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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.

The aim of the Life On Trees (LOT) program is to generate baseline knowledge about the number of eukaryotic species a single large aged tropical tree can host and to understand how these communities of organisms are assembled. The program is conducted in the Amazon and Andes biodiversity hotspots. Our first project, LOT-Amazon 2022, was performed on a spectacular Dussia tree (Fabaceae), which was 50 m high and 45 m wide. The sampling was 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 assigned observations into communities based on height zone or microhabitat and will examine similarities and nestedness in the composition of these communities. The first results show that a single tree can host a tremendous diversity (e.g., 42 orchids, 28 ferns, and more than 200 bryophytes, 180 lichen species identified, which are world records considering the 400m elevation). This confirms that large old tropical trees are important pools of biodiversity probably in relation with the variety of local microhabitats and tree age. Funding: Fonds de Dotation Biotope pour la Nature Web and/or Twitter account: www.lifeontrees.org