Pachyostosis, the increase in volume of bones, has been documented in a myriad of tetrapods that secondarily adapted to the aquatic environment. However, this convergent trait has hitherto only been quantified in complete ribs and long bones. This study presents a methodology designed to quantify the degree of pachyostosis in ribs and vertebrae for both complete and fragmentary skeletal elements. Using surface models, the volumes of costal and vertebral regions were measured and standardized using size proxies to devise a pachyostosis index. Sirenians are used as a case study, given the disparity in pachyostosis degrees that has already been documented in the clade, especially in their ribs. The new methodology can detect this disparity and further demonstrates that pachyostosis also affects their vertebrae. Milder cases of pachyostosis, such as in the extant Dugong dugon, are likely secondary specializations to a specific lifestyle. The craniocaudal distribution of the pachyostosis index values is also potentially helpful for further distinguishing finer lifestyle differences. This new method has the potential to clarify the causes of the disparity in pachyostosis degrees observed in the fossil record of sirenians, and it may also prove valuable in broader, non-mammalian contexts.

Investigations of bone microanatomy are commonly used to explore lifestyle strategies in vertebrates. While distinct microanatomical limb bone features have been established for exclusively aquatic and terrestrial lifestyles, identifying clear patterns for the semiaquatic lifestyle remains more challenging. Pinnipeds and mustelids provide an ideal framework for studying the diversity of semiaquatic adaptations. Here, we tested whether their humerus and femur showed significant differences in microanatomical and biomechanical parameters along the diaphysis relative to locomotor strategies. We μCT-scanned extant pinnipeds and included semiaquatic and generalist mustelids to build a reference for the amphibious lifestyle. Statistical analyses showed that comparisons away from the growth center distinguished species with non-weight-bearing hind limbs from those with all-limb weight-bearing. Among the latter, otariids and semiaquatic mustelids shared highly compact bones and a reduced medullary cavity, though otariids exhibited a more gradual medullocortical transition with more trabeculae. Contrary to the prevailing assumption, our findings indicate bone mass increase can also be associated with fast-swimming predators diving beyond shallow depths. Phocids, which cannot bear weight on their hind limbs, include species that spend 70%-80% of their lives at sea. These species exhibited extremely spongious bones: low compactness, thin cortex, and a trabeculae-filled medullary region-reminiscent of the osteoporotic-like condition seen in some extant cetaceans. Additionally, the shared humerus-femur pattern, regardless of locomotor type, suggests that skeletal adaptations can be systemic in carnivorans. This new comparative dataset broadens the spectrum of microanatomical patterns associated with an amphibious lifestyle and provides a foundation for better deciphering extinct species' locomotion.

Extant whales, dolphins, and porpoises result from a major macroevolutionary lifestyle transition that transformed land-dwelling cetaceans into fully aquatic species. This involved significant changes in sensory systems. The increase in brain size relative to body size (encephalization quotient) is an outstanding feature of modern cetaceans, especially toothed whales. Conversely, olfactory capabilities are assumed to have diminished along this transition, with airborne olfaction becoming less relevant. The extent and timing of olfactory reduction remain obscure due to challenges in accessing well-preserved fossil endocranial anatomy. This study shows that early cetaceans had already evolved an increased encephalization quotient, and that their olfactory apparatus was likely not yet under selective pressure leading to its reduction. We demonstrate this through an analysis of the extinct whale, Protocetus atavus, a member of the middle Eocene semiaquatic cetacean group Protocetidae. We provide the first documentation of its endocranial anatomy using high-resolution computed tomography and compare it to other early cetaceans as well as extant mammals. We conclude that cetaceans increased their brain size earlier than previously thought, while relying on a well-developed olfactory system at a time when they were still partly terrestrial.

The evolution of baleen whales (Mysticeti) comprises two main phases, namely, (i) a Paleogene phase, which saw the diversification of stem lineages, and (ii) a Neogene phase, dominated by modern-looking, toothless, baleen-bearing forms in the monophyletic group Chaeomysticeti. These two phases are separated by a global turnover event coinciding with a gap—or “dark age”—in the mysticete fossil record. This dark age occurred between 23 and ~18 Ma and is apparently detected worldwide, except in Zealandia. Here, we report on a new mysticete fossil from the Lower Miocene (Burdigalian: ~19.2 Ma) strata of the Chilcatay Formation cropping out at the newly discovered locality of Cerro Tiza (East Pisco Basin, Peru), which represents a limited but precious testament from the last phase of the baleen whale dark age. Two previously mentioned, slightly geologically younger fossils from the same formation are also reappraised herein, revealing the occurrence of at least another baleen whale taxon in the upper Chilcatay strata—one that belongs in the mysticete crown group. Although the Early Miocene remains a problematic time interval for the fossil record of baleen whales, our new results encourage the search for mysticete fossils in the Lower Miocene strata of the East Pisco Basin, whose basin fill preserves a cornucopia of extraordinarily informative marine vertebrate fossils of the Cenozoic age, as well as in coeval deposits worldwide.

Olfaction is a critical sense for tetrapods, playing a key role in survival and reproduction by aiding in food detection, predator avoidance, and social interactions. Olfactory performance has been experimentally tested in only a few taxa, so comparative analyses rely on anatomical and genomic proxies. Among anatomical proxies, the olfactory bulb endocast is widely used, particularly in extinct species, where it is often the only preserved proxy and can be reconstructed even in million-year-old fossils. While the functional significance of chemoreceptor genes has received attention, the extent to which the olfactory bulb endocast correlates with genomic proxies remains unclear. Using brain endocasts across all mammalian orders, we investigated the relationship between the absolute (absOB) and relative (relatOB) volumes of the olfactory bulb endocast and the number of intact chemoreceptor genes. While no clear correlations were found between absOB and the genomic proxies tested, we identified a significant correlation between relatOB and the total number of combined intact chemoreceptor genes (CombChemo), primarily driven by olfactory receptor genes (OR). Leveraging this correlation, and aiming to infer olfactory capabilities in taxa for which only the skull is available, we estimated OR numbers for three mammalian orders lacking genomic data, as well as for five extinct mammals. Building on studies that have established a link between intact OR and olfactory sensitivity and discrimination, we conclude that relatOB enables inference of olfactory capabilities in mammals. This provides a basis to investigate sensory evolution and opens perspectives for interpreting paleoecology and behavior of extinct mammals.

The fibula, despite being traditionally overlooked compared to the femur and the tibia, has recently received attention in primate functional morphology due to its correlation with the degree of arboreality (DOA). Highlighting further fibular features that are associated with arboreal habits would be key to improving palaeobiological inferences in fossil specimens. Here we present the first investigation on the trabecular bone structure of the primate fibula, focusing on the distal epiphysis, across a vast array of species. We collected μCT data on the distal fibula for 21 species of primates, with representatives from most of the orders, and we employed a recently developed approach implemented in the R package 'indianaBones' to isolate the entire trabecular bone underlying an epiphysis or articular facet. After extracting both traditional trabecular parameters and novel topological indices, we tested for the posited relationship between trabecular bone and DOA. To disentangle this effect from others related to body size and phylogenetic relationship, we included a body mass proxy as covariate and employed phylogenetic comparative methods. We ran univariate/multivariate and exploratory/inferential statistical analyses. The trabecular structure of the fibular distal epiphysis in primates does not appear to be associated with the DOA. Instead, it is strongly affected by body mass and phylogenetic relationships. Although we identified some minor trends related to human bipedalism, our findings overall discourage, at this stage, the study of distal fibula trabecular bone to infer arboreal behaviors in extinct primates. We further found that body size distribution is strongly related to phylogeny, an issue preventing us from unravelling the influence of the two factors and that we believe can potentially affect future comparative analyses of primates. Overall, our results add to previous evidence of how trabecular traits show variable correlation with locomotor aspects, size and phylogenetic history across the primate skeleton, thus outlining a complex scenario in which a network of interconnected factors affects the morphological evolution of primates. This work may represent a starting point for future studies, for example, focusing on the effect of human bipedalism on distal fibula trabecular bone, or aiming to better understand the effects of body size and phylogenetic history on primate morphological evolution.

Turbinals are bony or cartilaginous structures that are present in the nasal cavity of most tetrapods. They are involved in key functions such as olfaction, heat, and moisture conservation, as well as protection of the respiratory tract. Despite recent studies that challenged long-standing hypotheses about their physiological and genomic correlation, turbinals remain largely unexplored, particularly for non-mammalian species. Herein, we review and synthesise the current knowledge of turbinals using an integrative approach that includes comparative anatomy, physiology, histology and genomics. In addition, we provide synonyms and correspondences of tetrapod turbinals from about 80 publications. This work represents a first step towards drawing hypotheses of homology for the whole clade, and provides a strong basis to develop new research avenues.

The number of functional genes coding for olfactory receptors differs markedly between species and has repeatedly been suggested to be predictive of a species' olfactory capabilities. To test this assumption, we compiled a database of all published olfactory detection threshold values in mammals and used three sets of data on olfactory discrimination performance that employed the same structurally related monomolecular odour pairs with different mammal species. We extracted the number of functional olfactory receptor genes of the 20 mammal species for which we found data on olfactory sensitivity and/or olfactory discrimination performance from the Chordata Olfactory Receptor Database. We found that the overall olfactory detection thresholds significantly correlate with the number of functional olfactory receptor genes. Similarly, the overall proportion of successfully discriminated monomolecular odour pairs significantly correlates with the number of functional olfactory receptor genes. These results provide the first statistically robust evidence for the relationship between olfactory capabilities and their genomics correlates. However, when analysed individually, of the 44 monomolecular odourants for which data on olfactory sensitivity from at least five mammal species are available, only five yielded a significant correlation between olfactory detection thresholds and the number of functional olfactory receptors genes. Also, for the olfactory discrimination performance, no significant correlation was found for any of the 74 relationships between the proportion of successfully discriminated monomolecular odour pairs and the number of functional olfactory receptor genes. While only a rather limited amount of data on olfactory detection thresholds and olfactory discrimination scores in a rather limited number of mammal species is available so far, we conclude that the number of functional olfactory receptor genes may be a predictor of olfactory sensitivity and discrimination performance in mammals.

Turbinals are key bony elements of the mammalian nasal cavity, involved in heat and moisture conservation as well as olfaction. While turbinals are well known in some groups, their diversity is poorly understood at the scale of placental mammals, which span 21 orders. Here, we investigated the turbinal bones and associated lamellae for one representative of each extant order of placental mammals. We segmented and isolated each independent turbinal and lamella and found an important diversity of variation in the number of turbinals, as well as their size, and shape. We found that the turbinal count varies widely, from zero in the La Plata dolphin, (Pontoporia blainvillei) to about 110 in the African bush elephant (Loxodonta africana). Multiple turbinal losses and additional gains took place along the phylogeny of placental mammals. Some changes are clearly attributed to ecological adaptation, while others are probably related to phylogenetic inertia. In addition, this work highlights the problem of turbinal nomenclature in some placental orders with numerous and highly complex turbinals, for which homologies are extremely difficult to resolve. Therefore, this work underscores the importance of developmental studies to better clarify turbinal homology and nomenclature and provides a standardized comparative framework for further research.

Allometry, i.e., morphological variation correlated with size, is a major pattern in organismal evolution. Since size varies both within and among species, allometry occurs at different variational levels. However, the variability of allometric patterns across levels is poorly known since its evaluation requires extensive comparative studies. Here, we implemented a 3D geometric morphometric approach to investigate cranial allometry at three main variational levels-static, ontogenetic, and evolutionary-and two anatomical scales-entire cranium and cranial subunits-based on a dense intra- and interspecific sampling of extant armadillo diversity. While allometric trajectories differ among distantly related species, they hardly do so among sister families. This suggests that phylogenetic distance plays an important role in explaining allometric divergences. Beyond trajectories, our analyses revealed pervasive allometric shape changes shared across variational levels and anatomical scales. At the entire cranial scale, craniofacial allometry (relative snout elongation and braincase reduction) is accompanied notably by variations of nuchal crests and postorbital constriction. Among cranial subunits, the distribution of allometry was highly heterogeneous, with the frontal and petrosal bones showing the most pervasive shape changes, some of which were undetected at a more global scale. Evidence of widespread and superimposed allometric variations raises questions on their determinants and anatomical correlates and demonstrates the critical role of allometry in morphological evolution.

- Supplementary Data 1-2: raw data - Supplementary Tables 2-5: analyses results - Supplementary Data 3: time-tree - Supplementary Data 4: R code - Supplementary Information on the methods (Supplementary Table 1 and Notes 1-7), information, credits and copyright licenses for slow arboreal mammal images used in Figs.2 and 4 (Supplementary Note 8) - Details on Extended Data Figs. and Tables are provided in Extended Data Captions.doc

The study of how organisms grow is a fundamental aspect of palaeontology. Growth in teleosauroids is poorly understood and little studied, especially in an ontogenetic sense. We investigate growth rates of the most common and abundant teleosauroid, Macrospondylus bollensis , in which a large sample of multiple body sizes is available from the Posidonienschiefer Formation (Posidonia Shale) of southwestern Germany. We perform linear regression analyses on 62 specimens of Macrospondylus (16 juveniles, 7 subadults and 39 adults) using 21 cranial and postcranial measurements. Our results show that juvenile, subadult and adult individuals have near‐isometric or isometric growth throughout much of the body. Notably, we find that in Macrospondylus : (1) the femur grows at a faster rate than the skull and hindlimb zeugopodium; (2) the forelimb and hindlimb grow at the same rate; and (3) there is distinct ontogenetic signal in the growth of the orbit and supratemporal fenestra. We also find that limb scaling in Macrospondylus is somewhat comparable to that seen in the extant gavialids Gavialis gangeticus and Tomistoma schlegelii . Last, we examine evolutionary allometry in skull length relative to body size in Crocodylomorpha using femoral length as a proxy, which shows a near‐isometric trend. Non‐thalattosuchian crocodylomorphs (with the exception of one pholidosaurid and one dryosaurid taxon) are differentiated from thalattosuchians due to their shorter skulls, as previously suggested, but the scaling relationship with femur length remains unchanged.

Besides manatees, the suspensory extant 'tree sloths' are the only mammals that deviate from a cervical count (CC) of seven vertebrae. They do so in opposite directions in the two living genera (increased versus decreased CC). Aberrant CCs seemingly reflect neck mobility in both genera, suggesting adaptive significance for their head position during suspensory locomotion and especially increased ability for neck torsion in three-toed sloths. We test two hypotheses in a comparative evolutionary framework by assessing three-dimensional intervertebral range of motion (ROM) based on exhaustive automated detection of bone collisions and joint disarticulation while accounting for interacting rotations of roll, yaw and pitch. First, we hypothesize that the increase of CC also increases overall neck mobility compared with mammals with a regular CC, and vice versa. Second, we hypothesize that the anatomy of the intervertebral articulations determines mobility of the neck. The assessment revealed that CC plays only a secondary role in defining ROM since summed torsion (roll) capacity was primarily determined by vertebral anatomy. Our results thus suggest limited neck rotational adaptive significance of the CC aberration in sloths. Further, the study demonstrates the suitability of our automated approach for the comparative assessment of osteological ROM in vertebral series.

The present 3D Dataset contains the 3D models analyzed in Bianucci et al. 2023, A heavyweight early whale pushes the boundaries of vertebrate morphology, Nature.These include bones of the holotype of new species Perucetus colossus (MUSM 3248), as well as the articulated skeleton of Cynthiacetus peruvianus (holotype, MNHN.F.PRU10).The latter was used to estimate the total skeleton volume of P. colossus.

Eco-morphological convergence, i.e., similar phenotypes evolved in ecologically convergent taxa, naturally reproduces a common-garden experiment since it allows researchers to keep ecological factors constant, studying intrinsic evolutionary drivers. The latter may result in differential evolvability that, among individual anatomical parts, causes mosaic evolution. Reconstructing the evolutionary morphology of the humerus and femur of slow arboreal mammals, we addressed mosaicism at different bone anatomical spatial scales. We compared convergence strength, using it as indicator of evolvability, between bone external shape and inner structure, with the former expected to be less evolvable and less involved in convergent evolution, due to anatomical constraints. We identify several convergent inner structural traits, while external shape only loosely follows this trend, and we find confirmation for our assumption in measures of convergence magnitude. We suggest that future macroevolutionary reconstructions based on bone morphology should include structural traits to better detect ecological effects on vertebrate diversification.

The fossil record of cetaceans documents how terrestrial animals acquired extreme adaptations and transitioned to a fully aquatic lifestyle 1 , 2 . In whales, this is associated with a substantial increase in maximum body size. Although an elongate body was acquired early in cetacean evolution 3 , the maximum body mass of baleen whales reflects a recent diversification that culminated in the blue whale 4 . More generally, hitherto known gigantism among aquatic tetrapods evolved within pelagic, active swimmers. Here we describe Perucetus colossus —a basilosaurid whale from the middle Eocene epoch of Peru. It displays, to our knowledge, the highest degree of bone mass increase known to date, an adaptation associated with shallow diving 5 . The estimated skeletal mass of P. colossus exceeds that of any known mammal or aquatic vertebrate. We show that the bone structure specializations of aquatic mammals are reflected in the scaling of skeletal fraction (skeletal mass versus whole-body mass) across the entire disparity of amniotes. We use the skeletal fraction to estimate the body mass of P. colossus , which proves to be a contender for the title of heaviest animal on record. Cetacean peak body mass had already been reached around 30 million years before previously assumed, in a coastal context in which primary productivity was particularly high.

Examples of photoluminescence (PL) are being reported with increasing frequency in a wide range of organisms from diverse ecosystems. However, the chemical basis of this PL remains poorly defined, and our understanding of its potential ecological function is still superficial. Among mammals, recent analyses have identified free-base porphyrins as the compounds responsible for the reddish ultraviolet-induced photoluminescence (UV-PL) observed in the pelage of springhares and hedgehogs. However, the localization of the pigments within the hair largely remains to be determined. Here, we use photoluminescence multispectral imaging emission and excitation spectroscopy to detect, map, and characterize porphyrinic compounds in skin appendages in situ. We also document new cases of mammalian UV-PL caused by free-base porphyrins in distantly related species. Spatial distribution of the UV-PL is strongly suggestive of an endogenous origin of the porphyrinic compounds. We argue that reddish UV-PL is predominantly observed in crepuscular and nocturnal mammals because porphyrins are photodegradable. Consequently, this phenomenon may not have a specific function in intra- or interspecific communication but rather represents a byproduct of potentially widespread physiological processes.

Acquiring a subterranean lifestyle entails a substantial shift for many aspects of terrestrial vertebrates' biology. Although this lifestyle is associated with multiple instances of convergent evolution, the relative success of some subterranean lineages largely remains unexplained. Here, we focus on the mammalian transitions to life underground, quantifying bone microanatomy through high-resolution X-ray tomography. The true moles stand out in this dataset. Examination of this family's bone histology reveals that the highly fossorial moles acquired a unique phenotype involving large amounts of compacted coarse cancellous bone. This phenotype exceeds the adaptive optimum seemingly shared by several other subterranean mammals and can be traced back to some of the first known members of the family. This remarkable microanatomy was acquired early in the history of the group and evolved faster than the gross morphology innovations of true moles' forelimb. This echoes the pattern described for other lifestyle transitions, such as the acquisition of bone mass specializations in secondarily aquatic tetrapods. Highly plastic traits-such as those pertaining to bone structure-are hence involved in the early stages of different types of lifestyle transitions.