Bulk δ15N Research Articles

Investigating preservation of stable isotope ratios in subfossil deep-sea proteinaceous coral skeletons as paleo-recorders of biogeochemical information over multimillennial timescales

Paleoproxy records in deep-sea proteinaceous coral skeletons can reconstruct past ocean conditions on centennial to millennial time scales. Commonly recovered subfossil specimens could potentially extend these archives through the Holocene. However, protein matrix stability and integrity of stable isotope proxies over multi-millennial timescales in such specimens have never been examined. Here we compare amino acid (AA) composition together with bulk and AA compound-specific carbon (δ13C) and nitrogen (δ15N) isotopes in live-collected and subfossil (∼9.6–11.6 kyrs BP) Kulamanamana haumeaae deep-sea coral specimens from the central Pacific to understand the effects of long-duration benthic oxic exposure on primary coral chemistry. We find large coupled shifts in bulk δ15N (∼7‰) and δ13C (∼2‰) in the outermost portion (0–10 mm) of the subfossil coral, coincident with extensive alteration of the protein matrix. Microstructural changes in skeletal texture coincide with higher C/N ratios (+0.8) and isotope-based amino acid degradation parameters (e.g. ΣV ≥ 3), indicating extensive degradation of seawater-exposed gorgonin. However, interior gorgonin (>10 mm) retained amino acid molecular compositions (with exception of major Glycine loss) and bulk and amino acid-specific isotopic values that were similar to live-collected specimens. These results indicate that compound-specific isotope analysis of amino acids can reconstruct paleo-oceanographic biogeochemical and ecosystem information in subfossil corals beyond a clear diagenetic horizon, which is easily identifiable from an evaluation of C/N ratios together with the ΣV degradation proxy.

Tracking the nitrogen cycle in a vulnerable alluvial system using a multi proxy approach: Case study Varaždin alluvial aquifer, Croatia

At high concentrations nitrate is considered a serious environmental pollutant which degrades the quality of ground and surface waters. Such high nitrate concentrations (>50 mg NO3/L) have been observed for decades in the alluvial aquifer in the Varaždin region of Croatia. Here we employ a novel cross disciplinary approach (dual isotopes, chemical, bacteria diversity and mixing modelling) to determine sources of nitrate and processes that can influence nitrate concentration within this vulnerable alluvial aquifer. Ten groundwater wells were sampled across the region and in different hydrological conditions for basic chemical, stable isotopes (δ18O-H2O, δ2H-H2O, δ15N-NO3 and δ18O-NO3), and bacterial diversity analyses. In addition, solid samples, i.e. soil samples and fertilizers were collected and analysed for bulk δ15N. The primary nitrate sources were manure, sewage, soil organic N, and ammonia fertilizers, however we observe no clear evidence to indicate that synthetic fertilizers are a major contributor to groundwater nitrate concentrations. Whilst denitrification was observed in the parts of the study area with dissolved oxygen (DO) deficiency, i.e. anoxic conditions, nitrification has been identified as the major process responsible for nitrate behaviour within the aquifer system. Our results will facilitate the creation of a conceptual model of nitrate behaviour in the study area and from this, a numerical groundwater nitrate transport model. These data, understanding of nitrate dynamics and subsequent models will be critical for future sustainable water and agricultural management of the study area.

Using tissue cysteine to predict the trophic transfer of methylmercury and selenium in lake food webs

The biomagnification of toxic methylmercury (MeHg) and selenium (Se) through aquatic food webs using nitrogen stable isotopes (δ15N) varies among ecosystems but underlying mechanisms are yet unexplained. Given the strong links between MeHg and thiol-containing amino acids and proteins containing selenocysteine, our hypothesis was that cysteine content is a better predictor of MeHg and Se transfer through lake food webs than δ15N. Food web samples were collected from six lakes in Kejimkujik National Park, Nova Scotia, Canada, and the regression slopes of log MeHg or Se versus protein-bound cysteine or bulk δ15N were compared. Across all six lakes, MeHg varied by a factor of 10 among taxa and was significantly and positively related to both cysteine (R2 = 0.65–0.80, p < 0.001) and δ15N (R2 = 0.88–0.94, p < 0.001), with no among-system differences in these slopes. In contrast, total Se concentrations varied by less than a factor of 2 among taxa in four lakes and were significantly related to cysteine in only two food webs (R2 = 0.20 & 0.37, p = 0.014 & < 0.001); however, δ15N was not a predictor of Se in any lake (p = 0.052–0.777). Overall, these novel results indicate that cysteine content predicts MeHg, and sometimes Se, across trophic levels, providing a potential mechanism for among-system differences in their biomagnification.

Calibrating bulk and amino acid δ13C and δ15N isotope ratios between bivalve soft tissue and shell for paleoecological reconstructions

Ecological isotope proxies measured in ancient bivalve shell matrix protein have great promise for paleoecological reconstruction. Compound-specific isotopes of amino acids (CSI-AA) may be an ideal tool for developing paleoecological proxies, as initial work has indicated that CSI-AA is less subject to alteration under geologic conditions relative to bulk isotope values. While CSI-AA proxies have been applied in modern bivalve soft tissues and a few shell studies, they have yet to be systematically investigated in shell matrix protein. Here, we measured stable isotope values of carbon (δ13C) and nitrogen (δ15N) in both bulk and individual amino acids (AA), comparing soft tissue and shell organic fractions to test the fidelity of a suite of ecological isotope proxies in shell. We sampled three ubiquitous bivalve species in four seasons for one year from two common coastal environments: littoral rocky intertidal and estuarine delta ecosystems. Particulate organic matter (POM) was simultaneously collected to investigate relationships between tissue types and local POM isotope signatures. The ecological proxies tested include trophic niche breadth from bulk isotopes, and baseline δ13C and δ15N values, resource contribution, and trophic level from CSI-AA data. We found niche breadth corresponded well between tissue types, but that bulk isotope values were significantly higher in shell compared to soft tissue. In the CSI-AA record, there were no differences in baseline δ13C and δ15N proxies between tissue types. While no consistent seasonal trends were observed in the δ13C record, summer bulk and baseline δ15N values were lowest in both ecosystems in both tissues. Food resource contribution estimates from shell also closely matched soft tissue, however trophic level estimates were consistently higher in shell, attributed to a systematic offset in Glutamic acid δ15N values. We therefore propose here a new mollusk-specific trophic discrimination factor, and corresponding shell isotope enrichment factor to correct for isotopic routing, both required for accurate paleoecological reconstructions.

Amphipod Isotope Composition, Condition and Reproduction in Contrasting Sediments: A Reciprocal Transfer Experiment

Eutrophication is a process that results in excessive phytoplankton blooms, which sink to the sediment and enrich the organic matter (OM). This alters the available resources to benthic organisms and may have consequences for feeding ecology and reproduction strategies of marine populations. While effects of eutrophication on biodiversity are well documented, the more subtle effects of OM on population dynamics and diet plasticity are understudied. We performed a reciprocal transfer experiment with the benthic bioindicator amphipod Monoporeia affinis from two stations in the Baltic Sea with differing sediment OM (low and high) creating four treatments (low control, low transferred, high transferred, and high control). We investigated sediment OM effects on: i) the dietary niche and organism body condition of two different life stages of M. affinis utilizing bulk stable isotopes δ13C and δ15N, and C:N ratio; and ii) M. affinis fecundity and embryo viability. There was no initial significant differences between the females from different stations in terms of δ13C, δ15N, C:N, fecundity or viable embryos. However, we found that moving females from high OM to low OM (where the low OM sediment has higher δ15N and lower δ13C) significantly depleted their 13C values, while amphipods in low OM sediment had always significantly enriched 15N regardless of female origin indicating feeding on the new sediment. Although end-of-experiment females had lower C:N than initial females, individuals in low OM sediment presented significantly higher C:N (indicating higher body condition) than those in high OM sediment. Conversely to adult amphipods, no effects of OM were seen for juveniles δ13C or δ15N, but their individual biomass was larger in high OM sediment treatments and high OM transferred to low OM sediment. Our results indicate that the low range of sediment OM tested here altered female amphipod δ13C, δ15N and C:N ratios, with those in low OM treatments having a better body condition, but those in high OM treatments had a greater reproductive success in terms of offspring biomass. Our findings suggest a tradeoff between female condition and reproduction and indicates that even relatively small levels of sediment organic enrichment will impact female condition. Our study provides valuable information useful to interpret the effects of OM on amphipod populations used as bioindicators for anthropogenic impacts.

Microplastic loads within riverine fishes and macroinvertebrates are not predictable from ecological or morphological characteristics

Microplastics are a relatively new but important form of freshwater contamination that can be ingested by a range of different species, with particle counts thought to be predictable from species ecology and morphology. Here, we report levels of microplastics in a 26 μm-5 mm size range within the macroinvertebrate and fish community of a lowland river (Dorset Stour, SW England), and test the hypothesis that counts are predictable from characteristics such as feeding guild, body length and trophic position. Macroinvertebrates (n = 257, 12 taxa) and fish (n = 418, 9 species) were collected from distinct river reaches by kick sampling and rod and line angling, respectively. Batches of whole macroinvertebrates and individual fish gastrointestinal tracts were digested with 30% hydrogen peroxide before microplastic screening and FTIR polymer confirmation on a particle subset. Particles were found in 40% of pooled macroinvertebrate batches (taxa incidences: 14–75%) and 39% of fishes (species incidences: 29–47%). Dominant particle feature categories were ≤100 μm, blue/green, fragments and fibres identified as various polyolefins. Although particle counts in macroinvertebrates were highest in Ephemeroptera (mean of 0.74 particles per individual), the relationships between particle loads, batch number and guild were all non-significant. In fishes, particle counts were not significantly related to species, stomach structure, feeding guild or body length, with spatial differences also not apparent across the catchment. Individual fish particle counts were similarly not significantly associated with their trophic positions (calculated from bulk δ15N values for a subset of fishes) and parasite load of Pomphorhynchus tereticollis. Correlations between fish and macroinvertebrate particle counts within specific river reaches were also not significant. In entirety, these results indicated although loadings of microplastic particles were relatively consistent within the two communities, they were not predictable from any of their ecological or morphological characteristics.

Nitrogen sources (NO3− vs N2 fixation) inferred from bulk δ15N values of zooplankton from the deep water region of the Gulf of Mexico

Abstract Dinitrogen (N2) fixation is an important source of new N in oceanic oligotrophic waters, but its contribution has not been quantified in the central and southern Gulf of Mexico (GM), where mesoscale anticyclonic (ACE) and cyclonic eddies (CE) influence NO3− availability for phytoplankton. We investigated the temporal and spatial variation in zooplankton δ15N values as a proxy for baseline nitrogen sources and estimated the contribution of N2 fixation and subsurface nitrate to secondary production. Copepods, euphausiids and two size classes of zooplankton were collected from 0 to 200 m during five cruises. Zooplankton δ15N values were higher during winter due to wind-driven deepening of the mixed layer, reflecting NO3− transport toward the surface. There was a higher estimated contribution of N2 fixation during the highly stratified summer months. Zooplankton δ15N values from ACEs were significantly lower than in CE, associated with a deepening of the 25.5 isopycnal and lower depth-integrated [NO3−]. N2 fixation contributed 60–80% of the N in ACEs. Some stations within the Bay of Campeche showed higher δ15N values due to NO3− transport to the euphotic layer due to a semipermanent CE, regional upwelling and river discharge. Our results highlight the importance of fixed nitrogen over an extensive region of theGM.

Amino acid nitrogen stable isotopes as biomarkers of coastal phytoplankton assemblages and food web interactions

Marine phytoplankton and zooplankton face rapidly changing environments in the face of global warming and climate change. We investigated the effect of warmer water and lower pH conditions—projected for New Zealand coastal waters at the start of the next century—on both phytoplankton and zooplankton in a 20 d mesocosm experiment to determine whether amino acid stable isotopes could be used as biomarkers of environmental change. We also assessed whether key environmental drivers, such as those linked to climate change, altered the processing of amino acids at the base of the food web. Despite changes in phytoplankton biomass and community composition, we found no significant difference in either particulate organic matter (POM) bulk or amino acid-specific δ15N values, indicating that the trophic status of POM was not significantly influenced by lower pH and warming. Threonine δ15N values were the most sensitive to changes in the phytoplankton community and showed correlations with diatoms (positive) and small flagellates (negative), demonstrating potential as a biomarker for detecting changes related to these phytoplankton groups and thus making threonine a promising indirect indicator of climate change. Finally, δ15NPhe values tracked changes in the lower food web, likely due to faster turnover times, showing its valuable role as a tracer of the nitrogen baseline, even during accelerated metabolism in zooplankton.

Sidon on the breadth of the wild sea: Movement and diet on the Mediterranean coast in the Middle Bronze Age.

ObjectivesExcavations at Sidon (Lebanon) have revealed dual identities during the Middle Bronze Age (ca. 2000–1600 BCE): a maritime port and center for local distribution, as well as a settlement with a heavy subsistence dependence on the extensive inland hinterlands. We aim to investigate residential mobility at Sidon using isotopic analyses of 112 individuals from 83 burials (20 females, 26 males, and 37 subadults). Veneration and remembrance of the dead is evident from funerary offerings in and near the tombs. With marine fish a major component in funerary offerings, we predict major marine reliance in this coastal population.Materials and methodsNew isotopic evidence of paleomobility (87Sr/86Sr, δ18O) and diet (δ13Ccarbonate) is the focus of this research. Previous bulk bone collagen δ13C and δ15N analysis is strengthened by further sampling, along with δ34S where collagen yield was sufficient.ResultsThe five non‐locals identified (8.9% of the 56 analyzed) come from constructed tombs with high‐status grave goods except for one, which was heavily disturbed in antiquity. Dietary investigation of the population confirms reliance on terrestrial resources with no significant marine input. No significant differences in diet between the sexes or burial types are present.ConclusionsAlthough Sidon was part of a growing Mediterranean network evidenced through artefactual finds, relatively low immigration is evident. While religious feasts venerating the dead may have involved significant piscine components, no appreciable marine input in diet is observed. Fish may have been reserved for the deceased or only consumed on feast days alongside the dead rather than a regular part of the Bronze Age menu.

Physiology Drives Reworking of Amino Acid δ2H and δ13C in Butterfly Tissues

Studies of animal movement and migration over large geospatial scales have long relied on natural continental-scale hydrogen isotope (δ2H) gradients in precipitation, yet the physiological processes that govern incorporation of δ2H from precipitation into plant and then herbivore tissues remain poorly understood, especially at the molecular level. Establishing a biochemical framework for the propagation of δ2H through food webs would enable us to resolve more complicated regional-scale animal movements and potentially unlock new applications for δ2H data in animal ecology and eco-physiology. Amino acid δ2H analysis offers a promising new avenue by which to establish this framework. We report bulk tissue δ2H, δ13C, and δ15N data as well as amino acid δ2H and δ13C data from three Pipevine swallowtail (Battus philenor) tissues—caterpillars, butterfly bodies, and wings—as well as their obligate plant source: pipevine leaves (Aristolochia macrophylla). Insects are often dominant herbivores in terrestrial food webs and a major food source for many higher-level consumers, so it is particularly important to understand the mechanisms that influence insect tissue δ2H values. Our data reveal extensive δ2H variation within and among individuals of a relatively simple plant-herbivore system that cannot be explained by temporal or geospatial gradients of precipitation δ2H or dietary differences. Variations in essential amino acid δ2H and δ13C indicate thatB. philenoracquire these compounds from an additional source that is isotopically distinct from pipevine leaves, potentially gut microbes. We also found multiple isotopic carryover effects associated with metamorphosis. This study emphasizes the strong influence of physiology on consumer-diet δ2H discrimination in a local population of pipevines and swallowtails and provides a template that can be broadly applied to Lepidoptera—the second most diverse insect order—and other holometabolous insects. Understanding these physiological mechanisms is critical to interpreting the large degree of δ2H variation in consumer tissues often observed at a single collection site, which has implications for using δ2H isoscapes to study animal movement. Further investigation into amino acid δ2H holds promise to elucidate how subsets of amino acids may be best utilized to address specific ecological and physiological questions for which bulk tissue δ2H is insufficient.

Rewetting and Drainage of Nutrient-Poor Peatlands Indicated by Specific Bacterial Membrane Fatty Acids and a Repeated Sampling of Stable Isotopes (δ15N, δ13C)

Peatland degradation impairs soil functions such as carbon storage and the existence of biodiversity hotspots. Therefore, and in view of the ongoing climate change, an efficient method of evaluating peatland hydrology and the success of restoration efforts is needed. To understand the role of microbial groups in biogeochemical cycling, gaseous loss and isotopic fractionation that lead to specific isotopic depth patterns (δ13C, δ15N), we integrated previously published stable isotope data with a membrane fatty acid (mFA) analysis related to various microbial groups that are known to be common in peatlands. We performed two sampling campaigns to verify the observed stable isotope depth trends in nutrient-poor peatlands in Northern Europe. Cores were taken from adjacent drained (or rewetted) and undrained sites. Fungal-derived mFA abundance was highest in the uppermost part of the drained layer. We found increasing bacterial-derived mFA concentrations with depth peaking in the middle of the drained layers, which correlates with a δ15N peak of bulk material. The results support our hypothesis that changing peatland hydrology induce a shift in microbial community and metabolism processes and is therefore also imprinted in stable isotope values. Under waterlogged conditions overall levels of microbial-derived mFAs were generally low. Drained layers showed simultaneous changes in microbial abundance and composition and depth trends in stable isotope bulk values. Bacteria, particularly acidobacteria, can be expected to dominate increased denitrification with low oxygen saturation accompanied by increased δ15N bulk values in the remaining substrate. Interestingly, cores from recent rewetted peatlands show no depth trend of δ15N in the layers grown under rewetting conditions; this is congruent with relatively low concentrations of microbial-derived mFAs. Hence, we conclude that stable isotopes, especially δ15N values, reflect changing microbial metabolic processes, which differ between drained and undrained - and especially also for recent rewetted–peatlands. As today stable isotope measurements are routine measurements, these findings enable us to get cost- and time efficient reliable information of drainage and restoration success.

Quantifying Nitrate Formation Pathways in the Equatorial Pacific Atmosphere from the GEOTRACES Peru-Tahiti Transect

This study aims to better our collective understanding of the oxidative capacity and atmospheric chemistry over the equatorial Pacific. Bulk and size-segregated filter samples were collected during the GEOTRACES Eastern Tropical Pacific transect (4.1° S, 81.9° W to 10.5° S, 152.0° W; October–December 2013) and measured for aerosol concentration and complete isotopic composition of nitrate (δ15N, δ18O, Δ17O where Δ17O = δ17O – 0.52 × δ18O). Combined size-segregated filters produced data similar to that found in bulk filter samples, and notably neither δ15N nor δ18O showed any trends based on aerosol size. Similar to other studies, NO3– is concentrated (>80%) in the coarse size fractions (>1.5 μm). Bulk aerosol concentrations ranged from 6.6 to 89.8 nmol/m3. The bulk δ15N-, δ18O-, and Δ17O-nitrate ranged from −13.1 to −3.2‰, 68.5 to 79.3‰, and 23.5 to 28.4‰, respectively. Higher δ15N values near the coast are best explained by the influence of continental sources; lower δ15N values far from the coast may be associated with chemical fractionation during long-range transport or an oceanic source. Both Δ17O and δ18O are interpreted using kinetic analysis and gas concentrations from a global atmospheric chemical transport model (GEOS-Chem), which showed that nitrate production in this environment is dominated by OH oxidation (60%) and RONO2 hydrolysis (15%). To best match the δ18O and Δ17O observations in this study, the terminal oxygen isotopic values for ozone must be higher than those suggested by available observations and/or halogen-mediated chemistry must be more important than the models currently suggest.

Environmental Regulation of the Nitrogen Supply, Mean Trophic Position, and Trophic Enrichment of Mesozooplankton in the Mekong River Plume and Southern South China Sea

AbstractThe mean trophic position (TP) of mesozooplankton largely determines how much mass and energy is available for higher trophic levels like fish. Unfortunately, the ratio of herbivores to carnivores in mesozooplankton is difficult to identify in field samples. Here, we investigated changes in the mean TP of mesozooplankton in a highly dynamic environment encompassing four distinct habitats in the southern South China Sea: the Mekong River plume, coastal upwelling region, shelf waters, and offshore oceanic waters. We used a set of variables derived from bulk and amino acid nitrogen stable isotopes from particulate organic matter and four mesozooplankton size fractions to identify changes in the nitrogen source and TP of mesozooplankton across these habitats. We found clear indications of a shift in N sources for biological production from nitrate in near‐coastal waters with shallow mixed layer depths toward an increase in diazotroph‐N inputs in oceanic waters with deep mixed layer depths where diazotrophs shaped the phytoplankton community. The N source shift was accompanied by a lengthening of the food chain (increase in the TP). This may provide further support for the connection between diazotrophy and the indirect routing of N through the marine food web. Our combined bulk and amino acid δ15N approach also allowed us to estimate the trophic enrichment (TE) of mesozooplankton across the entire regional ecosystem. When put in the context of literature values, a high TE of 5.1‰ suggested a link between ecosystem heterogeneity and the less efficient transfer of mass and energy across trophic levels.

Carbon and Nitrogen Cycle Dynamic in Continental Late-Carboniferous to Early Permian Basins of Eastern Pangea (Northeastern Massif Central, France)

Late Carboniferous to early Permian organic-rich sedimentary successions of late-orogenic continental basins from the northeastern Massif Central (France) coincide with both the Variscan mountain dismantling and the acme of the long-lasting Late Paleozoic Ice Age. Here, we investigate the carbon and nitrogen cycles in the newly dated sedimentary successions of the Decize–La Machine and Autun basins during these geodynamic and climate upheavals. The sedimentary organic matter has been analyzed through Rock-Eval pyrolysis, palynofacies and elemental and isotope geochemistry along cored-wells and outcropping sections, previously accurately defined in terms of paleo-depositional environments. Rock-Eval and palynofacies data have evidenced two origins of organic matter: a phytoplanktonic/bacterial lacustrine origin (Type I organic matter, organic δ13C values around −23.5‰), and a terrestrial origin (vascular land plants, Type III organic matter, organic δ13C values around −20‰), mixed in the deltaic-lacustrine sediments during background sedimentation (mean organic δ13C values around −22‰). Episodes of high organic matter storage, reflected by black shales and coal-bearing deposits (total organic carbon up to 20 and 70%, respectively) are also recognized in the successions, and are characterized by large negative organic carbon isotope excursions down to −29‰. We suggest that these negative isotope excursions reflect secondary processes, such as organic matter remineralization and/or secondary productivity varying under strict local controls, or possibly larger scale climate controls. At times, these negative δ13C excursions are paired with positive δ15N excursions up to +10‰, reflecting water column denitrification and anammox during lake-water stratification episodes. Together, these isotopic signals (i.e., low sedimentary organic δ13C associated with high bulk δ15N values) indicate periods of high primary productivity of surface waters, where nitrogen and carbon cycles are spatially decoupled. These local processes on the sedimentary isotope archives may partially blur our ability to directly reconstruct paleoclimate variations in such continental settings using only C and N isotopes. At last, we explore an organic δ13C-based mixing model to propose ways to disentangle autochthonous versus allochthonous origin of organic matter in lacustrine continental settings.

Large‐scale patterns of green turtle trophic ecology in the eastern Pacific Ocean

AbstractTrophic position and niche width are fundamental components of a species’ ecology, reflecting resource use, and influencing key demographic parameters such as somatic growth, maturation, and survival. Concepts about a species’ trophic niche space have important implications for local management and habitat protection, and can shed light about resilience to changing climate for species occurring over broad spatial scales. For elusive marine animals such as sea turtles, trophic niche is challenging to study, and researchers often rely on other metrics, such as isotopic niche, as a proxy. Here, stable isotope analysis (δ13C and δ15N values) was conducted on bulk skin tissue of 718 green turtles (Chelonia mydas) distributed among 16 foraging areas in the eastern Pacific from the USA to Chile, a range spanning ~10,000 km. Compound‐specific nitrogen isotope analysis of amino acids (CSIA‐AA) was applied to 21 turtles among seven sites. Isotopic niche space was determined via Bayesian ellipse area (BEA) and convex hull area (CHA) analyses of bulk isotope values, which were also used along with amino acid δ15N values to determine trophic position (TP). Substantial variability in bulk tissue δ13C and δ15N values was found within and among sites, and amino acid δ15N values confirmed this was largely due to spatial differences in baseline nitrogen isotopic compositions, but also to a lesser extent from TP differences among the green turtle foraging populations. Isotope niche space varied among sites, influenced by the diversity of prey types and relative input of terrestrial‐ vs. marine‐derived nutrients; BEAs were the most suitable measurement of isotopic niche space due to the larger influence of outlying values with the CHA approach. Amino acid isotope‐derived TP estimates that accounted for local habitat conditions (e.g., mixed seagrass/macroalgae diet) performed the best among several approaches; TP ranged from 2.3 to 3.6, which indicates an omnivorous diet for most populations. In addition to providing additional spatial resolution for δ13C and δ15N isoscapes in the eastern Pacific, especially in coastal habitats, this study further establishes CSIA‐AA as an effective tool to study the trophic ecology of sea turtles across a variety of food webs and habitats.

High trophic plasticity in the mixotrophic Mastigias papua-Symbiodiniaceae holobiont: implications for the ecology of zooxanthellate jellyfishes

The trophic ecology of mixotrophic, zooxanthellate jellyfishes potentially spans a wide spectrum between autotrophy and heterotrophy. However, their degree of trophic plasticity along this spectrum is not well known. To better characterize their trophic ecology, we sampled the zooxanthellate medusa Mastigias papua in contrasting environments and sizes in Palau (Micronesia). We characterized their trophic ecology using isotopic (bulk δ13C and δ15N), elemental (C:N ratios), and fatty acid compositions. The different trophic indicators were correlated or anti-correlated as expected (Pearson’s correlation coefficient, rP &gt; 0.5 or &lt; -0.5 in 91.1% of cases, p &lt; 0.05), indicating good agreement. The sampled M. papua were ordered in a trophic spectrum between autotrophy and heterotrophy (supported by decreasing δ13C, C:N, proportion of neutral lipid fatty acids (NLFA:TLFA), n-3:n-6 and increasing δ15N, eicosapentaenoic acid to docosahexaenoic acid ratio (EPA:DHA)). This trophic spectrum was mostly driven by sampling location with little influence of medusa size. Moreover, previous observations have shown that in a given location, the trophic ecology of M. papua can change over time. Thus, the positions on the trophic spectrum of the populations sampled here are not fixed, suggesting high trophic plasticity in M. papua. The heterotrophic end of the trophic spectrum was occupied by non-symbiotic M. papua, whereas the literature indicates that the autotrophic end of the spectrum corresponds to dominant autotrophy, where more than 100% of the carbon requirement is obtained by photosynthesis. Such high trophic plasticity has critical implications for the trophic ecology and blooming ability of zooxanthellate jellyfishes.

Cooperative feeding in common dolphins as suggested by ontogenetic patterns in δ15 N bulk and amino acids.

Understanding the effect of stage-specific traits on species feeding habits can reveal how natural selection shapes life strategies. Amino acid (AA) nitrogen stable isotopes (δ15 N) provide multiple proxies of habitat baseline values and diet that can improve our understanding of species feeding strategies relative to their animal metabolism. We evaluated the effect of body length as a proxy for life stage and sex on the feeding habits of the common dolphin Delphinus delphis delphis using δ13 C and δ15 N in bulk tissue and AAs δ15 N from skin samples collected for almost two decades. For bulk δ13 C and δ15 N data, we used SIBER analysis to compare isotopic niches by sex and life stage. For AA δ15 N data, we developed a hierarchical Bayesian model (HBM) to estimate indices of trophic status (Δ15 N and trophic position). The model reflected the natural hierarchical structure of AA data by partitioning variability into three sources: between laboratory replicates, within dolphins and among dolphins. Estimates of Δ15 N based on all trophic and source AAs were more precise for each dolphin, less variable among dolphins and on average 2.4‰ higher than indices based on single trophic (Glx) and source (Phe) AAs. Precision was further increased when information was shared among individuals through random effects or regression models. Estimates of trophic position showed similar patterns. Both Δ15 N and δ15 Nbulk isotopic niches showed no difference by sex, suggesting that males and females have similar feeding habits and may not segregate. However, lower Δ15 N values for weaning calves and smaller juveniles discriminate them from adults, whereas δ15 N bulk isotopic niches do not. A trophic discrimination factor (TDFTro-Src ) of 3.1‰ was required for reasonable estimates of trophic position for these dolphins. Together, the lack of δ15 N differences between sexes, low variation between juveniles and adults and knowledge of common dolphins' social organization support intraspecific feeding cooperation as an important strategy to feed in the highly dynamic marine environment. Our study also presents an efficient way to analyse complex AA δ15 N data using HBM to investigate foraging behaviour in long-lived marine species difficult to study in the wild.

Interactions at surface-subterranean ecotones: structure and function of food webs within spring orifices.

Spring orifices are ecotones between surface and subterranean aquatic ecosystems. Invertebrates of different origins (e.g., surface, spring obligate, and subterranean) coexist in these spatially restricted environments, potentially competing for resources. However, processes that allow for population coexistence in these presumably low resource environments are not well understood. We examined invertebrate communities at two spring complexes in Texas, USA and assessed resource use and food web structure at spring orifices using stable isotopes of carbon (δ13C) and nitrogen (δ15N). Using bulk δ13C and δ15N of organisms and potential food sources, we elucidated dietary sources and found that invertebrate communities exhibited resource partitioning and contained two main food chains (periphyton versus terrestrial organic matter [OM]). In both spring complexes, several endemic spring orifice associated and subterranean taxa derived most of their diet from terrestrial OM. Analysis of compound-specific stable isotopes (i.e., δ13C of essential amino acids, EAAs) from two co-occurring elmid species indicated that the endemic spring orifice-associated species (Heterelmis comalensis) derived > 80% of its EAAs from bacteria, whereas the widespread surface species (Microcylloepus pusillus) derived its EAAs from a more equitable mix of bacteria, fungi, and algae. We additionally calculated niche overlap among of several taxonomically related groups (aquatic beetles and amphipods) that co-occur in spring ecotones and posterior probability estimates indicated little to no niche overlap among related species. Results indicate that invertebrates at subterranean-surface aquatic ecotones are partitioning food resources and highlight the importance of connections to riparian zones for persistence of several endemic invertebrates.

Palaeopathology and amino acid δ13C analysis: Investigating pre-Columbian individuals with tuberculosis at Pica 8, northern Chile (1050-500 BP)

Bulk δ15N and δ13C values of proteinaceous tissues are being increasingly used in bioarchaeological studies to elucidate the physio-pathological status of ancient individuals. This method has not always been successful.The present study aims to explore the novel use of single amino acid carbon isotope analysis in palaeopathology by investigating the effect of a serious infectious disease, tuberculosis (TB), on the isotope composition of two collagenous tissues (tendon, rib). This is achieved by comparing the bulk and amino acid stable isotope compositions of collagenous tissues collected from human remains with and without TB-like bone lesions. The sample set comprises twelve adult individuals (males = 6, females = 6), who were buried at Pica 8, an inland oasis situated on the mid-elevation plains of northern Chile (Late Intermediate Period, ~1050-500 BP). Similarity and consistency in the diet of these individuals are explored using amino acid carbon isotope analysis of 1-cm hair segments longitudinally cut along a single hair fibre.The results show that there is a difference between collagen δ13C serine values measured in the rib collagen of five individuals presenting TB-like bone lesions (3 males and 2 females) and in those five without lesions (3 males and 2 females; control individuals). The rib collagen δ13C serine values of pathological individuals are lower (more negative) than those of the control individuals. Within the group of pathological individuals, lower δ13C serine values in rib collagen are correlated with higher (more positive) δ15N values. Since only individuals (N = 10) with similar dietary intakes were included in the statistical analyses, it appears that the δ13C serine values could be linked to altered carbon metabolism, rather than induced by dietary factors. Serine accounts for less than 3% of the total carbon in bone collagen, and thus an altered serine metabolism would be masked within the averaged bulk carbon isotope value. In future studies, it is advised to undertake stable carbon isotope analysis of non-essential amino acids as a means of characterising pathologically altered body tissues.

Segmental isotope analysis of the vertebral centrum reveals the spatiotemporal population structure of adult Japanese flounder Paralichthys olivaceus in Sendai Bay, Japan

To identify the origin of various fishes and reconstruct their migration history at the individual level, isotope analysis is a powerful alternative to artificial tagging. We used a novel individual-based methodology to reconstruct individual migratory and/or trophic shifts associated with growth based on isotopic data in the vertebral centrum of adult Japanese flounder Paralichthys olivaceus in Sendai Bay. We measured carbon and nitrogen isotope ratios (δ13C and δ15N) in muscle tissues, and conducted a segmental isotope analysis of bulk δ13C (δ13Cbulk), bulk δ15N (δ15Nbulk), and δ15N of glutamic acid (δ15NGlu) and phenylalanine (δ15NPhe) in vertebral collagen. The δ15NGlu and δ15NPhe values for bone collagen revealed an increase in trophic position and a shift to lower trophic baselines (δ15NBase: indicative of δ15N values of primary trophic sources) for most individuals. For both δ13Cbulk and δ15Nbulk, we detected significant positive correlations between values for muscle and the outermost section of vertebral collagen. A nonlinear time-series analysis of δ13Cbulk and δ15Nbulk suggested that a combination of intrinsic (the timing of migration from the nursery to deep offshore areas in juveniles) and extrinsic (habitat and/or food qualities) factors influence the isotopic chronology. A segmental isotope analysis revealed the segregation of individuals among sampling sites at all life stages and changes in trophic positions and δ15NBase values during growth. Our results suggest that the P. olivaceus population in Sendai Bay has both temporal and spatial structure. The temporal structure may be caused by variation in the timing of migration from the nursery to the deep offshore area in juveniles, and the spatial structure may be explained by individual variation in habitat preferences.