Earliest evidence of primate captivity and translocation supports gift diplomacy between Teotihuacan and the Maya.

A multimethod archaeometry study (zooarchaeological, isotopic, ancient DNA, paleobotanical, and radiocarbon dating) of a spider monkey sacrificed in the ceremonial center of Teotihuacan, Mexico (1 to 550 CE) is interpreted as a diplomatic gift exchange with neighboring Maya. Not only does this spider monkey provide the earliest known instance of primate translocation and captivity in Mesoamerica, it helps date incipient modes of interregional diplomacy between two major powers during Early Classic Mesoamerica: Teotihuacan and the Maya. Details of human-primate interaction include age at capture and transport (before ∼3 y of age), captive duration (over 2 y), anthropogenic diet (staple was maize, though secondary resources unique to anthropogenic diet including arrowroot and chili pepper were also found), context of sacrifice (tethered and associated with complete golden eagle and an array of other statecrafts), and general site context (including presence of Maya vessels and Maya-style murals). The timing of the spider monkey's sacrifice (250 to 300 CE) and its life history suggest a reconsideration of epigraphically attested militaristic involvement of Teotihuacan at certain Maya sites. We propose that a period of more multilateral and fluid ritual exchange with Maya dignitaries preceded the Teotihuacan state's eventual ascent to prominence.

Historic trophic decline in New England's coastal marine ecosystem.

Overfishing is a worldwide occurrence that simplifies marine food webs, changes trophic patterns, and alters community structure, affecting not only the density of harvested species but also their trophic function. The northwestern Atlantic has a history of heavy fishing, and over the past century has also experienced destructive bottom fishing and harmful mobile fishing gear. After confirming that preservation solvent did not alter the nitrogen stable isotopes of preserved samples, we used museum specimens and modern samples to analyze nitrogen stable isotopes in tissues of two common demersal fishes pre-1950 (1850 to 1950) compared to 2021 to assess changes in trophic positions of coastal New England consumers over this time period. Both the mesopredator Centropristis striata (black sea bass) and the benthivore Stenotomus chrysops (scup) experienced significant declines in trophic position during this time. C. striata declined almost a full trophic level, S. chrysops declined half a trophic level, and these species are now occupying almost the same trophic position. Heavy fishing activities potentially shorten food chains, simplify trophic complexity, lessen the separation of trophic niches, and generally flatten food webs. The consequences of these within-species shifts are poorly investigated but could generate underappreciated cascading impacts on community structure and function. Archived natural-history collections are an invaluable resource for investigating ecological changes in natural communities through time. The evaluation of changing trophic positions via stable isotope analysis may allow fisheries managers to quantify large-scale effects of fishing on ecosystems and food webs over time.

Proteomic profile of bone "collagen" extracted for stable isotopes: Implications for bulk and single amino acid analyses.

RATIONALE: Protein studies in archaeology and paleontology have been dominated by stable isotope studies to understand diet and trophic levels, but recent applications of proteomic techniques have resulted in a more complete understanding of protein diagenesis than stable isotopes alone. In stable isotope analyses, samples are retained or discarded based on their properties. Proteomics can directly determine what proteins are present within the sample and may be able to allow previously discarded samples to be analyzed. METHODS: Protein samples that had been previously analyzed for stable isotopes, including those with marginal and poor sample quality, were characterized by liquid chromatography/mass spectrometry using an LTQ Orbitrap Velos mass spectrometer after separation on a Dionex Ultimate 3000 LC system. Data were analyzed using MetaMorpheus and custom R scripts. RESULTS: We found a variety of proteins in addition to collagen, although collagen I was found in the majority of the samples (most samples >80%). We also found a positive correlation between total deamidation and wt% N, suggesting that deamidation may impact the overall nitrogen signal in bulk analyses. The amino acid profiles of samples, including those of marginal or poor stable isotope quality, reflect the expected collagen I percentages, allowing their use in single amino acid stable isotope analyses. CONCLUSIONS: All the samples regardless of quality were found to have high concentrations of collagen I, making interpretations of dietary routing based on collagen I reasonably valid. The amino acid profiles on the marginal and poor samples reflect an expected collagen I profile and allow these samples to be recovered for single amino acid analyses.

Evaluation of two lipid removal methods for stable carbon and nitrogen isotope analysis in whale tissue.

RATIONALE: The presence of lipids in animal tissues can influence the interpretation of stable isotope data, particularly in lipid-rich tissues such as the skin and muscle of marine mammals. The traditionally employed chloroform-methanol delipidation protocol has the potential to alter δ15 N values in proteinaceous tissues. Our objective was to determine whether the use of cyclohexane could be an alternative extraction method, effectively removing lipids without altering δ15 N values. METHODS: Kidney, liver, muscle, and skin samples were collected from beach-cast Sowerby's beaked whales (Mesoplodon bidens). Control subsamples were processed without delipidation extraction, and duplicate subsamples were extracted with either chloroform-methanol or cyclohexane. δ13 C, δ15 N, and C:N values were determined by continuous-flow elemental analysis isotope ratio mass spectrometry. Paired Wilcoxon tests were used to evaluate the change in isotope ratios after extraction, and unpaired Wilcoxon tests were used to evaluate differences in isotope ratios between extractions. RESULTS: Use of cyclohexane is an effective delipidation technique for tissues with low and moderate lipid content. Chemical delipidation influenced δ15 N values; extracted samples generally showed an increase in δ15 N values which varied from 0.0‰ to 1.7‰. Chloroform-methanol extraction resulted in alterations to δ15 N values greater than the analytical precision for all analyzed tissues. Changes to δ15 N values after cyclohexane extraction were at or near the analytical precision for liver and muscle but greater than the analytical precision for kidney and skin. CONCLUSIONS: We recommend processing duplicate subsamples for stable isotope analysis, one with and one without extraction, in order to obtain accurate values for each isotope ratio. Prolonged chemical extractions are not necessary to effectively remove lipids. When samples are limited, we suggest using cyclohexane for tissues with low or moderate lipid content, and chloroform-methanol for lipid-rich tissues.

Stable isotopes from the African site of Elmina, Ghana and their usefulness in tracking the provenance of enslaved individuals in 18th- and 19th-century North American populations.

OBJECTIVES: Stable isotope values for historic period human remains from Elmina, Ghana, are compared to isotope data from 18th- and 19th-century North American sites as a test case for examining African origins and identifying first generation Africans in the Mid-Atlantic region of the United States. MATERIALS AND METHODS: Stable carbon, nitrogen, and oxygen isotope values were measured in skeletal remains. Values from the cosmopolitan port city of Elmina provide the first available reference data from Africa during this time period and region. These values serve as a proxy for West African groups in general which are statistically compared to Euro-Americans and African Americans. RESULTS: Elmina carbon isotope values are relatively higher than those of North Americans, and African Americans show greater statistical similarity to West Africans. Elmina nitrogen isotope values are higher than those of North Americans. Elmina oxygen isotope values are notably higher than those in all Mid-Atlantic North American sites in this study. DISCUSSION: Similarity in carbon isotope values between Elmina and African Americans suggests commonalities in food availability or food preferences between these groups. Elevated nitrogen isotope values in Elmina individuals support the documented reliance of the local population on marine dietary resources at this coastal port. While carbon and nitrogen isotopes provide insight into foodways, oxygen isotope data, sourced from drinking water, provide better geographical information. The higher oxygen values from Elmina not only differentiate this group from North American Mid-Atlantic sites, but also make it possible to identify outliers at these sites as potential recent arrivals from West Africa.

The effects of Paraloid B-72 and Butvar B-98 treatment and organic solvent removal on δ(13)C, δ(15)N, and δ(18)O values of collagen and hydroxyapatite in a modern bone.

Stable isotopes in bones are a powerful tool for diet, provenance, climate, and physiological reconstructions, but necessarily require well-preserved specimens unaltered by postmortem diagenesis or conservation practices. This study examines the effects of Paraloid B-72 and Butvar B-98, two common consolidants used in field and museum conservation, on δ(13)C, δ(15)N, and δ(18)O values from bone collagen and hydroxyapatite. The effects of solvent removal (100% acetone, 100% ethanol, 9:1 acetone:xylenes, 9:1 ethanol:xylenes) and drying methods (ambient air, vacuum, oven drying at 80°C) were also examined to determine if bones treated with these consolidants can successfully be cleaned and used for stable isotope analyses. Results show that introduction of Paraloid B-72 or Butvar B-98 in 100% acetone or 100% ethanol, respectively, with subsequent removal by the same solvents and drying at 80°C facilitates the most successful removal of consolidants and solvents. The δ(13)C values in collagen, δ(15)N in collagen, δ(18)O in hydroxyapatite phosphate, and δ(13)C in hydroxyapatite structural carbonate were unaltered by treatments with Paraloid or Butvar and subsequent solvent removal. The δ(18)O in hydroxyapatite structural carbonate showed nonsystematic variability when bones were treated with Paraloid and Butvar, which is hypothesized to be a result of hydroxyl exchange when bones are exposed to consolidants in solution. It is therefore recommended that δ(18)O in hydroxyapatite structural carbonate should not be used in stable isotope studies if bones have been treated with Paraloid or Butvar.

Understanding irregular shell formation of Nautilus in aquaria: chemical composition and structural analysis.

Irregular shell formation and black lines on the outside of live chambered nautilus shells have been observed in all adult specimens at aquariums and zoos soon after the organisms enter aquaria. Black lines have also been observed in wild animals at sites of broken shell, but continued growth from that point returns to a normal, smooth structure. In contrast, rough irregular deposition of shell continues throughout residence in aquaria. The composition and reasons for deposition of the black material and mitigation of this irregular shell formation is the subject of the current study. A variety of analytical techniques were used, including stable isotope mass spectrometry (SI-MS), inductively coupled plasma mass spectrometry (ICP-MS), micro x-ray fluorescence (µXRF), X-ray diffraction (XRD), and scanning electron microscopy (SEM) based X-ray microanalysis. Results indicate that the black material contains excess amounts of copper, zinc, and bromine which are unrelated to the Nautilus diet. The combination of these elements and proteins plays an important role in shell formation, growth, and strengthening. Further study will be needed to compare the proteomics of the shell under aquaria versus natural wild environments. The question remains as to whether the occurrence of the black lines indicates normal healing followed by growth irregularities that are caused by stress from chemical or environmental conditions. In this paper we begin to address this question by examining elemental and isotopic differences of Nautilus diet and salt water. The atomic composition and light stable isotopic ratios of the Nautilus shell formed in aquaria verses wild conditions are presented.

Dating human bone: is racemization dating species-specific?

Our recently developed dating technique based on the racemization rate of aspartic acid was applied to dating human bone, as well as that of other mammals, utilizing capillary electrophoresis mass spectrometry. First, several well-dated (mostly (14)C-dated and with strong archeological evidence) human bones ranging in age from 150 to ~10,000 years were used to develop a calibration curve for human bone. The D/L ratio of aspartic acid for these specimens ranged from 2.4% to ~10%, with a correlation coefficient of better than 0.99, indicating a strong linear relationship between the d/l ratio of aspartic acid and the age of the specimens. This calibration curve can now be used to date human archeological specimens of unknown age, up to ~10,000 years. However, when the technique was applied to well-dated mixed species of larger mammal bones such as bison, whale, llama, etc., the calibration curve showed a slower rate of racemization with a lower correlation (0.88). As additional large mammal bones with less certain age (i.e., using archeological evidence alone with no (14)C-dating) were dated the correlation coefficient decreased to 0.70. The correlation coefficient decreased further to 0.58 when the racemization data from all mammals (including human) were added to the calibration curve, indicating the importance of using well-dated, species-specific specimens for forming a calibration curve. This conclusion is consistent with our previously published calibration curve for a single species of silk (Bombyx mori), which followed the expected reversible first-order kinetics. These results support species specificity of amino acid racemization dating.

The history of Coast Salish "woolly dogs" revealed by ancient genomics and Indigenous Knowledge.

Ancestral Coast Salish societies in the Pacific Northwest kept long-haired "woolly dogs" that were bred and cared for over millennia. However, the dog wool-weaving tradition declined during the 19th century, and the population was lost. In this study, we analyzed genomic and isotopic data from a preserved woolly dog pelt from "Mutton," collected in 1859. Mutton is the only known example of an Indigenous North American dog with dominant precolonial ancestry postdating the onset of settler colonialism. We identified candidate genetic variants potentially linked with their distinct woolly phenotype. We integrated these data with interviews from Coast Salish Elders, Knowledge Keepers, and weavers about shared traditional knowledge and memories surrounding woolly dogs, their importance within Coast Salish societies, and how colonial policies led directly to their disappearance.

Collagen and carbonate isotope data of fauna from pre-Columbian Panama.

Raw isotope data of collagen (δ13Ccollagen and δ15Ncollagen) and carbonate (δ13Ccarbonate and δ18Ocarbonate) of bone, enamel, and dentine of 101 faunal samples from Parita Bay, Panama are presented. These samples were taken from four archeological sites that span a long termporal range beginning with early hamlet agriculture period marked by the introduction of agricultre (circa 6000 BCE), and extending into the time of Spanish contact (1521 CE). The collection represents twelve faunal species of secondary browsers (deer), potentially captive or habituated birds (waterfowl, parrots, guan, among others), and carnivores (ocelot and domesticated dog). One modern deer specimen was also taken to link archeological baselines with known modern environmental data. This data complements our argument, presented in the article "Domesticated landscapes of the Neotropics: Isotope signatures of human-animal relationships in pre-Columbian Panama" [1], that stable isotope analysis can be a useful proxy to document degrees to which human-plant/animal co-habitation has created anthropogenic ecosystems in the Neotropics.

Marking mosquitoes in their natural larval sites using 2H-enriched water: a promising approach for tracking over extended temporal and spatial scales.

BACKGROUND: 1.Tracking mosquitoes using current methods of mark-release-recapture are limited to small spatial and temporal scales exposing major gaps in understanding long-range movements and extended survival. Novel approaches to track mosquitoes may yield fresh insights into their biology which improves intervention activities to reduce disease transmission.Stable isotope enrichment of natural mosquito breeding sites allows large-scale marking of wild mosquitoes absent human handling. Mosquito larvae that develop in 2H-enriched water are expected to be detectable for over four months using tissue mass-fraction 2H measurements, providing opportunities for long-term mark-capture studies on a large scale. APPROACH: 2.A laboratory study followed by a field experiment of mosquito larval habitat 2H-enrichment was conducted in Mali, to evaluate potential labeling of wild mosquitoes. Twelve natural larval sites were enriched using [2H]-Deuterium-oxide (D2O, 99%). Enrichment level was maintained by supplementation following dilution by rains. Availability of 2H to mosquito larvae was enhanced by locally collected and cultured microorganisms (i.e. protozoa, algae and bacteria) reared in deuterated water, and provided as larval diet. Putative natural predators were removed from the larval sites and first instar larvae Anopheles gambiae s.l. larvae were added every other day. Emergence traps enabled collection of eclosing adults. Adult mosquitoes were kept at laboratory conditions for analysis of label attrition with age. RESULTS: 3.Deuterium enrichment of wild mosquitoes above background levels (maximum = 143.1 ppm) became apparent 5-6 days after initial exposure, after which 2H values increased steadily until ~24 days later (to a mean of approx. 220 ppm). Anopheles and Culex mosquitoes showed significantly different 2H values (211 and 194.2 ppm respectively). Both genera exhibited exponential label attrition (e (-x)) amounting to 21.6% by day 30 post emergence, after which attrition rate continuously decreased. Males of both taxa exhibited a higher mean 2H value compared to females. CONCLUSIONS: 4.Deuterium-oxide proved useful in marking mosquitoes in their natural larval sites and although costly, may prove valuable for studies of mosquitoes and other aquatic insects. Based on our field study, we provide a protocol for marking mosquito larval sites using deuterium-oxide.

Jaguar and puma captivity and trade among the Maya: Stable isotope data from Copan, Honduras.

From Moctezuma's zoo to animals kept in captivity at Teotihuacan, there is increasing evidence that Mesoamericans managed wild animals for a myriad of purposes. The present study situates ritualized animal management of highly symbolic fauna in the broader context of Classic Mesoamerica by examining another core site, the Maya center of Copan, Honduras (A.D. 426-822). In this study, we identify two animal populations among the faunal remains from public and private rituals spanning the Copan dynasty. One population, with diets heavily composed of atypically sourced C4 inputs indicative of artificial feeding, corresponds with the felids interred in Altar Q and Motmot caches. The second population is composed of felids and felid products bearing a predominance of C3 signatures indicative of a more natural dietary regime. As with Copan deer, species-specific δ18O variations within these felid populations further substantiates the postulation that an expansive faunal trade network operated throughout the greater Copan Valley and beyond. Animals routed from sites of capture into the mesh of this network would have been processed into pelts, venison and other secondary goods or delivered alive to centers of state power for ritual usage and display. Our data reveal that at Copan, wild animals were routinely brought into intimate contact with human settlements to be managed and physically manipulated in a variety of ways in order to fulfill ritual and symbolic purposes.