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To identify a novel optimized strategy for preventing fraudulent substitutions of squid species and origins, forty European squids (Loligo vulgaris) and forty flying squids (Todarodes sagittatus) from the Mediterranean Sea and Atlantic Ocean were analyzed for δ 13 C, δ 15 N, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, and Lu using isotope ratio mass spectrometry and inductively coupled plasma-mass spectrometry. While δ 13 C and δ 15 N variations were mainly species-related, they alone could not reliably distinguish samples. To address this issue, decision rules were developed using Classification and Regression Tree analysis. Threshold values for δ 13 C (-19.91‰), δ 15 N (14.87‰), and Pr (0.49 μg kg -1 ) enabled successful discrimination among Mediterranean European squids, Atlantic European squids, Mediterranean flying squids, and Atlantic flying squids, achieving over 90% accuracy, 81% precision, 80% sensitivity, and 93% specificity. This method holds promise for enhancing traceability and safety in the seafood industry, ensuring product integrity and consumer trust., 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.)

Oyster farming activities play a pivotal role in the biogeochemical cycles of coastal marine ecosystems, particularly in terms of sedimentary carbon cycling. To gain deep insights into the influence of expanding oyster culture on the sedimentary carbon cycle, surface sediments were collected from the Maowei Sea, which is the largest oyster farming bay in south China, based on six filed surveys between July 2010 and December 2022. The sediment samples were analyzed for total organic carbon (TOC), total nitrogen (TN), stable carbon and nitrogen isotopes (δ 13 C and δ 15 N) to evaluate the inter-annual variations in the source contribution to sedimentary organic matter (SOM). The results revealed that the average contents of sedimentary TOC and TN were 0.67 ± 0.41 % and 0.06 ± 0.03 %, respectively. Fluctuations in the C/N molar ratios ranged from 5.8 to 23.6, with an average of 12.6 ± 2.9, indicating a significant terrestrial input contribution to SOM in the study area. Furthermore, the integration of stable isotope analysis and Bayesian mixing model demonstrated a gradual increase in the mean proportion of shellfish biodeposition to SOM, from 12.0 ± 5.6 % in July 2010 to 21.1 ± 7.3 % in December 2022, consistent with the progressive expansion of oyster aquaculture along this coastal area, thereby emphasizing the substantial influence of oyster farming on SOM composition. With the anticipated expansion of oyster farming scale and production in the future, shellfish biodeposition is expected to assume a more important role in shaping SOM dynamics and sedimentary organic carbon cycling in coastal waters. Overall, this study provided an important perspective for better assessing the impact of expanding mariculture scale on coastal biogeochemical cycles, thereby making valuable contributions to future policy formulation concerning mariculture and ecological conservation., 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 B.V. All rights reserved.)

The ratio of net CO2 uptake (Anet) and stomatal conductance (gs) is an intrinsic measurement of leaf water use efficiency (WUEi); however, its measurement can be challenging for large phenotypic screens. Measurements of the leaf carbon isotope composition (δ13Cleaf) may be a scalable tool to approximate WUEi for screening because it in part reflects the competing influences of Anet and gs on the CO2 partial pressure (pCO2) inside the leaf over time. However, in C4 photosynthesis, the CO2-concentrating mechanism complicates the relationship between δ13Cleaf and WUEi. Despite this complicated relationship, several studies have shown genetic variation in δ13Cleaf across C4 plants. Yet there has not been a clear demonstration of if Anet or gs are the causal mechanisms controlling WUEi and δ13Cleaf. Our approach was to characterize leaf photosynthetic traits of two Zea mays recombinant inbred lines (Z007E0067 and Z007E0150) which consistently differ for δ13Cleaf even though they have minimal confounding genetic differences. We demonstrate that these two genotypes contrasted in WUEi driven by differences in the speed of stomatal responses to changes in pCO2 and light that lead to unproductive leaf water loss. These findings provide support that differences in δ13Cleaf in closely related genotypes do reflect greater WUEi and further suggest that differences in stomatal kinetic response to changing environmental conditions is a key target to improve WUEi., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

In order to evaluate mercury (Hg) accumulation patterns in Southern Ocean penguins, we measured Hg concentrations and carbon (δ 13 C) and nitrogen (δ 15 N) stable isotope ratios in body feathers of adult Adélie (Pygoscelis adeliae), gentoo (Pygoscelis papua), and chinstrap (Pygoscelis antarctica) penguins living near Anvers Island, West Antarctic Peninsula (WAP) collected in the 2010/2011 austral summer. With these and data from Pygoscelis and other penguin genera (Eudyptes and Aptenodytes) throughout the Southern Ocean, we modelled Hg variation using δ 13 C and δ 15 N values. Mean concentrations of Hg in feathers of Adélie (0.09 ± 0.05 μg g -1 ) and gentoo (0.16 ± 0.08 μg g -1 ) penguins from Anvers Island were among the lowest ever reported for the Southern Ocean. However, Hg concentrations in chinstrap penguins (0.80 ± 0.20 μg g -1 ), which undertake relatively broad longitudinal winter migrations north of expanding sea ice, were significantly higher (P < 0.001) than those in gentoo or Adélie penguins. δ 13 C and δ 15 N values for feathers from all three Anvers Island populations were also the lowest among those previously reported for Southern Ocean penguins foraging within Antarctic and subantarctic waters. These observations, along with size distributions of WAP krill, suggest foraging during non-breeding seasons as a primary contributor to higher Hg accumulation in chinstraps relative to other sympatric Pygoscelis along the WAP. δ 13 C values for all Southern Ocean penguin populations, alone best explained feather Hg concentrations among possible generalized linear models (GLMs) for populations grouped by either breeding site (AICc = 36.9, w i  = 0.0590) or Antarctic Frontal Zone (AICc = 36.9, w i  = 0.0537). Although Hg feather concentrations can vary locally by species, there was an insignificant species-level effect (w i  < 0.001) across the full latitudinal range examined. Therefore, feeding ecology at breeding locations, as tracked by δ 13 C, control Hg accumulation in penguin populations across the Southern Ocean., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A recent article by DePalma et al. reported that the season of the End-Cretaceous mass extinction was confined to spring/summer on the basis of stable isotope analyses and supplementary observations. An independent study that was concurrently under review reached a similar conclusion using osteohistology and stable isotope analyses. We identified anomalies surrounding the stable isotope analyses reported by DePalma et al. Primary data are not provided, the laboratory where the analyses were performed is not identified, and the methods are insufficiently specified to enable accurate replication. Furthermore, isotopic graphs for carbon and oxygen contain irregularities such as missing data points, duplicate data points, and identical-length error bars for both elements despite different scales, that appear inconsistent with laboratory instrument outputs. A close examination of such methodological omissions and data irregularities can help to raise the standards for future studies of seasonality and prevent inaccurate claims or confirmation bias., Competing Interests: The authors declare that they have no competing interests., (© 2024 During et al.)

Coastal sediments are the main deposition center for allochthonous and autochthonous organic carbon (OC). The discharge of terrestrial biomass, anthropogenic activities, oceanic primary productivity, and natural events contribute to this carbon pool. The OC buried in sediments undergoes alteration through physical, biological and chemical processes, becoming progressively refractory and more likely to be preserved on geological time scales. However, little is known about the rate of bulk OC alteration post weathering and bloom. We incubated coastal sediment slurries with isotopically distinct spikes of C4 corn leaves and cultured phytoplankton, individually and in 1:1 mixture. OC isotopic values and concentrations were probed at different time points to track degradation and incorporation in solid and liquid phases. Both amendments were composed of fresh OC with a high proportion of labile biochemicals (e.g. polysaccharides and proteins). Despite the small differences in their lability, corn leaves were incorporated into the sediments at a slower rate compared to phytoplankton. Following combined spiking of the terrestrial and marine amendments, no sign of synergistic effects was observed in system's response. Despite sediment sensitivity to OC input and the rapid alterations in its properties within the initial days of incubation, swiftly transitioning to a state of minimal change is indicative of a relatively stable system that retained the isotopic imprint of the OC spike for a long time (> 32 days). This isotopic remanence is likely due to heterotrophic bacteria that degrade OC to synthesize their biomass (food stock for successive generations) and incorporate its stable isotope characteristics. Hence, our work sheds light on the kinetics of biogeochemical changes, and recovery time of the system for returning to its pre-perturbation state., 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.)

This study explores changes in pastoral practices in the Jerusalem region (Iron Age II - Late Hellenistic) through a multi-isotope approach (strontium, carbon, oxygen, and nitrogen). Based on the analysis of 135 sheep, goat, and cattle teeth and bone samples from Givati Parking Lot we demonstrate the value of this method in reconstructing past animal husbandry, revealing adaptation and resilience of pastoral communities amidst environmental and socio-political changes. Isotopic analysis indicates local sourcing for most animals, with intriguing outliers from distant regions up to 150 km away, suggesting regional exchange networks. Notably, the Persian period (5th century BCE) exhibits a wider isotope range, implying increased flexibility and exploitation of diverse grazing lands, potentially driven by climate shifts and political upheavals. Conversely, Late Hellenistic (2nd century BCE) livestock display restricted movement, while showcasing a rise in desert caprines, indicative of increased import compared to the Persian era. These findings highlight the dynamism and adaptability of past pastoral communities, adjusting their strategies in response to various pressures. This study opens new avenues for understanding human-environment interactions in the Levant and underscores the power of multi-isotope approaches in unraveling intricate socio-economic and ecological dynamics of the past., (© 2024. The Author(s).)

Water-use efficiency (WUE) is affected by multiple leaf traits, including stomatal morphology. However, the impact of stomatal morphology on WUE across different ontogenetic stages of tree species is not well-documented. Here, we investigated the relationship between stomatal morphology, intrinsic water-use efficiency (iWUE) and leaf carbon isotope ratio (δ 13 C). We sampled 190 individuals, including juvenile and mature trees belonging to 18 temperate broadleaved tree species and 9 genera. We measured guard cell length (GCL), stomatal density (SD), specific leaf area (SLA), iWUE and bulk leaf δ 13 C as a proxy for long-term WUE. Leaf δ 13 C correlated positively with iWUE across species in both juvenile and mature trees, while GCL showed a negative and SD a positive effect on iWUE and leaf δ 13 C. Within species, however, only GCL was significantly associated with iWUE and leaf δ 13 C. SLA had a minor negative influence on iWUE and leaf δ 13 C, but this effect was inconsistent between juvenile and mature trees. We conclude that GCL and SD can be considered functional morphological traits related to the iWUE and leaf δ 13 C of trees, highlighting their potential for rapid phenotyping approaches in ecological studies., (© 2024 The Author(s). Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

The Atlantic bluefin tuna (ABFT) population ranges throughout the Atlantic Ocean and Mediterranean Sea, and consists of multiple contingents that use diverse habitats and show different movement patterns over the life cycle. Based on body size, elemental and isotopic data of C and N in muscle and liver, we analysed eastern-stock ABFT by comparing mid-sized individuals caught by hook-and-line gears with larger individuals harvested from traps in the Strait of Gibraltar (SoG). Our results show that trophic-related chemical markers have potential for separating temporarily sympatric contingents throughout the ABFT population range, reflecting size-dependent spatial distribution and differential patterns of residency and migration. We present evidence of long-term residency of ABFT in the SoG that persists until the estimated age of ∼5 years (size of ∼140 cm in straight fork length). This age apparently marks a turning point in the life history, where there occurs an ontogenetic switch in the migratory behaviour and distributional pattern. This study contributes new insight into our knowledge about size structure and residency-movement patterns in eastern ABFT. It shows distinct size-dependent migratory and spatial dynamics. The present results encourage further investigation on poorly studied ABFT contingents for a better understanding of the population dynamics towards more comprehensive management plans., 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.)

The aim of this study was to explore the effects of diet and habitat on the Hg levels of Japanese anchovy at different growth stages. We measured the amounts of Hg and stable carbon and nitrogen isotopes in the muscle and stable carbon and oxygen isotopes in the otoliths of 54 Japanese anchovy specimens obtained from the open seas of the Northwest Pacific Ocean (39°2'N ∼ 42°30'N, 154°02'E ∼ 161°29'E) between June and July 2021. Body length had a significant effect on δ 13 C bulk , δ 15 N bulk and M oto (P < 0.01). As individuals grew, δ 13 C bulk and δ 15 N bulk tended to gradually increase, but M oto tended to gradually decrease. The variation in the niches of Japanese anchovy at different growth stages showed that the standard ellipse-corrected area subsequently decreased with growth. It reached its smallest value in the 136-150 mm group, and there was no overlap between the 136-150 mm group and the other groups. The GAM results showed that Hg levels tended to decrease first and then increase with growth. There was a positive correlation between Hg levels and δ 18 O oto in fish. Hg levels increased gradually with increasing δ 15 N bulk . In our study, there may be a gradual shift in the diet of Japanese anchovy from phytoplankton to prey at higher trophic levels, and the depth of seawater in which the predators feed gradually increased with growth. Changes in diet and habitat were probably the main reasons for the increase in Hg levels., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Guanyu Hu reports was provided by Shanghai Ocean University College of Marine Living Resource Sciences and Management. Guanyu Hu reports a relationship with Shanghai Ocean University College of Marine Living Resource Sciences and Management that includes: funding grants. 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. If there are other authors, they 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.)

We employed Fourier infrared spectroscopy, laser infrared technology, and stable isotope analysis to investigate the relationships between characteristics of ingested plastics and size-related feeding ecology of a mesopelagic predator, longnose lancetfish Alepisaurus ferox. Plastics were detected in 81.48 % of specimens, up to 8.81 ± 8.29 items/individual and 0.59 ± 0.66 items/g wet weight of intestine, and were sized 20.00 μm to 6.50 cm. The majority were granules and fragments in shape, and polystyrene and acrylate copolymer in polymer type. The diversity indices exhibited a correlation with trophic position and body size groups, emphasizing that the ontogenetic dietary shift of A. ferox may influence the diversity of plastics ingested. This study provides new insights into the plastic pathways linking epipelagic and mesopelagic food webs and demonstrates that biochemical ecological tracers can effectively indicate the bioavailability of plastic correlated with growth in mesopelagic predator., 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.)

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.)

Carbon-13 isotopomics of triacylglycerol (TAG) fatty acids or free fatty acids in biological matrices holds considerable potential in food authentication, forensic investigations, metabolic studies, and medical research. However, challenges arise in the isotopic analysis of short- and medium-chain (C4 to C10) fatty acid methyl esters (SMCFAMEs) through gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The high volatility of these esters results in losses during their preparation, leading to isotopic fractionation. Moreover, the methoxy group added to acyl chains requires the correction of δ 13 C values, thereby increasing the uncertainty of the final results. Analyzing free fatty acids (FFAs) addresses both issues encountered with SMCFAMEs. To achieve this objective, we have developed a new protocol enabling the isotopomics of individual fatty acids (FAs) by GC-C-IRMS. The same experiment also provides the FA profile, i.e., the relative percentage of each FA in the TAG hydrolysate or its concentration in the studied matrix. The method exhibited high precision, as evidenced by the repeatability and within-lab reproducibility of results when tested on TAGs from both animal and vegetal origins. Compared to the analysis of FAMEs by GC-C-IRMS, the current procedure also brings several improvements in alignment with the principles of green analytical chemistry and green sample preparation. Thus, we present a two-in-one method for 13 C-isotopomic and metabolomic biomarker quantitation within quasi-universal TAG compounds, encompassing the short- and medium-acyl chains., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Nandrolone and its prohormones, including 19-norandrost-4-ene-3,17-dione and 19-norandrost-4-ene-3β,17β-diol, are anabolic steroids forbidden at all times in sports according to the World Anti-Doping Code Prohibited List and its metabolite 19-norandrosterone (19NA) is the preferred urinary target compound to identify their abuse. In recent years, an increasing number of 19NA isotope ratio mass spectrometry (IRMS) cases have arisen that, based on the initial testing procedure, were likely to result in an adverse analytical finding but were concluded negative after IRMS analysis. The current study was therefore set up to gain a better insight on the prevalence of nandrolone preparations with endogenous carbon isotope ratio values in Australia. Suitable workplace (non-athlete) urine samples that had previously been reported positive for 19NA were identified and analysed on IRMS. A total of 82% of the samples that were analysed were reported with enriched carbon isotope ratios of 19NA (i.e., 19NA greater than -26‰). This indicates that there is a high prevalence of nandrolone-containing anabolic androgenic steroid preparations in Australia that have 'endogenous' carbon isotope ratios which reduces the ability to identify exogenous nandrolone., (© 2024 John Wiley & Sons 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.)

Riverine sediments are important habitats for microbial activity in naturalised waterways to provide potential ecosystem services that improve stormwater quality. Yet, little is known about the sources of these sediment microbes, and the factors shaping them. This study investigated the dominant source of sediments in a tropical naturalised urban waterway, using two Bayesian methods for microbial and isotopic 13 C/ 15 N markers concurrently. Additionally, key factors shaping microbial communities from the surrounding landscape were evaluated. A comprehensive two-year field survey identified source land covers of interest based on topology and soil context. Among these land covers, riverbanks were the dominant source of sediments contribution for both edaphic and microbial components. The physico-chemical environment explains most of the variation in sediment communities compared to inter-location distances and microbial source contribution. As microbes provide ecosystem services important for rewilding waterways, management strategies that establish diverse sediment microbial communities are encouraged. Since riverbanks play a disproportionately important role in material contribution to sediment beds, management practices aimed at controlling soil erosion from riverbanks can improve overall functioning of waterway systems., 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 B.V.)

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.)

Large meteorite impacts must have strongly affected the habitability of the early Earth. Rocks of the Archean Eon record at least 16 major impact events, involving bolides larger than 10 km in diameter. These impacts probably had severe, albeit temporary, consequences for surface environments. However, their effect on early life is not well understood. Here, we analyze the sedimentology, petrography, and carbon isotope geochemistry of sedimentary rocks across the S2 impact event (37 to 58 km carbonaceous chondrite) forming part of the 3.26 Ga Fig Tree Group, South Africa, to evaluate its environmental effects and biological consequences. The impact initiated 1) a giant tsunami that mixed Fe 2+ -rich deep waters into the Fe 2+ -poor shallow waters and washed debris into coastal areas, 2) heating that caused partial evaporation of surface ocean waters and likely a short-term increase in weathering and erosion on land, and 3) injection of P from vaporization of the S2 bolide. Strata immediately above the S2 impact event contain abundant siderites, which are associated with organic matter and exhibit light and variable δ 13 C carb values. This is consistent with microbial iron cycling in the wake of the impact event. Thus, the S2 impact likely had regional, if not global, positive and negative effects on life. The tsunami, atmospheric heating, and darkness would likely have decimated phototrophic microbes in the shallow water column. However, the biosphere likely recovered rapidly, and, in the medium term, the increase in nutrients and iron likely facilitated microbial blooms, especially of iron-cycling microbes., Competing Interests: Competing interests statement:The authors declare no competing interest.

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).)

Biodegradability testing in soil helps to identify safe synthetic organic chemicals but is still obscured by the formation of soil-bound 'non-extractable' residues (NERs). Present-day methodologies using radiocarbon or stable ( 13 C, 15 N) isotope labeling cannot easily differentiate soil-bound parent chemicals or transformation products (xenoNERs) from harmless soil-bound biomolecules of microbial degraders (bioNERs). Hypothesizing a minimal retention of hydrogen in biomolecules, we here apply stable hydrogen isotope - deuterium (D) - labeling to unravel the origin of NERs. Soil biodegradation tests with D- and 13 C-labeled 2,4-D, glyphosate and sulfamethoxazole reveal consistently lower proportions of applied D than 13 C in total NERs and in amino acids, a quantitative biomarker for bioNERs. Soil-bound D thus mostly represents xenoNERs and not bioNERs, enabling an efficient quantification of xenoNERs by just measuring the total bound D. D or tritium (T) labeling could thus improve the value of biodegradability testing results for diverse organic chemicals forming soil-bound residues., (© 2024. The Author(s).)

The deep continental crust represents a vast potential habitat for microbial life where its activity remains poorly constrained. Organic acids like acetate are common in these ecosystems, but their role in the subsurface carbon cycle - including the mechanism and rate of their turnover - is still unclear. Here, we develop an isotope-exchange 'clock' based on the abiotic equilibration of H-isotopes between acetate and water, which can be used to define the maximum in situ acetate residence time. We apply this technique to the fracture fluids in Birchtree and Kidd Creek mines within the Canadian Precambrian crust. At both sites, we find that acetate residence times are <1 million years and calculated a rate of turnover that could theoretically support microbial life. However, radiolytic water-rock reactions could also contribute to acetate production and degradation, a process that would have global relevance for the deep biosphere. More broadly, our study demonstrates the utility of isotope-exchange clocks in determining residence times of biomolecules with possible applications to other environments., (© 2024. The Author(s).)

Historians and archaeologists have been debating the scale of animal husbandry in ancient Greece for decades. This study contributes to the debate by examining Classical and Hellenistic faunal assemblages from Magoula Plataniotki, New Halos, and Pherae through non-destructive zooarchaeological methods and a multi-isotopic (87Sr/86Sr, δ13C, and δ18O) approach. Zooarchaeological data suggest that small-scale sedentary animal husbandry focused on caprine production in Magoula Plataniotiki and New Halos, and small-scale and semi-specialised animal husbandry was practised in Pherae. Isotopic data show both sedentary and mobile management of livestock in all sites, indicating different levels of production intensity and variety of goals. Based on our results, we propose an economic model whereby semi-specialised and small-scale animal husbandry co-existed, confirming mixed husbandry models for ancient Greece., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Filioglou 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.)

We utilized amino acid (AA) and carbon stable isotope analyses to characterize phytoplankton-derived organic matter (OM) and trace the sources of organic carbon in the Amundsen Sea. Carbon isotope ratios of particulate organic carbon (δ 13 C-POC) range from -28.7‱ to -23.1‱, indicating that particulate organic matter originated primarily from phytoplankton. The dissolved organic carbon isotope (δ 13 C-DOC) signature (-27.1 to -21.0‱) observed in the sea-ice melting system suggests that meltwater contributes to the DOC supply of the Amundsen Sea together with OM produced by phytoplankton. A negative correlation between the degradation index and δ 13 C-POC indicates that the quality of OM significantly influences isotopic fractionation (r 2 = 0.59, p < 0.001). The AA distribution in the Amundsen Sea (5.43 ± 3.19 µM) was significantly larger than previously reported in the Southern Ocean and was associated with phytoplankton biomass (r 2 = 0.49, p < 0.01). Under conditions dominated by P. antarctica (DI = 2.29 ± 2.30), OM exhibited greater lability compared to conditions co-dominated by diatoms and D. speculum (DI = 0.04 ± 3.64). These results highlight the important role of P. antarctica in influencing the properties of OM, suggesting potential impacts on carbon cycling and microbial metabolic activity in the Amundsen Sea.

Global warming endangers reef-building corals as they lose their photosynthetic symbionts, which limits their ability to feed autotrophically. Consequently, heterotrophy, the capture of zooplankton, can become crucial for the energy budget of heat-stressed corals. However, it is difficult to assess the extent of the heterotrophic contribution in corals, as well as the dynamics of nutrient exchange between the host and its symbionts. In this pioneering study, we employed a suite of isotopic markers, including 13 C- and 15 N bulk tissue isotope measurements, compound-specific isotope analysis of amino acids (CSIA-AAs), and 13 C- and 15 N-labeled food incubations, to investigate nutrient acquisition and allocation in the coral Stylophora pistillata under controlled and heat-induced bleaching conditions. Bulk isotope values and inorganic carbon assimilation remained unchanged in the bleached corals compared to the control corals, overall indicating undisturbed autotrophic activity of the symbionts under heat stress. However, CSIA-AAs showed an increased dependence on heterotrophy for amino acid synthesis in both the host and the symbionts despite reduced assimilation of 15 N-labeled food. Overall, these results suggest that although S. pistillata reduces its assimilation of heterotrophic food under heat stress, the acquisition of amino acids by the coral host and symbionts still relies on heterotrophy. This study emphasizes the importance of using multiple indicators to gain a comprehensive understanding of coral nutrition. It shows that coral dependence on heterotrophy is not only associated with a decline in autotrophic availability. Rather, it demonstrates the ability of S. pistillata to adapt its utilization of food sources to the prevailing environmental conditions.IMPORTANCEThis work highlights that every isotopic marker displays a piece of different information concerning the diet of the model coral S. pistillata . By combining all markers, we observed that although S. pistillata exhibited reduced heterotrophic assimilation under heat stress, amino acid acquisition and synthesis remained dependent on heterotrophy. The findings emphasize the adaptability of corals in utilizing different food sources, which is vital for their resilience and recovery in changing environmental conditions. This research underscores the complexity of coral symbiosis and highlights the need for multiple indicators to understand dietary dynamics comprehensively., Competing Interests: The authors declare no conflict of interest.

This study aimed to improve the traceability of rice-producing areas to address the increasing demand for accurate methods to confirm food quality and safety. Compound-specific δ 13 C of fatty acids, δ 13 C of starch and bulk of rice were measured. PCA, PLS-DA and VIP value analysis of the obtained data were performed to track the source of rice from the six regions. The PLS-DA model established with bulk δ 13 C, starch δ 13 C, and fatty acid δ 13 C, which clearly separated the rice from six regions. The VIP graph showed the value of starch, C18:0 and C18:2 δ 13 C values (VIP > 1) were important to distinguish the origin of rice. Also, according to loading plots the contribution of starch δ 13 C was the largest. The findings indicate that the introduction of starch δ 13 C improves the precision of rice traceability and provides an effective method for identifying rice origin., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Liu Yu has patent pending to Dalian Maritime University., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The historical development of the vegetation of semi-dry grasslands in Central Europe is not satisfactorily understood. Long-term continuity of open vegetation or, conversely, deep-past forest phases are considered possible sources of the current extreme species diversity of these ecosystems. We aimed to reveal the trajectory of paleovegetation development in these ecosystems through detailed analysis of terrestrial in-situ soil geoarchives. We measured the bulk soil carbon and nitrogen contents, lipid molecular distribution, and compound-specific stable carbon and hydrogen isotopic signatures of mid- and long-chain n-alkanes extracted from soil and modern plant material tissues (i.e., deciduous and Pinus leaves and grass/herbaceous species). The C23-C33 n-alkane homologues were identified in soils with different abundances. Normally, C27 and C29 n-alkanes were the most abundant homologues in tree-leaf samples, while grass-derived n-alkanes were mostly C31 and C33 homologues. Soils were largely dominated by C29 and C31 n-alkanes. Odd-numbered C27-C33 soil n-alkane δ 13 C values ranged from -36.2‰ to -23.2‰, whereas their δ 2 H values showed a wider range of variability that fluctuated from -224‰ to -172‰. Molecular distribution in combination with radiocarbon analysis of soil organic matter (SOM) and δ 13 C and δ 2 H values of n-alkanes revealed a large contribution of C3 trees (both deciduous and coniferous trees/pine trees) as the main source of n-alkanes between the late Pleistocene and early Holocene (ca 15,000-8200 calibrated year before present/cal year BP). A clear shift toward more grassy/herbaceous vegetation was observed from the early Holocene (ca 11,700-8200 cal year BP) onwards. Distribution patterns of lipids and soil geochemical parameters showed that plants are the main source of SOM and that biodegradation and kinetic isotope fractionation are not the main reasons for 13 C enrichment in soil profiles. Past C3 vegetation shifts as well as paleoclimate changes (i.e., aridity) can have played a role in the observed 13 C depth profiles., 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 Inc.)

A comprehensive understanding of carbon cycling pathways in the soil-plant system is needed to develop models that accurately predict global carbon reservoir responses to anthropogenic perturbations. Honey is a carbon-rich natural food produced by wild and managed pollinating insects all over the world; the composition of a single sample is a function of millions of pollinator-plant interactions. We studied the 13 C/ 12 C and Δ 14 C of 121 honey samples sourced from the United States, and found a significant older carbon contribution. The effect is observed from 25 to 45° latitude, not correlated with 13 C/ 12 C, and consistent with a previously published study on European honeys. In specific cases, the measured values were up to 20 ‰ (Δ 14 C) higher than the expected atmospheric 14 CO 2 value for the given year, which shows a significant older carbon contribution. We hypothesize that the older carbon is from plant liquids derived in part from soil carbon or stored nonstructural carbohydrates from plants, which shifts the calibrated age of the sample by 5 years or more. Our work is the first to describe the widespread occurrence of older carbon in honey and shows that radiocarbon measurements can be a powerful tool to trace carbon allocations in terrestrial food webs and detect the atmosphere-soil-plant carbon cycle contributions., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Zsuzsa Lisztes-Szabó reports financial support was provided by Ministry for Culture and Innovation, Hungary. Edina Baranyai reports financial support was provided by Ministry for Culture and Innovation, Hungary. Ágota Ragyak reports financial support was provided by Ministry for Culture and Innovation, Hungary. Zsófi Sajtos reports financial support was provided by Ministry for Culture and Innovation, Hungary. Mihály Molnár reports financial support was provided by National Research, Development and Innovation Fund of Hungary. Szilárd Szabó reports financial support was provided by Széchenyi Plan Plus program. If there are other authors, they 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.)

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.)

The 1997/1998 El Niño event caused mass coral bleaching and mortality in many tropical and subtropical regions, including corals on Green Island, Taiwan, in the northwestern Pacific Ocean. This study analyzed coral carbon isotope ratios (δ 13 C), oxygen isotope ratios (δ 18 O), and Sr/Ca ratios for 29 years, including the 1997/1998 El Niño period, to examine how high water temperature events are recorded in coral geochemical indicators. Sr/Ca ratios in coral skeletons from Green Island show the lowest peak, means the highest temperature during the 1997/1998 El Niño period. However, we couldn't observe high-temperature events on δ 18 O. Furthermore, a negative δ 13 C shift was observed after El Niño events. The regime shift of δ 13 C might have been caused by temporal bleaching and/or a decrease in symbiotic algae due to high water temperature stress under the continuous decrease in δ 13 C in DIC due to the Suess effect., (© 2024. The Author(s).)

The successful utilization of stable carbon isotope approaches in investigating forest carbon dynamics has relied on the assumption that the carbon isotope compositions (δ13C) therein have detectable temporal variations. However, interpreting the δ13C signal transfer can be challenging, given the complexities involved in disentangling the effect of a single environmental factor, the isotopic dilution effect from background CO2 and the lack of high-resolution δ13C measurements. In this study, we conducted continuous in situ monitoring of atmospheric CO2 (δ13Ca) across a canopy profile in an old-growth temperate forest in northeast China during the normal year 2020 and the wet year 2021. Both years exhibited similar temperature conditions in terms of both seasonal variations and annual averages. We tracked the natural carbon isotope composition from δ13Ca to photosynthate (δ13Cp) and to ecosystem respiration (δ13CReco). We observed significant differences in δ13Ca between the two years. Contrary to in 2020, in 2021 there was a δ13Ca valley in the middle of the growing season, attributed to surges in soil CO2 efflux induced by precipitation, while in 2020 values peaked during that period. Despite substantial and similar seasonal variations in canopy photosynthetic discrimination (Δ13Ccanopy) in the two years, the variability of δ13Cp in 2021 was significantly lower than in 2020, due to corresponding differences in δ13Ca. Furthermore, unlike in 2020, we found almost no changes in δ13CReco in 2021, which we ascribed to the imprint of the δ13Cp signal on above-ground respiration and, more importantly, to the contribution of stable δ13C signals from soil heterotrophic respired CO2. Our findings suggest that extreme precipitation can impede the detectability of recent photosynthetic δ13C signals in ecosystem respiration in forests, thus complicating the interpretation of above- and below-ground carbon linkage using δ13CReco. This study provides new insights for unravelling precipitation-related variations in forest carbon dynamics using stable isotope techniques., (© The Author(s) 2024. Published by Oxford University Press.)

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 .

Preserved biological communities can provide baseline data about the historical ecosystems and environmental conditions that preceded recent anthropogenic alteration. Freshwater mussel shells show particularly good preservation, and the shell assemblages commonly found during archaeological excavations can offer insights into past ecosystems. We studied assemblages of Unio pictorum mussel shells from palaeochannel silts associated with the Late Bronze Age site of Must Farm in eastern England (c. 850 BC), on an ancient tributary of the modern-day River Nene. We compared archaeological shells from two sediment horizons (broadly 1300-700 BC) to live individuals collected from two analogous sites on the present-day Nene. Size and growth rate, interannual growth variability and stable isotope (δ 18 O and δ 13 C) composition were compared between the populations. Size and the von Bertalanffy growth parameter L ∞ differed among all four populations. Mean lengths and L ∞ were higher in the two modern populations (mean lengths 77.3 ± SE 0.8 and 73.8 ± SE 1.1 mm, L ∞ 91.8 ± 5.4 and 79.0 ± 8.1 mm) than the ancient populations (mean lengths 58.1 ± SE 1.6 mm and 68.4 ± SE 0.9 mm; L ∞ 71.5 ± 16.9 and 76.8 ± 6.2 mm). Modern individuals also showed greater variation in age-corrected year-to-year growth. δ 13 C was lower in modern shells (-11.8‰ for modern shells, -9.03‰ and -9.02‰ for ancient shell populations), potentially reflecting altered hydrological and nutrient regimes. δ 18 O and δ 13 C were positively correlated for all but one sampled ancient shell, but not modern shells. These results reflect changes in local environmental conditions, particularly the transition from a shallow, slow-flowing tributary to a deeper, canalised river with faster flow, as well as effects of anthropogenic nutrient enrichment. The findings demonstrate the importance of long-term data in studying anthropogenic ecosystem alteration and avoiding shifting baseline syndrome in conservation planning., (Global Change Biology© 2024 The Author(s). Global Change Biology published by John Wiley & Sons Ltd.)

The physical effects of climate warming have been well documented, but the biological responses are far less well known, especially at the ecosystem level and at large (intercontinental) scales. Global warming over the next century is generally predicted to reduce food web complexity, but this is rarely tested empirically due to the dearth of studies isolating the effects of temperature on complex natural food webs. To overcome this obstacle, we used 'natural experiments' across 14 streams in Iceland and Russia, with natural warming of up to 20°C above the coldest stream in each high-latitude region, where anthropogenic warming is predicted to be especially rapid. Using biomass-weighted stable isotope data, we found that community isotopic divergence (a universal, taxon-free measure of trophic diversity) was consistently lower in warmer streams. We also found a clear shift towards greater assimilation of autochthonous carbon, which was driven by increasing dominance of herbivores but without a concomitant increase in algal stocks. Overall, our results support the prediction that higher temperatures will simplify high-latitude freshwater ecosystems and provide the first mechanistic glimpses of how warming alters energy transfer through food webs at intercontinental scales., (© 2024 The Author(s). Global Change Biology published by John Wiley & Sons Ltd.)

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.)

Background: The use of stable carbon isotope ratios (δ13C) of sugar fractions of honey is a powerful tool to detect adulteration with sugar syrups. This is accomplished by calculating differences of the δ13C values between individual honey saccharides and comparing them to published purity criteria. A liquid chromatography-isotope ratio mass spectrometry (LC-IRMS) method for the determination of δ13C values of sugars in honey was previously validated by an interlaboratory comparison, but no further guidance was given how to include the obtained precision figures of the compound-specific δ13C values in the purity assessment of honey., Objective: To use existing data to estimate the standard deviation of the repeatability (sr) and reproducibility (sR) of differences (Δ δ13C) between the δ13C values of individual honey saccharides., Methods: Previously published δ13C values were used to calculate differences (Δ δ13C values) between δ13C fructose-δ13C glucose, δ13C glucose-δ13C disaccharides, etc. in a honey sample; sr and sR of Δ δ13C values were calculated according to ISO 5725-2:2019., Results: The Δ δ13C sr and sR values were essentially of the same magnitude as the sr and sR values of δ13C values of the sugar fractions. The precision of the Δ δ13C values was used to estimate the critical difference for comparing a test result with a reference value according to ISO 5725-6:1994. This varied between 0.26 and 1.10‰., Conclusion: The estimated critical differences can be used to determine whether a honey test result complies with published Δ δ13C purity criteria., Highlight: The proposed procedure will increase confidence in decisions based on compound-specific δ13C values regarding the conformity of honey with published purity criteria., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL.)

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.)

Using stable isotopes to detect and analyze the geographical origin of insects represents an important traceability technology, which requires a rich isotope database. In this study, we representatively sampled the Chinese provinces where flighted spongy moth complex (FSMC) has been reported and, for the first time, used co-kriging interpolation to predict the distribution patterns of FSMC δ 13 C values in the main distribution areas. From 2020 to 2022, we set up 60 traps in 12 provinces and cities in China and collected 795 FSMCs. Then, 6 main climatic factors were obtained by multi-collinearity screening from 21 types of meteorological data collected at the sample plots, and a correlation analysis was carried out by combining longitude, latitude, and altitude data with the δ 13 C values of FSMC. Next, we performed a co-kriging interpolation using the 2 climatic factors with the highest correlation (isothermality and altitude) and the δ 13 C values of FSMC. A cross-validation was performed to systematically test 11 candidate models and select the best semi-variogram model ("Exponential"), which was then used to build a co-kriging interpolation model. The geographical distribution patterns of the FSMC δ 13 C values obtained from the 2 interpolation models (i.e., interpolated with isothermality and altitude, respectively) were almost the same. Moreover, the δ 13 C values varied significantly at the regional scale, showing regular changes in spatial distribution. Overall, the reference indicator map of the δ 13 C values generated from stable isotopes can be used to greatly improve the efficiency of discrimination analyses on the geographical origin of FSMC., (© 2024 Institute of Zoology, Chinese Academy of Sciences.)