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Among others, dental microwear texture analysis (DMTA) is a proxy of dietary niches 1. Applied on herbivores, it allows to describe vegetal resources and assess environmental changes. However, up to now, there is no study correlating tooth microwear and tree cover. Here we propose to fill this gap by modelling this relation. As the diet of red deer (Cervus elaphus) varies from grass to browse depending on resource availabilities ², it is an ideal candidate to test such a model, which would be helpful in deciphering tree cover in the past. On the one hand, we created vector layers with QGIS for the habitats of nine European populations of extant wild-shot red deer 3 (N=283 adults). In each area, we randomly generated 5-km² circles assessing individual home ranges. We then superimposed these circular batches with the CORINE land Cover data (2018 update). DMTA was done using Scale-Sensitive Fractal Analysis 4 on the lower second molars of these deer. We found low pseudo-R² between DMTA and land cover when zooming in at the individual scale. At the population scale, we found that the textural fill volume is negatively correlated with the percentage of open habitats, wherein herbaceous dicots and monocots are more abundant than in forested habitats. Applied to early Pleistocene European cervids, notably the genus Eucladoceros, our results suggest similarities in tree cover between site of Chilhac with the forested area of Białowieża, Poland 5 and between the site of Senèze with the southern Iberic populations occupying open habitats 6. The land cover should be considered with caution, however. It does not directly reflect the preferred food resources of the red deer, but instead natural and agricultural landscapes, the categories of which need to be adapted to better address (palaeo)ecological issues. These preliminary results, however, provide interesting perspectives. [ABSTRACT FROM AUTHOR]

During the late Miocene Vallesian–Turolian transition, an important faunal turnover affected continental vertebrate faunas, and particularly primates with the replacement of hominids by cercopithecid monkeys in Europe. In order to better understand this turnover from a climatic perspective, stable oxygen and carbon isotope compositions of fossil equid tooth enamel contemporaneous with either hominids or cercopithecids have been analyzed. Hipparion teeth recovered from seven Greek localities spanning from the late Vallesian to the end of Turolian have δ18Op values of apatite phosphate suggesting an increase in mean air temperatures from 13±3°C at the beginning of the Vallesian to 17±2°C during the late Turolian. δ13C values of local plants have been estimated using the carbon isotope compositions of apatite carbonate. By applying to our data the relationship established between the δ13C value of modern C3 plants and mean annual precipitations, estimated average precipitations slightly decreased from 890 (+109 −100) mma−1 to 471 (+58 −54) mma−1 during this time span. According to Köppen's classification of climates, northern Greece was submitted to modern-like Mediterranean climates during the late Miocene. The change in climatic conditions, which took place during the late Vallesian–early Turolian transition, remained within the range of Mediterranean climate modes, thus being unlikely at the origin of a dramatic modification in local vegetation. This study strongly suggests that climate might not be the key factor of the Vallesian–Turolian major faunal turnover at least in the Eastern Mediterranean Basin. [ABSTRACT FROM AUTHOR]

The history and palaeoecology of the steppe bison ( Bison priscus ) remain incompletely understood despite its widespread distribution. Using dental microwear textural analysis (DMTA) and vegetation modelling, we reconstructed the diet and assessed the habitat of steppe bison inhabiting Eurasia and Alaska since the Middle Pleistocene. During the Late Pleistocene, steppe bison occupied a variety of biome types: from the mosaic of temperate summergreen forest and steppe/temperate grassland (Serbia) to the tundra biomes (Siberia and Alaska). Despite the differences in the identified biome types, the diet of steppe bison did not differ significantly among populations in Eurasia. DMTA classified it as a mixed forager in all populations studied. The DMTA of Bb1 bison-a recently identified genetically extinct sister-clade of Bison bonasus -was typical of a highly grazing bovid species and differed from all B. priscus populations. The results of the study temper the common perception that steppe bison were grazers in steppe habitats. The dietary plasticity of the steppe bison was lower when compared with modern European bison and may have played an important role in its extinction, even in the stable tundra biome of eastern Siberia, where it has survived the longest in all of Eurasia., Competing Interests: We declare we have no competing interests., (© 2024 The Author(s).)

The genus Macaca includes medium- to large-bodied monkeys and represents one of the most diverse primate genera, also having a very large geographic range. Nowadays, wild macaque populations are found in Asia and Africa, inhabiting a wide array of habitats. Fossil macaques were also present in Europe from the Late Miocene until the Late Pleistocene. Macaques are considered ecologically flexible monkeys that exhibit highly opportunistic dietary strategies, which may have been critical to their evolutionary success. Nevertheless, available ecological information regarding fossil European species is very sparse, limiting our knowledge of their evolutionary history in this geographic area. To further our understanding of fossil European macaque ecology, we investigated the dietary ecology of Macaca majori, an insular endemic species from Sardinia. In particular, we characterized the dental capabilities and potential dietary adaptations of M. majori through dental topographic and enamel thickness analyses of two M 2 s from the Early Pleistocene site of Capo Figari (1.8 Ma). We also assessed its diet through dental microwear texture analysis, while the microwear texture of M. majori was also compared with microwear textures from other European fossil macaques from mainland Europe. The dental topographic and enamel thickness analyses suggest that M. majori frequently consumes hard/mechanically challenging and/or abrasive foods. The results of the dental microwear analysis are consistent with this interpretation and further suggest that M. majori probably exhibited more durophagous dietary habits than mainland Plio-Pleistocene macaques. Overall, our results indicate that M. majori probably occupied a different dietary niche compared to its mainland fossil relatives, which suggests that they may have inhabited different paleoenvironments., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Theropithecus brumpti is a primate known from numerous craniodental specimens in the Plio-Pleistocene Shungura Formation (Lower Omo Valley, Ethiopia), but the anatomy of its hindlimb is documented only by a few associated and mostly incomplete postcranial specimens. The adaptations of T. brumpti are still debated, with its substrate preferences and its use of squatting postures recently discussed based on anatomical differences when compared with its extant representative, Theropithecus gelada. Here, we describe an associated femur and tibia (L 869-1 and L 869-2) of a presumed T. brumpti male and a partial foot (L 865-1r and L 865-1t) of a male T. brumpti, dated to ca. 2.6 Ma and ca. 2.32 Ma respectively. Based on univariate and bivariate morphometric analyses, we provide new data on the morphological correlates of substrate preferences and postural behaviors of this fossil species. Our results are in agreement with previous analyses and present T. brumpti as a predominantly terrestrial primate. We demonstrate the presence of osteological correlates associated with the use of squatting behaviors in T. brumpti but also point to significant anatomical differences between this paleontological species and T. gelada. These differences blur the functional value of characters previously identified as diagnostic of T. gelada and its postural behavior. We further document the postcranial distinctiveness of the Theropithecus clade in relation to the Papio clade. This study thus provides new insights into the postcranial anatomy and paleoecology of an abundant fossil primate from the Plio-Pleistocene of eastern Africa., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Modern humans have populated Europe for more than 45,000 years 1,2 . Our knowledge of the genetic relatedness and structure of ancient hunter-gatherers is however limited, owing to the scarceness and poor molecular preservation of human remains from that period 3 . Here we analyse 356 ancient hunter-gatherer genomes, including new genomic data for 116 individuals from 14 countries in western and central Eurasia, spanning between 35,000 and 5,000 years ago. We identify a genetic ancestry profile in individuals associated with Upper Palaeolithic Gravettian assemblages from western Europe that is distinct from contemporaneous groups related to this archaeological culture in central and southern Europe 4 , but resembles that of preceding individuals associated with the Aurignacian culture. This ancestry profile survived during the Last Glacial Maximum (25,000 to 19,000 years ago) in human populations from southwestern Europe associated with the Solutrean culture, and with the following Magdalenian culture that re-expanded northeastward after the Last Glacial Maximum. Conversely, we reveal a genetic turnover in southern Europe suggesting a local replacement of human groups around the time of the Last Glacial Maximum, accompanied by a north-to-south dispersal of populations associated with the Epigravettian culture. From at least 14,000 years ago, an ancestry related to this culture spread from the south across the rest of Europe, largely replacing the Magdalenian-associated gene pool. After a period of limited admixture that spanned the beginning of the Mesolithic, we find genetic interactions between western and eastern European hunter-gatherers, who were also characterized by marked differences in phenotypically relevant variants., (© 2023. The Author(s).)

Discoveries in recent decades indicate that the large papionin monkeys Paradolipopithecus and Procynocephalus are key members of the Late Pliocene - Early Pleistocene mammalian faunas of Eurasia. However, their taxonomical status, phylogenetic relationships, and ecological profile remain unclear. Here we investigate the two latter aspects through the study of the inner ear anatomy, as revealed by applying micro-CT scan imaging techniques on the cranium LGPUT DFN3-150 of Paradolichopithecus from the lower Pleistocene (2.3 Ma) fossil site Dafnero-3 in Northwestern Greece. Using geometric morphometric methods, we quantified shape variation and the allometric and phylogenetic signals in extant cercopithecines ( n  = 80), and explored the morphological affinities of the fossil specimen with extant taxa. LGPUT DFN3-150 has a large centroid size similar to that of baboons and their relatives. It shares several shape features with Macacina and Cercopithecini, which we interpret as probable retention of a primitive morphology. Overall, its inner ear morphology is more consistent with a stem Papionini more closely related to Papionina than Macacina, or to a basal crown Papionina. Our results, along with morphometrical and ecological features from previous studies, call into question the traditional hypothesis of a Paradolichopithecus -Macacina clade, and provide alternative perspectives in the study of Eurasian primate evolution during the late Neogene-Quaternary., Supplementary Information: The online version contains supplementary material available at 10.1007/s10764-022-00329-4., (© The Author(s) 2022.)

Dietary habits exert significant selective pressures on anatomical structures in animals, leading to substantial morphological adaptations. Yet, the relationships between the mandible and diet are still unclear, raising issues for paleodietary reconstructions notably. To assess the impact of food hardness and size on morphological structures, we used an experimental baseline using a model based on the domestic pig, an omnivorous mammal with bunodont, thick-enameled dentition, and chewing movements similar to hominids. We hypothesized that the consumption of different types of seeds would result in substantial differences in the morphology of the mandible despite similar overall diets. The experiment was conducted on four groups of juvenile pigs fed with mixed cereal and soy flours. The control group received only flours. We supplemented the four others with either 10 hazelnuts, 30 hazelnuts, 30% barley seeds, or 20% corn kernels per day. We investigated the shape differences between the controlled-fed groups using three-dimensional geometric morphometrics. Our results provide strong evidence that the supplemental consumption of a significant amount of seeds for a short period (95 days) substantially modify the mandibular morphology of pigs. Our analyses suggest that this shape differentiation is due to the size of the seeds, requiring high and repeated bite force, rather than their hardness. These results provide new perspectives for the use of mandibular morphology as a proxy in paleodietary reconstructions complementing dental microwear textures analyses., (© 2022 American Association for Anatomy.)

Currently, very little is known about the ecology of extinct Eurasian cercopithecids. Dietary information is crucial in understanding the ecological adaptations and diversity of extinct cercopithecids and the evolution of this family. For example, the colobine genus Dolichopithecus is represented by multiple large-bodied species that inhabited Eurasia during the Pliocene-Early Pleistocene. The available evidence, though limited, suggests semiterrestrial locomotion, which contrasts with most extant African and Asian colobines that exhibit morphological and physiological adaptations for arboreality and folivory. These differences raise questions regarding the dietary specialization of early colobine taxa and how/if that influenced their dispersion out of Africa and into Eurasia. To further our understanding of the ecology of Plio-Pleistocene cercopithecids, we characterized the dental capabilities and potential dietary adaptations of Dolichopithecus ruscinensis through dental topographic and enamel thickness analyses on an M 1 from the locality of Serrat d'en Vacquer, Perpignan (France). We also assessed the feeding behavior of D. ruscinensis through dental microwear texture analysis on a broad sample of fossil molars from fossil sites in France, Greece, Bulgaria, and Romania. Dental topographic and enamel thickness analyses suggest that D. ruscinensis could efficiently process a wide range of foods. Results of the dental microwear texture analysis suggest that its diet ranged from folivory to the consumption of more mechanically challenging foods. Collectively, this suggests a more opportunistic feeding behavior for Dolichopithecus than characteristic of most extant colobines., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Extant colobine monkeys are specialized leaf eaters. But during the late Miocene, western Eurasia was home to colobines that were less efficient at chewing leaves than they were at breaking seed shells. To understand the link between folivory and granivory in this lineage, the dietary niche of Mesopithecus delsoni and Mesopithecus pentelicus was investigated in southeastern Europe, where a major environmental change occurred during the late Miocene. We combined dental topographic estimates of chewing efficiency with dental microwear texture analysis of enamel wear facets. Mesopithecus delsoni was more efficient at chewing leaves than M. pentelicus, the dental topography of which matches an opportunistic seed eater. Concurrently, microwear complexity increases in M. pentelicus, especially in the northernmost localities corresponding to present-day Bulgaria. This is interpreted as a dietary shift toward hard foods such as seeds or tubers, which is consistent with the savanna and open mixed forest biomes that covered Bulgaria during the Tortonian. The fact that M. delsoni was better adapted to folivory and consumed a lower amount of hard foods than M. pentelicus suggests that colobines either adapted to folivory before their dispersal to Europe or evolved adaptations to leaf consumption in multiple occurrences., (© 2021 The Authors. Evolution © 2021 The Society for the Study of Evolution.)

We here report on fossil remains of the earliest known crown-Testudo, an extant clade of Mediterranean testudinid tortoises from the late Miocene (Vallesian, MN 10) from the hominoid locality Ravin de la Pluie (RPl) in Greece. The material studied is a small, nearly complete carapace with a clearly distinct hypo-xiphiplastral hinge. This supports the sensu stricto generic assignment. This new terrestrial testudinid specimen is characterized by a possible tectiform, narrow, elongated shell with a pentagonal pygal and a long, posteriorly elevated, lenticular and rounded dorsal epiplastral lip. These unique features differ from those of other known Mediterranean hinged forms and allow the erection of the new species Testudo hellenica sp. nov. This taxon is phylogenetically close to two Greek species, the extant T. marginata and the fossil T. marmorum (Turolian, around 7.3 Ma). This record provides evidence for the first appearance of the genus Testudo sensu stricto at a minimum age of 9 Ma., Competing Interests: The authors have declared that no competing interests exist.

A new study by Fraser et al (2018) urges the use of phylogenetic comparative methods, whenever possible, in analyses of mammalian tooth wear. We are concerned about this for two reasons. First, this recommendation may mislead the research community into thinking that phylogenetic signal is an artifact of some sort rather than a fundamental outcome of the evolutionary process. Secondly, this recommendation may set a precedent for editors and reviewers to enforce phylogenetic adjustment where it may unnecessarily weaken or even directionally alter the results, shifting the emphasis of analysis from common patterns manifested by large clades to rare cases.

This study aims to explore the feeding ecology of two terrestrial papionins, Papio and Theropithecus from the Shungura Formation in Ethiopia, the most complete stratigraphic and paleontological record of the African Plio-Pleistocene. Two aspects were evaluated using Dental Microwear Texture Analysis: differences in diet between the extinct genera and their extant relatives, and any potential dietary fluctuations over time. Amongst more than 2,500 cercopithecid dental remains, 154 Theropithecus molars and 60 Papio molars were considered. Thirty-nine extant wild baboons and 20 wild geladas were also considered. The results show that diets of extinct monkeys from Member G already differed between genera as it is the case for their extant representatives. The shearing facets on the Theropithecus molars display significant variations in microwear textures, suggesting several dietary shifts over time. Two events point to higher intakes of herbaceous monocots (tougher than dicots foliages), at about 2.91 Ma (between members B and C) and at 2.32 Ma (between members E and F). These two events are separated by an inverse trend at about 2.53 Ma (between members C and D). Some of these variations, such as between members E and F are supported by the enamel carbon isotopic composition of herbivorous mammals and with paleovegetation evidence.

A new fossil cranium of a large papionin monkey from the Lower Pleistocene site of Dafnero-3 in Western Macedonia, Greece, is described by means of outer and inner morphological and metric traits using high-resolution micro-computed tomography. Comparisons with modern cercopithecids and contemporaneous Eurasian fossil taxa suggest that the new cranium could equally be ascribed to either the Eurasian Paradolichopithecus or to the East Asian Procynocephalus. The combination of the available direct and indirect fossil evidence, including the new cranium from Dafnero, revives an earlier hypothesis that considers these two sparsely documented genera as synonyms. The timing and possible causes of the rise and demise of Paradolichopithecus - Procynocephalus are discussed., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Objectives: Dental microwear is a promising tool to reconstruct animals' diet because it reflects the interplay between the enamel surface and the food items recently consumed. This study examines the sources of inter-individual variations in dietary habits in a free-ranging population of mandrills (Mandrillus sphinx) using a combination of feeding monitoring and in vivo dental microwear textural analysis (DMTA)., Methods: We investigated the impact of seasonality and individual traits on four DMTA parameters. In parallel, we further studied the influence of the physical properties of the food items consumed on these four parameters, using three proxies (mechanical properties, estimates of phytolith and external grit contents)., Results: We found that seasonality, age, and sex all impact DMTA parameters but those results differ depending on the facet analyzed (crushing vs. shearing facets). Three DMTA parameters (anisotropy, complexity, and heterogeneity of complexity) appear sensitive to seasonal variations and anisotropy also differs between the sexes while textural fill volume tends to vary with age. Moreover, the physical properties of the food items consumed vary seasonally and also differ depending on individual sex and age., Conclusion: Considering the interplay between the tested variables and both dental microwear and diet, we reaffirm that food physical properties play a major role in microwear variations. These results suggest that DMTA parameters may provide valuable hints for paleoecological reconstruction using fragmentary fossil dental remains., (© 2017 Wiley Periodicals, Inc.)

Extant colobine monkeys have been historically described as specialized folivores. However, reports on both their behavior and dental metrics tend to ascribe a more varied diet to them. In particular, several species, such as Pygathrix nemaeus and Rhinopithecus roxellana, are dedicated seasonal seed eaters. They use the lophs on their postcanine teeth to crack open the hard endocarp that protects some seeds. This raises the question of whether the bilophodont occlusal pattern of colobine monkeys first evolved as an adaptation to folivory or sclerocarpic foraging. Here, we assess the sclerocarpic foraging ability of the oldest European fossil colobine monkey, Mesopithecus. We use computed microtomograpy to investigate the three-dimensional (3D) dental topography and enamel thickness of upper second molars ascribed to the late Miocene species Mesopithecus pentelicus from Pikermi, Greece. We compare M. pentelicus to a sample of extant Old World monkeys encompassing a wide range of diets. Furthermore, we combine classic dietary categories such as folivory with alternative categories that score the ability to crack, grind and shear mechanically challenging food. The 3D dental topography of M. pentelicus predicts an ability to crack and grind hard foods such as seeds. This is consistent with previous results obtained from dental microwear analysis. However, its relatively thin enamel groups M. pentelicus with other folivorous cercopithecids. We interpret this as a morphological trade-off between the necessity to avoid tooth failure resulting from hard food consumption and the need to process a high amount of leafy material. Our study demonstrates that categories evaluating the cracking, grinding or shearing ability, traditional dietary categories, and dental topography combine well to make a powerful tool for the investigation of diet in extant and extinct primates., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

GnpIS is an information system designed to help scientists working on plants and fungi to decipher the molecular and genetic architecture of trait variations by facilitating the navigation through genetic, genomic, and phenotypic information. The purpose of the present chapter is to illustrate how users can (1) explore datasets from phenotyping experiments in order to build new datasets for studying genotype × environment interactions in traits, (2) browse into the results of other genetic analysis data such as GWAS to generate or check working hypothesis about candidate genes or to identify important alleles and germplasms for breeding programs, and (3) explore the polymorphism in specific area of the genome using InterMine, JBrowse tools embedded in the GnpIS information system.

Both dust and silica phytoliths have been shown to contribute to reducing tooth volume during chewing. However, the way and the extent to which they individually contribute to tooth wear in natural conditions is unknown. There is still debate as to whether dental microwear represents a dietary or an environmental signal, with far-reaching implications on evolutionary mechanisms that promote dental phenotypes, such as molar hypsodonty in ruminants, molar lengthening in suids or enamel thickening in human ancestors. By combining controlled-food trials simulating natural conditions and dental microwear textural analysis on sheep, we show that the presence of dust on food items does not overwhelm the dietary signal. Our dataset explores variations in dental microwear textures between ewes fed on dust-free and dust-laden grass or browse fodders. Browsing diets with a dust supplement simulating Harmattan windswept environments contain more silica than dust-free grazing diets. Yet browsers given a dust supplement differ from dust-free grazers. Regardless of the presence or the absence of dust, sheep with different diets yield significantly different dental microwear textures. Dust appears a less significant determinant of dental microwear signatures than the intrinsic properties of ingested foods, implying that diet plays a critical role in driving the natural selection of dental innovations., (© 2016 The Author(s).)

While grazing as a selective factor towards hypsodont dentition on mammals has gained a lot of attention, the importance of fruits and seeds as fallback resources for many browsing ungulates has caught much less attention. Controlled-food experiments, by reducing the dietary range, allow for a direct quantification of the effect of each type of items separately on enamel abrasion. We present the results of a dental microwear texture analysis on 40 ewes clustered into four different controlled diets: clover alone, and then three diets composed of clover together with either barley, corn, or chestnuts. Among the seed-eating groups, only the barley one shows higher complexity than the seed-free group. Canonical discriminant analysis is successful at correctly classifying the majority of clover- and seed-fed ewes. Although this study focuses on diets which all fall within a single dietary category (browse), the groups show variations in dental microwear textures in relation with the presence and the type of seeds. More than a matter of seed size and hardness, a high amount of kernels ingested per day is found to be correlated with high complexity on enamel molar facets. This highlights the high variability of the physical properties of the foods falling under the browsing umbrella.

Plant-herbivore interactions are hypothesized to drive vole population cycles through the grazing-induced production of phytoliths in leaves. Phytoliths act as mechanical defences because they deter herbivory and lower growth rates in mammals. However, how phytoliths impair herbivore performance is still unknown. Here, we tested whether the amount of phytoliths changes tooth wear patterns. If confirmed, abrasion from phytoliths could play a role in population crashes. We applied dental microwear texture analysis (DMTA) to laboratory and wild voles. Lab voles were fed two pelleted diets with differing amounts of silicon, which produced similar dental textures. This was most probably due to the loss of food mechanical properties through pelletization and/or the small difference in silicon concentration between diets. Wild voles were trapped in Poland during spring and summer, and every year across a population cycle. In spring, voles feed on silica-rich monocotyledons, while in the summer they also include silica-depleted dicotyledons. This was reflected in the results; the amount of silica therefore leaves a traceable record in the dental microwear texture of voles. Furthermore, voles from different phases of population cycles have different microwear textures. We tentatively propose that these differences result from grazing-induced phytolith concentrations. We hypothesize that the high amount of phytoliths in response to intense grazing in peak years may result in malocclusion and other dental abnormalities, which would explain how these silicon-based plant defences help provoke population crashes. DMTA could then be used to reconstruct vole population dynamics using teeth from pellets or palaeontological material., (© 2016. Published by The Company of Biologists Ltd.)

Fossil-based estimates of diversity and evolutionary dynamics mainly rely on the study of morphological variation. Unfortunately, organism remains are often altered by post-mortem taphonomic processes such as weathering or distortion. Such a loss of information often prevents quantitative multivariate description and statistically-controlled comparisons of extinct species based on morphometric data. A common way to deal with missing data involves imputation methods that directly fill the missing cases with model estimates. Over the last years, several empirically-determined thresholds for the maximum acceptable proportion of missing values have been proposed in the literature, whereas other studies showed that this limit actually depends on various properties of the study data set and of the selected imputation method, and is by no way generalizable. We evaluate the relative performances of seven multiple imputation (MI) techniques through a simulation-based analysis under three distinct patterns of missing data distribution. Overall, Fully Conditional Specification and Expectation-Maximization algorithms provide the best compromises between imputation accuracy and coverage probability. MI techniques appear remarkably robust to the violation of basic assumptions such as the occurrence of taxonomically or anatomically biased patterns of missing data distribution, making differences in simulation results between the three patterns of missing data distribution much smaller than differences between the individual MI techniques. Based on these results, rather than proposing a new (set of) threshold value(s), we develop an approach combining the use of MIs with procrustean superimposition of principal component analysis results, in order to directly visualize the effect of individual missing data imputation on an ordinated space. We provide an R function for users to implement the proposed procedure.

Background: The equid Hippotherium primigenium, with moderately hypsodont cheek teeth, rapidly dispersed through Eurasia in the early late Miocene. This dispersal of hipparions into the Old World represents a major faunal event during the Neogene. The reasons for this fast dispersal of H. primigenium within Europe are still unclear. Based on its hypsodonty, a high specialization in grazing is assumed although the feeding ecology of the earliest European hipparionines within a pure C3 plant ecosystem remains to be investigated., Methodology/principal Findings: A multi-proxy approach, combining carbon and oxygen isotopes from enamel as well as dental meso- and microwear analyses of cheek teeth, was used to characterize the diet of the earliest European H. primigenium populations from four early Late Miocene localities in Germany (Eppelsheim, Höwenegg), Switzerland (Charmoille), and France (Soblay). Enamel δ(13)C values indicate a pure C3 plant diet with small (<1.4‰) seasonal variations for all four H. primigenium populations. Dental wear and carbon isotope compositions are compatible with dietary differences. Except for the Höwenegg hipparionines, dental microwear data indicate a browse-dominated diet. By contrast, the tooth mesowear patterns of all populations range from low to high abrasion suggesting a wide spectrum of food resources., Conclusions/significance: Combined dental wear and stable isotope analysis enables refined palaeodietary reconstructions in C3 ecosystems. Different H. primigenium populations in Europe had a large spectrum of feeding habits with a high browsing component. The combination of specialized phenotypes such as hypsodont cheek teeth with a wide spectrum of diet illustrates a new example of the Liem's paradox. This dietary flexibility associated with the capability to exploit abrasive food such as grasses probably contributed to the rapid dispersal of hipparionines from North America into Eurasia and the fast replacement of the brachydont equid Anchitherium by the hypsodont H. primigenium in Europe.