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Spatial differences in the isotope values of widely distributed marine apex consumers may reflect geographical differences in the isotopic composition of basal resources (e.g., phytoplankton) fueling food webs (bottom-up effects) or spatial differences in the trophic ecology of the taxon of interest (top-down effects). We examined spatial variation in δ 13 C and δ 15 N values from 264 South American sea lions (SASL, Otaria flavescens) of different age classes (adults, subadults and juveniles), their putative prey consisting of pelagic and benthic coastal fishes, and particulate organic matter (POM) measured from locations situated across >2300 km of the Chilean coast (between 18°42' and 39°17' S). We used generalized least squares (GLS) models to compare the form of the relationship between δ 13 C and δ 15 N and latitude between the three functional groups. Our results show that SASL from northern, central, and southern areas were isotopically distinct, with individuals from the north having lower δ 13 C and higher δ 15 N values in comparison to individuals from the south. When the relationship for each functional group was modelled individually using GLS, results indicated that for each degree of increasing latitude δ 15 N decreased on average by 0.12‰ (POM), 0.15‰ (prey), and 0.14‰ (SASL), while δ 13 C increased by 0.06‰ (POM) and 0.05‰ in both prey and SASL. We suggest that the latitudinal differences observed in SASL δ 13 C and δ 15 N values reflect baseline isotopic variation rather than marked differences in trophic ecology of these widely distributed consumers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Patterns of mother-embryo fractionation of 13 C and 15 N were assessed for their predictability across three species of batoids caught as by-catch in south-eastern Australia. Stable isotope analysis of 24 mothers and their litters revealed that isotope ratios of embryos were significantly different from their corresponding mothers and that the scale and direction of the difference varied within and across species. The range of variation across species was 3.5‰ for δ 13 C and 4‰ for δ 15 N, equivalent to a difference in trophic level. In one species (Urolophus paucimaculatus) litters could be significantly enriched or depleted in 13 C and 15 N relative to their mothers' isotope signatures. These results suggest that patterns of mother-embryo isotope fractionation vary within and between species and that these patterns may not be explained only by developmental mode. Contrasting patterns of fractionation between and within species make it difficult to adjust mother-embryo fractionation with broad-scale correction factors., (© 2022 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

Silver nanoparticles (AgNPs) are added as antibacterial and anti-odor agents to a wide range of textiles, with high potential for release into aquatic environments via domestic wastewater. Previous work demonstrating the negative impacts of AgNP exposure on periphyton production suggests benthic primary production could be reduced in aquatic ecosystems impacted by AgNP discharge. To evaluate the potential for AgNPs to alter benthic-pelagic coupling in aquatic ecosystems, tissue-stable isotope ratios of carbon and nitrogen from northern pike (Esox lucius) and yellow perch (Perca flavescens) were measured before, during, and after the addition of AgNPs to a whole-lake ecosystem, and compared to those collected from a nearby reference lake. A shift in carbon isotope ratios toward more negative values was observed in both P. flavescens and E. lucius collected from the lake where AgNPs were added, with no shift in similar magnitude observed in E. lucius from the reference lake. Consequently, Bayesian estimates of benthic energy consumed decreased by 32% for P. flavescens and by 40% for E. lucius collected after AgNP additions relative to pre-addition estimates, greater in magnitude or opposite in direction of trends observed in our reference lake. Analyses suggest no changes in fish nitrogen isotope ratios related to AgNP additions. We hypothesize that the observed reduction in littoral energy use of fish reported here is a response to AgNP settling in littoral benthic habitats-the main habitat in lakes supporting periphyton-as AgNP has been shown elsewhere to significantly reduce the rates of periphyton production. Further, our study highlights the need to broaden the scope of risk assessments for AgNPs and other emerging contaminants prone to settling to consider habitat-specific impacts on resource utilization by organisms after their release into aquatic ecosystems., (© 2023 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

Stable isotope analysis of fish muscle tissue has been used to quantify fish migratory behavior in many systems, and these chemical tracers are especially useful for euryhaline fishes that traverse significant salinities and gradients in baseline signatures. However, the removal of muscle tissues often requires lethal sampling. Fish scales may be removed non-lethally and offer potential alternatives to stable isotope ratios from muscle to indicate recent feeding histories. For this study, red drum Sciaenops ocellatus were collected in subtropical estuaries in the northwestern Gulf of Mexico and analyzed for stable isotope (𝛿 13 C and 𝛿 15 N) compositions of scale exteriors. Decalcification of scale samples was deemed unnecessary given the preservation of linearity and minimal offsets between paired decalcified and untreated scale samples. Stable isotope signatures of muscle tissue and scales were closely matched and indicated high degrees of residence within bays at fine spatial scales. This study also surveyed the existing body of literature comparing scale and muscle isotope values from the same individuals and found that the majority reports linear relationships with slopes close to unity, although species-specific offsets and intercepts vary. This work expands the body of literature indicating that scales are a viable non-lethal alternative for stable isotope assessments of dietary and habitat use histories for mobile fishes., (© 2023 Fisheries Society of the British Isles.)

Reef ecosystems are characterized by highly heterogenous habitats and functionally diverse fish communities. Few studies have examined how functional diversity differs among habitats within these communities, i.e., species associated with a specific habitat may have similar trophic ecologies meaning that the functional diversity within the community is driven by habitat diversity or, conversely, high functional diversity within each habitat would indicate that resource segregation also occurs at the habitat level. We used stable isotope ratios of carbon and nitrogen to estimate trophic position, resource use and ontogenetic niche shifts of 15 reef fishes associated with four distinct habitat types (cryptobenthic, epibenthic sand, epibenthic rock and hyperbenthic) on the Croatian coast of the Adriatic Sea. Trophic ecology was quite similar across fish assemblages, but there was strong evidence of niche segregation among fish species within each assemblage showing high functional diversity within each microhabitat. The sampled fish community contained benthic and pelagic resource users, along with multiple intermediate generalists. Consumer stable isotope ratios revealed considerable interspecific variation in resource use among fishes within each habitat type. The cryptobenthic fishes were a notable exception to this trend with the narrow range of resource use values, indicating reliance of these species on a single resource. The greatest diversity of trophic positions within a guild was observed in cryptobenthic and rock-associated epibenthic fishes. The majority of observed ontogenetic variation in studied fish species reflected an increase in benthic resource use and trophic position. However, the degree of ontogenetic variation in trophic ecology of studied species, if present, was generally low, showing no dramatic change in the ecology of any species. The size structuring among guilds was considerable, with cryptobenthic fishes the smallest on average and hyperbenthic fishes the largest, despite guilds having similar ranges of trophic positions., (© 2022 Fisheries Society of the British Isles.)

The combined use of otolith chemistry and tissue isotopes has the potential to reveal movements, habitat associations and food web interactions at a variety of spatial and temporal scales. Here, a combination of otolith Ba:Ca life-history transects with muscle tissue δ 13 C and δ 15 N values has been used to assess habitat use and oligohaline residence in red drum Sciaenops ocellatus in subtropical estuaries in the north-western Gulf of Mexico. Tissue isotopes were distinct among capture locations, particularly between bays with differing proximities to freshwater inflow sources. Otolith edge Ba:Ca values and tissue δ 13 C values were not correlated. These results indicated that fish were neither residing in nor feeding in oligohaline waters for significant periods of time within the tissue turnover window of several months prior to capture. Nonetheless, spatial differences in tissue isotope values indicated limited mixing among bays and relatively high site fidelity during estuarine occupancy. Lifetime otolith Ba:Ca transects revealed individual variability in the magnitude of residence in oligohaline waters. Using a mean oligohaline occupancy threshold, an estimated 82% of individuals used oligohaline waters at some point in their life. Nonetheless, 66% of individuals spent <20% of their life histories in oligohaline waters, suggesting intermittent and infrequent excursions into low salinity waters. Finally, a literature survey identified 56 peer-reviewed publications using combinations of otolith chemistry and tissue stable isotope ratios with a wide range of marker pairings and study aims. The diversity of ecological questions that can be asked with the combined use of these two approaches will provide valuable insight into fish ecology., (© 2022 Fisheries Society of the British Isles.)

Seasonal differences in the availability of resources potentially result in the food web architecture also varying through time. Stable isotope analyses are a logistically simple but powerful tool for inferring trophic interactions and food web structure, but relatively few studies quantify seasonal variations in the food web structure or nutrient flux across multiple trophic levels. We determined the temporal dynamics in stable isotope compositions (carbon, nitrogen and sulphur) of a fish community from a highly seasonal, temperate estuary sampled monthly over a full annual cycle. Sulphur isotope values in fish tissues discriminated among consumers exploiting pelagic and benthic resources but showed no seasonal variation. This implied limited change in the relative consumption of pelagic and benthic resources by the fish community over the study period despite major seasonal changes in phytoplankton biomass. Conversely, carbon and nitrogen isotope values exhibited seasonality marked by the commencement of the spring phytoplankton bloom and peak chlorophyll concentration, with δ 13 C values following expected trends in phytoplankton growth physiology and variation in δ 15 N values coinciding with changes in major nitrogen sources to plankton between nitrate and ammonium. Isotope shifts in fish muscle were detected within 2 weeks of the peak spring phytoplankton bloom, suggesting a rapid trophic transfer of carbon and nitrogen along food chains within the estuarine food web during periods of high production. Therefore we caution against the assumption that temporal averaging effectively dampens isotopic variability in tissues of higher trophic-level animals in highly dynamic ecosystems, such as temperate estuaries. This work highlights how stable isotope analyses can be combined with environmental data to gain a broader understanding of ecosystem functioning, while emphasising the need for temporally appropriate sampling in stable isotope-based studies., (© 2022 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

Although considered a hallmark in early ontogeny, weaning from breastmilk is difficult to monitor in wild primates and weaning ages remain unknown for wild bonobos (Pan Paniscus). Here, we calculated inter-birth intervals from demographic data and measured the isotopic offsets (Δ 15 N and Δ 13 C) between mother (n = 17) and offspring (n = 28) fecal sample pairs (n = 131, total n = 246) in the LuiKotale bonobos to assess nutritional weaning for the first time. We tested the effects of infant age, female parity, and sibling competition on Δ 15 N and Δ 13 C values. We found bonobo inter-birth intervals ranging from 2.2 to 7.3 years (x̄ = 4.7 ± 1.3 years) at LuiKotale. The Δ 15 N and Δ 13 C values suggested nutritional weaning on average by 6.6 and 7.0 years of age respectively, considerably exceeding weaning ages reported for chimpanzees (P. troglodytes) using the same approach. Our Δ 13 C data suggested that the number of offspring present affected nursing, with first-time mothers nursing more and possibly longer. The Δ 15 N and Δ 13 C values decreased with the arrival of the next sibling, suggesting sibling competition reduces milk access. Nevertheless, offspring may continue nursing 2.5-3 years after the birth of the next sibling, corresponding well with observations on low infant mortality. In conclusion, bonobo mothers provide remarkably enduring materna l support in the form of nursing concurrently to several offspring., (© 2024 Wiley Periodicals LLC.)

Trophic niche is the result of the long-term evolution of species and can reflect the pressures they experience in habitats. The whole-genome sequencing of giant pandas (Ailuropoda melanoleuca) has indicated that populations distributed in Qinling (QIN) and non-QIN probably diverged 300 ky ago. Although many studies regarding foraging strategy, habitat preference, and niche partition have been conducted on these populations, there is still a lack of precise quantification of trophic niches. Here, we calculated and compared isotopic trophic niche widths of giant pandas from Sichuan (SC) and QIN populations by measuring carbon and nitrogen isotopes of their hairs; combined with data from sympatric mammals, we explored the relative trophic positions of giant pandas in the ecosystem, respectively. The Stable Isotope Bayesian Ellipses in R (SIBER) model results showed the trophic niche width of QIN pandas was 3.44‰ 2 , which was significantly bigger than those of the SC population (2.03‰ 2 ), with an overlapping about 1.45‰ 2 ; and they both occupied a unique position in the context, almost one trophic level lower than herbivores. Then, we determined the isotopic ratios of the main foods from the habitats of these pandas; the results suggested that the isotopic difference between bamboo shoots and other parts plus the various feeding selections of pandas on them accounted for pandas' trophic niche widths. We considered the higher nutrition availability and digestible food resources giving QIN pandas a wider trophic niche than pandas from SC. This conclusion provides a new insight into the resource use and trophic ecology of giant pandas and is important to develop refined management plans for the two populations., (© 2023 International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

Patterns of feeding and growth of Atlantic salmon Salmo salar L. in the marine environment are critical to understanding how observed declines in recruitment may reflect warming or other oceanic drivers. The isotopic composition of scales can provide insight into differences in marine feeding location and possibly temperature regime. The authors used archived scale samples to measure δ 13 C and δ 15 N deposited in the scales of one sea-winter (1SW) salmon during their last season of growth at sea before they returned to five Irish rivers. δ 13 C values were related statistically to observed salmon body condition (Fulton's K), and fish with higher δ 13 C values tended to show significantly better condition. In contrast, δ 15 N values were negatively related to body condition. There was no important effect on condition of length at smolt migration, and the effect of duration of marine residence varied among rivers. It is likely that δ 13 C values partly reflected ambient ocean temperature and recent marine feeding environment before return migration, such that the observed relationship between higher δ 13 C values and increased body condition may express an advantage for adult fish feeding in warmer, potentially closer, waters. If greater body condition influences fitness, then a changing temperature regime in the Northeast Atlantic may drive shifts in salmon survival and reproduction. This study provides evidence that there is spatial and trophic variation at sea between salmon from rivers of origin that are located relatively close to each other, with potential consequences for body condition and, consequently, fitness and life history; this suggests that salmon populations from geographically proximate rivers within regions may exhibit differential responses to ocean-scale climatic changes across the Northeast Atlantic., (© 2021 Fisheries Society of the British Isles.)

Pulsed flows following heavy monsoon rain events alter riverine food webs, but their impact on headwater stream food webs across the continuum from forested canopy to open agricultural land use remains unclear. We investigated carbon and nitrogen stable isotopes in macroinvertebrates and fish in two tributaries of the Suyeung River, Korea, before and after heavy monsoon rains to assess changes in community trophic niches. Basal resources (leaf litter and biofilms) exhibited consistent δ 13 C and δ 15 N values across seasons, with biofilms showing higher δ 13 C values. δ 15 N values increased from forested to agricultural reaches, indicating varied nutrient inputs. Consumer isotope values remained stable over time but varied longitudinally, reflecting reliance on local resources. Trophic niches differed between watershed locations but overlapped seasonally. Despite a decrease in consumer δ 13 C ranges after heavy rainfalls, variations in their δ 15 N ranges and the ellipse centroid (SEAc) of isotopic niches between sites resulted in broadly consistent SEAc across locations and seasons. This indicates limited evidence for directional reshaping of food-web properties across channel reaches following monsoon rains. Downstream isotopic shifts suggest substantial agricultural influences on food webs. Overall, our findings highlight that monsoon rains may have minimal effects on the community trophic niches of stream food webs., (© 2024. The Author(s).)

Rationale: Natural variations in the abundance of the stable isotopes of nitrogen (δ 15 N) and carbon (δ 13 C) offer valuable insights into metabolic fluxes. In the wake of strong interest in cancer metabolism, recent research has revealed δ 15 N and δ 13 C variations in cancerous compared to non-cancerous tissues and cell lines. However, our understanding of natural isotopic variations in cultured mammalian cells, particularly in relation to metabolism, remains limited. This study aims to start addressing this gap using metabolic modulations in cells cultured under controlled conditions., Methods: Prostate cancer cells (PC3) were cultured in different conditions and their δ 15 N and δ 13 C were measured using isotope ratio mass spectrometry. Isotopic variations during successive cell culture passages were assessed and two widely used cell culture media (RPMI and DMEM) were compared. Metabolism was modulated through glutamine deprivation and hypoxia., Results: Successive cell culture passages generally resulted in reproducible δ 15 N and δ 13 C values. The impact of culture medium composition on δ 15 N and δ 13 C of the cells highlights the importance of maintaining a consistent medium composition across conditions whenever possible. Glutamine deprivation and hypoxia induced a lower δ 13 C in bulk cell samples, with only the former affecting δ 15 N. Gaps between theory and experiments were bridged and the lessons learned throughout the process are provided., Conclusions: Exposing cultured cancer cells to hypoxia allowed us to further investigate the relation between metabolic modulations and natural isotopic variations, while mitigating the confounding impact of changing culture medium composition. This study highlights the potential of natural δ 13 C variations for studying substrate fluxes and nutrient allocation in reproducible culture conditions. Considering cell yield and culture medium composition is pivotal to the success of this approach., (© 2024 The Author(s). Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Rationale: Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (δ 13 C) and nitrogen (δ 15 N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone., Methods: Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the δ 13 C and δ 15 N values of powdered killer whale (Orcinus orca) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics., Results: No significant differences in the δ 15 N values were observed for any of the experimental protocols. However, the δ 13 C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics., Conclusions: If only the δ 15 N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ 13 C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally., (© 2024 The Author(s). Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

The economic, socio-political, and cultural significance of camelids in the Andean region is well-recognized, yet an understanding of their management evolution over pre-historical periods remains limited. This study aims to fill this gap by conducting the first cross-regional assessment of camelid pastoralism in Peru from 900 BCE to 1470 CE, using stable carbon and nitrogen isotopic compositions from the bone collagen and fibers of 577archaeological camelids across 21 sites. This research investigates the spatio-temporal shifts in camelid dietary habits, focusing on how the rise of intensive agriculture may have influenced change and led to the evolution of distinct roles for camelids in coastal versus non-coastal Andean economies. Our analysis indicates an increase in δ13C values over time on the coast, suggesting a shift towards maize-based camelid diets. Conversely, δ13C values decrease over time in highland environments, suggesting camelids consumed relatively more wild C3 forage and/or cultivated crops such as tubers. The study also reveals a significant positive relationship between latitude and δ15N values, suggesting increasing environmental aridity enriches δ15N in bone collagen. After controlling for this latitudinal effect, we observe a rise in δ15N values in both coastal and non-coastal camelids, suggesting that in later periods camelids may have been foddered in agricultural fields that were enriched with guano or dung fertilizer used to intensify production. Importantly, this research uncovers a distinct dietary divergence between coastal and inland camelids. The observed divergence in diets suggests contrasting socio-economic uses of camelids, where coastal camelids were predominantly involved in ceremonial and political activities, while those in non-coastal areas were crucial to the subsistence economy., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Noe 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.)

Uniform enrichment of 15 N and 13 C in proteins is commonly employed for 2D heteronuclear NMR measurements of the three-dimensional protein structure. Achieving a high degree of enrichment of both elements is important for obtaining high quality data. Uniform labeling of proteins and glycoproteins expressed in higher organisms (yeast or mammalian cell lines) is more challenging than expression in Escherichia coli , a prokaryote that grows on simple, chemically defined media but does not provide appropriate eukaryotic modifications. It is difficult to achieve complete incorporation of both heavy isotopes, and quality control measures are important for quantitating the level of their enrichment. Mass spectrometry measurements of the isotopic distribution of the intact protein or its proteolytic fragments provide the means to assess the enrichment level. A mass accuracy of 1 ppm or better is shown to be required to distinguish the correct combination of 13 C and 15 N enrichment due to subtle shifts in peak centroids with differences in the underlying, but unresolved, isotopic fine structure. A simple computer program was developed to optimize the fitting of experimental isotope patterns to statistically derived distributions. This method can determine the isotopic abundance from isotope patterns and isotopologue masses in conventional MS data for peptides, intact proteins, and glycans. For this purpose, MATLAB's isotope simulator, isotopicdist, has been modified to permit the variation of 15 N and 13 C enrichment levels and to perform a two-dimensional grid search of enrichment levels of both isotopes. We also incorporated an alternate isotope simulator, js-emass, into MATLAB for use in the same fitting program. Herein we benchmark this technique on natural abundance ubiquitin and uniformly [ 15 N, 13 C]-labeled ubiquitin at both the intact and peptide level, outline considerations for data quality and mass accuracy, and report several improvements we have made to the previously reported analysis of the [ 15 N, 13 C]-enriched human IgG Fc domain, a glycoprotein that has been expressed in Saccharomyces cerevisiae .

Rapid coagulation of reptile blood often hinders its use in studies in remote and difficult-to-access areas, necessitating chemical preservation. Therefore, understanding the potential effects of anticoagulants on the isotopic compositions of blood is essential to avoid issues in interpreting the results for ecological studies. In this study we aimed to verify whether the storage time of the blood tissue in anticoagulants can influence its isotopic compositions of the broad-snouted caiman ( Caiman latirostris ), an ectothermic top predator from eastern South America. Blood samples were obtained from ten adult females of C. latirostris from a commercial breeding facility in 2015. Samples were stored in vials containing ethylenediaminetetraacetic acid (EDTA) and sodium heparin (SH) and centrifuged after 2 and 8 h to separate red blood cells and plasma. No effect of time was found on the δ 13 C and δ 15 N of whole blood, plasma, and red blood cells in contact with the two types of anticoagulants, EDTA and SH. The findings have practical implications for researchers in this field, as they suggest that anticoagulants can be used effectively for at least eight hours under refrigeration.

Estuaries exhibit high physicochemical variability and the properties of estuaries and the constituent segments are not yet systematically understood. This study aims to reveal the spatial heterogeneity of predominant organic sources using carbon and nitrogen stable isotope ratios (δ 13 C and δ 15 N, respectively) of Japanese eels (Anguilla japonica), one of the ideal natural samplers. In the Miyakoda River, Japan, our study site, the effectiveness of eel δ 13 C and δ 15 N values as standard indices of predominant organic sources was tested by employing the River Continuum Concept. This study then extended the application of these indices to the estuary, i.e., the Hamana Lake system, into which the Miyakoda River flows. Once in the upper estuary, eel δ 13 C and δ 15 N values became high, with the latter peaking in this river-estuary continuum, indicating that artificial labile subsidies (nutrients and organic matter) with high δ 15 N values were rapidly assimilated into the food web. Eel δ 15 N values decreased again in the middle estuary. Nevertheless, the influence of terrestrial organic subsidies extended into this segment, as evidenced by the low δ 13 C values of eels. These results suggest that refractory organic matter with low δ 15 N values, such as plant-derived ones, is slowly assimilated into the food web in the downstream estuarine segments. The higher δ 13 C values in the lower estuary suggested that the contribution of eelgrass or macroalgae occurred in addition to benthic microalgae. Thus, our results emphasize the need to consider the multiple energy flows to understand the estuary as a continuum., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The addition of Navicula sp. to shrimp nurseries can improve the growth of Penaeus vannamei reared in biofloc systems. However, the contribution of microalgae to the biofloc formation and the effective contribution to shrimp nutrition remain unknown. In this study, Navicula sp. was added to biofloc nursery systems of P. vannamei at distinct time frequencies for evaluating its nutritional contribution to shrimp growth. Nursery rearing was carried out in bioflocs for 35 days at a stocking density of 3000 post-larvae m -3 . Shrimp were fed using a commercial feed plus fresh culture of Navicula sp. at different frequencies: no addition of Navicula sp. (WN - control), the addition of 10 × 10 4 cells mL -1 of the diatom every 5, 10 and 15 days (N5, N10 and N15, respectively). Food sources relative contribution to P. vannamei development was estimated using a Bayesian mixture model. The isotopic discrimination factor (Δ 15 N and Δ 13 C) for each food source was determined experimentally. After 35 days of culture, survival (∼93 %) was similar across all treatments but there was a significant difference in weight gain and feed conversion ratio. The N10 treatment (0.50 ± 0.05 g, 0.99 ± 0.01) exhibited better growth parameters when compared to the WN treatment (0.33 ± 0.07 g, 11.46 ± 0.30). Biofloc was the food source most assimilated by shrimp followed by Navicula sp. and commercial feed. Contribution of Navicula sp. was higher in the N5 treatment. In the treatments with diatom addition, an inverse correlation was observed between the relative contributions of biofloc and Navicula sp., indicating that Navicula sp. is not in the biofloc composition, but it is directly consumed by P. vannamei post-larvae. Biofloc and Navicula sp. exhibited larger contributions to the growth of shrimp, reinforcing the importance of natural food sources to the aquaculture of P. vannamei post-larvae.

Knowledge of predator-prey interactions is key in ecological studies and understanding ecosystem function, yet this is still poorly explored in the deep-sea environment. Carbon (δ 13 C: 13 C/ 12 C) and nitrogen (δ 15 N: 15 N/ 14 N) stable isotope ratios of a deep-diving species, the short-finned pilot whale (Globicephala macrorhynchus), were used to explore knowledge gaps on its ecological niche and foraging habitats in the Webbnesia marine ecoregion (Tenerife Island, n = 27 animals vs. Madeira, n = 31; 500 km apart) where animals display distinct levels of site fidelity. Specifically, we tested whether intraspecific isotopic variation results from differences between geographic areas (due to possible foraging plasticity between regions), sexes, and/or years (2015-2020) using Generalized Linear Models. In general, significant differences (p < 0.05) were found in the stable isotope profiles of pilot whales between the two archipelagos, which were also reflected in their isotopic niche. The higher mean and wider range of δ 15 N values in Tenerife suggest that pilot whales consume prey of higher trophic levels and more diverse than Madeira. The higher mean and wider range of δ 13 C values in Madeira suggest that in that island, pilot whales rely on prey from more diverse habitats. There was significant variation between some years, but not between sexes. Finally, we discuss pilot whales' foraging strategies worldwide and infer the reliance on benthic or benthopelagic food sources in the Webbnesia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: To evaluate the potential of compound-specific isotope analysis of amino acids (CSIA-AA) for investigating infant feeding practices, we conducted a long-term study that compared infant and maternal amino acid (AA) nitrogen isotope ratios., Materials and Methods: Fingernail samples were collected from a single mother-infant dyad over 19 months postpartum. Carbon and nitrogen stable isotope ratios were measured in the bulk keratin of the fingernail samples. Selected samples were then hydrolyzed and derivatized for compound-specific nitrogen isotope analysis of keratin AAs., Results: As in previous studies, infant bulk keratin nitrogen isotope values increased during exclusive breastfeeding and fell with the introduction of complementary foods and eventual cessation of breastfeeding. Infant trophic AAs had elevated nitrogen isotope values relative to the mother, while the source AAs were similar between the mother and infant. Proline and threonine appeared to track the presence of human milk in the infant's diet as the isotopic composition of these AAs remained offset from maternal isotope values until the cessation of breastfeeding., Discussion: Although CSIA-AA is costly and labor intensive, it appears to hold potential for estimating the duration of breastfeeding, even after the introduction of complementary foods. Through the analysis of a full suite of AAs, it may also yield insights into infant physiology and AA synthesis., (© 2024 The Author(s). American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

Golden cuttlefish play a significant role in the food web of the East and Yellow Seas and are a valuable fishery resource in Chinese coastal waters. Samples of golden cuttlefish were obtained from the northern East China Sea between September 2021 and March 2022, and stable isotope methods were utilized in this study to examine the variations in the forage ecology of golden cuttlefish. Our findings reveal dynamic shifts in carbon and nitrogen stable isotopes (δ 13 C and δ 15 N), highlighting intricate foraging strategies tailored to growth and environmental changes. A notable trend emerges: an initial growth-linked rise in δ 13 C and δ 15 N enrichment, followed by seasonal fluctuations mirroring seasonal food availability. The ontogenetic niche evolution displays striking habitat shifts and trophic level escalation in small mantle length stages, transitioning to niche overlap and subtle trophic shifts later on. Sex-specific differences emerge, with females occupying higher trophic levels than males in most samples. This comprehensive study underscores the complexity and adaptability of golden cuttlefish feeding ecology, inviting further inquiry into their intricate relationships within the marine ecosystem., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Rationale: Understanding the migration of marine animals is hindered by the limitations of traditional tracking methods. It is therefore crucial to develop alternative methods. Stable isotope-based tracking has proven useful for this task, although it requires detailed isoscapes in the focal area. Here, we present predator-based isoscapes of the coastal zone of the Patagonian Shelf Large Marine Ecosystem (PSLME), which offers a novel tool for geolocation., Methods: Whole-blood samples from breeding Magellanic penguins nesting at 11 colonies were used to create δ 15 N and δ 13 C isoscapes. Isotopic values were assigned to random positions inside their corresponding foraging area. Spatial analysis and data interpolation resulted in δ 15 N and δ 13 C isoscapes for the coastal zone of the PSLME, which were validated through cross-validation., Results: The isoscapes mean standard error ranged from 0.05 to 0.41 for δ 15 N and from 0.07 to 0.3 for δ 13 C, similar to the error range of the mass spectrometer used for measuring isotope ratios. Predictive surfaces reflected the latitudinal trends, with δ 13 C and δ 15 N values increasing northwards. δ 13 C values showed a strong latitudinal gradient, while δ 15 N values had two distinct domains, with higher values in the north. The error surface indicated the highest certainty within 130 km from the shore and within the reported Magellanic penguin foraging areas., Conclusions: Both isoscapes revealed strong spatial variation. The δ 13 C isoscape showed a latitudinal gradient, consistent with patterns in other oceans. The δ 15 N isoscape clearly separated northern and southern colonies, likely influenced by nitrogen sources. The error obtained fell within the measurement error ranges, adding credibility to the models., (© 2024 John Wiley & Sons Ltd.)

Reservoir construction promotes many environmental impacts, including the enhancement of mercury concentrations in fish. The processes that can influence mercury concentrations in fish in Amazonian reservoirs are still little explored in depth, especially when we consider the possible particularities of the ecosystems in question. This study aims to investigate how mercury concentrations in fish could be influenced by the Tucuruí dam, considering possible changes in their feeding and trophic position according to the dam position (up or downstream). Fish were sampled upstream and downstream of the Tucuruí reservoir, and total mercury (THg) and stable isotopes of carbon and nitrogen (δ 15 N and δ 13 C) were measured in muscles. We observed three different Hg bioaccumulation patterns influenced by the dam. These differences occurred due to species trophic niche changes corroborated by the isotope analysis. Higher THg concentrations downstream compared to those upstream ones were only observed for Geophagus proximus. On the contrary, Plagioscion squamosissimus, from downstream, presented lower concentrations than upstream ones. The isotopic niche of these two species presented different changes according to the sampled site. THg biomagnification was higher upstream compared to downstream, considering that the regression slope was approximately two times higher upstream versus downstream. THg concentrations in fish were explained by the differences in their feeding habits according to their location in relation to the dam. The difference in THg biomagnification was able to reflect differences in structure of the food web chain in ecosystems under the dam's influence., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Understanding the mechanism by which non-native fish species integrate into native communities is crucial for evaluating the possibility of their establishment success. The genus Pangasianodon, comprising Pangasianodon gigas and Pangasianodon hypophthalmus, has been introduced into reservoirs, which are non-native habitats, for fishery stock enhancement. P. gigas and P. hypophthalmus often successfully establish and co-occur in several Thai reservoirs, but there is little information on differences in food resource use between the two species. To investigate the trophic niche width of P. gigas and P. hypophthalmus in a Thai reservoir, we conducted stable carbon and nitrogen ratio (δ 13 C and δ 15 N) analyses. We examined the degree of individual specialization in both species using the δ 13 C and δ 15 N values of muscle and liver tissues, which provides long- and short-term diet information. The isotopic niches did not overlap between P. gigas and P. hypophthalmus. The δ 15 N value of P. gigas was significantly higher than that of P. hypophthalmus, whereas the δ 13 C value did not significantly differ between the two species. The isotopic niche sizes were larger in P. hypophthalmus than in P. gigas. Individual specialization was observed in P. hypophthalmus but not in P. gigas, indicating that intraspecific variation in food resource use was larger in P. hypophthalmus compared to P. gigas. These findings suggest that trophic niche partitioning was one of the factors facilitating the establishment success of P. gigas and P. hypophthalmus in a reservoir, but the establishment process may differ between the two species., (© 2024 Fisheries Society of the British Isles.)

To evaluate the nutritional modes of orchids associated with 'rhizoctonia' fungi, analyses of hydrogen (δ 2 H), carbon (δ 13 C), and nitrogen (δ 15 N) stable isotope ratios are usually adopted. However, previous studies have not fully accounted for exchangeable hydrogens, which could affect these evaluations. Here, we performed standard δ 13 C, δ 15 N, and δ 2 H analyses on bulk samples. Additionally, we conducted δ 2 H analysis on α-cellulose and cellulose nitrate samples to investigate whether the heterogeneity of exchangeable hydrogens among plant species influences the assessment of nutritional modes. The δ 2 H of orchids were consistently higher than those of surrounding autotrophic plants, irrespective of the three pretreatments. Although the rhizoctonia-associated orchid exhibited lower δ 13 C, its δ 2 H was higher than those of the autotrophs. Notably, among all response variables, δ 15 N and δ 2 H exhibited high abilities for discriminating the nutritional modes of rhizoctonia-associated orchids. These results indicate that a time-efficient bulk sample analysis is an effective method for evaluating plant nutritional modes, as the heterogeneity of exchangeable hydrogens does not significantly impact the estimation. Using δ 15 N and δ 2 H benefits the assessment of partial mycoheterotrophy among rhizoctonia-associated orchids., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Objectives: Justinian plague and its subsequent outbreaks were major events influencing Early Medieval Europe. One of the affected communities was the population of Saint-Doulchard in France, where plague victim burials were concentrated in a cemetery enclosure ditch. This study aimed to obtain more information about their life-histories using the tools of isotope analysis., Materials and Methods: Dietary analysis using carbon and nitrogen isotopes was conducted on 97 individuals buried at Le Pressoir in Saint-Doulchard, with 36 of those originating from the enclosure ditch. This sample set includes all individuals analyzed for plague DNA in a previous study. Mobility analysis using strontium isotope analysis supplements the dietary study, with 47 analyzed humans. The results are supported by a reference sample set of 31 animal specimens for dietary analysis and 9 for mobility analysis., Results: The dietary analysis results showed significantly different dietary behavior in individuals from the ditch burials, with better access to higher quality foods richer in animal protein. 87 Sr/ 86 Sr ratios are similar for both studied groups and indicate a shared or similar area of origin., Discussion: The results suggest that the ditch burials contain an urban population from the nearby city of Bourges, which overall had a better diet than the rural population from Saint-Doulchard. It is implied that city's population might have been subjected to high mortality rates during the plague outbreak(s), which led to their interment in nearby rural cemeteries., (© 2024 The Author(s). American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

Rationale: Carbon (δ 13 C) and nitrogen (δ 15 N) stable isotope analysis is a powerful tool to investigate diverse questions in fish ecology, such as their trophic position or migration strategies. These questions appear particularly important to protect endangered European eel. However, elevated lipid content in eel muscle can bias δ 13 C values, as lipids are 13 C-depleted compared to proteins and carbohydrates., Methods: We measured δ 13 C and δ 15 N values of bulk and lipid-free samples of eel muscle. Lipid-free samples were obtained after the extraction of lipids with cyclohexane. Lipid-corrected δ 13 C values, using five different mathematical equations based on bulk δ 13 C values, were compared to lipid-free δ 13 C values. We also evaluated the effect of lipid extraction on δ 15 N values. The analyses were based on linear regression performed on 333 individuals captured in nine lagoons and four rivers., Results: Independently to the capture site or habitat (river or lagoon), the predicted lipid-corrected δ 13 C values were highly consistent with the measured lipid-free δ 13 C values (R 2  > 0.90). The application of specific equations for each habitat or capture site only slightly increases these R 2 (1.5% or less). The lipid extraction treatment significantly decreased by 0.2‰ the δ 15 N values compared to bulk samples., Conclusions: Given the excellent prediction of mathematical equations and the small decrease of δ 15 N values after lipids extraction, we propose to use mathematical correction to estimate δ 13 C values of eel muscle. As the habitats or sites did not strongly influence the results, the coefficients from our study can be applied to other studies on European eel., (© 2024 John Wiley & Sons Ltd.)

Rationale: Hair is known to preserve diet history and other physiological information during its growth period and is often used in chemical analyses. However, the growth patterns and turnover of hair vary according to the animal species or habitat, so understanding these patterns in the target animal is necessary for interpreting the results of hair analyses. In this study, we aimed to elucidate the growth pattern and dietary information of winter coat hair in captive sika deer (Cervus nippon)., Methods: Experiments involving hair-staining and shaving were conducted to elucidate the growth pattern of sika deer hair. A diet-switching experiment was conducted to ascertain what dietary information is reflected in the carbon and nitrogen stable isotope values of the deer winter coat. Hair samples collected from each body site (head, shoulder, back, and hip) were analyzed using an elemental analyser interfaced with an isotope-ratio mass spectrometer., Results: The winter coat grows from early September to early November, and then stops after that. During the growth period of the winter coat, the hair of the shoulder and back grew at a constant rate. The carbon and nitrogen stable isotope values of hair reflected the deer's feeding history during hair growth, but there seemed to be a time lag in the hip hair., Conclusions: The results suggest that the guard hair of the shoulder is suitable for hair analysis in sika deer. The obtained information on growth patterns and isotopic change of hair from captive sika deer according to diet be useful for interpreting the results of future analyses using hair samples of wild deer., (© 2024 John Wiley & Sons Ltd.)

Climate variability impacts the structure and functioning of marine ecosystems and can trigger behavioural responses in organisms. We investigated whether such variability modulates diet and migration in the North Atlantic fin whale (Balaenoptera physalus). To reconstruct the dietary and migratory behaviours over time, we conducted stable isotope analysis of nitrogen (δ 15 N) and carbon (δ 13 C) along baleen plates from 29 fin whales sampled off southwestern (SW) Iceland in summer. We estimated a baleen growth rate of 16.1 ± 2.5 cm per year from the stable isotope oscillations observed along the baleens. We also assigned a deposition date for each baleen segment, thus obtaining isotopic sequential time series. We then assessed the potential association of these time series with the main climate patterns of the North Atlantic basin. Baleen δ 15 N and δ 13 C values are associated with the North Atlantic Oscillation (NAO) and the Atlantic Multidecadal Oscillation (AMO). During high AMO and low NAO periods, which tend to decrease krill abundance, there is an increase in both the mean and standard deviation of baleen δ 15 N values, suggesting that fin whales shift to higher trophic resources and expand their dietary niche. Additionally, high AMO periods, which relate to positive temperature anomalies, lead to a decrease in baleen δ 13 C values, suggesting that fin whales adjust their migratory routes and destinations towards higher latitudes. Significant variation in isotopic niche width between years also reflected these dietary and migratory behavioural shifts. This highlights the plasticity of the North Atlantic fin whale behaviour, a trait likely to strengthen the resilience of the species within the current context of rapid and intense climate variability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Objectives: Based on the analysis of stable carbon and nitrogen isotopes of bone collagen, stable carbon isotopes of bone apatite and an extensive AMS dating series (~10,000-299 years cal BP), the human paleodiets of 34 individuals from the Central Pampean Dunefields (Argentina, South America) are evaluated., Materials and Methods: These data are interpreted from the isotopic ecology of animals with archaeofaunal evidence of consumption and isotopic models of human diet. Multivariate carbon and nitrogen stable isotope model and Bayesian stable isotope ellipses were used to interpret human diets., Results: Analysis of isotopic values indicates intake of enriched lipids and/or carbohydrates in relation to the proteins consumed throughout the Holocene. The isotopic values of Middle Holocene humans in relation to the values of exploited resources point out that individuals obtained protein mainly from guanaco. Subsequently, there was an increase in the human breadth diet during the Late Holocene, with a greater relevance of small prey of high trophic levels and vegetables. This contrasts with zooarchaeological information indicating generalist human diets during the Middle Holocene and specialized human diets in guanaco during the Late Holocene., Conclusions: It is proposed that during the Middle Holocene arid period, the combination of low human population density and high residential mobility in wide foraging ranges allowed the guanaco to be the main source of protein. During the Late Holocene humid period, there was an increase in human population density and a decrease in residential mobility, which caused greater pressure on foraging territories and increased dietary breadth., (© 2024 Wiley Periodicals LLC.)

Refining the role of apex predators in marine food webs is a necessary step in predicting the consequences of their global decline under the footprint of fishing activities. White sharks (Carcharodon carcharias) are vulnerable predators, performing large migrations and able to forage on a variety of prey in different habitats. In the Northeast Pacific, juvenile and adult white sharks are found seasonally at the same aggregation sites, such as Guadalupe Island off Mexico. While adults are thought to target local pinniped colonies, very few predator-prey interactions have been documented and the diet of juveniles in this area remains poorly understood. Here we used carbon/nitrogen stable isotopes and fatty acids to characterize the trophic ecology of white sharks at Guadalupe Island. In contrast to the ontogenetic trophic shift paradigm, we detected no influence of size on muscle stable isotope and fatty acid composition, revealing no significant dietary variation between juvenile and adult sharks. Stable isotopes did not allow definitive conclusions to be drawn regarding the diet of white sharks at Guadalupe Island, due to significant variability in the contribution of different potential prey depending on the trophic discrimination factors used. However, most sharks were rich in polyunsaturated fatty acids (such as long-chain omega 3), suggesting a local diet of mainly pelagic prey (potentially large fish or cephalopods). A few individuals appeared to show recent consumption of pinnipeds, with higher proportions of saturated and monounsaturated fatty acids. These individual differences in fatty acid composition could reflect an ecological trade-off between consumption of prey rich in fat (marine mammals) versus prey rich in polyunsaturated fatty acids (pelagic prey), respectively meeting the energetic and physiological needs of white sharks. Although ontogenetic trophic changes were not able to be discerned, our results provide new insights into the physiological drivers of predator-prey interactions, which can benefit the definition of conservation strategies in a changing ocean., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Nitrogen (N)-fixing symbiosis is critical to terrestrial ecosystems, yet possession of this trait is known for few plant species. Broader presence of the symbiosis is often indirectly determined by phylogenetic relatedness to taxa investigated via manipulative experiments. This data gap may ultimately underestimate phylogenetic, spatial, and temporal variation in N-fixing symbiosis. Still needed are simpler field or collections-based approaches for inferring symbiotic status. N-fixing plants differ from non-N-fixing plants in elemental and isotopic composition, but previous investigations have not tested predictive accuracy using such proxies. Here we develop a regional field study and demonstrate a simple classification model for fixer status using nitrogen and carbon content measurements, and stable isotope ratios (δ 15 N and δ 13 C), from field-collected leaves. We used mixed models and classification approaches to demonstrate that N-fixing phenotypes can be used to predict symbiotic status; the best model required all predictors and was 80-94% accurate. Predictions were robust to environmental context variation, but we identified significant variation due to native vs. non-native (exotic) status and phylogenetic affinity. Surprisingly, N content-not δ 15 N-was the strongest predictor, suggesting that future efforts combine elemental and isotopic information. These results are valuable for understudied taxa and ecosystems, potentially allowing higher-throughput field-based N-fixer assessments., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Measuring stable isotopes in different tissues offers the opportunity to provide insight into the foraging ecology of a species. This study aimed to assess how diet varies between yellow females, yellow males, and dull individuals of a Saffron Finch (Sicalis flaveola) population. We measured δ13C and δ15N in blood over a year, and in different feathers, to estimate seasonal consistency of resource use for each category. We conducted this study in a private farm in the Central Brazilian savannas. We sampled 195 individuals in seven field samplings between January 2017 and March 2018. The mean blood δ13C values were similar among yellow females, yellow males and dull individuals, indicating that this population of Saffron Finch predominantly accesses similar resources throughout the year, with a predominant C4 signal. Although Saffron Finch is considered a granivorous species, the mean δ15N values found indicate that both adults and juveniles also incorporate in their tissues some invertebrate. The slight isotope-tissue difference between feathers and blood is similar to the reported in previous studies and may reflect tissue-to-tissue discrimination. The isotopic space of yellow males was greater than that of yellow females and dull individuals, indicating greater dietary diversity due to greater inter-individual variation in diet. In Saffron Finch, which delays plumage maturation, competition-driven partitioning of food resources seems essential in driving carotenoid-based plumage coloration between age classes and sexes.

This study aimed to reconstruct the environmental conditions and the crop management practices and plant characteristics when agriculture appeared in western Europe. We analyzed oak charcoal and a large number of cereal caryopsides recovered from La Draga (Girona, Spain), an early (5300 to 4800 cal. BC) agricultural site from the Iberian Peninsula. The carbon isotope discrimination (Δ 13 C) values of oak, the dominant forest species in the region, indicates prevalence of a wet climate at the site. Further, we reconstructed crop management conditions, achievable yield, and crop characteristics through the analysis of Δ 13 C, nitrogen isotope composition (δ 15 N), nitrogen content, and the reconstructed weight of wheat and barley caryopsides, following protocols developed by our team [Araus et al. , Nat. Commun. 5 , 3953 (2014)] and comparison of these parameters with present-day organic agriculture in the region. In parallel, a regional perspective was achieved through the study of wheat and barley grains of seventeen Neolithic sites from the western Mediterranean. The results suggest that rather than small-garden cultivation, a more extensive agriculture was practiced under good water availability and moderate manuring. Moreover, results from La Draga evidence that grain weight and spike morphology were comparable to contemporary cereals. Growing conditions and the prevalence of improved crop traits indicate that agriculture was fairly consolidated at the time it reached the western edge of Europe., Competing Interests: Competing interests statement:The authors declare no competing interest.

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.)

Recent droughts have strongly impacted forest ecosystems and are projected to increase in frequency, intensity, and duration in the future together with continued warming. While evidence suggests that tree diversity can regulate drought impacts in natural forests, few studies examine whether mixed tree plantations are more resistant to the impacts of severe droughts. Using natural variations in leaf carbon (C) and nitrogen (N) isotopic ratios, that is δ 13 C and δ 15 N, as proxies for drought response, we analyzed the effects of tree species richness on the functional responses of tree plantations to the pan-European 2018 summer drought in seven European tree diversity experiments. We found that leaf δ 13 C decreased with increasing tree species richness, indicating less drought stress. This effect was not related to drought intensity, nor desiccation tolerance of the tree species. Leaf δ 15 N increased with drought intensity, indicating a shift toward more open N cycling as water availability diminishes. Additionally, drought intensity was observed to alter the influence of tree species richness on leaf δ 15 N from weakly negative under low drought intensity to weakly positive under high drought intensity. Overall, our findings suggest that dual leaf isotope analysis helps understand the interaction between drought, nutrients, and species richness., (© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.)

Rationale: Concerns exist over observed shifts in value and variance of nitrogen isotopes following physicochemical extraction of lipids from organic matter. The mechanisms behind these apparent changes in bulk tissue δ 15 N values are not fully understood yet have major implications for analytical costs and integrity of data interpretations., Methods: Changes in proximate analysis, amino acid composition, C:N ratios, bulk tissue and amino acid δ 13 C and δ 15 N values, and resulting isotope-based food web metrics were compared between lipid-intact and lipid-extracted muscle tissue of fishes spanning <1% to >20% muscle fat content to identify mechanisms of nitrogen isotope fractionation associated with physicochemical lipid extraction., Results: Bulk δ 13 C and δ 15 N values increased and %N, C:N ratios and crude protein content decreased following lipid extraction. Resulting bulk isotope niche spacing and overlap varied significantly between lipid-intact and lipid-extracted tissues. While amino acid composition significantly changed during lipid extraction, particularly for lipid-associated amino acids (e.g., Glu, Lys, Ser), individual amino acid δ 13 C and δ 15 N values, and their associated compound-specific isotope analysis of amino acids (CSIA-AA)-based food web metrics, did not., Conclusions: Physicochemical lipid extraction caused significant tissue composition changes (e.g., leaching of amino acids and 15 N-deplete nitrogenous waste) that affected δ 13 C and δ 15 N values and tissue %C and %N beyond simply removing lipids. However, lipid extraction did not alter individual amino acid δ 13 C or δ 15 N values or their associated CSIA-AA-based food web metrics., (© 2024 The Author(s). Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Killer whales (Orcinus orca) occur seasonally in the eastern Canadian Arctic (ECA), where their range expansion associated with declining sea ice have raised questions about the impacts of increasing killer whale predation pressure on Arctic-endemic prey. We assessed diet and distribution of ECA killer whales using bulk and compound-specific stable isotope analysis (CSIA) of amino acids (AA) of 54 skin biopsies collected from 2009 to 2020 around Baffin Island, Canada. Bulk ECA killer whale skin δ 15 N and δ 13 C values did not overlap with potential Arctic prey after adjustment for trophic discrimination, and instead reflected foraging history in the North Atlantic prior to their arrival in the ECA. Adjusted killer whale stable isotope (SI) values primarily overlapped with several species of North Atlantic baleen whales or tuna. Amino acid (AA)-specific δ 15 N values indicated the ECA killer whales fed primarily on marine mammals, having similar glutamic acid δ 15 N-phenylalanine δ 15 N (δ 15 N Glx-Phe ) and threonine δ 15 N (δ 15 N Thr ) as mammal-eating killer whales from the eastern North Pacific (ENP) that served as a comparative framework. However, one ECA whale grouped with the fish-eating ENP ecotype based δ 15 N Thr . Distinctive essential AA δ 13 C of ECA killer whale groups, along with bulk SI similarity to killer whales from different regions of the North Atlantic, indicates different populations converge in Arctic waters from a broad source area. Generalist diet and long-distance dispersal capacity favour range expansions, and integration of these insights will be critical for assessing ecological impacts of increasing killer whale predation pressure on Arctic-endemic species., (© 2024 The Author(s). Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.)

Aquatic food webs are spatially complex, potentially contributing to intraspecific variability in production pathway reliance of intermediate trophic level consumers. Variation in trophic reliance may be described by well-established trophic indicators, like stable isotope ratios (δ13C, δ15N), along with emerging trophic indicators, such as fatty acid composition. We evaluated stable isotope ratios and fatty acid profiles of European smelt (Osmerus eperlanus) among and within distinct regions of three large Swedish lakes (Hjälmaren, Mälaren, Vättern) which differed in trophic status. We expected that smelts in more oligotrophic lakes and regions would be characterized by distinct stable isotope signatures and fatty acid profiles, with particularly high polyunsaturated fatty acid (PUFA) relative levels. However, we acknowledge that frequent movement of smelts among regions may serve to spatially integrate their diet and lead to limited within-lake variation in stable isotope ratios and fatty acid composition. As expected, in comparison with more productive lakes (i.e., Hjälmaren and Mälaren), smelts from ultra-oligotrophic Vättern were characterized by low δ15N, high δ13C and high percent of a dominant PUFA, docosahexaenoic acid (DHA). Smelts from different regions of the morphometrically complex Mälaren displayed differential stable isotope ratios and fatty acid relative concentrations, which were consistent with within-lake differences in productivity and water residence times, suggesting that smelts in this lake forage locally within distinct regions. Finally, at the individual smelt level there were particularly strong and consistent associations between a well-established trophic indicator (δ13C) and percent DHA, suggesting that the relative concentration of this fatty acid may be a useful additional trophic indicator for smelt., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Höök 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.)

Rationale: Carbon, nitrogen and sulphur stable isotopes in feathers grown by seabirds while breeding reflect the local isoscape and diet in the vicinity of the colony, so may make it possible to discriminate individual birds from different colonies., Methods: Black-legged kittiwake Rissa tridactyla inner primary feathers from two colonies about 350 km apart in the North Sea were used to test whether δ 13 C, δ 15 N and δ 34 S differed between individuals from the two colonies. Feather tips cut from breeding birds caught at nests were compared with tips of moulted feathers (grown 1 year earlier) found on the ground., Results: Isotopic compositions showed no overlap between the two colonies in δ 13 C, δ 15 N or δ 34 S in tips of newly-grown feathers sampled from breeding adult kittiwakes. There was some overlap in δ 13 C, δ 15 N and δ 34 S from moulted feathers, but discriminant analysis allowed >90% of individuals to be assigned to their colony. In five of six comparisons, mean isotopic compositions were the same in new and moulted feathers but not for δ 34 S at one of the two colonies., Conclusions: This study has demonstrated for the first time that stable isotopes in inner primary feathers of kittiwakes can allow accurate identification of the breeding colony of individual birds from two different colonies within the North Sea. Further research is required to determine if this method can be applied with greater spatial resolution and to a larger number of colonies., (© 2024 The Authors. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Blooms of the Rhizostomeae Nemopilema nomurai and the Semaeostomeae Cyanea nozakii have become more prominent in the coastal waters of China since the end of the 20th century. However, the trophic ecology of these jellyfish species remain incompletely understood. In this study, the trophic characterizations of N. nomurai and C. nozakii populations were assessed using stable isotope analysis (SIA), with a focus on the important bloom area offshore of the Yangtze Estuary. Our results indicated obvious trophic differences between two scyphomedusae. The higher trophic position of the C. nozakii population in the coastal planktonic food web was reflected by its relatively large δ 15 N value compared to that of N. nomurai. The MixSIAR model indicated that small copepods (<1000 μm) and seston were important food sources for N. nomurai, and showed a stable dietary, irrespective of N. nomurai size. Conversely, C. nozakii exhibited a more diverse diet composition, and gelatinous organisms also were an important part of the diet of C. nozakii. Moreover, a pronounced ontogenetic shift in the diet of C. nozakii was observed, consisting of an increase in the proportion of zooplanktonic prey (excluding seston) in the C. nozakii diet with diameter. This study provides isotopic evidence of the substantial difference in trophic ecology between N. nomurai and C. nozakii, which resulted from the variations in SI values and diet compositions. Inconsistent size-based variation patterns were observed in trophic ontogenetic shifts within the N. nomurai and C. nozakii groups, highlighting a need for further investigation. These results will give insights into the characteristics of trophic ecology and functional roles of Rhizostomeae and Semaeostomeae, and indicate the need for a more careful consideration of the representations of Rhizostomeae and Semaeostomeae in coastal ecosystems, so as not to underestimate the knowledge of taxon-specific ecological effects on energy flow., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The stepwise oxygenation of Earth's surficial environment is thought to have shaped the evolutionary history of life. Microfossil records and molecular clocks suggest eukaryotes appeared during the Paleoproterozoic, perhaps shortly after the Great Oxidation Episode at ca. 2.43 Ga. The mildly oxygenated atmosphere and surface oceans likely contributed to the early evolution of eukaryotes. However, the principal trigger for the eukaryote appearance and a potential factor for their delayed expansion (i.e., intermediate ocean redox conditions until the Neoproterozoic) remain poorly understood, largely owing to a lack of constraints on marine and terrestrial nutrient cycling. Here, we analyzed redox-sensitive element contents and organic carbon and nitrogen isotope compositions of relatively low metamorphic-grade (greenschist facies) black shales preserved in the Flin Flon Belt of central Canada to examine open-marine redox conditions and biological activity around the ca. 1.9 Ga Flin Flon oceanic island arc. The black shale samples were collected from the Reed Lake area in the eastern part of the Flin Flon Belt, and the depositional site was likely distal from the Archean cratons. The black shales have low Al/Ti ratios and are slightly depleted in light rare-earth elements relative to the post-Archean average shale, which is consistent with a limited contribution from felsic igneous rocks in Archean upper continental crust. Redox conditions have likely varied between suboxic and euxinic at the depositional site of the studied section, as suggested by variable U/Al and Mo/Al ratios. Organic carbon and nitrogen isotope compositions of the black shales are approximately -23‰ and +13.7‰, respectively, and these values are systematically higher than those of broadly coeval continental margin deposits (approximately -30‰ for δ 13 C org and +5‰ for δ 15 N bulk ). These elevated values are indicative of high productivity that led to enhanced denitrification (i.e., a high denitrification rate relative to nitrogen influx at the depositional site). Similar geochemical patterns have also been observed in the modern Peruvian oxygen minimum zone where dissolved nitrogen compounds are actively lost from the reservoir via denitrification and anammox, but the large nitrate reservoir of the deep ocean prevents exhaustion of the surface nitrate pool. Nitrogen must have been widely bioavailable in the ca. 1.9 Ga oceans, and its supply to upwelling zones must have supported habitable environments for eukaryotes, even in the middle of oceans around island arcs., (© 2024 John Wiley & Sons Ltd.)

Objectives: Early colonial documents from central Mesoamerica detail raising and planting of European livestock and crops alongside native ones. The extent to which Indigenous people, especially of the rural commoner class, consumed newly introduced foods is less known. This gap in knowledge is addressed through stable isotope analysis and comparison to published archaeological botanical, human, and faunal data., Materials and Methods: Stable isotope analysis of bone collagen and bioapatite is applied to 74 skeletal samples of Indigenous human remains representing Colonial period individuals from El Japón-a farming hamlet in the Xochimilco area-to provide insight into long-term individual dietary practices in the context of a rapidly transforming Mesoamerican world., Results: Carbon isotope ratios in collagen (δ 13 C collagen ) average -8.1 0 / 00 VPDB (SD 0.55), while δ 15 N averages 8.9 0 / 00 AIR (SD 0.50). δ 13 C bioapatite averages -2.9 0 / 00 VPDB (SD 0.60). Modest increase in carbon isotopic diversity is observed among more recent males from El Japón when compared to earlier males and females., Discussion: Based on the isotopic results, it is estimated that the individuals of El Japón consumed maize or other C 4 plants as a central source of carbohydrates. Dietary protein was largely supplied through domestic maize-fed fauna but potentially supplemented by wild terrestrial and aquatic fauna and fowl. Similarity in skeletal isotopic composition between precontact Mesoamericans from other sites and El Japón individuals of both earlier and later stratigraphy is interpreted as continuity in local diets and foodways despite potentially available European alternatives. Colonial taxation demands on preexisting agricultural regimes may have incentivized maize production, thus indirectly contributing to the maize-centered aspect of local foodways., (© 2024 The Authors. American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

The determination of δ 13 C and δ 15 N values is a common method in archaeological isotope analysis-in studying botanical and human remains, dietary practices, and less typically soils (to understand methods of agricultural cultivation, including fertilization). Stable isotope measurements are also commonly used in ecological studies to distinguish different ecosystems and to trace diachronic processes and biogeochemical mechanisms, however, the application of this method in geochemical prospection, for determining historic land-use impact, remains unexplored. The study at hand focuses on a deserted site of a Cistercian manor, dating from the thirteenth to fifteenth centuries. Isotopic measurements of anthropogenically influenced soils have been compared to approximately 400 archaeobotanical, soil, and sediment samples collected globally. The results reveal the potential of isotope measurements in soil to study the impact of past land use as isotope measurements identify specific types of agricultural activities, distinguishing crop production or grazing. δ 13 C and δ 15 N ratios also likely reflect fertilization practices and-in this case-the results indicate the presence of cereal cultivation (C 3 cycle plants) and fertilization and that the site of the medieval manor was primarily used for grain production rather than animal husbandry., (© 2024. Charles University, Faculty of Arts, 2024.)

The volume of human carbon (δ13C) and nitrogen (δ15N) isotope data produced in archaeological research has increased markedly in recent years. However, knowledge of bone remodelling, its impact on isotope variation, and the temporal resolution of isotope data remains poorly understood. Varied remodelling rates mean different elements (e.g., femur and rib) produce different temporal signals but little research has examined intra-element variability. This study investigates human bone remodelling using osteon population density and the relationship with carbon and nitrogen isotope data at a high resolution, focusing on variation through femoral cross-sections, from periosteal to endosteal surfaces. Results demonstrate considerable differences in isotope values between cross-sectional segments of a single fragment, by up to 1.3‰ for carbon and 1.8‰ for nitrogen, illustrating the need for standardised sampling strategies. Remodelling also varies between bone sections, occurring predominantly within the endosteal portion, followed by the midcortical and periosteal. Therefore, the endosteal portion likely reflects a shorter period of life closer to the time of death, consistent with expectations. By contrast, the periosteal surface provides a longer average, though there were exceptions to this. Results revealed a weak negative correlation between osteon population density and δ15N or δ13C, confirming that remodelling has an effect on isotope values but is not the principal driver. However, a consistent elevation of δ15N and δ13C (0.5‰ average) was found between the endosteal and periosteal regions, which requires further investigation. These findings suggest that, with further research, there is potential for single bone fragments to reconstruct in-life dietary change and mobility, thus reducing destructive sampling., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 de Gruchy 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.)

Discerning assimilated diets of wild animals using stable isotopes is well established where potential dietary items in food webs are isotopically distinct. With the advent of mixing models, and Bayesian extensions of such models (Bayesian Stable Isotope Mixing Models, BSIMMs), statistical techniques available for these efforts have been rapidly increasing. The accuracy with which BSIMMs quantify diet, however, depends on several factors including uncertainty in tissue discrimination factors (TDFs; Δ) and identification of appropriate error structures. Whereas performance of BSIMMs has mostly been evaluated with simulations, here we test the efficacy of BSIMMs by raising domestic broiler chicks (Gallus gallus domesticus) on four isotopically distinct diets under controlled environmental conditions, ideal for evaluating factors that affect TDFs and testing how BSIMMs allocate individual birds to diets that vary in isotopic similarity. For both liver and feather tissues, δ13C and δ 15N values differed among dietary groups. Δ13C of liver, but not feather, was negatively related to the rate at which individuals gained body mass. For Δ15N, we identified effects of dietary group, sex, and tissue type, as well as an interaction between sex and tissue type, with females having higher liver Δ15N relative to males. For both tissues, BSIMMs allocated most chicks to correct dietary groups, especially for models using combined TDFs rather than diet-specific TDFs, and those applying a multiplicative error structure. These findings provide new information on how biological processes affect TDFs and confirm that adequately accounting for variability in consumer isotopes is necessary to optimize performance of BSIMMs. Moreover, results demonstrate experimentally that these models reliably characterize consumed diets when appropriately parameterized., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Gurney 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.)

For many species, the relationship between space use and diet composition is complex, with individuals adopting varying space use strategies such as territoriality to facilitate resource acquisition. Coyotes ( Canis latrans ) exhibit two disparate types of space use; defending mutually exclusive territories (residents) or moving nomadically across landscapes (transients). Resident coyotes have increased access to familiar food resources, thus improved foraging opportunities to compensate for the energetic costs of defending territories. Conversely, transients do not defend territories and are able to redirect energetic costs of territorial defense towards extensive movements in search of mates and breeding opportunities. These differences in space use attributed to different behavioral strategies likely influence foraging and ultimately diet composition, but these relationships have not been well studied. We investigated diet composition of resident and transient coyotes in the southeastern United States by pairing individual space use patterns with analysis of stable carbon (δ 13 C) and nitrogen (δ 15 N) isotope values to assess diet. During 2016-2017, we monitored 41 coyotes (26 residents, 15 transients) with GPS radio-collars along the Savannah River area in the southeastern United States. We observed a canopy effect on δ 13 C values and little anthropogenic food in coyote diets, suggesting 13 C enrichment is likely more influenced by reduced canopy cover than consumption of human foods. We also observed other land cover effects, such as agricultural cover and road density, on δ 15 N values as well as reduced space used by coyotes, suggesting that cover types and localized, resident-like space use can influence the degree of carnivory in coyotes. Finally, diets and niche space did not differ between resident and transient coyotes despite differences observed in the proportional contribution of potential food sources to their diets. Although our stable isotope mixing models detected differences between the diets of resident and transient coyotes, both relied mostly on mammalian prey (52.8%, SD = 15.9 for residents, 42.0%, SD = 15.6 for transients). Resident coyotes consumed more game birds (21.3%, SD = 11.6 vs 13.7%, SD = 8.8) and less fruit (10.5%, SD = 6.9 vs 21.3%, SD = 10.7) and insects (7.2%, SD = 4.7 vs 14.3%, SD = 8.5) than did transients. Our findings indicate that coyote populations fall on a feeding continuum of omnivory to carnivory in which variability in feeding strategies is influenced by land cover characteristics and space use behaviors., Competing Interests: The authors declare that they have no competing interests., (© 2024 Webster et al.)

The influences of oceanographic changes on diet composition and trophic level for pollock (Gadus chalcogrammus) inhabiting the East Sea off the Korean coast were examined based on stomach content and stable isotope analyses during 2016 and 2017. The diets of pollock consisted mainly of benthic crustaceans (particularly carid shrimps and euphausiids) and cephalopods, with a predominance of teleosts in the diets of larger individuals in deeper habitats. In 2016, amphipods, carid shrimps and cephalopods featured strongly in pollock diets, and the contribution of amphipods decreased in the diets of larger individuals and deeper depths. In 2017, euphausiids dominated at shallower depths, whereas the contributions of carid shrimps and teleosts increased in deeper habitats. Body-size-related differences in carbon stable isotope (δ 13 C) values were present in both 2016 and 2017, but size-related differences in nitrogen stable isotope (δ 15 N) values were only observed in 2017. The increased contribution of euphausiids during 2017 resulted in a distinct decrease in the trophic level of pollock compared to co-occurring higher trophic level predators, which can be linked to changes in habitat water temperature. Combined stomach contents and isotopic analyses provide a more comprehensive understanding of how fish diets and trophic levels fluctuate with changes in the type and abundance of prey resources in response to environmental changes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This paper reports the first record of total mercury (THg) concentrations in albacore (Thunnus alalunga), one of the main tuna species caught from the Western Equatorial Atlantic Ocean and presents a preliminary comparison with other regions and tuna species. Mean, standard deviation and range of concentrations in T. alalunga (515 ± 145 ng g -1 ww; 294-930 ng g -1 ww) with 92 % being of methyl-Hg, are higher than in albacore from other Atlantic Ocean subregions despite their smaller body size. These concentrations are similar to those from the Pacific and Indian oceans, but lower than in the Mediterranean. Compared to other sympatric tuna species, concentrations are higher than those in T. albacares and similar to T. obesus. These results are discussed considering the potential differences in stable isotope values ( 13 C and 15 N) of T. alalunga populations from multiple oceanic areas and compared to other tuna species worldwide., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. All data used in this paper is available as supplementary material., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

We studied δ 13 C, δ 15 N and δ 18 O values, and total mercury (THg) concentrations in muscle samples from deep-sea predators - five beaked whale species and sperm whales - stranded along the coast of Hokkaido, in the north of Japan in 2010 and 2019. The δ 13 C, δ 15 N and δ 18 O values, THg concentrations, and body length (BL) of Stejneger's beaked whales were similar to those of Hubbs' beaked whales, which belong to the same genus. In contrast, δ 13 C values, THg concentrations, and BL of Sato's beaked whales were markedly different from those of Baird's beaked whales, which belong to the same genus. Stejneger's and Hubbs' beaked whales living around Hokkaido may compete in their ecological niches, whereas Sato's and Baird's beaked whales may segregate their ecological niches. Although Cuvier's beaked whales and sperm whales belong to different genera and their BLs were significantly different, their δ 13 C and δ 15 N values were similar, probably because they can dive and stay in deeper waters than other beaked whale species. The δ 13 C values in combined samples from all whales increased with increasing BL, probably owing to the larger whale species' dietary preference for squid. The δ 13 C values in combined samples from all whales were positively correlated with THg concentrations, whereas the δ 15 N values in the combined samples were negatively correlated. The δ 18 O values in combined samples from most whales tended to be positively correlated with THg concentrations. These correlations may be explained by a higher THg load from deep-sea feeding than from pelagic feeding and by a feeding shift towards lower trophic levels.