Simultaneous observation of 2H and 13C enrichment of methyl phosphonic acid via Orbitrap-IRMS with applications to nerve agent forensics.

Quantification of the stable isotopes within a compound aids forensic investigations as it provides a fingerprint which can determine that compound's source substrates, synthetic route, and possible mechanisms of degradation. Previous stable isotope studies have explored 13C and 2H measurements of the sarin precursors methylphosphonic dichloride (DC) and methylphosphonic difluoride (DF) as forensic signatures. However, these measurements required different sample preparations and measurement techniques. Orbitrap isotope ratio mass spectrometry (Orbitrap-IRMS) is a developing technique which can characterize multiple stable isotopes simultaneously. Here, we apply Orbitrap-IRMS to simultaneously observe the 13C and 2H content of methylphosphonic acid (MPA), the hydrolysis product of DC and DF, which can be used as a proxy for the isotopic content of DC and DF. Our method requires 20 min analyses and consumes ≈60 nmol of sample, with precisions of ≈0.9 ‰ (13C) and ≈3.6 ‰ (2H). We apply our method to both commercially acquired MPA and MPA obtained from the hydrolysis of commercially acquired DC. We validate our methods via comparison to elemental-analyzer isotope ratio mass spectrometry (EA-IRMS). The combined 13C and 2H measurement creates a more robust forensic tool than either isotope individually. Our results demonstrate the viability of Orbitrap-IRMS for chemical forensic measurements.

Chemometric Discrimination of the Geographical Origin of Rheum tanguticum by Stable Isotope Analysis.

Rheum tanguticum is one of the primary rhubarb species used for food and medicinal purposes, and it has recently been gaining more attention and recognition. This research represents the first attempt to use stable isotopes and elemental analysis via IRMS to identify the geographical origin of Rheum tanguticum. A grand total of 190 rhubarb samples were gathered from 38 locations spread throughout the provinces of Gansu, Sichuan, and Qinghai in China. The carbon content showed a decreasing trend in the order of Qinghai, followed by Sichuan, and then Gansu. Nitrogen content was notably higher, with Qinghai and Sichuan displaying similar levels, while Gansu had the lowest nitrogen levels. Significant differences were noted in the δ13C (-28.9 to -26.5‱), δ15N (2.6 to 5.6‱), δ2H (-120.0 to -89.3‱), and δ18O (16.0‱ to 18.8‱) isotopes among the various rhubarb cultivation areas. A significant negative correlation was found between %C and both longitude and humidity. Additionally, δ13C and δ15N isotopes were negatively correlated with longitude, and δ15N showed a negative correlation with humidity as well. δ2H and δ18O isotopes exhibited a strong positive correlation with latitude, while significant negative correlations were observed between δ2H and δ18O isotopes and temperature, precipitation, and humidity. The LDA, PLS-DA, and k-NN models all exhibited strong classification performance in both the training and validation sets, achieving accuracy rates between 82.1% and 91.7%. The combination of stable isotopes, elemental analysis, and chemometrics provides a reliable and efficient discriminant model for accurately determining the geographical origin of R. tanguticum in different regions. In the future, the approach will aid in identifying the geographical origin and efficacy of rhubarb in other studies.

Detection of adulteration in Iranian grape molasses added glucose/fructose/sugar beet syrups with 13C/12C isotope ratio analysis method.

Grape molasses (GM), produced from grapes, is a traditional Iranian food and is widely consumed in Iran. However, GM adulteration is among the most widespread illegitimate procedures involving contamination of food with foreign materials, such as adding sugar-water solution, date syrup, sugar beet syrup, and grape sauce. This study used stable carbon 13C/12C isotope ratio analysis method to detect adulteration of GM samples with glucose syrups (GS), fructose syrups (FS), and beet sugar syrups (BS) at the ratio of 0%, 10%, 30%, and 50% (by weight). Physicochemical properties of GM including °Brix, conductivity, specific gravity, pH, moisture content, ash content, hydroxymethyl furfural, sugar content, and rheological properties of samples were investigated. The δ13C isotope ratio of the GM was determined as -26.61%, that of the GS as -13.23%, that of the FS as -13.42%, and that of the BS as -16.58%. The δ13C isotope ratio increased by the addition of adulterant syrups to GM. The addition of each adulterant syrup had a different effect on the physicochemical parameters; however, the °Brix and specific gravity had a positive correlation with the δ13C isotope ratio results. The magnitudes of G' and G" increase with an increase in frequency representing the viscoelastic behavior of samples. The obtained results of this study suggest the use of δ13C isotope ratio method as a fast and accurate method to investigate the adulteration of grape molasses.

Isotopic Composition of C, N, and S as an Indicator of Endometrial Cancer.

OBJECTIVES: The metabolic pathway of cancerous tissue differs from healthy tissue, leading to the unique isotopic composition of stable isotopes at their natural abundance. We have studied if these changes can be developed into diagnostic or prognostic tools in the case of endometrial cancer. METHODS: Measurements of stable isotope ratios were performed using isotope ratio mass spectrometry for nitrogen, carbon, and sulfur isotopic assessment. Uterine tissue and serum samples were collected from patients and the control group. RESULTS: At a natural abundance, the isotopic compositions of all three of the studied elements of uterus cancerous and healthy tissues are different. However, no correlation of the isotopic composition of the tissues with that of serum was found. CONCLUSIONS: Differences in the isotopic composition of the tissues might be a potential prognostic tool. However, the lack of a correlation between the differences in the isotopic composition of the tissues and serum seems to exclude their application as diagnostic biomarkers, which, however, might be possible if a position-specific isotopic analysis is performed.

Use of stable isotope ratio analysis to investigate the biology and clinical significance of seal parasites.

Stranded seals are often infected with a broad range of parasites, although whether they are the cause of significant morbidity or an incidental finding is usually unclear. In this study we used stable isotope ratio analysis, a method frequently used to investigate food webs, to explore the extent to which common seal parasites feed directly on host tissue and fluids or absorb host-derived metabolites, which in turn may give an indication of their potential impact on the host's health. The trophic discrimination factor Δ15N for the nasal mite, Halarachne halichoeri, was mostly positive, ranging between −0.015 and 3.2‰ (n = 6), while for the Acanthocephalan worm, Corynosoma strumosum and the anisakid nematode, Pseudoterranova decipiens, Δ15N ranged between −4.2 and −2.0‰ (n = 7), and between −6.7 and −0.8‰ (n = 5) respectively. In the case of the lungworm, Otostrongylus circumlitus, Δ15N measured between −5.6 and 0‰ for worms collected in the stomach (n = 5), between −1.1 and 0.2‰ for worms collected from the heart (n = 3), between −0.7 and 1.9‰ for worms situated in the lungs (n = 4). Based on Δ15N, parasites could be clearly divided into those that were on a higher trophic level than their host suggesting a predator­prey-like relationship, and those that were not. It is hypothesized that Δ15N may be indicative of the clinical significance of parasite­host associations.

The carbon isotope ratio of breath is elevated by short-term and long-term added sugar and animal protein intake in a controlled feeding study.

BACKGROUND: The breath carbon isotope ratio (CIR) was recently identified as a noninvasive candidate biomarker of short-term added sugars (AS) intake. OBJECTIVES: This study aimed to better understand the potential of the breath CIR as a dietary biomarker. We evaluated the effects of short-term and long-term intakes of AS, animal protein (AP), and related variables on breath CIR, in the context of typical dietary intake patterns. METHODS: We conducted a 15-d controlled feeding study of 100 adults (age 18-70 y, 55% females) in Phoenix, AZ. Participants were provided individualized diets that approximated habitual food intakes and recorded the timing of food consumption. Three breath samples (fasting, midday, and evening) were collected on each of 3 nonconsecutive study days. We modeled the effects of dietary intake in each of 8 h preceding collection of the breath sample on breath CIR with a linear mixed model, which also included 15-d mean intakes, sex, age, and BMI. RESULTS: Median (IQR) intakes of AS and AP in our study were 65 (38) and 67 (33) g/d, respectively. Midday and evening breath CIRs correlated strongly with each other (0.80) and with fasting breath CIR (0.77 and 0.68, respectively). In our linear mixed models, breath CIR increased by AS consumed 1-4 h before sample collection, AP consumed 3-6 h before sample collection, and 15-d intakes of AS and AP, all with similar effect sizes. The breath CIR was also inversely associated with 15-d intakes of intrinsic sugars and plant protein; thus, associations with 15-d intakes were particularly strong when expressed proportionally as the AS ratio (added sugars/total sugars) and AP ratio (animal protein/total protein). CONCLUSIONS: The breath CIR is a promising measure of long-term intakes of AS and AP, especially as proportional intakes. Approaches to increase specificity would benefit the further development of this biomarker.

Geochemical fingerprinting of Norway spruce from the Eastern Carpathians: Sr isotopic and multi-elemental signatures.

Ensuring efficient wood traceability within procurement chains is essential for establishing sustainable forest management and minimizing environmental damage in countries that produce and export timber. While some progress has been made with key legislative reforms to tackle this issue, the effectiveness of law enforcement still relies on the availability of appropriate analytical tools to determine the provenance of wood. This study documents the Sr isotopic and multi-elemental signatures of Norway spruce trees in the Eastern Carpathians, Romania - an area known for intensive forest logging. The research focused on trees from Rodna, Rarau, and Calimani, analyzing tree rings from 1970 to 2020 to assess temporal variability and develop site-specific geochemical fingerprints. In conjunction with tree samples, corresponding soil, groundwater, and river samples were analyzed, providing a comprehensive view of the bioavailable 87Sr/86Sr ratios in these ecosystems. The study found a strong correlation between the Sr isotope ratios in the wood and those in corresponding water and soil samples, confirming that the wood accurately mirrors local geochemical signatures. The Sr isotope ratios were consistent across most of the cores from trees within the same site but varied among sites within the same region. Principal Component Analysis of the Sr isotopic and multi-elemental data highlighted closer clustering of samples from the Calimani area, though distinguishing the samples from the Rodna and Rarau regions proved more difficult due to overlapping data points. This study broadens our understanding of geochemical variability and its implications. The insights gained provide a valuable foundation for future research aimed at enhancing wood traceability methods.

Machine Learning Models to Classify Shiitake Mushrooms (Lentinula edodes) According to Their Geographical Origin Labeling.

The shiitake mushroom has gained popularity in the last decade, ranking second in the world for mushrooms consumed, providing consumers with a wide variety of nutritional and healthy benefits. It is often not clear the origin of these mushrooms, so it becomes of great importance to the consumers. In this research, different machine learning algorithms were developed to determine the geographical origin of shiitake mushrooms (Lentinula edodes) consumed in Korea, based on experimental data reported in the literature (δ13C, δ15N, δ18O, δ34S, and origin). Regarding the origin of shiitake in three categories (Korean, Chinese, and mushrooms from Chinese inoculated sawdust blocks), the random forest model presents the highest accuracy value (0.940) and the highest kappa value (0.908) for the validation phase. To determine the origin of shiitake mushrooms in two categories (Korean and Chinese, including mushrooms from Chinese inoculated sawdust blocks in the latter ones), the support vector machine model is chosen as the best model due to the high accuracy (0.988) and kappa (0.975) values for the validation phase. Finally, to determine the origin in two categories (Korean and Chinese, but this time including the mushrooms from Chinese inoculated sawdust blocks in the Korean ones), the best model is the random forest due to its higher accuracy value (0.952) in the validation phase (kappa value of 0.869). The accuracy values in the testing phase for the best selected models are acceptable (between 0.839 and 0.964); therefore, the predictive capacity of the models could be acceptable for their use in real applications. This allows us to affirm that machine learning algorithms would be suitable modeling instruments to determine the geographical origin of shiitake.

Kinetic Modeling of In Vivo K+ Distribution and Fluxes with Stable K+ Isotopes: Effects of Dietary K+ Restriction.

Maintaining extracellular potassium (K+) within narrow limits, critical for membrane potential and excitability, is accomplished through the internal redistribution of K+ between extracellular fluid (ECF) and intracellular fluid (ICF) in concert with the regulation of renal K+ output to balance K+ intake. Here we present evidence from high-precision analyses of stable K+ isotopes in rats maintained on a control diet that the tissues and organs involved in the internal redistribution of K+ differ in their speed of K+ exchange with ECF and can be grouped into those that exchange K+ with ECF either rapidly or more slowly ("fast" and "slow" pools). After 10 days of K+ restriction, a compartmental analysis indicates that the sizes of the ICF K+ pools decreased but that this decrease in ICF K+ pools was not homogeneous, rather occurring only in the slow pool (15% decrease, p < 0.01), representing skeletal muscles, not in the fast pool. Furthermore, we find that the dietary K+ restriction is associated with a decline in the rate constants for K+ effluxes from both the "fast" and "slow" ICF pools (p < 0.05 for both). These results suggest that changes in unidentified transport pathways responsible for K+ efflux from ICF to ECF play an important role in buffering the internal redistribution of K+ between ICF and ECF during K+ restriction. Thus, the present study introduces novel stable isotope approaches to separately characterize heterogenous ICF K+ pools in vivo and assess K+ uptake by individual tissues, methods that provide key new tools to elucidate K+ homeostatic mechanisms in vivo.

Improving the Determination of Carbon Isotope Ratios of Endogenous Steroids Found in Human Serum.

The determination of serum concentrations of testosterone (T) and 4-androstenedione (A4) was implemented into the steroidal module of the Athlete Biological Passport in 2023. Monitoring T, A4, and the ratio of T/A4 in a longitudinal manner enables the detection of the misuse of low-dose T administrations especially in female athletes, whereas urinary markers of the steroid profile may not be influenced significantly. In contrast to the urinary steroid profile, knowledge on confounding factors regarding serum concentrations of T and A4 is yet comparably scarce, and corroborating exogenous sources of the target analytes by isotope ratio mass spectrometry (IRMS) is desirable. In a recent study, it was demonstrated that carbon isotope ratios (CIRs) of serum steroids can be determined if analyte concentrations permit. The therein-employed method utilized 2D-GC/IRMS, and only a limited number of potential endogenous reference compounds were available. The here-presented approach uses complementary analyte purification strategies, addressing previous limitations. A high-performance liquid chromatography cleanup was developed and fully validated for serum steroids in order to enable all doping control laboratories to potentially implement this method alongside existing protocols for urinary steroids. Besides the already-investigated endogenous steroids cholesterol, dehydroepiandrosterone sulfate, androsterone sulfate, and epiandrosterone sulfate, two additional steroids were included in the test menu, namely, pregnenolone sulfate and 5-androstene-3ß,17ß-diol sulfate. Serum steroid concentrations down to 25 ng/mL were found to allow robust CIR determinations, and a reference population encompassing 124 male and female athlete samples was investigated to enable the calculation of population-based thresholds for all relevant steroid combinations.

Investigation of isotopic linkages between raw materials and black powder.

The stable isotope analysis of black powder (BP) is of great significance for its comparison and source inference. Previous studies have verified the feasibility of distinguishing different BP samples through stable isotopes. However, the impact of raw materials and synthesis processes on the stable isotopes of BP remains unclear. On the one hand, the raw materials of BP are widely sourced, and whether stable isotopes can distinguish different source materials remains to be studied. On the other hand, the synthesis of BP involves the physical mixing of raw materials, and whether this process leads to isotope fractionation also needs further investigation. To address these problems, stable isotope ratios of 27 charcoals, 15 potassium nitrates, 6 self-made and 10 commercial BP samples were analyzed. The results showed that the stable isotope ratios can be utilized to distinguish charcoals and potassium nitrates from different manufacturers and batches. No significant differences in the nitrogen and oxygen stable isotope ratios between the self-made BP and its raw materials were observed, indicating that the physical mixing process does not induce significant fractionation of stable isotopes. However, the carbon stable isotope ratios of charcoal increased (within 2SD) after being synthesized into BP. Due to the utilization of additives and variations in the synthesis process, the correlation between the stable isotope ratios of commercial BP and its raw materials was complex. The findings of this study provide a scientific reference for tracing the source of BP.

Carbon-13-isotopomics and metabolomics of fatty acids from triacylglycerols: overcoming the limitations of GC-C-IRMS for short- and medium-acyl chains.

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 δ13C 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 13C-isotopomic and metabolomic biomarker quantitation within quasi-universal TAG compounds, encompassing the short- and medium-acyl chains.

A multi-analytical approach for the identification of pollutant sources on black crust samples: Stable isotope ratio of carbon, sulphur, and oxygen.

This study is focused on the identification of pollutant sources on black crust (BC) samples from the Monumental Cemetery of Milan (Italy), through a multi-analytical approach based on the determination of stable isotope ratios of carbon, sulphur, and oxygen. Six black crust samples, mainly developed on marble sculptures over a time span of 100-150 years, were analysed. For the first time, δ13C was measured for BC samples: δ13C values of the pulverized samples (from -1.2 to +1.3 ‰) are very close to the values obtained from the carbonate matrix, whereas after the removal of the matrix through acidification, δ13C values of BC samples from Milan range from -27.2 to -22.1 ‰, with no significant variation between samples with different ratios of organic carbon to elemental carbon. In sum, the δ13C values obtained for all BC samples fall within the range of anthropogenic emissions such as vehicle traffic, coal combustion and industrial emissions. δ34S and δ18O values of sulphate from BC samples range from -6.3 to +7.0 ‰ and from +7.6 to +10.5 ‰, respectively. Coupling the analysis of the oxygen isotope ratio with that of sulphur enables a more precise identification of the origin of sulphates: the observed isotopic composition falls in the range typical for anthropogenic emission of sulphur dioxide. Overall, in this study, C, S and O isotopes were combined for the first time to assess pollutant sources on black crust samples: this multi-stable isotope approach allowed to show that the BC formation on monuments from the Monumental Cemetery of Milan mostly results from anthropogenic emissions from fossil fuels combustion by road vehicles and factories, as well as domestic heating.

First Report on the Trophic Transfer and Priority List of Liquid Crystal Monomers in the Pearl River Estuary.

Liquid crystal monomers (LCMs) are emerging organic pollutants due to their potential persistence, toxicity, and bioaccumulation. This study first characterized the levels and compositions of 19 LCMs in organisms in the Pearl River Estuary (PRE), estimated their bioaccumulation and trophic transfer potential, and identified priority contaminants. LCMs were generally accumulated in organisms from sediment, and the LCM concentrations in all organisms ranged from 32.35 to 1367 ng/g lipid weight. The main LCMs in organisms were biphenyls and analogues (BAs) (76.6%), followed by cyanobiphenyls and analogues (CBAs) (15.1%), and the least were fluorinated biphenyls and analogues (FBAs) (11.2%). The most abundant LCM monomers of BAs, FBAs, and CBAs in LCMs in organisms were 1-(4-propylcyclohexyl)-4-vinylcyclohexane (15.1%), 1-ethoxy-2,3-difluoro-4-(4-(4-propylcyclohexyl) cyclohexyl) benzene (EDPBB, 10.1%), and 4'-propoxy-4-biphenylcarbonitrile (5.1%), respectively. The niche studies indicated that the PRE food web was composed of terrestrial-based diet and marine food chains. Most LCMs exhibited biodilution in the terrestrial-based diet and marine food chains, except for EDPBB and 4,4'-bis(4-propylcyclohexyl) biphenyl (BPCHB). The hydrophobicity, position of fluorine substitution of LCMs, and biological habits may be important factors affecting the bioaccumulation and trophic transfer of LCMs. BPCHB, 1-(prop-1-enyl)-4-(4-propylcyclohexyl) cyclohexane, and EDPBB were characterized as priority contaminants. This study first reports the trophic transfer processes and mechanisms of LCMs and the biomonitoring in PRE.

Evaluating Source Complexity in Blended Milk Cheese: Integrated Strontium Isotope and Multi-Elemental Approach to PDO Graviera Naxos.

Graviera Naxos, a renowned cheese with Protected Designation of Origin status, is crafted from a blend of cow, goat, and sheep milk. This study focused on assessing the Sr isotopic and multi-elemental composition of both the processed cheese and its ingredients, as well as the environmental context of Naxos Island, including samples of milk, water, soil, and feed. The objective was to delineate the geochemical signature of Graviera Naxos cheese and to explore the utility of Sr isotopes as indicators of geographic origin. The 87Sr/86Sr values for Graviera Naxos samples ranged from 0.70891 to 0.70952, indicating a relatively narrow range. However, the Sr isotopic signature of milk, heavily influenced by the feed, which originates from geologically distinct areas, does not always accurately reflect the local breeding environment. Multi-elemental analysis revealed variations in milk composition based on type and season; yet, no notable differences were found between raw and pasteurized milk. A mixing model evaluating the contributions of milk, sea salt, and rennet to the cheese's Sr isotopic signature suggested a significant average contribution of 73.1% from sea salt. This study highlights the complexities of linking dairy products with their geographical origins and emphasizes the need for sophisticated geochemical authentication methods.

Analytical strategies for sensitive and precise determination of 87Sr/86Sr in olive oil through ion extraction, chromatographic separation, and multicollector inductively-coupled plasma mass-spectrometry.

Several food regulatory bodies regard olive oil as highly susceptible to food fraud, largely due to its substantial economic worth. Precise analytical tools are being developed to uncover these types of fraud. This study examines an innovative approach to extract strontium (Sr) from the olive oil matrix (via EDTA complexation and ion-exchange chromatography) and to determine its isotope composition by MC-ICP-MS. This technique was compared to a commonly used technique (i.e. acid extraction and extraction chromatography), and then validated. Three olive oils that are sold in France were prepared and analyzed by two methods: 1) acid extraction prior to Sr purification by Sr-spec resin and 2) complexation by EDTA prior to Sr purification by AG50W-X8. These methods were applied for the determination of the 87Sr/86Sr isotope ratio of 23 olive oils from various countries. We also demonstrated the feasibility of the method for the detection of olive oil mixtures.

The marriage between stable isotope ecology and plant metabolomics - new perspectives for metabolic flux analysis and the interpretation of ecological archives.

Even though they share many thematical overlaps, plant metabolomics and stable isotope ecology have been rather separate fields mainly due to different mass spectrometry demands. New high-resolution bioanalytical mass spectrometers are now not only offering high-throughput metabolite identification but are also suitable for compound- and intramolecular position-specific isotope analysis in the natural isotope abundance range. In plant metabolomics, label-free metabolic pathway and metabolic flux analysis might become possible when applying this new technology. This is because changes in the commitment of substrates to particular metabolic pathways and the activation or deactivation of others alter enzyme-specific isotope effects. This leads to differences in intramolecular and compound-specific isotope compositions. In plant isotope ecology, position-specific isotope analysis in plant archives informed by metabolic pathway analysis could be used to reconstruct and separate environmental impacts on complex metabolic processes. A technology-driven linkage between the two disciplines could allow us to extract information on environment-metabolism interaction from plant archives such as tree rings but also within ecosystems. This would contribute to a holistic understanding of how plants react to environmental drivers, thus also providing helpful information on the trajectories of the vegetation under the conditions to come.

Stable Isotope Ratio Analysis for the Geographic Origin Discrimination of Greek Beans "Gigantes-Elefantes" (Phaseolus coccineus L.).

Adulteration of high-value agricultural products is a critical issue worldwide for consumers and industries. Discrimination of the geographical origin can verify food authenticity by reducing risk and detecting adulteration. Between agricultural products, beans are a very important crop cultivated worldwide that provides food rich in iron and vitamins, especially for people in third-world countries. The aim of this study is the construction of a map of the locally characteristic isotopic fingerprint of giant beans, "Fasolia Gigantes-Elefantes PGI", a Protected Geographical Indication product cultivated in the region of Kastoria and Prespes, Western Macedonia, Greece, with the ultimate goal of the discrimination of beans from the two areas. In total, 160 samples were collected from different fields in the Prespes region and 120 samples from Kastoria during each cultivation period (2020-2021 and 2021-2022). The light element (C, N, and S) isotope ratios were measured using Isotope Ratio Mass Spectrometry (IRMS), and the results obtained were analyzed using chemometric techniques, including a one-way ANOVA and Binomial logistic regression. The mean values from the one-way ANOVA were δ15NAIR = 1.875‱, δ13CV-PDB = -25.483‱, and δ34SV-CDT = 4.779‱ for Kastoria and δ15NAIR = 1.654‱, δ13CV-PDB = -25.928‱, and δ34SV-CDT = -0.174‱ for Prespes, and showed that stable isotope ratios of C and S were statistically different for the areas studied while the Binomial logistic regression analysis that followed correctly classified more than 78% of the samples.

Using isotopic fingerprints in gastropod shells to validate commercial production pathway and geographic provenance.

Growing demand for high-value seafood is fuelling provenance fraud, which threatens the sustainability of wild fisheries while posing biosecurity and human health risks. Here, we investigated carbon (δ 13C) and oxygen (δ 18O) isotopes in abalone shells (Haliotis sp.) to determine the production method and geographical provenance. Using X-ray diffraction and isotope ratio mass spectrometry, we found that shell mineralogy did not influence isotope values. Isotope values between wild and farmed sectors were statistically different, with 64% of individuals correctly classified as farmed or wild. Subsequently, we successfully distinguished the provenance of abalone collected from farms (with 83% of individuals correctly classified), as well as wild-caught abalone collected from four state jurisdictions (with 88% correctly classified). Carbon isotopes were strongly correlated to longitude, with both isotopes correlated to latitude. Overall, this study demonstrates the potential of isotopic fingerprints in gastropod shells to track the provenance of commercially valuable species.

Additive positive effect of warming and elevated nitrogen deposition on Sphagnum biomass production at mid-latitudes.

Global warming and increased atmospheric nitrogen (N) deposition can adversely impact Sphagnum moss populations and ecological functions in peatlands. Based on the anticipated increases in temperature and N levels at global scale, we investigated the effects of simultaneous warming and N treatment on growth and ecophysiological activity of Sphagnum papillosum, a predominant moss at mid-latitudes, utilizing a growth chamber experiment. Warming treatments increased the maximum yield of photosystem II (Fv/Fm) of S. papillosum while decreasing the stable carbon isotope ratio. However, warming treatment alone did not cause significant changes in the biomass increase from that of the control. Regarding N treatment, the low N treatment decreased Fv/Fm under the current temperature but did not affect the biomass increase. In contrast to these results, a simultaneous warming and high N treatment significantly enhanced the biomass production compared to that of the control, exhibiting additive effect of warming and high N treatment on Sphagnum biomass production. These responses were attributed to the improved photosynthetic performances by warming and N treatment. The results of this study contribute to the prediction of Sphagnum responses to warming and changes in N deposition.