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Belgium represents a key region for studying the Middle to Upper Palaeolithic transition (MUPT) in North-West Europe. This area sits at the crossroads between Palaeolithic cultural facies with influences from eastern, western and southern Europe intermingling during the Late Middle Palaeolithic and the MUPT. Until recently, a temporal gap believed to be around 4ka (ca 42-38 ky calBP) existed between the Late Mousterian and the earliest dated Aurignacian settlements in the region [1, 2]. The dates obtained on Neanderthal remains from Spy fell into this gap, making them the latest Neanderthals in the region [3]. Including the dates from Spy, a gap of two millennia remained between the dates on Neanderthals and the beginning of the Aurignacian. Based on this chronological evidence, the transition from Neanderthals to Anatomically Modern Humans (AMH) in this region was believed to have been without contact between species. AMH would have settled in an area Neanderthals abandoned long before. As part of the PalaeoChron project, we have redated the Neanderthal specimens from Spy (tooth, maxilla and scapula), Engis 2 (skull and tooth) and Fond-de-Forêt (femur), using the compound specific radiocarbon dating method in place at the Oxford Radiocarbon Accelerator Unit. This method is based on the extraction of the amino acid hydroxyproline that occurs in mammalian collagen using preparative liquid chromatography. This method is more efficient than others in eliminating modern carbon contamination such as conservation materials. In this presentation, we report the new radiocarbon dates obtained on the Belgian Neanderthal specimens. These results show how much impact sample preparation can have on the AMS measurement when specimens have been heavily preserved with conservation materials, which is often the case for human remains. These results also now place the Belgian Neanderthal remains from Spy, Engis and Fond-de-Forêt in their proper chronometric context and allow us to refine our understanding of the disappearance of Neanderthals in north-western Europe and integrate this with other evidence for the human occupation of this region during the Palaeolithic.

Remains of the foot, upper and lower limb, thorax and cranium of Homo naledi present a mosaic of primitive and Homo-like traits. These include curved phalanges in the hands, although a human like wrist and palm, an ape-like thorax with Homo-like vertebrae, and a shoulder girdle indicative of climbing competency. The individual bones of the foot and lower limbs largely seem to show an individual compatible with obligate bipedalism, although the pedal phalanges also show curvature [1,2]. Despite the remarkably voluminous assemblages, the H. naledi remains do not include a complete lower limb confidently ascribed to a single individual. A complete lower limb of H. naledi would be informative in what it could tell us regarding potential locomotion. The aim of this study was to reconstruct the lower limbs of the H. naledi skeleton and analyse the potential gait of H. naledi whilst also reviewing recent work on the functional morphology of H. naledi and how this pertains to inferences about bipedal locomotion. The H. naledi lower limb was constructed using estimated femoral, tibial and fibular lengths from the most complete remains of H. naledi currently available [2,3]. Pelvis remains were too fragmented to reconstruct the pelvis. All transformations were performed in ‘LhpFusionBox’, which is a musculoskeletal software primarily used to analyse gait in clinical contexts, but recently adapted for paleoanthropologists [4]. Estimated lengths were used as a reference to reconstruct the individual bones by using anatomical land-marks (ALs) to scale other H. naledi material to the size of the estimated bones. Both a modern human model and new lower limb associations of a juvenile skeleton found in the Dinaledi chamber were used as guides to reconstruct a complete lower limb. Reconstructions of individual isolated and scaled limb and foot bones were then placed together to have a complete lower limb re- construction taking into account ligaments, muscles and following the orientation of joint surfaces. The entire limb was then fused to a modern human walking motion to analyse potential locomotion. The reconstruction and biomechanical analysis of the H. naledi lower limb largely demonstrates a morphology compatible with obligate bipedalism, with a medial arch (although reduced), elongated limbs, marked bicondylar angle and joint surfaces compatible with bipedal gait. The elongation of the lower limb is generally seen as a marker of obligate bipedalism although the H. naledi limbs are exceptionally elongated relative to the diaphyseal diameter of the long bones and preserved joint proportions. Longer limbs are generally thought of as more energy efficient, but they also require a greater moment of inertia, which increases energy costs. Whilst the longer tibia (and leg) may have necessitated a longer swing phase, low limb mass (as evidenced by long bone gracility and small joints) may have offset this energetically. Other unique traits such as the flaring ilium, flattened femoral lower neck and overall general mix of primitive and Homo-like traits may not have impeded obligate bipedalism, but they may have been advantageous for climbing. Primitive traits are often thought of as vestiges of an ancient past on the way to the modern human form of obligate bipedalism, however, Homo erectus and other skeletons from the genus Homo are largely thought to be fully obligate bipedals from approximately 1.5 million years onwards, and as H. naledi has been dated between 335 and 236ka [5], it is therefore curious why this particular branch of the hominid tree would ‘hang on’ to the ‘primitive traits’ for at least a million years longer. It is therefore likely that this hominin engaged in both arboreal climbing and bipedal walking.

Over the last few years, the recovery and the analyses of genomes of ancient modern humans, Neandertals, and Denisovans have changed our understanding of the origins, the movements, and the relatedness of archaic and modern human populations. How- ever, in many cases endogenous DNA represents such a small fraction of the DNA extracted from specimens that sequencing of the complete ancient genomes is economically infeasible. Thus, to date, only three Neandertal genomes have been sequenced to high coverage [1-3]. Even though Neandertal genome sequences of low coverage [4] can be used to reconstruct various aspects of Neandertal genetic history, many analyses, for example estimation of population size and levels of inbreeding, rely on the reliable diploid genotypes. Recent studies have shown that certain skeletal elements, such as the inner part of the petrous bone and the ce- mentum layer in teeth [5 and references therein], may preserve DNA better over time. There is also evidence that the preservation of endogenous DNA may vary substantially even within a few millimeters distance in a single specimen [2, 4]. Due to the value and scarcity of ancient hominin remains, it is critical that the smallest possible amount of destructive sampling is involved in the recovery of genetic material. A usual sampling strategy typically involves taking around 50 mg of powder from a single location of a given bone or tooth. We investigated here whether taking multiple smaller samples in a step-wise manner of the Neandertal specimens from the Mezmaiskaya Cave in Russia and the Troisième caverne of Goyet in Belgium may improve the yield of an- cient human DNA. We removed between 8.5 and 27.2 mg of bone powder from a Mezmaiskaya 1 rib fragment, between 2.5 and 35.1 mg from a Mezmaiskaya 2 skull fragment, and between 5.8 and 53.8 mg from the Goyet Q56-1 femur fragment, amounting to between 15 and 38 powder subsets per specimen and an average input of 16.6 mg of powder per extraction. Importantly, to minimize the impact of contamination, we treated each powder aliquot with 0.5\% sodium hypochlorite solution prior to DNA extraction. The DNA extracts from the same specimen varied by several orders of magnitude in their proportion of endogenous DNA (between 0.07\% and 54.7\%), their content of nuclear genomes (between 0.01 and 78-fold coverage), as well as in the levels of present-day human contamination (0.2-50.3\%). There was no significant correlation between the amount of powder used for the extraction and the overall amount of the endogenous DNA or the levels of present-day human DNA contamination. Thus, these results indicate that ancient DNA preservation varies greatly within one specimen and that the removal of multiple, small sub-samples instead of one larger sample, here coupled with a decontamination procedure, can drastically improve the likelihood of isolating large enough amounts of DNA to make whole genome sequencing feasible. This approach allowed us to identify extracts with exceptionally high endogenous DNA content and low levels of present-day human DNA contamination (2\%), enabling us to generate three additional high-coverage Neandertal genomes. The high-quality genome sequences of multiple Neandertals form a unique reference resource for the scientific community and are valuable for analyses that require reliable diploid genotypes and haplotype information. For example, these data open new opportunities to investigate Neandertal population history, to identify genetic variants that arose uniquely on the Neandertal lineage and might have changed through time, and to determine those that may underlie archaic-specific traits or adaptations.

The main excavations at the Troisième caverne of Goyet in Belgium were conducted by Edouard Dupont in 1868 who identified Palaeolithic human occupations later attributed to the Middle and Upper Palaeolithic. These are represented by an archaeologi- cal record that spans the Mousterian, Lincombian-Ranisian-Jerzmanowician, Aurignacian, Gravettian, and Magdalenian, and then extends into the Neolithic and historic periods. Due to the lack of detailed documentation of the excavated materials, their asso- ciation to a specific chronocultural context has been challenging. Morphometric and taphonomic analyses, combined with direct radiocarbon dating as well as isotopic and genetic analyses, were used to assign human remains to either late Neanderthals or an- cient modern humans from different chronocultural groups. In 2016 the first palaeogenetic investigation of Neanderthal specimens from Goyet was published [1]. Taxonomic assignment was confirmed by performing hybridization capture of the mitochondrial DNA (mtDNA) and later inspecting diagnostic mutations at nucleotide positions that distinguish modern humans from Nean- derthals. Moreover, a phylogenetic reconstruction placed seven nearly complete mtDNA sequences from Goyet within the diver- sity of late Neanderthal mtDNA. An around two-fold coverage nuclear genome was later sequenced from one of those individuals (Goyet Q56-1) [2], revealing a high genetic similarity to other late Neanderthals that is well correlated to their geographical dis- tance. Analyzing modern human remains retrieved at Goyet, mtDNA genomes were initially reported for two specimens directly dated to the Aurignacian, five to the Gravettian, and one to the Magdalenian [3]. Aurignacian-related individuals were particu- larly intriguing as they were found to carry mtDNA haplogroup M, which is almost entirely absent in present-day Europeans. For Gravettian- to Magdalenian-related individuals, the shift from U2/U5 to U8 haplogroups was detected locally - as in other regions of Central Europe - likely influenced by the genetic bottleneck during the Last Glacial Maximum (LGM). Furthermore, nuclear sequences of five modern human individuals from Goyet were produced through genome-wide targeted enrichment [4] revealing local replacement between Aurignacian- and Gravettian-related populations. However, the genetic component associated with a 35,000-year-old individual (Goyet Q116-1) reappeared after the LGM, first in Spain and then in other European regions includ- ing in a Magdalenian-related individual from Goyet (Goyet Q-2). This individual was later found to be the best proxy for a genetic component that was largely displaced in Europe from around 14,000 years ago onwards while surviving in high proportion among Mesolithic individuals from Iberia [5]. Here we present new palaeogenetic data of Neanderthal and modern human individuals from this iconic site. First, we expand the molecular taxonomic identifications with three additional Neanderthal specimens and reconstruct their partial mtDNA genomes. Those confirm the general picture of a limited genetic diversity for late Neanderthals, which is also apparent among the Goyet Neanderthals. Second, working on modern human remains, we produced new mtDNA and nuclear data from four Gravettian specimens. They belong to mtDNA haplogroups U2 and U5, further extending the observa- tion of both mtDNA types being largely present in pre-LGM Europe. Moreover, their nuclear genomes provide additional evidence for the genetic affinity between Gravettian-related groups across Europe, from the present-day regions of the Czech Republic to Belgium and Southern Italy. In conclusion, the deep temporal range covered by the human remains from the Troisième caverne of Goyet provides the unique opportunity to describe within a single archaeological site the major genetic transformations that took place in Europe throughout the Middle and Upper Palaeolithic.

Trou Magrite is a cave site located at Pont-à-Lesse in the Lesse Valley, commune of Dinant, Belgium. It has been known since E. Dupont conducted excavations at the site in 1867 [1]. The most recent fieldwork was done by L. Straus and M. Otte in 1991-92 [2]. Trou Magrite yielded rich lithic assemblages, osseous artifacts, mobiliary art, and numerous faunal remains. Several human re- mains were also recovered and identified as Palaeolithic humans by E. Dupont but have been only partially published thus far. The archaeological record covers a broad time range spanning from the Middle and Upper Palaeolithic to the Mesolithic, Neolithic, and Iron Age. An important Middle Palaeolithic collection is present, probably representing several occupation phases during the Late Pleistocene [2]. Unfortunately, although E. Dupont conducted excavations that can be characterized as modern for that time, the materials from the different so-called “fauna-bearing levels” that he defined in the field were mixed post-excavation [3]. In 2015, we initiated a multidisciplinary re-assessment of the human and faunal collections from Trou Magrite in order to update the inven- tory of human remains already identified, check for the presence of human remains that may have been previously overlooked, and verify their chronocultural context. We revised the already known human collection, conducted a systematic sorting of the faunal material, and combined the use of morphometrics, taphonomy, stable isotopes, dating, and genetic analyses to perform taxonomic and chronocultural identifications. Here we present two previously unidentified Neandertal fossils that we isolated from the Trou Magrite faunal material excavated by E. Dupont in the 19th century. They represent two different individuals: an adult/adolescent, represented by an upper right permanent canine, and a neonate, represented by the diaphysis of a left femur. Whereas no endoge- nous DNA was recovered from the tooth, the palaeogenetic analyses of the neonate femur confirmed its Neandertal status and indicate its sex to be male. We will present the biological characteristics and mitochondrial DNA phylogenetic position of the Trou Magrite Neandertals, in particular with regard to the other Northern European Neandertals. Our project adds Trou Magrite to the list of Belgian sites that have yielded Neandertal fossils and helps to emphasize the importance of the Mosan Basin in Neandertal studies.

Subsistence strategies are key paleoecological features of Paleolithic hunter-gatherers and their deeper understanding provides crit- ical insights into essential aspects of human evolution. In this study, we discuss new collagen stable isotopic values (C, N, S) rep- resenting seven Gravettian individuals from the Troisième caverne of Goyet in Belgium. The dietary strategies of the Gravettian humans from Goyet are in line with the general trends observed among Western European Gravettian populations. These pop- ulations show both a low intake of mammoth and a high consumption of other terrestrial mammals as well as aquatic resources, such as at the sites Arene Candide and La Rochette. This is different for more eastern Gravettian hunter-gatherers, for example in Kostenki, Brno-Francouzska, Mal’ta, Předmostí, and Dolní Věstonice where the dietary contribution of mammoth meat was sig- nificantly higher. The stable isotopic data of the Gravettian humans from Goyet indicate that their dietary ecology was essentially based on terrestrial resources like reindeer, horse, and, to a lesser extent, mammoth. However, they yielded δ15N values that are substantially lower than those of the earlier modern humans and Neandertals from the same site [1-2]. We hypothesize that the Gravettian humans had much less mammoth in their diet than all earlier humans from the same region. It was previously shown that in northwestern Europe a decline of mammoth, a key prey species, could already be detected at the onset of the Upper Paleolithic [2]. This trend appears to continue into the Gravettian, despite the persistence of the typical mammoth ecological niche, which is represented by a grassland with high δ15N values. Interestingly, through isotopic analysis, we are able to track the spread of the horse from the local ecosystem (represented by specimens from Walou Cave, Belgium) into this niche now under-occupied by the mammoth. Radiocarbon dates obtained from several mammoth skeletal remains from the Troisième caverne of Goyet showed that this megaherbivore was indeed part of the ecosystem during pre-LGM periods. However, from the Gravettian in Goyet and the surrounding region we have only one mammoth specimen represented by a long bone, and interestingly, its sulphur isotopic signal indicates that this individual was not of local origin. We propose that the local mammoth population was under intensive hunting pressure or may even have been no longer present in the region. Instead, single individuals from other regions may have made it into the area and ended up as prey animals. While the δ15N values of all Goyet Gravettian humans are relatively homogeneous, their δ13C values are variable. This indicates significant dietary differences among the seven individuals, an observation that has not been described before for hunter-gatherers pre-dating the Gravettian. The human δ34S values also support substantial differences in life mobility history between different individuals, which were not observed for the Goyet Neandertals. The result that different mem- bers of the same chrono-group had various individual mobility histories has implications for land use procurement strategies of those hunter-gatherer groups. In conclusion, our new isotopic results demonstrate a broad ecological flexibility among Gravettian humans, which can be seen in different human ecosystem interactions across Europe. The Goyet individuals contribute substan- tially to a more complete understanding of hunter-gatherer’s ecology during this particular phase of the European Late Pleistocene. Our study shows that the Gravettian cannot be depicted as a uniform entity from an ecological perspective. It instead indicates that during this period, and not earlier, both inter- and intra-group diversity in subsistence strategies can be tracked through stable isotopic analysis.

Association for Tropical Biology and Conservation annual meeting https://www.atbc2024.org Large old tropical trees as keystone biodiversity structures: the Life on Trees program Leponce Maurice1, Basset Yves2, Aristizábal-Botero Ángela1, Albán Castillo Joaquina3, Aguilar Rengifo Guillermo4, Barbut Jérôme5, Buyck Bart5, Butterill Phil6, Calders Kim7, Carrias Jean-François8, Catchpole Damien9, D’hont Barbara7, Delabie Jacques10, Drescher Jochen11, Ertz Damien12, Heughebaert André13, Hofstetter Valérie14, Leroy Céline15, Leveque Antoine16, Macedo Cuenca Victor4, Melki Frédéric17, Michaux Johan18, Ocupa Horna Luis19, Pillaca Huacre Luis3, Poirier Eddy20, Ramage Thibault21, Rougerie Rodolphe5, Rouhan Germinal5, Rufray Vincent17, Salas Guererro Marcos4, Scheu Stefan11, Schmidl Jürgen22, Silva Dávila Diana3, Valenzuela Gamarra Luis23, Vanderpoorten Alain18, Villemant Claire5, Youdjou Nabil1, Pascal Olivier17 1 Royal Belgian Institute of Natural Sciences, Vautier st. 29, Brussels, 1000, Belgium; 2 Smithsonian Tropical Research Institute, Panama; 3 Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima, Peru; 4 Servicio Nacional de Áreas Naturales Protegidas por el Estado, Ministerio del Ambiente, Peru; 5 Muséum national d'Histoire naturelle, Paris, France; 6 Biology Centre, Czech Academy of Sciences, České Budějovice,Czech Republic; 7 Ghent University, Belgium; 8 Université Clermont-Auvergne, Clermont-Ferrand, France; 9 Independent Consultant, Lima, Peru; 10 Centro de Pesquisas do Cacau – CEPEC, Itabuna, Brasil; 11 Göttingen University, Germany; 12 Meise Botanic Garden, Belgium; 13 Belgian Biodiversity Platform, Brussels, Belgium; 14 AGROSCOPE, Nyon, Switzerland; 15 AMAP (Univ. Montpellier, CIRAD, CNRS, INRAE, IRD), Montpellier, France; 16 PatriNat (OFB-CNRS-MNHN), Paris, France; 17 Fonds de Dotation Biotope Pour La Nature, Mèze, France; 18 Université de Liège, Belgique; 19 Centro de Investigación en Biología Tropical y Conservación, Piura, Perú ; 20 Independent entomologist, Cayenne, Guyane ; 21 Independent entomologist, Concarneau, France ; 22 Universität Erlangen-Nürnberg, Germany. ; 22 Jardín Botánico de Missouri, Peru E-mail: (presenting author): mleponce@naturalsciences.be The aim of the Life on Trees (LOT, www.lifeontrees.org) program is to generate baseline knowledge about the number of eukaryotic species that a single large mature tropical tree can host and to understand how these communities of organisms are assembled. The program is being undertaken in the Andean Amazon biodiversity hotspot. Our first project, LOT01 in the Andean foothills in 2022, located at 400m a.s.l., involved the study of a spectacular Dussia tessmannii tree (Fabaceae), towering at 50 meters in height and 45m wide. Our second project, LOT02 in the Andes in 2023, at 2450m a.s.l., focused on a 32-meter-tall Ficus americana subsp. andicola. Surveys were carried out by professional climbers, guided by experts of the different eukaryotic groups studied (plants, fungi, animals, protists). To better understand the contribution of different tree components (bark, leaves, fruits, flowers, living and dead wood) to overall tree biodiversity, we partitioned observations into communities based on vertical strata or microhabitat and will examine similarities and nestedness in the composition of these communities. Initial findings indicate that significant diversity is harbored by the individual tree at both locations (e.g., LOT01 vs LOT02: 42 vs 114 orchid species, 28 vs 28 fern species, 200+ vs 300+ bryophyte species, and 180 vs 100+ lichen species identified). These figures set world records for their respective elevations. This confirms that large old tropical trees are important pools of biodiversity, probably related to the variety of local microhabitats and tree age.

Canopy laser scanning to study the complex architecture of large old trees Barbara D'hont1 , Professor Kim Calders1 , Professor Alexandre Antonelli6 , Dr. Thomas Berg7 , Dr. Karun Dayal1 , Dr. Leonard Hambrecht5 , Dr. Maurice Leponce2,3, Prof. Arko Lucieer5 , Olivier Pascal4 , Professor Pasi Raumonen8, Professor Hans Verbeeck1 1Q-ForestLab, Department of Environment, Ghent University, Ghent, Belgium, 2Royal Belgian Institute of Natural Sciences, Brussels, Belgium, 3Université Libre de Bruxelles, Brussels, Belgium, 4Fonds de Dotation Biotope Pour La Nature, France, 5School of Geography, Planning, and Spatial Sciences, University of Tasmania, , Australia, 6Royal Botanic Gardens, Kew, Richmond, Surrey, United Kingdom, 7ARAÇÁ Project, Nova Friburgo, Rio de Janeiro, Brazil, 8Faculty of Information Technology and Communication Sciences, Tampere University, Tampere, Finland Large trees are keystone structures providing multiple ecosystem functions in forests all around the world: they disproportionately contribute to forest biomass and biodiversity. Large trees can have an extremely complex structure, housing many epiphytes on their stem and branches. High point-density 3D point clouds, in which leaves and epiphytes in the tree can be distinguished, are useful to make the link between the distribution of organisms on the tree, the tree architecture and its microclimate. In addition, a comprehensive branching model can improve above ground biomass (AGB) estimates. Highly detailed, complete point clouds of large trees are, however, exceptionally difficult to derive. With terrestrial laser scanning, the state-of-the-art method to capture 3D tree structure, the plant material blocks the view of (or, occludes) the top part of the dense crown. Drone or airborne laser scanning data on the other hand, lacks detail in the subcanopy. Combining these two methods minimises occlusion; however, increased distance from the tree to the scanner still leads to a relatively low resolution of the canopy point clouds. To improve the level of precision of the tree point clouds, we introduce a new concept, called canopy laser scanning (CLS). With CLS, a laser scanner is operated statically inside the tree canopy, reducing the distance between the area of interest and the instrument. We lifted a high-end laser scanner (RIEGL vz-400(i)) inside the canopy of six large emergent trees. Four of these trees are located in different types of tropical rainforests in Colombia, Brazil and Peru. They are part of biodiversity programs in which organisms and their spatial distributions are studied (Life On Trees, Araçá). The two other trees are famous giants located in the wet temperate eucalypt forests of southern Tasmania. We will present the practical aspects of CLS, evaluate the extra value of using canopy scans, looking at occlusion and point cloud precision, estimate epiphyte cover and AGB. We demonstrate that canopy laser scanning opens up new opportunities in sciences in which multi-disciplinary teams perform in depth research on large individual trees.