Forelimb muscle activation patterns in American alligators: Insights into the evolution of limb posture and powered flight in archosaurs.

The evolution of archosaurs provides an important context for understanding the mechanisms behind major functional transformations in vertebrates, such as shifts from sprawling to erect limb posture and the acquisition of powered flight. While comparative anatomy and ichnology of extinct archosaurs have offered insights into musculoskeletal and gait changes associated with locomotor transitions, reconstructing the evolution of motor control requires data from extant species. However, the scarcity of electromyography (EMG) data from the forelimb, especially of crocodylians, has hindered understanding of neuromuscular evolution in archosaurs. Here, we present EMG data for nine forelimb muscles from American alligators during terrestrial locomotion. Our aim was to investigate the modulation of motor control across different limb postures and examine variations in motor control across phylogeny and locomotor modes. Among the nine muscles examined, m. pectoralis, the largest forelimb muscle and primary shoulder adductor, exhibited significantly smaller mean EMG amplitudes for steps in which the shoulder was more adducted (i.e., upright). This suggests that using a more adducted limb posture helps to reduce forelimb muscle force and work during stance. As larger alligators use a more adducted shoulder and hip posture, the sprawling to erect postural transition that occurred in the Triassic could be either the cause or consequence of the evolution of larger body size in archosaurs. Comparisons of EMG burst phases among tetrapods revealed that a bird and turtle, which have experienced major musculoskeletal transformations, displayed distinctive burst phases in comparison to those from an alligator and lizard. These results support the notion that major shifts in body plan and locomotor modes among sauropsid lineages were associated with significant changes in muscle activation patterns.

A three-dimensional geometric morphometric analysis of the morphological transformation of Caiman lower jaw during post-hatching ontogeny.

Shape ontogenetic changes of the lower jaw in crocodylians are poorly understood. In order to answer some questions related to the inter- and intraspecific morphological variation of the mandible of two extant Caiman species, we performed a three-dimensional geometric morphometric approach. For this purpose, we used landmarks and semilandmarks on two ontogenetic mandibular series of 48 and 15 post-hatching specimens of C. yacare and C. latirostris, respectively. We have also examined the relationship between these anatomical transformations and ontogenetic shifts in diet. We performed a principal component analysis (PCA) for the two species, and regression and partial least squares (PLS) analyses for each species, separately. As a result, species were segregated along the PC1 with specimens of C. yacare showing more gracile mandibles, and specimens of C. latirostris more robust ones. The PC2 and regression analyses showed an age gradient and represented ontogenetic shape changes. Adult caiman mandibles are higher and wider than juvenile ones, and shape changes are more conspicuous in C. latirostris. The PLS analyses showed a significant relationship between shape and diet. Morphological changes of the PLS1 of block-1 match with those of the regression analysis for both species. We have detected morphological transformations in areas where the musculature in charge of mandibular movements is attached. Common morphological changes occurring during ontogeny seem to reflect the same mechanical properties required for crushing and killing in both species, driven by an ontogenetic shift in the diet from invertebrates to vertebrates. Additionally, interspecific differences were also found to be correlated to ontogenetic changes in diet and could be related to dissimilar feeding mechanical requirements (e.g., stiffness and toughness of the item consumed), and to different habitat preferences. Robust mandibles would be more suitable for shallow and fully vegetated environments, as it can be seen in C. latirostris, whereas slender jaws seem to be more suitable for more aquatic species such as C. yacare.

Cross-species chromosome painting and repetitive DNA mapping illuminate the karyotype evolution in true crocodiles (Crocodylidae).

Crocodilians have maintained very similar karyotype structures and diploid chromosome numbers for around 100 million years, with only minor variations in collinearity. Why this karyotype structure has largely stayed unaltered for so long is unclear. In this study, we analyzed the karyotypes of six species belonging to the genera Crocodylus and Osteolaemus (Crocodylidae, true crocodiles), among which the Congolian endemic O. osborni was included and investigated. We utilized various techniques (differential staining, fluorescence in situ hybridization with repetitive DNA and rDNA probes, whole chromosome painting, and comparative genomic hybridization) to better understand how crocodile chromosomes evolved. We studied representatives of three of the four main diploid chromosome numbers found in crocodiles (2n = 30/32/38). Our data provided new information about the species studied, including the identification of four major chromosomal rearrangements that occurred during the karyotype diversification process in crocodiles. These changes led to the current diploid chromosome numbers of 2n = 30 (fusion) and 2n = 38 (fissions), derived from the ancestral state of 2n = 32. The conserved cytogenetic tendency in crocodilians, where extant species keep near-ancestral state, contrasts with the more dynamic karyotype evolution seen in other major reptile groups.

Discovery of facultative parthenogenesis in a new world crocodile.

Over the past two decades, there has been an astounding growth in the documentation of vertebrate facultative parthenogenesis (FP). This unusual reproductive mode has been documented in birds, non-avian reptiles-specifically lizards and snakes-and elasmobranch fishes. Part of this growth among vertebrate taxa is attributable to awareness of the phenomenon itself and advances in molecular genetics/genomics and bioinformatics, and as such our understanding has developed considerably. Nonetheless, questions remain as to its occurrence outside of these vertebrate lineages, most notably in Chelonia (turtles) and Crocodylia (crocodiles, alligators and gharials). The latter group is particularly interesting because unlike all previously documented cases of FP in vertebrates, crocodilians lack sex chromosomes and sex determination is controlled by temperature. Here, using whole-genome sequencing data, we provide, to our knowledge, the first evidence of FP in a crocodilian, the American crocodile, Crocodylus acutus. The data support terminal fusion automixis as the reproductive mechanism; a finding which suggests a common evolutionary origin of FP across reptiles, crocodilians and birds. With FP now documented in the two main branches of extant archosaurs, this discovery offers tantalizing insights into the possible reproductive capabilities of the extinct archosaurian relatives of crocodilians and birds, notably members of Pterosauria and Dinosauria.

Neuroanatomy of the crocodylomorph Portugalosuchus azenhae from the late cretaceous of Portugal.

We present the first detailed braincase anatomical description and neuroanatomical study of Portugalosuchus azenhae, from the Cenomanian (Late Cretaceous) of Portugal. This eusuchian crocodylomorph was originally described as a putative Crocodylia and one of the oldest representatives of this clade; however, its phylogenetic position remains controversial. Based on new data obtained from high resolution Computed Tomography images (by micro-CT scan), this study aims to improve the original description of this taxon and also update the scarce neuroanatomical knowledge of Eusuchia and Crocodylia from this time interval, a key period to understand the origin and evolution of these clades. The resulting three-dimensional models from the CT data allowed a detailed description of its well-preserved neurocranium and internal cavities. Therefore, it was possible to reconstruct the cavities of the olfactory region, nasopharyngeal ducts, brain, nerves, carotid arteries, blood vessels, paratympanic sinus system and inner ear, which allowed to estimate some neurosensorial capabilities. By comparison with other crocodylomorphs, these analyses showed that Portugalosuchus, back in the Cenomanian, already displayed an olfactive acuity, sight, hearing and cognitive skills within the range of that observed in other basal eusuchians and crocodylians, including extant species. In addition, and in order to test its disputed phylogenetic position, these new anatomical data, which helped to correct and complete some of the original observations, were included in one of the most recent morphology-based phylogenies. The position of Portugalosuchus differs slightly from the original publication since it is now located as a "thoracosaurid" within Gavialoidea, but still as a crocodylian. Despite all this, to better contrast these results, additional phylogenetic analyses including this new morphological character coding together with DNA data should be performed.

Ontogenetic variability of the intertympanic sinus distinguishes lineages within Crocodylia.

The phylogenetic relationships within crown Crocodylia remain contentious due to conflicts between molecular and morphological hypotheses. However, morphology-based datasets are mostly constructed on external characters, overlooking internal structures. Here, we use 3D geometric morphometrics to study the shape of the intertympanic sinus system in crown crocodylians during ontogeny, in order to assess its significance in a taxonomic context. Intertympanic sinus shape was found to be highly correlated with size and modulated by cranial shape during development. Still, adult sinus morphology distinguishes specimens at the family, genus and species level. We observe a clear distinction between Alligatoridae and Longirostres, a separation of different Crocodylus species and the subfossil Malagasy genus Voay, and a distinction between the Tomistoma and Gavialis lineages. Our approach is independent of molecular methods but concurs with the molecular topologies. Therefore, sinus characters could add significantly to morphological datasets, offering an alternative viewpoint to resolve problems in crocodylian relationships.

Cranial anatomy of the mekosuchine crocodylian Trilophosuchus rackhami Willis, 1993.

One of the best-preserved crocodylian fossil specimens from the Cenozoic of Australia is the holotype of the mekosuchine Trilophosuchus rackhami, from the middle Miocene (13.56 ± 0.67 Ma) Ringtail Site at Riversleigh, northwestern Queensland. Although lacking most of the snout, the holotype skull of T. rackhami (QMF16856) has an exceptionally well-preserved cranium. Micro-CT scanning of the holotype has allowed for all the preserved cranial bones to be digitally disarticulated, facilitating an unprecedented insight into the cranial anatomy of not just T. rackhami, but any mekosuchine. Trilophosuchus rackhami was a small-bodied crocodylian and one of the most morphologically distinct mekosuchines, characterized by a unique combination of cranial characteristics several of which are exclusive to the species. Fossil material that is definitively referrable to the species T. rackhami is currently known solely from the middle Miocene Ringtail Site. However, an isolated parietal from Hiatus Site at Riversleigh demonstrates that Trilophosuchus also occurred during the late Oligocene (~25 Ma), extending the range of the genus by more than 10 million years. The new description of T. rackhami also allowed for a reevaluation of its phylogenetic relationships. Our results reaffirm the placement of T. rackhami as a member of Mekosuchinae within the subclade Mekosuchini. In all analyses, Mekosuchinae was consistently found to be monophyletic and part of the larger crocodylian clade Longirostres. However, the assignment of Mekosuchinae as a subset of Crocodylidae is brought into question, suggesting that the status of Mekosuchinae as a subfamily should be reconsidered.

Orbital soft tissues, bones, and allometry: Implications for the size and position of crocodylian eyes.

Although the visual system of crocodylians has attracted interest regarding optical parameters and retinal anatomy, fundamental questions remain about the allometry of the eyeball and whether such scaling is the same across all crown groups of crocodylians. In addition, anatomy and identities of adnexal soft tissues that interact with the visual system are not well understood in many cases. We used contrast-enhancing iodine stain and high-resolution micro-computed tomography to assess the anatomy of orbital soft tissues, including extraocular muscles and glands, in crocodylians. We also used regression analysis to estimate the allometric relationship between the bony orbit and eyeball across Alligator mississippiensis and Crocodylus niloticus for the first time. Results revealed tight, negatively allometric relationships between the bony orbit and eyeball. Notably, the eyes of C. niloticus were larger for a given orbit size than the eyes of A. mississippiensis, although the slope of the relationship was no different between these two crown crocodylian groups. Among the findings from our anatomical study, new details were uncovered about the homologies of muscles of the abducens complex. In particular, M. rectus lateralis of crocodylians is revealed to have a more complex form than previously appreciated, being adhered to the tendon of the nictitating membrane, which may be apomorphic for Crocodylia. Our calculation of the orbit-eyeball allometric relationship and study of the adnexal soft tissues of the crocodylian visual system, in combination with previous work by other teams in other crown saurian clades, is a critical, formerly missing, piece in the Extant Phylogenetic Bracket for restoring the visual apparatus of extinct crocodyliforms and other archosauriform groups.

Re-evaluation of the morphology and phylogeny of Diplocynodon levantinicum Huene & Nikoloff, 1963 and the stratigraphic age of the West Maritsa coal field (Upper Thrace Basin, Bulgaria).

Diplocynodon levantinicum Huene & Nikoloff, 1963 was described based on few bone fragments from the West-Maritsa lignite basin of Central Bulgaria. Huene & Nikoloff, 1963 assumed a late Pliocene age, implying that this species represents the stratigraphically youngest crocodilian of Europe. In this current study, we re-evaluate the stratigraphy of the West-Maritsa Basin and conclude a late Oligocene age of ~26 Ma for the Kipra coal-seam, the fossiliferous horizon. Furthermore, topotypical and undescribed D. levantinicum specimens are accessible now and allowed for a deeper taxonomic and phylogenetic analysis. A comparison with other Diplocynodon species reveals D. levantinicum as a valid species, having (1) a long suborbital fenestra, (2) a very short dentary symphysis, (3) a large gap between the first and second dentary alveolus, (4) an occlusion pit in line with the tooth row posterior to the 14th dentary alveolus, (5) a sulcus lateral to the glenoid fossa and, (6) a lingual foramen for the articular artery situated entirely on the surangular. The phylogenetic analyses find D. levantinicum deeply nested inside the Diplocynodontinae subfamily. After the disappearance of the Paratethyan influence (Solenovian regional stage) in the Upper Thrace Basin this species has roamed during the late Oligocene extensive freshwater lake and swamp ecosystems represented by the Maritsa Formation. Diplocynodon levantinicum represents the only nominal Diplocynodon taxon of late Oligocene (Chattian) age.

Dispatches from the age of crocodiles: New discoveries from ancient lineages.

Crocodilians inspire researchers and the public alike with their explosive hunting methodologies, distinct craniofacial and dental morphology, and resplendent fossil record. This special issue highlights recent advances in the biology and paleontology of this fascinating lineage of vertebrates. The authors in this volume bring crocodylians and their extinct ancestors to life using a variety of approaches including fieldwork, imaging, 3D modeling, developmental biology, physiological monitoring, dissection, and a host of other comparative methods. Our journey begins with early crocodylomorphs from the Triassic, carries us through the radiation of crocodyliforms during the rest of the Mesozoic Era, and finally celebrates the diversification development and biology of extant crocodylians. Crocodyliform science has grown appreciably the past few decades. New fossil species and genetic evidence continue to keep phylogenies and our understanding of relationships wavering in key places of the tree such as the relationships of the extinct marine thalattosuchians as well as still living species like gharials. The application of imaging approaches and 3D modeling to both preserved tissues as well as living specimens is now revealing patterns in brain and lung evolution and function, growth strategies, and feeding and locomotor behaviors across the lineage. Comparative anatomical studies are offering new data on genitals, cephalic venous drainage and thoracoabdominal pressures. The new discoveries found here only reveal there is far more work to be done to understand the biology and behavior responsible for the great radiation extinct suchians and their crocodylian descendants experienced during their conquest of Mesozoic and Tertiary ecosystems.

Neuroanatomy of the mekosuchine crocodylian Trilophosuchus rackhami Willis, 1993.

Although our knowledge on crocodylomorph palaeoneurology has experienced considerable growth in recent years, the neuroanatomy of many crocodylomorph taxa has yet to be studied. This is true for Australian taxa, where thus far only two crocodylian crocodylomorphs have had aspects of their neuroanatomy explored. Here, the neuroanatomy of the Australian mekosuchine crocodylian Trilophosuchus rackhami is described for the first time, which significantly increases our understanding on the palaeoneurology of Australian crocodylians. The palaeoneurological description is based on the taxon's holotype specimen (QMF16856), which was subjected to a µCT scan. Because of the exceptional preservation of QMF16856, most neuroanatomical elements could be digitally reconstructed and described in detail. Therefore, the palaeoneurological assessment presented here is hitherto the most in-depth study of this kind for an extinct Australian crocodylomorph. Trilophosuchus rackhami has a brain endocast with a distinctive morphology that is characterized by an acute dural peak over the hindbrain region. While the overall morphology of the brain endocast is unique to T. rackhami, it does share certain similarities with the notosuchian crocodyliforms Araripesuchus wegeneri and Sebecus icaeorhinus. The endosseous labyrinth displays a morphology that is typical for crocodylians, although a stand-out feature is the unusually tall common crus. Indeed, the common crus of T. rackhami has one of the greatest height ratios among crocodylomorphs with currently known endosseous labyrinths. The paratympanic pneumatic system of T. rackhami is greatly developed and most similar to those of the extant crocodylians Osteolaemus tetraspis and Paleosuchus palpebrosus. The observations on the neuroanatomy of T. rackhami are also discussed in the context of Crocodylomorpha. The comparative palaeoneurology reinforces previous evaluations that the neuroanatomy of crocodylomorphs is complex and diverse among species, and T. rackhami has a peculiar neuromorphology, particularly among eusuchian crocodyliforms.

Giant dwarf crocodiles from the Miocene of Kenya and crocodylid faunal dynamics in the late Cenozoic of East Africa.

We describe two new osteolaemine crocodylids from the Early and early Middle Miocene of Kenya: Kinyang mabokoensis tax. nov. (Maboko, 15 Ma) and Kinyang tchernovi tax. nov. (Karungu and Loperot, 18 Ma). Additional material referable to Kinyang is known from Chianda and Moruorot. The skull was broad and dorsoventrally deep, and the genus can be diagnosed based on the combined presence of a partial overbite, a subdivided fossa for the lateral collateral ligament on the surangular, and a maxilla with no more than 13 alveoli. Phylogenetic analyses based on morphological and combined morphological and molecular data support a referral of Kinyang to Osteolaeminae, and morphological data alone put the new taxon at the base of Euthecodontini. Some Kinyang maxillae preserve blind pits on the medial caviconchal recess wall. Kinyang co-occurs with the osteolaemine Brochuchus at some localities, and together, they reinforce the phylogenetic disparity between early Neogene osteolaemine-dominated faunas and faunas dominated by crocodylines beginning in the Late Miocene in the Kenya Rift. The causes of this turnover remain unclear, though changes in prevailing vegetation resulting from tectonic and climatic drivers may provide a partial explanation.

The effects of skull flattening on suchian jaw muscle evolution.

Jaw muscles are key features of the vertebrate feeding apparatus. The jaw musculature is housed in the skull whose morphology reflects a compromise between multiple functions, including feeding, housing sensory structures, and defense, and the skull constrains jaw muscle geometry. Thus, jaw muscle anatomy may be suboptimally oriented for the production of bite force. Crocodylians are a group of vertebrates that generate the highest bite forces ever measured with a flat skull suited to their aquatic ambush predatory style. However, basal members of the crocodylian line (e.g., Prestosuchus) were terrestrial predators with plesiomorphically tall skulls, and thus the origin of modern crocodylians involved a substantial reorganization of the feeding apparatus and its jaw muscles. Here, we reconstruct jaw muscles across a phylogenetic range of crocodylians and fossil suchians to investigate the impact of skull flattening on muscle anatomy. We used imaging data to create 3D models of extant and fossil suchians that demonstrate the evolution of the crocodylian skull, using osteological correlates to reconstruct muscle attachment sites. We found that jaw muscle anatomy in early fossil suchians reflected the ancestral archosaur condition but experienced progressive shifts in the lineage leading to Metasuchia. In early fossil suchians, musculus adductor mandibulae posterior and musculus pterygoideus (mPT) were of comparable size, but by Metasuchia, the jaw musculature is dominated by mPT. As predicted, we found that taxa with flatter skulls have less efficient muscle orientations for the production of high bite force. This study highlights the diversity and evolution of jaw muscles in one of the great transformations in vertebrate evolution.

New frontiers in imaging, anatomy, and mechanics of crocodylian jaw muscles.

New imaging and biomechanical approaches have heralded a renaissance in our understanding of crocodylian anatomy. Here, we review a series of approaches in the preparation, imaging, and functional analysis of the jaw muscles of crocodylians. Iodine-contrast microCT approaches are enabling new insights into the anatomy of muscles, nerves, and other soft tissues of embryonic as well as adult specimens of alligators. These imaging data and other muscle modeling methods offer increased accuracy of muscle sizes and attachments without destructive methods like dissection. 3D modeling approaches and imaging data together now enable us to see and reconstruct 3D muscle architecture which then allows us to estimate 3D muscle resultants, but also measurements of pennation in ways not seen before. These methods have already revealed new information on the ontogeny, diversity, and function of jaw muscles and the heads of alligators and other crocodylians. Such approaches will lead to enhanced and accurate analyses of form, function, and evolution of crocodylians, their fossil ancestors and vertebrates in general.

Morphometrics of the Spinal Cord and Surrounding Structures in Alligator mississippiensis.

Understanding the fluid dynamics of the cerebrospinal fluid requires a quantitative description of the spaces in which it flows, including the spinal cord and surrounding meninges. The morphometrics of the spinal cord and surrounding tissues were studied in specimens of the American alligator (Alligator mississippiensis) ranging from hatchlings through adults. Within any size class of alligators (i.e., hatchlings), along the axial length there are significant differences in the size of the spinal cord, meninges, and vertebral canal; these differences can be used to define discrete cervical, thoracic, lumbar and caudal regions. When compared across the range of body sizes in Alligator, every structure in each spinal region had a distinctive growth rate; thus, the physical arrangements between the structures changed as the alligator grew. The combination of regional differentiation and differential growth rates was particularly apparent in the lumbar meninges where a unique form of lumbar cistern could be identified and shown to decrease in relative size as the alligator ages. This analysis of the spinal cord and surrounding tissues was undertaken to develop a data set that could be used for computational flow dynamics of the crocodilian cerebrospinal fluid, and also to assist in the analysis of fossil archosaurs.

Saurochory in crocodiles does not favor seed dispersai and viability / Saurocoría en cocodrilos no favorece la dispersión y la viabilidad de semillas

RESUMEN La dispersión de semillas por reptiles (saurocoría) ha recibido atención recientemente y se ha reportado el consumo de frutos y semillas en cocodrilianos, actuando como potenciales dispersores de semillas. Evaluamos si la saurocoría en Crocodylus actus y C. moreletii afecta la viabilidad de tres especies de plantas Delonixregia, Inga sp. y Citrullus lanatus. Se Utilizaron tres individuos juveniles de cada especie de cocodrilo y se alimentaron con 22 semillas por especie de planta, para un total de 66 semillas por recinto (132 para ambos). Las semillas se mezclaron con la dieta habitual cada semana, las semillas no consumidas y las excretadas se recolectaron y sembraron en suelo tratado con composta para evaluar la tasa de germinación relativa. Un total de 99 semillas fueron consumidas pero solo se recuperaron de las heces 14 semillas de C. lanatus, y germinando solo una de ellas (7, 14 %) con respecto al 50 % en el grupo control. Los resultados indican que la saurocoría de C. acutus y C. moreletii tiene un efecto negativo sobre la viabilidad de las semillas de las especies vegetales estudiadas, concordando con otros estudios realizados en diferentes especies.

ABSTRACT Seed dispersal by reptiles (saurochory) has recently received attention, and the consumption of fruits and seeds has been reported in crocodilians despite being mainly carnivores, acting as potential seed dispersers. We evaluate whether saurochory by Crocodylus acutus and C. moreletii affect the seed viability of three species of plants (Delonix regia, Inga sp., and Citrullus lanatus). We performed feeding trials, using three juvenile individuals of each species of crocodile, and fed them 22 seeds per plant species for a total of 66 seeds per enclosure (132 for both species). Seeds were combined with the usual diet each week. The unconsumed and excreted seeds were collected and planted in soil treated with compost to evaluate the relative germination rate. A total of 99 seeds were consumed, of which only 14 seeds of C. lanatus were recovered from the faeces, and only one of those germinated (7.14 %) with respect to 50 % in the control group. The results indicate that saurochory by C. acutus and C. moreletii has a negative effect on seed viability and germination of the plant species studied, as found in other studies using different species.

An intermediate crocodylian linking two extant gharials from the Bronze Age of China and its human-induced extinction.

A solid phylogenetic framework is the basis of biological studies, yet higher level relationships are still unresolved in some major vertebrate lineages. One such group is Crocodylia, where the branching pattern of three major families (Alligatoridae, Crocodylidae and Gavialidae) has been disputed over decades due to the uncertain relationship of two slender-snouted lineages, gavialines and tomistomines. Here, we report a bizarre crocodylian from the Bronze Age of China, which shows a mosaic of gavialine and tomistomine features across the skeleton, rendering support to their sister taxon relationship as molecular works have consistently postulated. Gavialine characters of the new Chinese crocodylian include a novel configuration of the pterygoid bulla, a vocal structure known in mature male Indian gharials. Extinct gavialines have repeatedly evolved potentially male-only acoustic apparatus of various shapes, illuminating the deep history of sexual selection on acoustic signalling in a slender-snouted group of crocodylians. Lastly, a cutmark analysis combined with accelerator mass spectrometry (AMS) radiocarbon dating of bone remains demonstrated that two individuals from Shang and Zhou dynasties in Guangdong, China, suffered head injuries and decapitation. Archaeological evidence together with historical accounts suggests the human-induced extinction of this unique crocodylian only a few hundred years ago.

'Ear stones' in crocodylians: a cross-species comparative and ontogenetic survey of otolith structures.

The vestibular system of the inner ear is a crucial sensory organ, involved in the sensation of balance and equilibrium. It consists of three semicircular canals that sense angular rotations of the head and the vestibule that detects linear acceleration and gravity. The vestibule often contains structures, known as the otoliths or 'ear stones'. Otoliths are present in many vertebrates and are particularly well known from the fossil record of fish, but surprisingly have not been described in detail in most tetrapods, living or extinct. Here, we present for the first time a survey of the otoliths of a broad sample of extant crocodylian species, based on computed tomography scans. We find that otoliths are present in numerous crocodylian species of different growth stages, and they continue to increase in size during ontogeny, with positive allometry compared to skull length. Our results confirm that otoliths are a common component of the crocodylian vestibular system, and suggest they play an important role in sensory detection. Otoliths are likely common, but overlooked, constituents of the inner ear in tetrapods, and a broader study of their size, shape and distribution promises insight into sensory abilities.

A "hotspot" within a hotspot: the reptiles of the Estação Ecológica and Área de Proteção Ambiental de Murici, Atlantic Forest of northeastern Brazil / Um "hotspot" dentro de um hotspot: os répteis da Estação Ecológica e Área de Proteção Ambiental de Murici, Mata Atlântica do nordeste do Brasil

Abstract: Currently the Atlantic Forest hotspot has less than 11% of its original coverage. However approximately 300 species of reptiles are known to inhabit this ecoregion, of which 34% are endemic. The creation of protected areas represents a strategy for preserving ecosystems and managing land use, and for attaining the proper management of these protected areas, information on local biodiversity is essential. Herein we provide the first list of reptile species for the Estação Ecológica and Área de Proteção Ambiental de Murici, two overlapping protected areas located in the state of Alagoas, one of the most important sets of forest remnants for the conservation of the Atlantic Forest in the northeast of Brazil. The species list was constructed based on expeditions and occasional encounters in the area between 1994 and 2022. A total of 89 reptile species were obtained during the 28 years of collection, being two species of Crocodylia, three species of Testudines and 84 species of Squamata. This richness is by far one of the greatest ever documented for the Atlantic Forest. Additionally, two species registered in the area are considered threatened according to the national list (Amerotyphlops paucisquamus and Bothrops muriciensis) and six are defined as data deficient for the assessment of their conservation status. Three species are recorded for the first time in the Atlantic Forest north of the São Francisco River: Dipsas indica, Trilepida salgueiroi and Cercophis auratus. We claim that the species list provided here will serve as a starting point for further studies in this rich reptile "hotspot" within the Atlantic Forest.

Resumo: Atualmente o hotspot da Mata Atlântica tem menos de 11% de sua cobertura original. Porém aproximadamente 300 espécies de répteis são conhecidas por habitar esta ecorregião, das quais 34% são endêmicas. A criação de unidades de conservação representa uma estratégia de preservação de ecossistemas e gestão do uso do solo, e para o manejo adequado dessas áreas protegidas é fundamental a informação sobre a biodiversidade local. Apresentamos aqui a primeira lista de espécies de répteis para a Estação Ecológica e Área de Proteção Ambiental de Murici, duas unidades de conservação parcialmente sobrepostas localizadas no estado de Alagoas, um dos conjuntos de remanescentes florestais mais importantes para a conservação da Mata Atlântica no nordeste do Brasil. A lista de espécies foi construída com base em expedições e encontros ocasionais na área entre 1994 e 2022. Um total de 89 espécies de répteis foram registradas durante os 28 anos de coleta, sendo duas espécies de Crocodylia, três espécies de Testudines e 84 espécies de Squamata. Essa riqueza é de longe uma das maiores já documentadas para a Mata Atlântica. Além disso, duas espécies registradas na área são consideradas ameaçadas segundo a lista nacional (Amerotyphlops paucisquamus e Bothrops muriciensis) e seis são definidas como dados insuficientes para a avaliação de seu status de conservação. Três espécies são registradas pela primeira vez na Mata Atlântica ao norte do rio São Francisco: Dipsas indica, Trilepida salgueiroi e Cercophis auratus. Afirmamos que a lista de espécies aqui fornecida servirá como ponto de partida para novos estudos neste rico "hotspot" de répteis dentro da Mata Atlântica.

The impact of molecular data on the phylogenetic position of the putative oldest crown crocodilian and the age of the clade.

The use of molecular data for living groups is vital for interpreting fossils, especially when morphology-only analyses retrieve problematic phylogenies for living forms. These topological discrepancies impact on the inferred phylogenetic position of many fossil taxa. In Crocodylia, morphology-based phylogenetic inferences differ fundamentally in placing Gavialis basal to all other living forms, whereas molecular data consistently unite it with crocodylids. The Cenomanian Portugalosuchus azenhae was recently described as the oldest crown crocodilian, with affinities to Gavialis, based on morphology-only analyses, thus representing a potentially important new molecular clock calibration. Here, we performed analyses incorporating DNA data into these morphological datasets, using scaffold and supermatrix (total evidence) approaches, in order to evaluate the position of basal crocodylians, including Portugalosuchus. Our analyses incorporating DNA data robustly recovered Portugalosuchus outside Crocodylia (as well as thoracosaurs, planocraniids and Borealosuchus spp.), questioning the status of Portugalosuchus as crown crocodilian and any future use as a node calibration in molecular clock studies. Finally, we discuss the impact of ambiguous fossil calibration and how, with the increasing size of phylogenomic datasets, the molecular scaffold might be an efficient (though imperfect) approximation of more rigorous but demanding supermatrix analyses.