Reference Collection Research Articles

Assessing DNA for fish identifications from reference collections: the good, bad and ugly shed light on formalin fixation and sequencing approaches.

Natural history collections are repositories of biodiversity and are potentially used by molecular ecologists for comparative taxonomic, phylogenetic, biogeographic and forensic purposes. Specimens in fish collections are preserved using a combination of methods with many fixed in formalin and then preserved in ethanol for long-term storage. Formalin fixation damages DNA, thereby limiting genetic analyses. In this study, the authors compared the DNA barcoding and identification success for frozen and formalin-fixed tissues obtained from specimens in the CSIRO Australian National Fish Collection. They studied 230 samples from fishes (consisting of >160 fish species). An optimized formalin-fixed, paraffin-embedded DNA extraction method resulted in usable DNA from degraded tissues. Four mini barcoding assays of the mitochondrial DNA (mtDNA) were characterized with Sanger and Illumina amplicon sequencing. In the good quality DNA (without exposure to formalin), up to 88% of the specimens were correctly matched at the species level using the cytochrome oxidase subunit 1 (COI) mini barcodes, whereas up to 58% of the specimens exposed to formalin for less than 8 weeks were correctly identified to species. In contrast, 16S primers provided higher amplification success with formalin-exposed tissues, although the COI gene was more successful for identification. Importantly, the authors found that DNA of a certain size and quality can be amplified and sequenced despite exposure to formalin, and Illumina sequencing provided them with greater power of resolution for taxa identification even when there was little DNA present. Overall, within parameter constraints, this study highlights the possibilities of recovering DNA barcodes for identification from formalin-fixed fish specimens, and the authors provide guidelines for when successful identification could be expected.

Mitigating the potential contamination risk incurred in sampling herbarium specimens for phytolith reference collection

Phytoliths are emerging as a critical paleoenvironmental and archaeological proxy record in a variety of landscapes. These non-organic objects preserve well in both terrestrial and lacustrine sedimentary contexts. Creating a local reference collection is a crucial step for contending with the multiplicity and diversity of phytolith forms, but previous authors have noted a potential problem with one of the most common methods for extracting phytoliths from fresh and herbaria specimens, the dry-ashing method. This method incurs a contamination risk in the muffle furnace during ashing. The standard weight of plant tissue for extraction is relatively high (200 mg) when considering the amount of material accessible in herbaria collections. We devised the following experimental study to test the contamination potential of our muffle furnace, and the possibility of using a smaller weight of plant tissue (100 mg) to derive the same results and reduce potential risk. The results of this study indicate that reducing the initial plant material by half the weight will result in a slightly lower contamination risk and equivalent diversity of morphotypes in the final phytolith sample. Therefore, we argue that reducing the plant tissue will enable a less costly use of herbaria material and a lower chance of inter-sample contamination.

Classification of ground-truth fire debris samples using artificial neural networks

Neural networks are a class of biologically inspired machine learning models that are used for classification and regression problems. This work assesses the classification performance of neural networks on ground-truth fire debris samples using ions that are representative of several compound classes that are typically present in ignitable liquids. An optimal neural network model was selected from a subset of candidate models that were trained on in-silico mixed fire debris samples from the National Center for Forensic Science Substrate and Ignitable Liquid Reference Collection databases. An optimal decision threshold was determined using a defined ratio of misclassification costs. A cost ratio corresponding to a false positive classification having a cost that is 10 times greater than a false negative classification resulted in a decision threshold of log likelihood ratio of 0.966. This decision threshold resulted in a false positive rate of 0.07 and a true positive rate of 0.59 for the ground-truth validation data. This study demonstrates the selection of an optimal decision threshold using ROC analysis and exhibits the potential of neural network models for the evaluation of fire debris evidence.

Evaluation of microscopy techniques and ATR-FTIR spectroscopy on textile fibers from the Vesuvian area: A pilot study on degradation processes that prevent the characterization of bast fibers

The unambiguous identification of bast fibers represents one of the main challenges for archaeologists and conservators dealing with archaeological textiles, motivated by the difficulties of their investigation through the morphological features only. Therefore, in this work, a combined approach of microscopy and spectroscopy techniques is proposed. The research aims to test FT-IR spectroscopy in Attenuated Total Reflectance (ATR) mode, as a valid tool for the characterization of archaeological fibers, as already demonstrated for the modern ones, and additionally, the application of the same technique to evaluate the effect of degradation on the molecular composition of archaeological bast fibers, that may prevent their identification. To this end, a set of archeological samples, from Pompeii and the Vesuvian area, previously investigated using morphological analyses, have been studied. The results obtained showed that the preservation state of the fibres does not make their characterization simple. To understand how the ageing process influences the IR spectra of archeological fibers, a reference collection was acquired and analyzed. Moreover, two type of selected bast fibers samples were artificially aged using a climatic chamber. The outcomes achieved confirmed that the degradation process has influenced the IR spectra of the fibers to the point of making them spectrally indistinguishable, highlighting useful information to understand the role of the degradation process in the ATR FT-IR spectra of archaeological bast fibers.

Seed morphology uncovers 1500\xa0years of vine agrobiodiversity before the advent of the Champagne wine

A crucial aspect of viticulture is finally unveiled as the historical dynamics of its agrobiodiversity are described in the Champagne region for the first time. Outline analyses were carried out to compare the morphology of archaeological grape seeds from Troyes and Reims (first c. AD to fifteenth c. AD) with that of a reference collection of modern seeds, including wild vines and traditional grape varieties, believed to be ancient and characteristic of the French vine heritage. This allows us to document the chronological dynamics of the use of the wild Vitis type and of the diversity of the varieties used, based on morphological disparity. After showing the existence of morphological types corresponding to geographical groups, we highlight a geochronological dynamic. Our results show that the wild type is used throughout the series, up to the Middle Ages. In addition, domestic forms, morphologically related to southern varietal groups, are very early involved in the Champagne grape agrodiversity. The groups corresponding to the typical grape varieties of today do not appear until the second millennium. These previously unsuspected dynamics are discussed in light of the social, societal and climatic changes documented for the period.

Modernizing Medical Museums Through the 3D Digitization of Pathological Specimens.

The anatomical collections at the National Museum of Health and Medicine (NMHM) contain skeletal specimens that highlight the history of military and civilian medicine dating from the American Civil War and the founding of the museum as the Army Medical Museum in 1862. Today, NMHM curates over 6400 gross skeletal specimens consisting primarily of pathological or anomalous single bone elements that display a variety of pathological conditions, including congenital anomalies, neoplasms, healed and unhealed trauma and infectious diseases, and surgical interventions such as amputations and excisions. In an effort to increase accessibility to these pathological specimens, NMHM is collaborating with Virginia Commonwealth University (VCU) and the Laboratory Division of the Federal Bureau of Investigation (FBI) to digitize and disseminate high-quality 3D models via online portals, enabling scholars and educators to manipulate, analyze, and 3D print the models from anywhere in the world. Many institutions with courses in paleopathology and forensic anthropology do not have reference collections or access to museum collections for hands-on teaching. Therefore a digital repository of osteological specimens can provide an unprecedented and unique resource of exemplars for scholars and educators. The sharing of these military medical assets improves historical knowledge and diagnostic capabilities in the fields of medicine and anthropology. This chapter outlines the digitization processes that are being utilized to increase access to these pathological skeletal specimens through multimodal 3D capture.

A Preliminary Checklist of Flowering Plants in Pangkor Selatan Forest Reserve, Perak, Peninsular Malaysia

A preliminary botanical survey was conducted in the coastal hill forest in Pangkor Selatan Forest Reserve, Perak, Peninsular Malaysia. Standard methods were used to record the species found around this forest. The first method involved the collection of herbarium specimens which are complete with fruit and flowers (fertile specimens). Unfortunately during this survey many species did not bear fruits or flowers. The second method involved the collection of sterile specimens (without fruits and flowers) to be used as voucher specimens and the third method was by random observation where the plants were observed without any specimen collection. The fourth method involved the establishment of small plots where all plants were enumerated. In the first and second methods, the specimens were collected and processed. The fertile specimens were kept in the Herbarium of Universiti Kebangsaan Malaysia (UKMB) as a reference in the future. Identification was done by comparing with those reference collections in herbarium UKMB and herbarium of Forest Research Institute (KEP) plus referring to local botanical books. A total of 237 taxa belonging to 170 genera and 64 families were recorded. Dicotyledons are represented by 59 families while the monocotyledons are represented by only 5 families. The family Euphorbiaceae is the most speciose with 21 species and followed by Dipterocarpaceae with 14 species. Both the genera <i>Diospyros </i>and<i> Syzygium </i>are the largest with 8 species each. The species that are commonly found in these forests include <i>Cnestis palala</i>,<i> Rourea minor</i>, <i>Hopea beccariana, Diospyros clavigera</i>, <i>Hydnocarpus filipes</i>,<i> Fordia unifoliata</i>,<i> Memecylon pubescens</i>, <i>Pternandra coerulescens</i>,<i> Ficus variegata</i>,<i> F</i>.<i> vasculosa</i>, <i>Knema stenophylla</i>, <i>Syzygium zeylanicum</i>,<i> Strombosia javanica</i>,<i> Eugeissona tristis</i>,<i> Orania sylvicola</i>, <i>Xanthophyllum affine</i>, <i>Gynotroches axillaris</i>,<i> Aidia densiflora</i>,<i> Diplospora malaccensis</i>,<i> Gardenia carinata</i>,<i> Psydrax </i>sp. 10, <i>Paramignya </i>sp.,<i> Grewia laevigata</i>,<i> Schoutenia accrescens </i>ssp.<i> accrescens </i>and<i> Rinorea anguifera</i>. A total of 25 endemic taxa to Peninsular Malaysia has been recorded in this study including <i>Fordia unifoliata</i>, a small tree of ca. 10 m tall, endemic to Perak and restricted to coastal hill forest. In addition, a total of 48 species of flowering plants in Pangkor Selatan Forest Reserve was listed in 2020 IUCN Red List Categories and related reference. It is clear that this coastal hill forest has supported many species of flowering plants including endemic taxa and this forest need to be conserved.

AJOM new records and collections of fungi: 101-150

This article is the second in the Asian Journal of Mycology Notes series, wherein we report 50 new fungal collections distributed in two phyla, six classes, 23 orders and 38 families. The present study provides descriptions and illustrations for three new species (Acolium yunnanense, Muyocopron cinnamomi and Thyrostroma ulmeum), 44 new host records and new geographical distributions and three new reference collections. All these introductions are supported by morphological data as well as the multi-gene phylogenetic analyses. This article provides a venue to publish fungal collections with new sequence data, which is important for future studies. An accurate and timely report of new fungus-host or fungus-county records are essential for diagnostics, identification and management of economically significant fungal groups, especially the phytopathogens.

Qualitative and Quantitative Analyses of Surface Alterations of Stone Tools Made of Raw Materials from the Carpathian Basin

Wear analysis performed on archaeological stone tools from prehistory aims at characterizing their surface alterations in order to determine their functions and describe past human technical behaviors. However, the reliability and repeatability of the method is questioned and there is a major difficulty facing scientists analyzing wear, which is related to proposing quantified and repeatable analyses and interpretations of taphonomic and anthropogenic (related to manufacture and use) evidence. We propose the creation of an experimental reference collection of surface alterations focused primarily on selected stone types commonly found at Hungarian archaeological sites. This collection will allow the characterization of the topographic signatures of manufacture, use, and taphonomic processes. The qualitative (through observation using optical microscopy) and quantitative (through surface measurements using confocal microscopy) analyses will aim at examining: 1) the variability of surface alterations, 2) protocols that can allow the identification and characterization of surface topographic signatures, and 3) the appropriate metrological setups that will allow for each of the alteration processes to be differentiated. We present here the results of the preliminary experiments, which involved flakes for bone and reed processing made of six different raw materials. It has been confirmed that evidence of use can be identified on each one of them. Beyond the presentation of the first images from the reference collection, we briefly present the future phases of the project.

Charcoal anatomy of Brazilian species. II. 15 native species occurring in Atlantic or Amazon rainforest.

Anthracology is the identification of charcoal remains through wood anatomy. Paleoecological and paleoethnobotanical evidence from anthracological studies provides information on past environments as well as the fuel economy and use of plants by those living in ancient societies. Historical ecology and forest conservation can also accrue from findings in anthracological studies. Charcoal identification must rely on adequate reference material, in particular reference collections and descriptions of charcoal anatomy. This paper presents charcoal anatomy descriptions of fifteen Brazilian native species that occur in the Atlantic Forest and the Amazon Rainforest. The charcoal anatomy of six of these species is here described for the first time. Samples were analyzed under a reflected light microscope; the descriptions followed the procedures and terminology recommended by the International Association of Wood Anatomists. Increased knowledge of the charcoal and wood anatomy of native tropical species may improve taxonomic identification, thereby increasing accordingly the amount and quality of data for sociocultural inferences about past societies. In addition, it contributes to a better knowledge of the native flora, which helps to prevent deforestation and to drive more sustainable charcoal production chains.

Starchy food residue on a potsherd from a late Holocene hunter-gatherer site in Argentine Patagonia: towards the visibility of wild underground storage organs

The aim of this paper is to present recent advances in the microbotanical analyses of an organic residue on a potsherd from a late Holocene hunter-gatherer site in Argentine Patagonia, which is the first evidence of this kind for the processing of starchy food. Standard methods were carried out for starch grain recovery and morphometric analysis, care being taken in the prevention of cross-contamination and evaluation of post-depositional factors. Diagnostic features of the ancient starch grains recovered were compared against those from a list, available in the bibliography, of plants potentially processed in pottery, but no match was found. An anatomical and starch grain reference collection was established with three of the most frequently consumed traditional wild underground storage organs (USOs) of Patagonia, Alstroemeria aurea Graham, Tropaeolum porifolium Cav. and Diposis patagonica Skottsb., in order to compare these against the archaeobotanical record. We suggest that T. aff. porifolium (and probably also A. aff. aurea) were processed in the pot from which the sherd came, and discuss these results in terms of a better understanding of the role of wild USOs in subsistence and the possible cooking methods used in hunter-gatherer societies in Argentine Patagonia.

Results of excavations of the settlement of Galik 11 in 2020

This paper discusses the results of excavations of the settlement-site of Galik 11, situated on the lower Luga River. The materials from the excavations represent three complexes of different periods with finds of the late Neo- lithic — Bronze Age related to the cultures of the pit-comb ware, corded ware, and textile ceramics. The findings of dif- ferent groups are separated planigraphically. The excavations have resulted in obtaining reference collections of materials of the corded ware and textile ceramics of the Bronze Age in the region under consideration.

Species Composition and Ecological Distribution of the Subfamily Cicindelinae Latreille, 1801 (Coleoptera: Carabidae) Based on Voucher Specimens in Sarawak

The subfamily Cicindelinae which belongs to the family Carabidae are commonly known as tiger beetles. Despite the recent studies conducted on this taxon, current ecological knowledge on tiger beetles is limited. Thus, this study aimed to provide a current checklist of tiger beetles as well as their species distribution, abundances and habitat preferences in Sarawak based on voucher specimens from Universiti Malaysia Sarawak Insect Reference Collection (UIRC). A total of 76 specimens of tiger beetles were examined in which comprising of seven genera from eight species. The genus Neocollyris was represented with two species, namely Neocollyris (s. str.) ermaginata and Neocollyris (Stenocollyris) sarawakensis macrodera, while the other genera were represented by only one species, respectively. The most abundant species with 64 individuals (84.21%) were represented by Cosmodela aurulenta, followed by Abroscelis tenuipes araneipes and Therates labiatus with three individuals (3.95%) and N. ermaginata with two individuals (2.63%). In this study, there were eight ecological habitats being identified, namely heath forest, limestone forest, littoral forest, mixed dipterocarp forest, oil palm plantation, peat swamp forest, riverine forest and urban area. The data indicates that C. aurulenta being recorded at seven ecological habitats suggesting this species to have a wide distribution capability while species such as A. tenuipes araneipes, Callytron doriai, Cicindela sp., Myriochila (s. str.) specularis brevipennis, N. ermaginata and N. s. macrodera suggesting these species to have a very narrow habitat specialisation. The results of this preliminary study provide evidence on the need to conduct further studies on the ecological aspects of tiger beetles in Sarawak.&#x0D; &#x0D; Keywords: Cicindelinae, ecological distribution, Sarawak, tiger beetles, voucher specimens

Species Composition and Distribution of Tropical Marine Macroalgae In The Pari Island Reef Cluster, Jakarta

The objective of this study was to analyze the species composition and distribution of marine macroalgae at Pari Island reef cluster-Seribu Islands reef system Jakarta based on specimen information available in Reference Collection of Research Center for Oceanography (RCO-LIPI). The Macroalgae specimens were collected irregularly by researchers during marine expedition projects from 1973 until 2014. Taxonomic identification of specimens was conducted in 2014, validation of species names was updated in May 2020 through www.macroalgaebase.org. The floristic composition value of macroalgae was analyzed using the C/P, R/P, and (R+C)/P ratio. Bray-Curtis similarity index analyzes were performed to visualize differences in macroalgae species from the different islands. We recorded a total of 41 species from 25 genus and 18 families. Rhodophyta represented the most significant number of taxa with 20 species (49%), followed by Chlorophyta with 11 species (27%) and Phaeophyta with ten species (24%). In this study, we found that Sargassum polycystum, Gracilaria salicornia, Amphiroa fragilisima, and Halimeda opuntia are dominant species in the study area. The highest macroalgae diversity found in Kongsi Island with 33 species, while the lower diversity found in Pari Island with five species. Bray-Curtis Similarity index showed that Kongsi Island and Burung Island have the highest similarity value with 29,26%, while Tikus Island and Pari Island have the lowest similarity value with 11,76%. The macroalgae distribution in the Pari island reef cluster influenced by the difference of substrate profile and environmental pressure derived from human activities. Proper management of macroalgae resources is necessary to preserve the sustainability of the macroalgae ecosystem. Keywords: macroalgae, composition, distribution, Pari Island

Wood identification of charcoal with Confocal Laser Scanning Microscopy

Wood identification of barbecue charcoal from commercial packages of three retailers (B1, B2, B3) in Slovenia and Croatia was performed with help of Confocal Laser Scanning Microscopy (CLSM). CLSM enabled us to image key identification features of charcoal wood that were compared with light micrographs of wood from the reference collection. Product B1 contained charcoal made exclusively of beech wood (Fagus sylvatica) and the declaration indicated the address of the producer, in Serbia which allowed traceability of the wood. The selection of wood species in product B2, consisted of red oak (Quercus cerris or Q. rubra), black locust (Robinia pseudoacacia), and cherry (Prunus avium), which could originate from Serbia, and it did not contain tropical wood as stated on the package. Product B3 contained wood from at least four (sub)topical species which could not be exactly identified to species/genus level. The declaration on the product did not allow traceability of wood. As the risks of illegal logging are high for wood of (sub)tropical origin, our results support the initiative that the monitoring of the charcoal trade should be covered by the EUTR - European Timber Regulations.

Investigating Animal Remains in Estonia

In Estonia, faunal remains have been an important part of archaeological material since the 19th century. During the 20th century, the interest in faunal history was rather volatile, but gained some stability during the 1990s. Since then, zooarchaeology in Estonia has developed substantially, focusing on a variety of topics. Together with methods from traditional zooarchaeology, interdisciplinary methods like the studies of ancient DNA and stable isotopes are increasingly used. However, despite the growing understanding of the importance of faunal remains in archaeological and historical research, there are still problems with collecting animal remains during the fieldwork and documenting and organising them. On the other hand, interest in scientific methods and destructive sampling of the osseous remains have become increasingly popular in science projects and international collaboration. In order to use osteological collections reasonably and ethically, proper systemisation is essential.In Estonia, there are two research centres for zooarchaeology, where scientific collections are administered – Tallinn University and the University of Tartu. Tallinn collections comprise material mostly from the northern part of the country, plus an extensive reference collection for fish has been developed there. In Tartu, mostly material from southern Estonia is managed, together with continuously expanding reference collection of mammals and birds. To improve the gathering and management of the osteological material in Estonia and reduce the shortage for storage space, a new central repository for osteological collections (both human and animal) was established in 2019. Concurrently, a new central database for the osteological data was created.In this paper, we introduce the zooarchaeological collections and some of the latest research topics in Estonia with an aim to broaden the understanding and potential of zooarchaeology in the Baltic region.

Boiled, fried, or roasted? Determining culinary practices in Medieval France through multidisciplinary experimental approaches

While culinary practices evolved during the Middle Ages (historically dated between A.D. 476 and 1492), these changes are difficult to detect as they are seldom studied. Around the 12th and 13th centuries A.D., changes can be seen in the types of utensils and cooking hearths, but also new animal and plant varieties were introduced, along with recipes used by people in the upper echelons of society being put in writing. These novelties could be the result of changing tastes and modes of preparing food, but to ascertain this we need to study the archaeological materials to determine the cooking methods that may have been used and compare these to their context and time period. To this end, experiments were designed and carried out to determine the traces left behind by different cooking methods. Our results have made it possible to create a reference collection consisting of ceramics as well as faunal and botanical remains. The study of cooking traces, as well as the morphological changes visible on botanical elements, in particular starch grains, have allowed us to establish criteria that can be employed in use-wear analysis. We then compared the experimental reference collection with archaeological materials recovered from Couzan, a Medieval site in Forez, France. Our results support the value of using starch grain analysis when studying the function of dark paste ceramics, and clearly demonstrate the interest of bringing together different sub-disciplines in archaeology when using an experimental approach.

A step forward in tropical anthracology: understanding woodland vegetation and wood uses in ancient Sri Lanka based on charcoal records from Mantai, Kirinda and Kantharodai

The aim of this study is to present the anthracological results from three archaeological sites located in the North, North West and South East of Sri Lanka. The study is based on the observation and analysis of 1689 charcoal fragments using for support the reference collection of South Indian wood at the Institute of Archaeology ( UCL), Inside Wood (2004-onwards) and several wood anatomy atlases. Mantai (200 BCE-850 CE), an urban site, has yielded 25 taxa with significant presence of cf. Cocos nucifera among other taxa. Kantharodai (400-170- BCE), an urban site, has yielded 19 taxa from arid zones (Fabaceae, Rubiaceae), mangroves (Rhizophoraceae) and dune zones (cf. Cocos nucifera). Kirinda (500–900 CE), a fishing settlement, has yielded 24 taxa including Fabaceae (Dalbergia, Acacia) and Rubiaceae, belonging to dry deciduous forest and open savannas. This collective data set allows for the identification of discernible patterns related to the use of ecological interfaces between the forest and the open plains, used and actively managed by humans, and the possibility to identify if this changed with an increase in maritime trade and/or changes in agriculture over time. This study provides evidence of the differences in the vegetation present as well as use of wood fuel and other specific uses of wood for each site examined. It also sheds new light on tropical anthracology regarding quantification and accuracy in taxa identification.

Novo método de extração de amidos do cálculo dentário e criação de uma coleção identificada para a reconstrução das dietas do passado

A identificação de amidos vegetais embebidos no cálculo dentário tem vindo a potenciar os estudos paleodietéticos. Este trabalho apresenta um novo protocolo de extração e tratamento de amidos vegetais e divulga a criação duma coleção identificada para a subsequente identificação.&#x0D; O protocolo de extração consta de três fases – extração, preparação e identificação –, com uma duração de 4 dias, onde o cálculo dentário é sujeito à dissolução ácida ligeira da matriz fosfocálcica, à precipitação dos amidos, e à observação e identificação morfométrica em microscópica ótica ou eletrónica. Para tal, foi criada uma coleção identificada, a partir de alimentos habituais na dieta em território Português em fresco, secos ou em farinha. Adicionou-se uma chave dicotómica de consulta rápida para facilitar o processo de identificação.&#x0D; Esta coleção conta com 19 espécies, estando prevista a sua ampliação caso surjam novos projetos de investigação.&#x0D; Esperamos que novos estudos beneficiem desta metodologia para a reconstrução das dietas do passado.

Use-wear and residue analysis of pounding tools used by wild capuchin monkeys (Sapajus libidinosus) from Serra da Capivara (Piauí, Brazil)

Bearded capuchin monkeys (Sapajus libidinosus) from Serra da Capivara National Park (Brazil), perform the widest range of activities using stone tools of all the non-human tool-using primates. The behaviours behind this range of tool-use have been closely documented, but little is known about the characteristics of the tools themselves. Here we redress this imbalance and adopt an archaeological perspective to the analysis of capuchin pounding tools. We apply, for the first time, systematic microscopic techniques to the analysis of capuchin stone tools used for digging, cracking cashew nuts and seed processing to characterise their damage patterns combined with residue spatial distribution and micro-remains analysis. This work presents a standardized methodology for future primate archaeological use-wear studies as well as forming a reference collection which can be used to identify different activities within the primate archaeological record. Furthermore, understanding the archaeologically visible traces of primate percussive behaviours represents an initial step in developing a methodology to investigate if similar activities were practiced by early hominins and to help identify these activities in the Plio-Pleistocene archaeological record.