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.
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Comparison of powdered enamel sample pretreatment methods for strontium isotope analysis
Most researchers assume minimal impact of pretreatment on strontium isotope ratios ( 87 Sr/ 86 Sr) for bones and teeth, and methods vary tremendously. We compared 14 pretreatment methods, including no prep other than powdering enamel, ashing, soaking in water, an oxidizing agent (bleach or hydrogen peroxide) or acetic acid (0.1 M, 1.0 M, and 1.0 M buffered with calcium acetate), and a combination of these steps. We prepared and analyzed aliquots of powdered molar enamel from three proboscideans (one modern captive Indian elephant, Elephas maximus indicus ; one Pleistocene mastodon, Mammut americanum ; and one Miocene gomphothere, Afrochoerodon kisumuensis ). Each pretreatment was performed in triplicate and we measured 87 Sr/ 86 Sr, Sr concentration, and uranium (U) concentration, using the same lab space and instrumentation for all samples. Variability in 87 Sr/ 86 Sr and Sr and U concentrations was considerable across pretreatments. Mean 87 Sr/ 86 Sr across methods ranged from 0.70999 to 0.71029 for the modern tooth, 0.71458 to 0.71502 for the Pleistocene tooth, and 0.70804 to 0.70817 for the Miocene tooth. The modern tooth contained the least Sr and negligible U. The Pleistocene tooth contained slightly more Sr and measurable amounts of U, and the Miocene tooth had approximately 5x more Sr and U than the Pleistocene tooth. For all three teeth, variance in 87 Sr/ 86 Sr, Sr concentrations, and U concentrations among replicates was statistically indistinguishable across pretreatments, but there were apparent differences among pretreatments for the modern and Pleistocene teeth. Both contained relatively little Sr, and it is possible that small amounts of exogenous Sr from reagents, building materials or dust affected some replicates for some pretreatments. For the modern tooth, median 87 Sr/ 86 Sr varied considerably (but statistically insignificantly) across pretreatments. For the Pleistocene tooth, variability in median 87 Sr/ 86 Sr was also considerable; some pretreatments were statistically distinct but there were no obvious patterns among methods. For the Miocene tooth, variability in median 87 Sr/ 86 Sr was much smaller, but there were significant differences among pretreatments. Most pretreatments yielded 87 Sr/ 86 Sr and Sr concentrations comparable to, or lower than, untreated powder, suggesting selective removal of exogenous material with high 87 Sr/ 86 Sr. Further evaluation of the mechanisms driving isotopic variability both within and among pretreatment methods is warranted. Researchers should clearly report their methods and avoid combining data obtained using different methods. Small differences in 87 Sr/ 86 Sr could impact data interpretations, especially in areas where isotopic variability is low.
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- Award ID(s):
- 1749676
- PAR ID:
- 10415106
- Date Published:
- Journal Name:
- Frontiers in Environmental Chemistry
- Volume:
- 4
- ISSN:
- 2673-4486
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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