Integrating archaeology and ancient DNA analysis to address invasive species colonization in the Gulf of Alaska

The intentional and unintentional movement of plants and animals by humans has transformed ecosystems and landscapes globally. Assessing when and how a species was introduced are central to managing these transformed landscapes, particularly in island environments. In the Gulf of Alaska, there is considerable interest in the history of mammal introductions and rehabilitating Gulf of Alaska island environments by eradicating mammals classified as invasive species. The Arctic ground squirrel (Urocitellus parryii) is of concern because it affects vegetation and seabirds on Gulf of Alaska islands. This animal is assumed to have been introduced by historic settlers; however, ground squirrel remains in the prehistoric archaeological record of Chirikof Island, Alaska, challenge this timeline and suggest they colonized the islands long ago. We used 3 lines of evidence to address this problem: direct radiocarbon dating of archaeological squirrel remains; evidence of prehistoric human use of squirrels; and ancient DNA analysis of dated squirrel remains. Chirikof squirrels dated to at least 2000 years ago, and cut marks on squirrel bones suggested prehistoric use by people. Ancient squirrels also shared a mitochondrial haplotype with modern Chirikof squirrels. These results suggest that squirrels have been on Chirikof longer than previously assumed and that the current population of squirrels is closely related to the ancient population. Thus, it appears ground squirrels are not a recent, human‐mediated introduction and may have colonized the island via a natural dispersal event or an ancient human translocation.We thank T. Rick, D. Grayson, R. Fleischer, M. Hawkins, A. West, and C. Mikeska for their contributions to this research. We also thank 3 reviewers and the editors of Conservation Biology who greatly improved this paper. This work was funded by the National Geographic Society, the University of Maine, the Smithsonian Institution, and Boston University. The findings and conclusions in this article are those of the authors and do not necessarily represent the views of the U.S. Fish and Wildlife Service. (National Geographic Society; University of Maine; Smithsonian Institution; Boston University)Published versio

Sedimentary DNA and molecular evidence for early human occupation of the Faroe Islands

The Faroe Islands, a North Atlantic archipelago between Norway and Iceland, were settled by Viking explorers in the mid-9th century CE. However, several indirect lines of evidence suggest earlier occupation of the Faroes by people from the British Isles. Here, we present sedimentary ancient DNA and molecular fecal biomarker evidence from a lake sediment core proximal to a prominent archaeological site in the Faroe Islands to establish the earliest date for the arrival of people in the watershed. Our results reveal an increase in fecal biomarker concentrations and the first appearance of sheep DNA at 500 CE (95% confidence interval 370-610 CE), pre-dating Norse settlements by 300 years. Sedimentary plant DNA indicates an increase in grasses and the disappearance of woody plants, likely due to livestock grazing. This provides unequivocal evidence for human arrival and livestock disturbance in the Faroe Islands centuries before Viking settlement in the 9th century

Sedimentary DNA and Molecular Evidence for Early Human Occupation of the Faroe Islands

The Faroe Islands, a North Atlantic archipelago between Norway and Iceland, were settled by Viking explorers in the mid-9th century CE. However, several indirect lines of evidence suggest earlier occupation of the Faroes by people from the British Isles. Here, we present sedimentary ancient DNA and molecular fecal biomarker evidence from a lake sediment core proximal to a prominent archaeological site in the Faroe Islands to establish the earliest date for the arrival of people in the watershed. Our results reveal an increase in fecal biomarker concentrations and the first appearance of sheep DNA at 500 CE (95% confidence interval 370-610 CE), pre-dating Norse settlements by 300 years. Sedimentary plant DNA indicates an increase in grasses and the disappearance of woody plants, likely due to livestock grazing. This provides unequivocal evidence for human arrival and livestock disturbance in the Faroe Islands centuries before Viking settlement in the 9th century

Isotopic evidence for long-distance connections of the AD thirteenth century Promontory caves occupants

Las cuevas Promontory (Utah) y la Cueva Franktown (Colorado) contienen registros de alta fidelidad en cuanto a las ocupaciones de corto término de grupos humanos con cultura material conectada al Subártico/Planicies del Norte. Esta investigación utiliza excremento, pelo, piel de cuero y colágeno del hueso de bisonte provenientes de los sitios Promontory y Franktown para establecer una base de datos isotópicos locales y variabilidad isotópica de carbono e identifica la piel de cuero de procedencia lejana. La envoltura de tobillo de un mocasín recuperado de la Cueva 1 de Promontory tuvo un valor δ13C que indica un componente C4 considerable en la dieta del animal, lo que es un resultado distinto a las dietas C3 que se identificaron a partir del análisis de 171 muestras de bisonte provenientes de Promontory y el norte de Utah. Aplicamos una combinación única de análisis isotópico de tejidos múltiples, mapas isotópicos (“isoscapes”) de carbono, ADN antiguo (identificación de sexo y especie), tasa de remodelación de tejidos, contextos arqueológicos y la ecología del bisonte para demonstrar que el valor alto δ13C probablemente no es un resultado debido al consumo de plantas locales, movilidad del bisonte o comercio. En cambio, la piel de cuero del bisonte fue probablemente adquirida a través de viajes de larga distancia hacia zonas con pastos C4 abundantes muy al sur o al este. El conocimiento extenso sobre el paisaje obtenido a través de asociaciones de larga distancia debió permitir a los habitantes de las cuevas Promontory tomar decisiones bien informadas sobre las direcciones y rutas de movimiento para realizar un cambio territorial, el cual parece haber ocurrido a finales del sigo XIII.The Promontory caves (Utah) and Franktown Cave (Colorado) contain high-fidelity records of short-term occupations by groups with material culture connections to the Subarctic/Northern Plains. This research uses Promontory and Franktown bison dung, hair, hide, and bone collagen to establish local baseline carbon isotopic variability and identify leather from a distant source. The ankle wrap of one Promontory Cave 1 moccasin had a δ13C value that indicates a substantial C4 component to the animal's diet, unlike the C3 diets inferred from 171 other Promontory and northern Utah bison samples. We draw on a unique combination of multitissue isotopic analysis, carbon isoscapes, ancient DNA (species and sex identification), tissue turnover rates, archaeological contexts, and bison ecology to show that the high δ13C value was not likely a result of local plant consumption, bison mobility, or trade. Instead, the bison hide was likely acquired via long-distance travel to/from an area of abundant C4 grasses far to the south or east. Expansive landscape knowledge gained through long-distance associations would have allowed Promontory caves inhabitants to make well-informed decisions about directions and routes of movement for a territorial shift, which seems to have occurred in the late thirteenth century

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Studying Past Ecosystems and Human Behaviors Using Environmental and Ancient DNA

Isolating and studying degraded DNA from preserved organismal remains and environmental samples allows new inferences about past ecosystem compositions, population dynamics, and, in the context of archaeological remains, human interactions with their environment. In this dissertation, I addressed how sequencing depth and stochasticity of metabarcoding PCR influences various measures of biodiversity. I found that sequencing depth and stochasticity between PCR replicates significantly influence estimates of alpha but not beta diversity. In my second chapter, I used eDNA isolated from permafrost cores spanning the last 50,000 years in the Klondike, Canada to characterize community composition and turnover of plant and mammalian communities. In this chapter, I characterized floral and faunal change over the last 50,000 years, with clear shifts from steppe to boreal forest habitat delineated with the presence and absence of arctic ground squirrels and woody plants. Finally, I isolated ancient DNA from archaeological moccasins to observe hunting patterns of Bison used by occupants of the Promontory Caves of Utah, an archaeological site occupied 1240-1290 AD. I found the majority (87%) of moccasins were constructed from female bison, supporting prior hypotheses of hunting strategies targeting cow-calf herds at the end of fall preparing for overwintering. My dissertation highlights some of the many questions that degraded DNA present in soil, bone, and preserved hides can contribute towards answering

Studying Past Ecosystems and Human Behaviors Using Environmental and Ancient DNA

Isolating and studying degraded DNA from preserved organismal remains and environmental samples allows new inferences about past ecosystem compositions, population dynamics, and, in the context of archaeological remains, human interactions with their environment. In this dissertation, I addressed how sequencing depth and stochasticity of metabarcoding PCR influences various measures of biodiversity. I found that sequencing depth and stochasticity between PCR replicates significantly influence estimates of alpha but not beta diversity. In my second chapter, I used eDNA isolated from permafrost cores spanning the last 50,000 years in the Klondike, Canada to characterize community composition and turnover of plant and mammalian communities. In this chapter, I characterized floral and faunal change over the last 50,000 years, with clear shifts from steppe to boreal forest habitat delineated with the presence and absence of arctic ground squirrels and woody plants. Finally, I isolated ancient DNA from archaeological moccasins to observe hunting patterns of Bison used by occupants of the Promontory Caves of Utah, an archaeological site occupied 1240-1290 AD. I found the majority (87%) of moccasins were constructed from female bison, supporting prior hypotheses of hunting strategies targeting cow-calf herds at the end of fall preparing for overwintering. My dissertation highlights some of the many questions that degraded DNA present in soil, bone, and preserved hides can contribute towards answering

Revisiting the effect of PCR replication and sequencing depth on biodiversity metrics in environmental DNA metabarcoding.

Environmental DNA (eDNA) metabarcoding is an increasingly popular tool for measuring and cataloguing biodiversity. Because the environments and substrates in which DNA is preserved differ considerably, eDNA research often requires bespoke approaches to generating eDNA data. Here, we explore how two experimental choices in eDNA study design-the number of PCR replicates and the depth of sequencing of PCR replicates-influence the composition and consistency of taxa recovered from eDNA extracts. We perform 24 PCR replicates from each of six soil samples using two of the most common metabarcodes for Fungi and Viridiplantae (ITS1 and ITS2), and sequence each replicate to an average depth of ~84,000 reads. We find that PCR replicates are broadly consistent in composition and relative abundance of dominant taxa, but that low abundance taxa are often unique to one or a few PCR replicates. Taxa observed in one out of 24 PCR replicates make up 21-29% of the total taxa detected. We also observe that sequencing depth or rarefaction influences alpha diversity and beta diversity estimates. Read sampling depth influences local contribution to beta diversity, placement in ordinations, and beta dispersion in ordinations. Our results suggest that, because common taxa drive some alpha diversity estimates, few PCR replicates and low read sampling depths may be sufficient for many biological applications of eDNA metabarcoding. However, because rare taxa are recovered stochastically, eDNA metabarcoding may never fully recover the true amplifiable alpha diversity in an eDNA extract. Rare taxa drive PCR replicate outliers of alpha and beta diversity and lead to dispersion differences at different read sampling depths. We conclude that researchers should consider the complexity and unevenness of a community when choosing analytical approaches, read sampling depths, and filtering thresholds to arrive at stable estimates