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Sulfur isotope composition of organic tissues is a commonly used tool for gathering information about provenance and diet in archaeology and paleoecology. However, the lack of maps predicting sulfur isotope variations on the landscape limits the possibility to use this isotopic system in quantitative geographic assignments. We compiled a database of 2,680 sulfur isotope analyses in the collagen of archaeological human and animal teeth from 221 individual locations across Western Europe. We used this isotopic compilation and remote sensing data to apply a multivariate machine-learning regression, and to predict sulfur isotope variations across Western Europe. The resulting model shows that sulfur isotope patterns are highly predictable, with 65% of sulfur isotope variations explained using only 4 variables representing marine sulfate deposition and local geological conditions. We used this novel sulfur isoscape and existing strontium and oxygen isoscapes of Western Europe to apply triple isotopes continuous-surface probabilistic geographic assignments to assess the origin of a series of teeth from local animals and humans from Brittany. We accurately and precisely constrained the origin of these individuals to limited regions of Brittany. This approach is broadly transferable to studies in archaeology and paleoecology as illustrated in a companion paper (Colleter et al. 2021).

Estimates suggest billions of dollars are lost annually in the US due to fuel tax fraud. One method of fuel fraud is called “cocktailing” and involves blending products that are non-taxed, lower value, taxed at a lower rate, or unwanted/less-refined petroleum to diesel fuels. The goal of this study was to investigate compound specific isotope analysis (CSIA) using isotope ratio mass spectrometry (IRMS) for small aromatics contained in diesel fuel to determine whether this approach could be used to identify cocktailing and potentially fingerprint possible sources. However, the high chemical complexity of diesel fuels complicates CSIA owing to the need to fully separate individual compounds for effective isotope analysis. Therefore, different methods were investigated to selectively isolate aromatics for CSIA and evaluate these methods for isotopic fractionation. Analyses indicate that there is enough variability in isotopic ratios (δ2H and δ13C) between toluene samples obtained from different sources to use CSIA to differentiate/identify the origin of potential fuel adulterants. Three isolation methods were identified that provided sufficiently pure aromatic fractions for CSIA: selective solvent extraction, ionic liquid coated solid phase microextraction (SPME), and a combination of the two. However, due to the labor-intensive nature of selective solvent extraction, ionic liquid coated SPME represents the best method to quickly isolate aromatics from diesel fuel, without sacrificing selectivity or sensitivity. All methods tested can result in isotopic fractionation, but this can be compensated for by applying a correction factor. Furthermore, the chemical composition of a sample appeared to be important in the degree to which fractionation occurred during isolation. While the tested approaches for aromatic extraction from diesel showed promise, additional studies are required to refine and validate the methods prior to routine use in fuel cocktailing investigations. [Display omitted] •Toluene from diverse sources has adequate δ2H and δ13C variability for isotopic differentiation from other chemicals in a fuel.•Four different isolation methods coupled with CSIA of small aromatics in diesel fuel were evaluated.•Three isolation methods provided sufficiently pure aromatic fractions for CSIA, ionic liquid coated SPME was the best.•All isolation methods resulted in some isotopic fractionation; however, this is amenable to mathematical correction

The outbreak of certain bacteria can trigger severe diseases, even posing a threat to public safety, leading to significant social panic and economic losses. Thus, tracing the origin of bacteria is of great significance. Stable isotope analysis technology offers a new way to determine the geographical information of bacteria, yet related research still fails to meet the application requirements of this technology in practical cases of bacterial traceability. In this study, a systematic stable isotope analysis method for bacteria and their culture conditions, based on practical geographical environments, was established for the first time. Escherichia coli and Staphylococcus aureus were cultured with water from five regions and different culture media, and the stable isotope ratios of H/O/C/N in the two bacteria and the culture media were measured to explore the relationship between bacteria and their cultivation site. The results showed that there were linear relationships between the hydrogen and oxygen stable isotopes of the two bacteria and the culture water. The combined discriminant model constructed using multi-isotope (H/O/C/N) characteristics achieved a 100% accuracy rate in identifying the types of culture media. These results indicate that research on the isotope association between bacteria and their culture water can be used to infer the cultivation region, and the specific source of bacteria can be further inferred through the multi-isotope combination discriminant model. This study can provide a relatively complete research idea for bacterial geographical traceability research, and improve the efficiency and accuracy of bacterial traceability work in practical investigations. Key points Established a systematic stable isotope analysis method for bacteria. E. coli and S. aureus have linear H/O isotope correlations with culture water. Multi-isotope discriminant model achieves 100% accuracy in identifying culture media.

Megalithism has been repetitively tied to specialised herding economies in Iberia, particularly in the mountainous areas of the Basque Country. Legaire Sur, in the uplands of Álava region, is a recently excavated passage tomb (megalithic monument) that held a minimum number of 25 individuals. This study analysed the carbon, nitrogen, oxygen, and strontium isotope ratios of 18 individuals, in a multi-tissue sampling study (successional tooth enamel sampling, incremental dentine sampling, and bulk bone collagen sampling). The results provide a high-resolution reconstruction of individual mobility, weaning, and dietary lifeways of those inhumed at the site. Oxygen and strontium isotope analysis suggest all individuals come from a similar, likely local, geological region, aside from one biological female who presents a notably different geographical birthplace, weaning, and dietary life history than the rest of the burial population. Comparisons to other nearby megalithic sites (∼35km as the crow flies), located in a valley area, reveal that, whilst sharing the same mortuary practices, these individuals held notably different lifeways. They highlight notably earlier ages of cessation of nursing (≤2 years at Legaire Sur vs. ≥4 years in other megalithic tombs), and a greater dependence on pastoralism than previously observed in lowland megalithic graves. The results from Legaire Sur reveal the complexity of the Late Neolithic-Chalcolithic transition in north-central Iberia, categorising yet another separate socio-economic group with distinctive lifeways inhabiting the region.

The characterization of Neandertals’ diets has mostly relied on nitrogen isotope analyses of bone and tooth collagen. However, few nitrogen isotope data have been recovered from bones or teeth from Iberia due to poor collagen preservation at Paleolithic sites in the region. Zinc isotopes have been shown to be a reliable method for reconstructing trophic levels in the absence of organic matter preservation. Here, we present the results of zinc (Zn), strontium (Sr), carbon (C), and oxygen (O) isotope and trace element ratio analysis measured in dental enamel on a Pleistocene food web in Gabasa, Spain, to characterize the diet and ecology of a Middle Paleolithic Neandertal individual. Based on the extremely low δ66Zn value observed in the Neandertal’s tooth enamel, our results support the interpretation of Neandertals as carnivores as already suggested by δ15N isotope values of specimens from other regions. Further work could help identify if such isotopic peculiarities (lowest δ66Zn and highest δ15N of the food web) are due to a metabolic and/or dietary specificity of the Neandertals.

Maya archaeologists have long been interested in understanding ancient diets because they provide information about broad-scale economic and societal transformations. Though paleodietary studies have primarily relied on stable carbon ( δ 13 C) and nitrogen ( δ 15 N) isotopic analyses of human bone collagen to document the types of food people consumed, stable sulfur ( δ 34 S) isotope analysis can potentially provide valuable data to identify terrestrial, freshwater, or marine/coastal food sources, as well as determine human mobility and migration patterns. Here we assess applications of δ 34 S for investigating Maya diet and migration through stable isotope analyses of human bone collagen ( δ 13 C, δ 15 N, and δ 34 S) from 114 individuals from 12 sites in the Eastern Maya lowlands, temporally spanning from the Late Preclassic (300 BCE—300 CE) through Colonial periods (1520–1800 CE). Results document a diet dominated by maize and other terrestrial resources, consistent with expectations for this inland region. Because δ 34 S values reflect local geology, our analyses also identified recent migrants to the Eastern lowlands who had non-local δ 34 S signatures. When combined with other indicators of mobility (e.g., strontium isotopes), sulfur isotopic data provide a powerful tool to investigate movement across a person’s lifespan. This study represents the largest examination of archaeological human δ 34 S isotope values for the Maya lowlands and provides a foundation for novel insights into both subsistence practices and migration.

The major components of human diet both past and present may be estimated by measuring the carbon and nitrogen isotope ratios (δ13C and δ15N) of the collagenous proteins in bone and tooth dentine. However, the results from these two tissues differ substantially: bone collagen records a multi-year average whilst primary dentine records and retains time-bound isotope ratios deriving from the period of tooth development. Recent studies harnessing a sub-annual temporal sampling resolution have shed new light on the individual dietary histories of our ancestors by identifying unexpected radical short-term dietary changes, the duration of breastfeeding and migration where dietary change occurs, and by raising questions regarding factors other than diet that may impact on δ13C and δ15N values. Here we show that the dentine δ13C and δ15N profiles of workhouse inmates dating from the Great Irish Famine of the 19th century not only record the expected dietary change from C3 potatoes to C4 maize, but when used together they also document prolonged nutritional and other physiological stress resulting from insufficient sustenance. In the adults, the influence of the maize-based diet is seen in the δ13C difference between dentine (formed in childhood) and rib (representing an average from the last few years of life). The demonstrated effects of stress on the δ13C and δ15N values will have an impact on the interpretations of diet in past populations even in slow-turnover tissues such as compact bone. This technique also has applicability in the investigation of modern children subject to nutritional distress where hair and nails are unavailable or do not record an adequate period of time.

The rise of atmospheric oxygen fundamentally changed the chemistry of surficial environments and the nature of Earth’s habitability 1 . Early atmospheric oxygenation occurred over a protracted period of extreme climatic instability marked by multiple global glaciations 2 , 3 , with the initial rise of oxygen concentration to above 10 −5 of the present atmospheric level constrained to about 2.43 billion years ago 4 , 5 . Subsequent fluctuations in atmospheric oxygen levels have, however, been reported to have occurred until about 2.32 billion years ago 4 , which represents the estimated timing of irreversible oxygenation of the atmosphere 6 , 7 . Here we report a high-resolution reconstruction of atmospheric and local oceanic redox conditions across the final two glaciations of the early Palaeoproterozoic era, as documented by marine sediments from the Transvaal Supergroup, South Africa. Using multiple sulfur isotope and iron–sulfur–carbon systematics, we demonstrate continued oscillations in atmospheric oxygen levels after about 2.32 billion years ago that are linked to major perturbations in ocean redox chemistry and climate. Oxygen levels thus fluctuated across the threshold of 10 −5 of the present atmospheric level for about 200 million years, with permanent atmospheric oxygenation finally arriving with the Lomagundi carbon isotope excursion at about 2.22 billion years ago, some 100 million years later than currently estimated. Sulfur isotope and iron–sulfur–carbon systematics on marine sediments indicate that permanent atmospheric oxygenation occurred around 2.22 billion years ago, about 100 million years later than currently estimated.

The widespread importance of variable types of primary production, or energy channels, to consumer communities has become increasingly apparent. However, the mechanisms underlying this “multichannel” feeding remain poorly understood, especially for aquatic ecosystems that pose unique logistical constraints given the diversity of potential energy channels. Here, we use bulk tissue isotopic analysis along with carbon isotope (δ13C) analysis of individual amino acids to characterize the relative contribution of pelagic and benthic energy sources to a kelp forest consumer community in northern Chile. We measured bulk tissue δ13C and δ15N for >120 samples; of these we analyzed δ13C values of six essential amino acids (EAA) from nine primary producer groups (n = 41) and 11 representative nearshore consumer taxa (n = 56). Using EAA δ13C data, we employed linear discriminant analysis (LDA) to assess how distinct EAA δ13C values were between local pelagic (phytoplankton/particulate organic matter), and benthic (kelps, red algae, and green algae) endmembers. With this model, we were able to correctly classify nearly 90% of producer samples to their original groupings, a significant improvement on traditional bulk isotopic analysis. With this EAA isotopic library, we then generated probability distributions for the most important sources of production for each individual consumer and species using a bootstrap-resampling LDA approach. We found evidence for multichannel feeding within the community at the species level. Invertebrates tended to focus on either pelagic or benthic energy, deriving 13–67% of their EAA from pelagic sources. In contrast, mobile (fish) taxa at higher trophic levels used more equal proportions of each channel, ranging from 19% to 47% pelagically derived energy. Within a taxon, multichannel feeding was a result of specialization among individuals in energy channel usage, with 37 of 56 individual consumers estimated to derive >80% of their EAA from a single channel. Our study reveals how a cutting-edge isotopic technique can characterize the dynamics of energy flow in coastal food webs, a topic that has historically been difficult to address. More broadly, our work provides a mechanism as to how multichannel feeding may occur in nearshore communities, and we suggest this pattern be investigated in additional ecosystems.

Plants commonly live in a symbiotic association with arbuscular mycorrhizal fungi (AMF). They invest photosynthetic products to feed their fungal partners, which, in return, provide mineral nutrients foraged in the soil by their intricate hyphal networks. Intriguingly, AMF can link neighboring plants, forming common mycorrhizal networks (CMNs). What are the terms of trade in such CMNs between plants and their shared fungal partners? To address this question, we set up microcosms containing a pair of test plants, interlinked by a CMN of Glomus intraradices or Glomus mosseae. The plants were flax (Linum usitatissimum; a C₃ plant) and sorghum (Sorghum bicolor; a C₄ plant), which display distinctly different ¹³C/¹²C isotope compositions. This allowed us to differentially assess the carbon investment of the two plants into the CMN through stable isotope tracing. In parallel, we determined the plants' \"return of investment\" (i.e. the acquisition of nutrients via CMN) using ¹⁵N and ³³P as tracers. Depending on the AMF species, we found a strong asymmetry in the terms of trade: flax invested little carbon but gained up to 94% of the nitrogen and phosphorus provided by the CMN, which highly facilitated growth, whereas the neighboring sorghum invested massive amounts of carbon with little return but was barely affected in growth. Overall biomass production in the mixed culture surpassed the mean of the two monocultures. Thus, CMNs may contribute to interplant facilitation and the productivity boosts often found with intercropping compared with conventional monocropping.