Regional heterothermies recorded in the oxygen isotope composition of harbour seal skeletal elements.

Regional heterothermy is a strategy used by marine mammals to maintain a high and stable core body temperature, but its identification needs in situ measurements difficult to set up in extant wild organisms and inapplicable to extinct ones. We have analysed the oxygen isotope composition of bioapatite phosphate (δ18Op) from one permanent tooth and from thirty-six skeletal elements of one adult male harbour seal (Phoca vitulina) from the Baie de Somme (Hauts-de-France, France). We propose that the observed intra-skeletal δ18Op variability reflects tissue temperature heterogeneities typical of the pinniped regional heterothermy strategy. Our δ18Op data indicate that bone hydroxylapatite from harbour seal autopod skeletal elements (metacarpals, metatarsals, and phalanxes) mineralises at a lower temperature than that of the bone from the axial skeleton (e.g. vertebrae, ribs, and girdle bones). The results suggest that it is possible to locate a history of regional heterothermies in amphibious marine vertebrates using the δ18Op values of their mineralised tissues. This enables direct evaluation of the thermophysiology of both modern and fossil Pinnipedia from their skeletons opening perspectives on understanding their thermal adaptation to the marine environment in the fossil record. In addition to thermophysiology, oxygen isotope data from the permanent teeth of Pinnipedia, which are formed during the in utero phase from body fluid of the mother and at a stable temperature, could be valuable for locating the geographical areas inhabited by existing Pinnipedia females during their gestation period.

Determination of water balance maintenance in Orcinus orca and Tursiops truncatus using oxygen isotopes.

The secondary adaptation of Cetacea to a fully marine lifestyle raises the question of their ability to maintain their water balance in a hyperosmotic environment. Cetacea have access to four potential sources of water: surrounding salt oceanic water, dietary free water, metabolic water and inhaled water vapour to a lesser degree. Here, we measured the 18O/16O oxygen isotope ratio of blood plasma from 13 specimens belonging to two species of Cetacea raised under human care (four killer whales Orcinus orca, nine common bottlenose dolphins Tursiops truncatus) to investigate and quantify the contribution of preformed water (dietary free water, surrounding salt oceanic water) and metabolic water to Cetacea body water using a box-modelling approach. The oxygen isotope composition of Cetacea blood plasma indicates that dietary free water and metabolic water contribute to more than 90% of the total water input in weight for cetaceans, with the remaining 10% consisting of inhaled water vapour and surrounding water accidentally ingested or absorbed through the skin. Moreover, the contribution of metabolic water appears to be more important in organisms with a more lipid-rich diet. Beyond these physiological and conservation biology implications, this study opens up questions that need to be addressed, such as the applicability of the oxygen isotope composition of cetacean body fluids and skeletal elements as an environmental proxy of the oxygen isotope composition of present and past marine waters.

Determination of groundwater origins and vulnerability based on multi-tracer investigations: New contributions from passive sampling and suspect screening approach.

Knowledge about groundwater origins and their interactions with surface water is fundamental to assess their vulnerability. In this context, hydrochemical and isotopic tracers are useful tools to investigate water origins and mixing. More recent studies examined the relevance of contaminants of emerging concern (CECs) as co-tracers to distinguish sources contributing to groundwater bodies. However, these studies focused on known and targeted CECs a priori selected regarding their origin and/or concentrations. This study aimed to improve these multi-tracer approaches using passive sampling and qualitative suspect screening by exploring a larger variety of historical and emerging concern contaminants in combination with hydrochemistry and water molecule isotopes. With this objective, an in-situ study was conducted in a drinking water catchment area located in an alluvial aquifer recharged by several water sources (both surface and groundwater sources). CECs determined by passive sampling and suspect screening allowed to provide in-depth chemical fingerprints of groundwater bodies by enabling the investigation of >2500 compounds with an increased analytical sensitivity. Obtained cocktails of CECs were discriminating enough to be used as chemical tracer in combination with hydrochemical and isotopic tracers. In addition, the occurrence and type of CECs contributed to a better understanding of groundwater-surface water interactions and highlighted short-time hydrological processes. Furthermore, the use of passive sampling with suspect screening analysis of CECs lead to a more realistic assessment and mapping of groundwater vulnerability.

Multi-element stable isotope geochemistry and arsenic speciation of hydrothermal vent fauna (Alviniconcha sp., Ifremeria nautilei and Eochionelasmus ohtai manusensis), Manus Basin, Papua New Guinea.

Deep-sea hydrothermal vent communities, revealing patterns of niche partitioning, live in a limited area characterised by sharp physico-chemical gradients. In this study, we investigated carbon, sulfur, nitrogen stable isotopes as well as arsenic (As) speciations and concentrations for two snails (Alviniconcha sp. and Ifremeria nautilei) and a crustacean, (Eochionelasmus ohtai manusensis), occupying distinct niches in the hydrothermal vent field of the Vienna Woods, Manus Basin, Western Pacific. δ13C values of Alviniconcha sp. (foot), I. nautilei (foot and chitin) and E. o. manusensis (soft tissue) are similar, from -28 to -33‰ (V-PDB). The δ15N values of Alviniconcha sp. (foot and chitin), I. nautilei (foot and chitin) and E. o. manusensis (soft tissue) range from 8.4 to 10.6‰. The δ34S values of Alviniconcha sp. (foot and chitin), I. nautilei (foot) and E. o. manusensis (soft tissue) range from 5.9 to 11.1‰. Using stable isotopes, for the first time, we inferred a Calvin-Benson (RuBisCo) metabolic pathway for Alviniconcha sp. along with the presence of γ-Proteobacteria symbionts for the Vienna Woods communities. For I. nautilei, a feeding pattern is proposed with γ-Proteobacteria symbiosis and a Calvin-Benson-Bassham diet with mixotrophic feeding. E. ohtai manusensis is filtering bacteria with a CBB feeding strategy, with δ15N values indicating possible higher position in the trophic chain. Arsenic concentrations in the dry tissue of Alviniconcha (foot), I. nautilei (foot) and E. o. manusensis (soft tissue) are high, from 4134 to 8478 µg/g, with inorganic As concentrations of 607, 492 and 104 µg/g, respectively and dimethyl arsenic (DMA) concentrations of 11.12, 0.25 and 11.2 µg/g, respectively. Snails occurring in a vent proximal position have higher As concentration than barnacles, a pattern not observed for S concentrations. Arsenosugars were not put in evidence indicating that the available organic material for the vent organisms are not surface derived.

Isotopic systematics point to wild origin of mummified birds in Ancient Egypt.

Millions of mummified birds serving for religious purpose have been discovered from archeological sites along the Nile Valley of Egypt, in majority ibises. Whether these birds were industrially raised or massively hunted is a matter of heavy debate as it would have a significant impact on the economy related to their supply and cult, and if hunted it would have represented an ecological burden on the birds populations. Here we have measured and analysed the stable oxygen, carbon and radiogenic strontium isotope compositions as well as calcium and barium content of bones along with the stable carbon, nitrogen and sulfur isotope composition of feathers from 20 mummified ibises and birds of prey recovered from various archeological sites of Ancient Egypt. If these migratory birds were locally bred, their stable oxygen, radiogenic strontium and stable sulfur isotopic compositions would be similar to that of coexisting Egyptians, and their stable carbon, nitrogen and oxygen isotope variance would be close, or lower than that of Egyptians. On one hand, isotopic values show that ibises ingested food from the Nile valley but with a higher isotopic scattering than observed for the diet of ancient Egyptians. On the other hand, birds of prey have exotic isotopic values compatible with their migratory behaviour. We therefore propose that most mummified ibises and all the birds of prey analysed here were wild animals hunted for religious practice.

Simultaneous δ2 H and δ18 O analyses of water inclusions in halite with off-axis integrated cavity output spectroscopy.

Stable isotope compositions of ancient halite fluid inclusions have been recognized to be valuable tools for reconstructing past environments. Nevertheless, in order to better understand the genesis of halite deposits, it could be of great interest to combine both δ2 H and δ18 O measurements of the water trapped as inclusions in the defects of the mineral lattice. We developed a method combining off-axis integrated cavity output spectroscopy (OA-ICOS) connected on line with a modified elemental analyzer (EA-OA-ICOS) to perform those measurements. The first step was to test the method with synthetic halite crystals precipitated in the laboratory from isotopically calibrated waters. Water isotopic signatures have been measured with conventional techniques, equilibration for δ18 O and chromium reduction for δ2 H. Then, we modified and optimized a conventional EA to connect it online with an OA-ICOS instrument for H2 O measurements. The technique is first evaluated for calibrated free water samples. The technique is also evaluated for salt matrix effect, accuracy, and linearity for both isotopic signatures. Then, the technique is used to measure simultaneously δ2 H and δ18 O values of halite water inclusions precipitated from the evaporation experiments. Data generated with this new technique appeared to be comparable with those inferred from prior off-line technique studies. The advantages offered by the OA-ICOS technique are the simultaneous acquisition of both isotopic ratios and the substantial reduction of data acquisition time and sample aliquot size. Natural halite samples have been analyzed with this method. Natural halite samples as old as Precambrian have also been analyzed with this method.

Combined oxygen and sulphur isotope analysis-a new tool to unravel vertebrate (paleo)-ecology.

Reconstructing the living environment of extinct vertebrates is often challenging due to the lack of proxies. We propose a new proxy to the living environment based on the combined oxygen and sulphur stable isotope analysis of vertebrate hydroxyapatite. We tested this isotopic proxy to 64 biogenic apatite (bones) samples that represent a wide spectrum of the extant vertebrate phylogenetic diversity including crocodiles, snakes, turtles, mammals, birds, lizards, fish and amphibians. We show that the combination of these two isotopic systems allows the living environment of all these vertebrates to be unambiguously distinguished between freshwater (aquatic vs semi-aquatic), seawater (aquatic vs semi-aquatic) and terrestrial. The main goal of this study is to provide a present-day isotopic reference frame and to discuss methodological issues that will serve to interpret future oxygen and sulphur isotope results obtained either from fossil or modern skeletal material. This new isotopic approach of combined oxygen and sulphur isotope analysis will be particularly useful to document major aquatic-terrestrial transitions in the fossil record but also to better constrain the living environment of some present-day species.

δ15 N values in plants are determined by both nitrate assimilation and circulation.

Nitrogen (N) assimilation is associated with 14 N/15 N fractionation such that plant tissues are generally 15 N-depleted compared to source nitrate. In addition to nitrate concentration, the δ15 N value in plants is also influenced by isotopic heterogeneity amongst organs and metabolites. However, our current understanding of δ15 N values in nitrate is limited by the relatively small number of compound-specific data. We extensively measured δ15 N in nitrate at different time points, in sunflower and oil palm grown at fixed nitrate concentration, with nitrate circulation being varied using potassium (K) conditions and waterlogging. There were strong interorgan δ15 N differences for contrasting situations between the two species, and a high 15 N-enrichment in root nitrate. Modelling shows that this 15 N-enrichment can be explained by nitrate circulation and compartmentalisation whereby despite a numerically small flux value, the backflow of nitrate to roots via the phloem can lead to a c. 30‰ difference between leaves and roots. Accordingly, waterlogging and low K conditions, which down-regulate sap circulation, cause a decrease in the leaf-to-root isotopic difference. Our study thus suggests that plant δ15 N can be used as a natural tracer of N fluxes between organs and highlights the potential importance of δ15 N of circulating phloem nitrate.

Thermophysiologies of Jurassic marine crocodylomorphs inferred from the oxygen isotope composition of their tooth apatite.

Teleosauridae and Metriorhynchidae were thalattosuchian crocodylomorph clades that secondarily adapted to marine life and coexisted during the Middle to Late Jurassic. While teleosaurid diversity collapsed at the end of the Jurassic, most likely as a result of a global cooling of the oceans and associated marine regressions, metriorhynchid diversity was largely unaffected, although the fossil record of Thalattosuchia is poor in the Cretaceous. In order to investigate the possible differences in thermophysiologies between these two thalattosuchian lineages, we analysed stable oxygen isotope compositions (expressed as δ18O values) of tooth apatite from metriorhynchid and teleosaurid specimens. We then compared them with the δ18O values of coexisting endo-homeothermic ichthyosaurs and plesiosaurs, as well as ecto-poikilothermic chondrichthyans and osteichthyans. The distribution of δ18O values suggests that both teleosaurids and metriorhynchids had body temperatures intermediate between those of typical ecto-poikilothermic vertebrates and warm-blooded ichthyosaurs and plesiosaurs, metriorhynchids being slightly warmer than teleosaurids. We propose that metriorhynchids were able to raise their body temperature above that of the ambient environment by metabolic heat production, as endotherms do, but could not maintain a constant body temperature compared with fully homeothermic ichthyosaurs and plesiosaurs. Teleosaurids, on the other hand, may have raised their body temperature by mouth-gape basking, as modern crocodylians do, and benefited from the thermal inertia of their large body mass to maintain their body temperature above the ambient one. Endothermy in metriorhynchids might have been a by-product of their ecological adaptations to active pelagic hunting, and it probably allowed them to survive the global cooling of the Late Jurassic, thus explaining the selective extinction affecting Thalattosuchia at the Jurassic-Cretaceous boundary. This article is part of the theme issue 'Vertebrate palaeophysiology'.

Improved online hydrogen isotope analysis of halite aqueous inclusions.

We demonstrate an improved method based on continuous-flow elemental analyser pyrolysis isotopic ratio mass spectrometry (CF-EA-PY-IRMS) to measure the 2 H/1 H ratios of water trapped in halite crystals. Two challenges to overcome are the low hydrogen concentration of samples (10-50 µmol H2 ·g-1 ) and the high chloride concentration released when reacting halite in an elemental analyser. We describe an optimization procedure for determining the 2 H/1 H ratio of this trapped water with an acceptable accuracy. This technique involves the use of a high-temperature Cr reactor to quantitatively convert H2 O into H2 . The initial step was performed on halite crystals precipitated from a water reservoir where 2 H/1 H ratios were monitored from its initial stage until the end of evaporation. The 2 H/1 H isotopic analyses were automated online in continuous-flow mode. Precision of the method was determined for those "synthetic" samples with hydrogen concentrations ranging from 0.2 to 0.5 wt%. 2 H/1 H isotopic ratios of evaporating waters bracket the compositions of water inclusions. The formation of fluid inclusions is not instantaneous and records the isotopic signature of the residual waters across a time range during which the isotopic values of the water still evolve. This property explains why the δ2 HVSMOW standard deviation of ±5‰ (2σ) observed for 10-mg aliquots of halite exceeds the instrumental error (about ±1.5‰ 2σ) determined on the basis of IAEA-CH7, NBS 30, and NBS 22 references along with calibrated waters with and without added halite crystals. We also applied this method to Mesoproterozoic (1.4 Ga) and Neoproterozoic (0.8 Ga) halite samples with relatively low hydrogen concentrations (300-1500 ppm). The measured δ2 HVSMOW values for Precambrian waters range from -89‰ to -54‰. We propose that this technique offers a new perspective and great potential for palaeoenvironmental reconstructions based on the 2 H/1 H analyses of water trapped in halite.

Euryhaline ecology of early tetrapods revealed by stable isotopes.

The fish-to-tetrapod transition-followed later by terrestrialization-represented a major step in vertebrate evolution that gave rise to a successful clade that today contains more than 30,000 tetrapod species. The early tetrapod Ichthyostega was discovered in 1929 in the Devonian Old Red Sandstone sediments of East Greenland (dated to approximately 365 million years ago). Since then, our understanding of the fish-to-tetrapod transition has increased considerably, owing to the discovery of additional Devonian taxa that represent early tetrapods or groups evolutionarily close to them. However, the aquatic environment of early tetrapods and the vertebrate fauna associated with them has remained elusive and highly debated. Here we use a multi-stable isotope approach (δ13C, δ18O and δ34S) to show that some Devonian vertebrates, including early tetrapods, were euryhaline and inhabited transitional aquatic environments subject to high-magnitude, rapid changes in salinity, such as estuaries or deltas. Euryhalinity may have predisposed the early tetrapod clade to be able to survive Late Devonian biotic crises and then successfully colonize terrestrial environments.

2 H/1 H measurements of amphiboles and nominally anhydrous minerals (clinopyroxene, garnet and diamond) using high-temperature continuous flow elemental analyser/pyrolysis/isotope ratio mass spectrometry.

RATIONALE: We have used a high-precision, high-efficiency method for the measurement of the 2 H/1 H ratios of hydrous silicates (amphiboles) and nominally anhydrous minerals (NAM) such as clinopyroxene, garnet and diamond, which are usually extremely resistant to pyrolysis. This opens up new fields of investigation to better understand the conditions of formation for deep-Earth minerals. METHODS: The technique described here involves Isotope Ratio Mass Spectrometry (IRMS) on-line in continuous flow mode with an Elemental Analyser (EA) using "purge and trap" technology rather than separation by conventional packed column gas chromatography (GC). The system is equipped with a special high-temperature furnace reaching 1500°C, with a longer hot zone and improved temperature stability. Emphasis is put on the efficiency of the system to reliably pyrolyse refractory minerals difficult to analyse with other conventional systems. RESULTS: While conventional systems usually fail to generate hydrogen suitable for isotopic analyses, with the technique presented here we were able to measure 2 H/1 H ratios from four diamond samples (δ2 H = -60, -77, -84 and -79‰ V-SMOW; average SD = 4.5‰; n = 2), three garnet samples (δ2 H from -70 to -63‰), and nine clinopyroxenes (δ2 H from -92 to -58‰) associated with seven amphiboles (δ2 H from -76 to -27‰) from single mantle rock. CONCLUSIONS: The possibility of using such a system to reliably measure 2 H/1 H ratios from refractory minerals, which are usually extremely difficult to analyse, offers a new tool of investigation for providing us with unrivaled clues to study the deep interiors of Earth.

CO2 and temperature decoupling at the million-year scale during the Cretaceous Greenhouse.

CO2 is considered the main greenhouse gas involved in the current global warming and the primary driver of temperature throughout Earth's history. However, the soundness of this relationship across time scales and during different climate states of the Earth remains uncertain. Here we explore how CO2 and temperature are related in the framework of a Greenhouse climate state of the Earth. We reconstruct the long-term evolution of atmospheric CO2 concentration (pCO2) throughout the Cretaceous from the carbon isotope compositions of the fossil conifer Frenelopsis. We show that pCO2 was in the range of ca. 150-650 ppm during the Barremian-Santonian interval, far less than what is usually considered for the mid Cretaceous. Comparison with available temperature records suggest that although CO2 may have been a main driver of temperature and primary production at kyr or smaller scales, it was a long-term consequence of the climate-biological system, being decoupled or even showing inverse trends with temperature, at Myr scales. Our analysis indicates that the relationship between CO2 and temperature is time scale-dependent at least during Greenhouse climate states of the Earth and that primary productivity is a key factor to consider in both past and future analyses of the climate system.

Oxygen isotopes suggest elevated thermometabolism within multiple Permo-Triassic therapsid clades.

The only true living endothermic vertebrates are birds and mammals, which produce and regulate their internal temperature quite independently from their surroundings. For mammal ancestors, anatomical clues suggest that endothermy originated during the Permian or Triassic. Here we investigate the origin of mammalian thermoregulation by analysing apatite stable oxygen isotope compositions (δ18Op) of some of their Permo-Triassic therapsid relatives. Comparing of the δ18Op values of therapsid bone and tooth apatites to those of co-existing non-therapsid tetrapods, demonstrates different body temperatures and thermoregulatory strategies. It is proposed that cynodonts and dicynodonts independently acquired constant elevated thermometabolism, respectively within the Eucynodontia and Lystrosauridae + Kannemeyeriiformes clades. We conclude that mammalian endothermy originated in the Epicynodontia during the middle-late Permian. Major global climatic and environmental fluctuations were the most likely selective pressures on the success of such elevated thermometabolism.

Carbon, nitrogen and oxygen isotope fractionation during food cooking: Implications for the interpretation of the fossil human record.

OBJECTIVES: Stable isotope data provide insight into the reconstruction of ancient human diet. However, cooking may alter the original stable isotope compositions of food due to losses and modifications of biochemical and water components. METHODS: To address this issue, carbon, nitrogen and oxygen isotope ratios were measured on meat aliquots sampled from various animals such as pork, beef, duck and chicken, and also from the flesh of fishes such as salmon, European seabass, European pilchard, sole, gilt-head bream, and tuna. For each specimen, three pieces were cooked according to the three most commonly-known cooking practices: boiling, frying and roasting on a barbecue. RESULTS: Our data show that cooking produced isotopic shifts up to 1.8‰, 3.5‰, and 5.2‰ for δ13 C, δ15 N, and δ18 O values, respectively. Such variations between raw and cooked food are much greater than previously estimated in the literature; they are more sensitive to the type of food rather than to the cooking process itself, except in the case of boiling. CONCLUSIONS: Reconstructions of paleodietary may thus suffer slight bias in cases of populations with undiversified diets that are restrained toward a specific raw or cooked product, or using a specific cooking mode. In cases of oxygen isotope compositions from skeletal remains (bones, teeth), they not only constitute a valuable proxy for reconstructing past climatic conditions, but they could also be used to improve our knowledge of past human diet.

Oxygen isotope fractionation between bird bone phosphate and drinking water.

Oxygen isotope compositions of bone phosphate (δ18Op) were measured in broiler chickens reared in 21 farms worldwide characterized by contrasted latitudes and local climates. These sedentary birds were raised during an approximately 3 to 4-month period, and local precipitation was the ultimate source of their drinking water. This sampling strategy allowed the relationship to be determined between the bone phosphate δ18Op values (from 9.8 to 22.5‰ V-SMOW) and the local rainfall δ18Ow values estimated from nearby IAEA/WMO stations (from -16.0 to -1.0‰ V-SMOW). Linear least square fitting of data provided the following isotopic fractionation equation: δ18Ow = 1.119 (±0.040) δ18Op - 24.222 (±0.644); R 2 = 0.98. The δ18Op-δ18Ow couples of five extant mallard ducks, a common buzzard, a European herring gull, a common ostrich, and a greater rhea fall within the predicted range of the equation, indicating that the relationship established for extant chickens can also be applied to birds of various ecologies and body masses. Applied to published oxygen isotope compositions of Miocene and Pliocene penguins from Peru, this new equation computes estimates of local seawater similar to those previously calculated. Applied to the basal bird Confuciusornis from the Early Cretaceous of Northeastern China, our equation gives a slightly higher δ18Ow value compared to the previously estimated one, possibly as a result of lower body temperature. These data indicate that caution should be exercised when the relationship estimated for modern birds is applied to their basal counterparts that likely had a metabolism intermediate between that of their theropod dinosaur ancestors and that of advanced ornithurines.

Record of Nile seasonality in Nubian neonates.

The oxygen isotope compositions of bones (n = 11) and teeth (n = 20) from 12 Sudanese individuals buried on Sai Island (Nubia) were analysed to investigate the registration of the evolution of the Nile environment from 3700 to 500 years BP and the potential effects of ontogeny on the oxygen isotope ratios. The isotopic compositions were converted into the composition of drinking water, ultimately originating from the Nile. δ18O values decrease during ontogeny; this is mainly related to breastfeeding and physiology. Those of neonates present very large variations. Neonates have a very high bone turnover and are thus able to record seasonal δ18O variations of the Nile waters. These variations followed a pattern very similar to the present one. Nile δ18O values increased from 1.4 to 4.4 ‰ (Vienna Standard Mean Ocean Water) from the Classic Kerma (∼3500 BP) through the Christian period (∼1000 BP), traducing a progressive drying of Northeast Africa.

Oxygen isotope fractionation between bird eggshell calcite and body water: application to fossil eggs from Lanzarote (Canary Islands).

Oxygen and carbon isotope compositions of fossil bird eggshell calcite (δ(18)Ocalc and δ(13)Ccalc) are regularly used to reconstruct paleoenvironmental conditions. However, the interpretation of δ(18)Ocalc values of fossil eggshells has been limited to qualitative variations in local climatic conditions as oxygen isotope fractionations between calcite, body fluids, and drinking water have not been determined yet. For this purpose, eggshell, albumen water, and drinking water of extant birds have been analyzed for their oxygen and carbon isotope compositions. Relative enrichments in (18)O relative to (16)O between body fluids and drinking water of +1.6 ± 0.9 ‰ for semi-aquatic birds and of +4.4 ± 1.9 ‰ for terrestrial birds are observed. Surprisingly, no significant dependence to body temperature on the oxygen isotope fractionation between eggshell calcite and body fluids is observed, suggesting that bird eggshells precipitate out of equilibrium. Two empirical equations relating the δ(18)Ocalc value of eggshell calcite to the δ(18)Ow value of ingested water have been established for terrestrial and semi-aquatic birds. These equations have been applied to fossil eggshells from Lanzarote in order to infer the ecologies of the Pleistocene marine bird Puffinus sp. and of the enigmatic giant birds from the Pliocene. Both δ(13)Ccalc and δ(18)Ocalc values of Puffinus eggshells point to a semi-aquatic marine bird ingesting mostly seawater, whereas low δ(13)Ccalc and high δ(18)Ocalc values of eggshells from the Pliocene giant bird suggest a terrestrial lifestyle. This set of equations can help to quantitatively estimate the origin of waters ingested by extinct birds as well as to infer either local environmental or climatic conditions.

High-precision (34) S/(32) S measurements in vertebrate bioapatites using purge-and-trap elemental analyser/isotope ratio mass spectrometry technology.

RATIONALE: In ecological studies, the sulfur isotope compositions (δ(34) S values) of soft tissues (e.g. hair, skin, nail, muscle, collagen) allow the determination of both past and present-day living environments of organisms. However, technical limitations have so far prevented reliable sulfur isotope analyses of minerals having low sulfur content, such as bioapatite, which is the crystalline component of skeletal tissues. The development of 'purge-and-trap' technology in elemental analysers recently demonstrated new possibilities to solve some of those technical difficulties. METHODS: We have used a VarioPYROcube elemental analyser (EA) equipped with 'purge-and-trap' technology, interfaced in continuous flow mode to an Isoprime 100 isotope ratio mass spectrometer, to measure the sulfur isotope compositions of bioapatite samples. We first characterised a working calibrated material of chemical composition close to those of our samples, the low-S bearing phosphorite BCR32, against the two δ(34) SV-CDT reference-calibrated materials, NBS-127 and IAEA-SO-5. We have confirmed a δ(34) SV-CDT value of +18.4‰ (1σ = 0.5; n = 18) in agreement with the previously published value. Using BCR32 as a compositional and isotopic reference material, we have then measured the δ(34) SV-CDT values of various bioapatite tissues (bone, dentine and enamel) from both modern and fossil vertebrates living in different environments (marine, freshwater and terrestrial). RESULTS: Our results demonstrate the capacity of this analytical setup to measure the δ(34) SV-CDT values of low-S bioapatite samples (0.14 to 1.19 wt%) with a good analytical precision (1σ = 0.5; n = 14). Our results also show that the δ(34) SV-CDT values of modern and fossil vertebrate bioapatites allow discrimination between marine environments and freshwater or terrestrial ones. CONCLUSIONS: Sulfur isotope analysis of bioapatite has great potential to track the living environment of extinct vertebrates for which only fossilised bones or teeth have been preserved. Copyright © 2016 John Wiley & Sons, Ltd.

Natural variations of copper and sulfur stable isotopes in blood of hepatocellular carcinoma patients.

The widespread hypoxic conditions of the tumor microenvironment can impair the metabolism of bioessential elements such as copper and sulfur, notably by changing their redox state and, as a consequence, their ability to bind specific molecules. Because competing redox state is known to drive isotopic fractionation, we have used here the stable isotope compositions of copper ((65)Cu/(63)Cu) and sulfur ((34)S/(32)S) in the blood of patients with hepatocellular carcinoma (HCC) as a tool to explore the cancer-driven copper and sulfur imbalances. We report that copper is (63)Cu-enriched by ∼0.4‰ and sulfur is (32)S-enriched by ∼1.5‰ in the blood of patients compared with that of control subjects. As expected, HCC patients have more copper in red blood cells and serum compared with control subjects. However, the isotopic signature of this blood extra copper burden is not in favor of a dietary origin but rather suggests a reallocation in the body of copper bound to cysteine-rich proteins such as metallothioneins. The magnitude of the sulfur isotope effect is similar in red blood cells and serum of HCC patients, implying that sulfur fractionation is systemic. The (32)S-enrichment of sulfur in the blood of HCC patients is compatible with the notion that sulfur partly originates from tumor-derived sulfides. The measurement of natural variations of stable isotope compositions, using techniques developed in the field of Earth sciences, can provide new means to detect and quantify cancer metabolic changes and provide insights into underlying mechanisms.