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Variability of oxygen isotopes in environmental water is recorded in tooth enamel, providing a record of seasonal change, dietary variability, and mobility. Physiology dampens this variability, however, as oxygen passes from environmental sources into blood and forming teeth. We showcase two methods of high resolution, 2-dimensional enamel sampling, and conduct modeling, to report why and how environmental oxygen isotope variability is reduced in animal bodies and teeth. First, using two modern experimental sheep, we introduce a sampling method, die-saw dicing, that provides high-resolution physical samples (n = 109 and 111 sample locations per tooth) for use in conventional stable isotope and molecular measurement protocols. Second, we use an ion microprobe to sample innermost enamel in an experimental sheep (n = 156 measurements), and in a Pleistocene orangutan (n = 176 measurements). Synchrotron and conventional μCT scans reveal innermost enamel thicknesses averaging 18 and 21 μm in width. Experimental data in sheep show that compared to drinking water, oxygen isotope variability in blood is reduced to 70–90 %; inner and innermost enamel retain between 36 and 48 % of likely drinking water stable isotope range, but this recovery declines to 28–34 % in outer enamel. 2D isotope sampling suggests that declines in isotopic variability, and shifted isotopic oscillations throughout enamel, result from the angle of secretory hydroxyapatite deposition and its overprinting by maturation. This overprinting occurs at all locations including innermost enamel, and is greatest in outer enamel. These findings confirm that all regions of enamel undergo maturation to varying degrees and confirm that inner and innermost enamel preserve more environmental variability than other regions. We further show how the resolution of isotope sampling — not only the spatial resolution within teeth, but also the temporal resolution of water in the environment — impacts our estimate of how much variation teeth recover from the environment. We suggest inverse methods, or multiplication by standard factors determined by ecology, taxon, and sampling strategy, to reconstruct the full scale of seasonal environmental variability. We advocate for combined inverse modeling and high-resolution sampling informed by the spatiotemporal pattern of enamel formation, and at the inner or innermost enamel when possible, to recover seasonal records from teeth. 

AGU Fall Meeting, Chicago, Il, December 12-16, 2022; Presentation PP42D-1141 

Variability in resource availability is hypothesized to be a significant driver of primate adaptation and evolution, but most paleoclimate proxies cannot recover environmental seasonality on the scale of an individual lifespan. Oxygen isotope compositions (δ 18 O values) sampled at high spatial resolution in the dentitions of modern African primates ( n = 2,352 near weekly measurements from 26 teeth) track concurrent seasonal precipitation, regional climatic patterns, discrete meteorological events, and niche partitioning. We leverage these data to contextualize the first δ 18 O values of two 17 Ma Afropithecus turkanensis individuals from Kalodirr, Kenya, from which we infer variably bimodal wet seasons, supported by rainfall reconstructions in a global Earth system model. Afropithecus ’ δ 18 O fluctuations are intermediate in magnitude between those measured at high resolution in baboons ( Papio spp.) living across a gradient of aridity and modern forest-dwelling chimpanzees ( Pan troglodytes verus ). This large-bodied Miocene ape consumed seasonally variable food and water sources enriched in 18 O compared to contemporaneous terrestrial fauna ( n = 66 fossil specimens). Reliance on fallback foods during documented dry seasons potentially contributed to novel dental features long considered adaptations to hard-object feeding. Developmentally informed microsampling recovers greater ecological complexity than conventional isotope sampling; the two Miocene apes ( n = 248 near weekly measurements) evince as great a range of seasonal δ 18 O variation as more time-averaged bulk measurements from 101 eastern African Plio-Pleistocene hominins and 42 papionins spanning 4 million y. These results reveal unprecedented environmental histories in primate teeth and suggest a framework for evaluating climate change and primate paleoecology throughout the Cenozoic. 

Blood water oxygen isotope compositions can provide valuable insights into physiological processes and ecological patterns. While blood samples are commonly drawn for medical or scientific purposes, blood fractions are infrequently measured for oxygen isotopic compositions (δ¹⁸O) because such measurements are time consuming and expensive.

We sampled blood from sheep, goats, and iguanas raised in field and animal laboratories into serum, EDTA, heparin, and uncoated plastic vials commonly used in medical and scientific research, then separated red blood cell (RBC) and plasma or serum blood fractions. These were injected into helium‐flushed Exetainer tubes where they naturally outgassed endogenous CO₂(goat blood), or into He‐ and CO₂‐flushed tubes (iguana blood). The CO₂gas was sampled on a GasBench II system, and δ¹⁸O was measured by an isotope ratio mass spectrometer (IRMS).

Repeated δ¹⁸O measurements were stable over multiple days. The addition of desiccated blood solids to water standards had little impact on their δ¹⁸O measurements, suggesting that organic molecular constituents within blood serum and plasma do not interfere with blood water δ¹⁸O values. We observed slight but statistically significant δ¹⁸O offsets between plasma, serum and RBC fractions. Mass‐dependent body water turnover times for iguanas were derived from the data.

We demonstrate that a simple blood‐CO₂equilibration method using the GasBench can quickly, reliably and accurately characterize water δ¹⁸O in the plasma, RBC, and whole blood fractions of mammalian and reptilian blood samples (precision ≤ 0.1‰). This method will expand the application of blood stable isotope analysis in physiological and medical research.