In the last few decades, the field of ancient DNA has taken a new direction towards using sedimentary ancient DNA (sedaDNA) for studying human and mammalian population dynamics as well as past ...ecosystems. However, the screening of numerous sediment samples from archaeological sites remains a time-consuming and costly endeavor, particularly when targeting hominin DNA. Here, we present a novel high-throughput method that facilitates the fast and efficient analysis of sediment samples by applying a pooled testing approach. This method combines multiple extracts, enabling early parallelization of laboratory procedures and effective aDNA screening. Pooled samples with detectable aDNA signals undergo detailed analysis, while empty pools are discarded. We have successfully applied our method to multiple sediment samples from Middle and Upper Paleolithic sites in Europe, Asia, and Africa. Notably, our results reveal that an aDNA signal remains discernible even when pooled with four negative samples. We also demonstrate that the DNA yield of double-stranded libraries increases significantly when reducing the extract input, potentially mitigating the effects of inhibition. By embracing this innovative approach, researchers can analyze large numbers of sediment samples for aDNA preservation, achieving significant cost reductions of up to 70% and reducing hands-on laboratory time to one-fifth.
Previous dating of the Vi-207 and Vi-208 Neanderthal remains from Vindija Cave (Croatia) led to the suggestion that Neanderthals survived there as recently as 28,000–29,000 B.P. Subsequent dating ...yielded older dates, interpreted as ages of at least ∼32,500 B.P. We have redated these same specimens using an approach based on the extraction of the amino acid hydroxyproline, using preparative high-performance liquid chromatography (Prep-HPLC). This method is more efficient in eliminating modern contamination in the bone collagen. The revised dates are older than 40,000 B.P., suggesting the Vindija Neanderthals did not live more recently than others across Europe, and probably predate the arrival of anatomically modern humans in Eastern Europe. We applied zooarchaeology by mass spectrometry (ZooMS) to find additional hominin remains. We identified one bone that is Neanderthal, based on its mitochondrial DNA, and dated it directly to 46,200 ± 1,500 B.P. We also attempted to date six early Upper Paleolithic bone points from stratigraphic units G₁, Fd/d+G₁ and Fd/d, Fd. One bone artifact gave a date of 29,500 ± 400 B.P., while the remainder yielded no collagen. We additionally dated animal bone samples from units G₁ and G₁–G₃. These dates suggest a co-occurrence of early Upper Paleolithic osseous artifacts, particularly split-based points, alongside the remains of Neanderthals is a result of postdepositional mixing, rather than an association between the two groups, although more work is required to show this definitively.
The Assimilation Model (AM) was developed to explain the presence of evidence for small, but not insignificant, anatomical contributions of Eurasian archaic humans (including Neandertals) to modern ...Eurasians within a framework of the documented African origin of the majority of modern Eurasian ancestry. The AM was based entirely on morphological evidence for introgression in Eurasia and was initially not well supported by genetic studies, including ancient DNA. However, beginning with the publication of the first Neandertal draft genome in 2010, genomic data have consistently demonstrated low levels of archaic human gene flow into modern Eurasian populations and recently have shown modern gene flow into a Siberian Neandertal. Current data also indicate the complexity of archaic–modern human population dynamics during the late Pleistocene, another tenet of the AM. We conclude that the AM is strongly supported by current genetic and genomic data, as well as morphological evidence that has been documented elsewhere.
•First systematically explored underwater Middle Paleolithic site in the central Mediterranean and southeastern Europe.•Improved underwater field investigation methodology for Paleolithic sites ...applied.•Comparative approach to lithic material from underwater, open-air and cave sites.•Importance of underwater research for further development of Paleolithic archaeology.
This paper presents the investigation methodology and results of analyses of lithic material from Kaštel Štafilić – Resnik (Dalmatia, Croatia), the first systematically explored underwater Middle Paleolithic site in the central Mediterranean and southeastern Europe. The present site contains lithic elements from one or several open-air habitation sites from the time when the sea level was considerably lower than today. The archeological assemblage (Mousterian industry) and relative chronology indicate that almost all finds are contemporary with the Neandertals. The obtained results were compared to other Middle Paleolithic sites from the eastern Adriatic region. An integrative approach to lithic material from different types of sites (underwater, open-air, cave) provided better understanding of formation processes, a more complete picture of the region occupied by the Mousterian people, and clearer insight into their mobility patterns. Given this new evidence, it has become clear that the Neandertals of the Adriatic region controlled a much larger range of territory than previously documented.
Beginning with excavations during the 1970s, Vindija Cave (Croatia) has yielded significant Middle and Upper Paleolithic fossil and archaeological finds. We report on seven recently identified ...hominid fossils, a newly associated partial hominid cranial vault from level G
3, nine possible bone retouchers, and a revised interpretation of the Mousterian artifact assemblage from the site. This new information reinforces our knowledge of the complex biocultural phenomena revealed in unit G and earlier deposits at Vindija. Six of the new hominid fossils derive from stratigraphic units G and I, while one lacks exact provenience. All specimens preserving diagnostic anatomy are from Neandertals. One of the postcranial remains, a radius fragment which exhibits Neandertal-like anatomy, comes from level G
1and is congruent with the previously established association of Neandertals with an early Upper Paleolithic industry at the site. The partial cranial vault represents the most complete Neandertal from Vindija. The possible retouchers derive from unit G. Our analysis of these artifacts suggests that both percussion and pressure techniques may have been used by Neandertals in the final stage of tool production (retouching).
This paper also presents a revision of the artifact analysis for late Mousterian level G
3. We separated raw materials into two main groups due to the differing ways that the materials fracture and the differing morphology of the debitage. The use of raw material in level G
3is different from earlier Middle Paleolithic levels at Vindija. This indicates that the G
3late Neandertals were making choices regarding source material somewhat more like the Upper Paleolithic people at the site. When interpreted within a larger regional framework, the Vindija archaeological and hominid fossil remains demonstrate a complex, mosaic pattern of biocultural change in the Late Pleistocene of south-central Europe.
ABSTRACT
This paper presents palaeoecological reconstructions and late Neanderthal responses to environmental conditions in two different palaeoecological zones (continental south‐central Europe and ...the eastern Adriatic) during MIS 3. Our interpretations are based on the available records of climatic conditions, characteristics of Mousterian industries and site characteristics. A variety of data obtained by different methods of analyses (sediments and biological remains) were used for palaeoecological reconstructions, while hominin behaviour was examined from various perspectives based on lithic remains (technology, typology, raw material use), subsistence strategies, land use and site structures. In both regions, Neanderthals practised a broad exploitation of different faunal and lithic resources available to them and successfully adapted to local environments. Based on a variety of evidence, we offer a new synthesis and examine whether continental south‐central Europe and the Adriatic really acted as refugia during the time of late Neanderthal occupations. We also consider the possible causes and/or timing of Neanderthal disappearance in these regions.
During the Last Glacial Maximum (LGM, 30 to 16.5 ka ago), the Great Adriatic-Po Region (GAPR) was deeply affected by the spread of glaciers from the Alps to the southern foreland and by the dropping ...of the sea level to ~ -120 m amsl. The combination of these two events triggered the aggradation of the Great Po Plain (GPP), a vast flat area between the Alpine chain, the Italian Peninsula and the north-western Balkan Peninsula, physically and ecologically featured through a range of palaeogeographic and palaeoecological conditions. The low-elevated Prealpine sectors and the Alpine foothills supported more extensive forest stands, due to increased orographic rainfall. These were open boreal forests which persisted throughout the LGM, while open woodlands, steppes, semideserts and wetlands occupied the lowlands. A complex ecogradient, including both an Alpine and a continental timberline, is documented by the fossil records at the NE Alpine border, with a larch-pine forest-steppe belt, in contact with steppes and loess areas extending in the plain, on the dry extreme of the gradient. Still, edaphic wetlands occupied the waterlogged silty soils in the lowlands. Other areas, marked by active geodynamic processes, supported semideserts, i.e. grooves of xerophytic herbs and shrubs. Enhanced aridity and the development of deflation areas, prompted the accretion of loess cover at the northern and southern margins of the GPP. Fauna recorded the gradual disappearance of mammoth, woolly rhino and giant deer, together with cave bear. Gravettian and Epigravettian hunter-gatherer groups inhabited the GPP, although their presence and settlement dynamics at the margins and across this region has long been questioned. As a matter of fact, a handful of archaeological sites composes a patchy record of the peopling of the plain itself. At the northern rim of the GAPR, characterized by a well-developed karst region, several caves and rock shelters record the presence of hunters of bisons and horses at the margins of the GPP and ibexes and cave bears in some hilly landscapes. Nonetheless, evidence of contacts across this area is provided by the exploitation of chert sources and by stylistic and technical similarities in the lithic industries. The work resumes the currently available multidisciplinary data and adds new petroarchaeological evidence for reconstructing the settlement dynamics of the Gravettian - Epigravettian hunter-gatherers in this vast region up to the early Late Glacial, when the Prealpine and the Apennine foothills, along with the Dinarids, were persistently settled.
The 1998/1999 direct dating of two Neandertal specimens from level G¹ of Vindija Cave in Croatia to ≈28,000 and ≈29,000 radiocarbon ($^{14}C$) years ago has led to interpretations concerning the late ...survival of Neandertals in south-central Europe, patterns of interaction between Neandertals and in-dispersing early modern humans in Europe, and complex biocultural scenarios for the earlier phases of the Upper Paleolithic. Given improvements, particularly in sample pretreatment techniques for bone radiocarbon samples, especially ultrafiltration of collagen samples, these Vindija G₁ Neandertal fossils are redated to ≈32,000-33,000$^{14}C$years ago and possibly earlier. These results and the recent redating of a number of purportedly old modern human skeletal remains in Europe to younger time periods highlight the importance of fine chronological control when studying this biocultural time period and the tenuous nature of monolithic scenarios for the establishment of modern humans and earlier phases of the Upper Paleolithic in Europe.
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