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.

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.

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.

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.