotope analysis provided new insights into the origins of wool textiles, and demonstrates that isotopic provenancing of keratin preserved in anoxic waterlogged contexts is feasible. We also demonstrate the utility of δ2H analysis to understand the location of origin of archaeological protein samples., Competing Interests: The authors have declared that no competing interests exist.

Background: An understanding of year-round habitat use is essential for determining how carry-over effects shape population dynamics in long-distance migratory songbirds. The recent discovery of long-term migratory staging sites in many species, prior to arrival at final wintering sites, adds complexity to efforts to decipher non-breeding habitat use and connections between sites. We investigated whether habitat conditions during migratory staging carry over to influence great reed warbler (Acrocephalus arundinaceus) body condition at final wintering sites in Zambia. We asked whether the presence/absence and strength of such carry-over effects were modified by contrasting rainfall conditions during 2 years., Results: First, we found that individuals staging in a dry year had higher corticosterone (CORT f ) and stable nitrogen isotope values (suggesting higher aridity) than birds staging in a wet year, indicating that regional weather affected staging conditions. Second, we found that carry-over effects from staging habitat conditions (measured via carbon and nitrogen isotopes) to final winter site body condition (measured via scaled mass index and β-hydroxybutyrate) were only present in a dry year, suggesting that environmental factors have consequences for the strength of carry-over effects. Our results also suggest that wet conditions at final winter sites may buffer the effects of poor staging conditions, at least in the short term, since individuals that staged in a dry year had higher scaled mass indices in Zambia than individuals that staged in a wet year., Conclusions: This study provides a first insight into the connections between long-term migratory staging sites and final wintering sites, and suggests that local environmental factors can modify the strength of carry-over effects for long-distance migratory birds.

Despite a number of studies in areas of focused methane seepage, the extent of transitional sediments of more diffuse methane seepage, and their influence upon biological communities is poorly understood. We investigated an area of reducing sediments with elevated levels of methane on the South Georgia margin around 250 m depth and report data from a series of geochemical and biological analyses. Here, the geochemical signatures were consistent with weak methane seepage and the role of sub-surface methane consumption was clearly very important, preventing gas emissions into bottom waters. As a result, the contribution of methane-derived carbon to the microbial and metazoan food webs was very limited, although sulfur isotopic signatures indicated a wider range of dietary contributions than was apparent from carbon isotope ratios. Macrofaunal assemblages had high dominance and were indicative of reducing sediments, with many taxa common to other similar environments and no seep-endemic fauna, indicating transitional assemblages. Also similar to other cold seep areas, there were samples of authigenic carbonate, but rather than occurring as pavements or sedimentary concretions, these carbonates were restricted to patches on the shells of Axinulus antarcticus (Bivalvia, Thyasiridae), which is suggestive of microbe-metazoan interactions.

Trophic assimilation efficiency (conversion of resource biomass into consumer biomass) is thought to be a limiting factor for food chain length in natural communities. In host-parasitoid systems, which account for the majority of terrestrial consumer interactions, a high trophic assimilation efficiency may be expected at higher trophic levels because of the close match of resource composition of host tissue and the consumer's resource requirements, which would allow for longer food chains. We measured efficiency of biomass transfer along an aphid-primary-secondary-tertiary parasitoid food chain and used stable isotope analysis to confirm trophic levels. We show high efficiency in biomass transfer along the food chain. From the third to the fourth trophic level, the proportion of host biomass transferred was 45%, 65% and 73%, respectively, for three secondary parasitoid species. For two parasitoid species that can act at the fourth and fifth trophic levels, we show markedly increased trophic assimilation efficiencies at the higher trophic level, which increased from 45 to 63% and 73 to 93%, respectively. In common with other food chains, δ(15)N increased with trophic level, with trophic discrimination factors (Δ(15)N) 1.34 and 1.49‰ from primary parasitoids to endoparasitic and ectoparasitic secondary parasitoids, respectively, and 0.78‰ from secondary to tertiary parasitoids. Owing to the extraordinarily high efficiency of hyperparasitoids, cryptic higher trophic levels may exist in host-parasitoid communities, which could alter our understanding of the dynamics and drivers of community structure of these important systems., (© 2016 The Authors.)

Many established models of animal foraging assume that individuals are ecologically equivalent. However, it is increasingly recognized that populations may comprise individuals who differ consistently in their diets and foraging behaviors. For example, recent studies have shown that individual foraging site fidelity (IFSF, when individuals consistently forage in only a small part of their population's home range) occurs in some colonial breeders. Short-term IFSF could result from animals using a win-stay, lose-shift foraging strategy. Alternatively, it may be a consequence of individual specialization. Pelagic seabirds are colonial central-place foragers, classically assumed to use flexible foraging strategies to target widely dispersed, spatiotemporally patchy prey. However, tracking has shown that IFSF occurs in many seabirds, although it is not known whether this persists across years. To test for long-term IFSF and to examine alternative hypotheses concerning its cause, we repeatedly tracked 55 Northern Gannets (Morus bassanus) from a large colony in the North Sea within and across three successive breeding seasons. Gannets foraged in neritic waters, predictably structured by tidal mixing and thermal stratification, but subject to stochastic, wind-induced overturning. Both within and across years, coarse to mesoscale (tens of kilometers) IFSF was significant but not absolute, and foraging birds departed the colony in individually consistent directions. Carbon stable isotope ratios in gannet blood tissues were repeatable within years and nitrogen ratios were also repeatable across years, suggesting long-term individual dietary specialization. Individuals were also consistent across years in habitat use with respect to relative sea surface temperature and in some dive metrics, yet none of these factors accounted for IFSF. Moreover, at the scale of weeks, IFSF did not decay over time and the magnitude of IFSF across years was similar to that within years, suggesting that IFSF is not primarily the result of win-stay, lose-shift foraging. Rather, we hypothesize that site familiarity, accrued early in-life, causes IFSF by canalizing subsequent foraging decisions. Evidence from this and other studies suggests that IFSF may be common in colonial central-place foragers, with far-reaching consequences for our attempts to understand and conserve these animals in a rapidly changing environment.

Background: Asthma, a chronic inflammatory condition defined by episodic shortness of breath with expiratory wheezing and cough, is a serious health concern affecting more than 250 million persons. Genome-wide association studies have identified ORM (yeast)-like protein isoform 3 (ORMDL3) as a gene associated with susceptibility to asthma. Although its yeast ortholog is a negative regulator of de novo ceramide biosynthesis, how ORMDL3 contributes to asthma pathogenesis is not known., Objectives: We sought to decipher the molecular mechanism for the pathologic functions of ORMDL3 in asthma and the relationship to its evolutionarily conserved role in regulation of ceramide homeostasis., Methods: We determined the relationship between expression of ORMDL3 and ceramide in epithelial and inflammatory cells and in asthma pathogenesis in mice., Results: Although small increases in ORMDL3 expression decrease ceramide levels, remarkably, higher expression in lung epithelial cells and macrophages in vitro and in vivo increased ceramide production, which promoted chronic inflammation, airway hyperresponsiveness, and mucus production during house dust mite-induced allergic asthma. Moreover, nasal administration of the immunosuppressant drug FTY720/fingolimod reduced ORMDL3 expression and ceramide levels and mitigated airway inflammation and hyperreactivity and mucus hypersecretion in house dust mite-challenged mice., Conclusions: Our findings demonstrate that overexpression of ORMDL3 regulates ceramide homeostasis in cells in a complex manner and suggest that local FTY720 administration might be a useful therapeutic intervention for the control of allergic asthma., (Copyright © 2015 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

Introduction: Anticoagulation, fibrinogen consumption, fibrinolytic activation, and platelet dysfunction all interact to produce different clot formation responses after trauma. However, the relative contributions of these coagulation components to overall clot formation remain poorly defined. We examined for sources of heterogeneity in clot formation responses after trauma., Methods: Blood was sampled in the emergency department from patients meeting trauma team activation criteria at an urban trauma center. Plasma prothrombin time of 18 s or longer was used to define traumatic coagulopathy. Mean kaolin-activated thrombelastography (TEG) parameters were calculated and tested for heterogeneity using analysis of means. Discriminant analysis and forward stepwise variable selection with linear regression were used to determine if prothrombin time, fibrinogen, platelet contractile force (PCF), and D-dimer concentration, representing key mechanistic components of coagulopathy, each contribute to heterogeneous TEG responses after trauma., Results: Of 95 subjects, 16% met criteria for coagulopathy. Coagulopathic subjects were more severely injured with greater shock and received more blood products in the first 8 h compared with noncoagulopathic subjects. Mean (SD) TEG maximal amplitude (MA) was significantly decreased in the coagulopathic group (57.5 [SD, 4.7] mm vs. 62.7 [SD, 4.7], t test P < 0.001). The MA also exceeded the ANOM predicted upper decision limit for the noncoagulopathic group and the lower decision limit for the coagulopathic group at α = 0.05, suggesting significant heterogeneity from the overall cohort mean. Fibrinogen and PCF best discriminated TEG MA using discriminant analysis. Fibrinogen, PCF, and D-dimer were primary covariates for TEG MA using regression analysis., Conclusions: Heterogeneity in TEG-based clot formation in emergency department trauma patients was linked to changes in MA. Individual parameters representing fibrin polymerization, PCFs, and fibrinolysis were primarily associated with TEG MA after trauma and should be the focus of early hemostatic therapies.

Jellyfish are highly topical within studies of pelagic food-webs and there is a growing realisation that their role is more complex than once thought. Efforts being made to include jellyfish within fisheries and ecosystem models are an important step forward, but our present understanding of their underlying trophic ecology can lead to their oversimplification in these models. Gelatinous zooplankton represent a polyphyletic assemblage spanning >2,000 species that inhabit coastal seas to the deep-ocean and employ a wide variety of foraging strategies. Despite this diversity, many contemporary modelling approaches include jellyfish as a single functional group feeding at one or two trophic levels at most. Recent reviews have drawn attention to this issue and highlighted the need for improved communication between biologists and theoreticians if this problem is to be overcome. We used stable isotopes to investigate the trophic ecology of three co-occurring scyphozoan jellyfish species (Aurelia aurita, Cyanea lamarckii and C. capillata) within a temperate, coastal food-web in the NE Atlantic. Using information on individual size, time of year and δ (13)C and δ (15)N stable isotope values, we examined: (1) whether all jellyfish could be considered as a single functional group, or showed distinct inter-specific differences in trophic ecology; (2) Were size-based shifts in trophic position, found previously in A. aurita, a common trait across species?; (3) When considered collectively, did the trophic position of three sympatric species remain constant over time? Differences in δ (15)N (trophic position) were evident between all three species, with size-based and temporal shifts in δ (15)N apparent in A. aurita and C. capillata. The isotopic niche width for all species combined increased throughout the season, reflecting temporal shifts in trophic position and seasonal succession in these gelatinous species. Taken together, these findings support previous assertions that jellyfish require more robust inclusion in marine fisheries or ecosystem models.

Avian studies of endocrine disruption traditionally have focused on reproductive impairment, given that many environmental contaminants affect sex steroid hormones. There is also increasing interest in altered thyroid function, and associated early development, particularly in altricial species with extended developmental windows. Both types of effect are relevant under the complex pollutant conditions created in streams draining urban areas, but case studies are scarce. Therefore, the authors measured breeding performance, as well as nestling growth, condition, and plasma thyroid hormones, in 87 Eurasian dipper (Cinclus cinclus) nests on 36 urban and rural streams in south and mid-Wales (UK); invertebrate prey data were also collected. The objective of the present study was to evaluate whether urban stream pollution or food scarcity might affect reproduction or development in this specialized aquatic songbird. Clutch sizes and egg fertility were similar on rural and urban streams, whereas nest success was actually higher at urban sites and food abundance was not significantly reduced. However, subtle but important differences were apparent. Urban nestlings were significantly lighter than rural nestlings for their body size (condition index), and brood sex ratios were increasingly male biased with increasing urbanization. The nestling thyroid hormone profile closely reflected urban land use, whereas depressed triiodothyronine (T3) hormones and poorer body condition were associated with higher exposure to polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) at urbanized sites. These data suggest that PCBs, PBDEs, and/or accompanying contaminants in urban streams could be affecting dipper nestling development, with potential consequences for the birds' fitness., (© 2014 SETAC.)

Rationale: We describe an analytical procedure that allows sample collection and measurement of carbon isotopic composition (δ(13)C(V-PDB) value) and dissolved inorganic carbon concentration, [DIC], in aqueous samples without further manipulation post field collection. By comparing outputs from two different mass spectrometers, we quantify with the statistical rigour uncertainty associated with the estimation of an unknown measurement. This is rarely undertaken, but it is needed to understand the significance of field data and to interpret quality assurance exercises., Methods: Immediate acidification of field samples during collection in evacuated, pre-acidified vials removed the need for toxic chemicals to inhibit continued bacterial activity that might compromise isotopic and concentration measurements. Aqueous standards mimicked the sample matrix and avoided headspace fractionation corrections. Samples were analysed using continuous-flow isotope-ratio mass spectrometry, but for low DIC concentration the mass spectrometer response could be non-linear. This had to be corrected for., Results: Mass spectrometer non-linearity exists. Rather than estimating precision as the repeat analysis of an internal standard, we have adopted inverse linear calibrations to quantify the precision and 95% confidence intervals (CI) of the δ(13)C(DIC) values. The response for [DIC] estimation was always linear. For 0.05-0.5 mM DIC internal standards, however, changes in mass spectrometer linearity resulted in estimations of the precision in the δ(13)C(VPDB) value of an unknown ranging from ± 0.44‰ to ± 1.33‰ (mean values) and a mean 95% CI half-width of ±1.1-3.1‰., Conclusions: Mass spectrometer non-linearity should be considered in estimating uncertainty in measurement. Similarly, statistically robust estimates of precision and accuracy should also be adopted. Such estimations do not inhibit research advances: our consideration of small-scale spatial variability at two points on a small order river system demonstrates field data ranges larger than the precision and uncertainties. However, without such statistical quantification, exercises such as inter-lab calibrations are less meaningful., (© 2014 The Authors. Rapid Communications in Mass Spectrometry Published by John Wiley & Sons Ltd.)