Ontogenetic and interelemental study of appendicular bones of Caiman latirostris Daudin, 1802 sheds light on osteohistological variability in crocodylians.

The osteohistology of vertebrates provides a reliable source to deduce biological information, particularly regarding growth and development. Although osteohistological studies in Neosuchia (Crocodyliformes, Mesoeucrocodylia) are relatively numerous, the number of species studied within the group is still small. Extant crocodilians are known to exhibit intraspecific variability linked to environmental conditions, habitat, feeding, and other intrapopulation factors. Here, we analyzed the osteohistology of the living South American Caiman latirostris throughout posthatching ontogeny. The histology of several appendicular bones of 13 different-sized captive and wild individuals were examined. Although some thin sections revealed the classic lamellar, parallel-fibered, or woven bone matrices, others showed a variation and a mix between the organization of the bone tissue. These histological differences are likely related to variability in the growth dynamics of caimans. In some bones of the juveniles studied, remnants of embryonic bone were observed. Osteohistological variation related to prevailing environmental conditions is documented. Furthermore, our results show ontogenetic variation in the type of bone tissues deposited throughout the development of C. latirostris. This study offers a broad framework for life history interpretations for C. latirostris and provides insight into the evolutionary history and ontogenetic growth of extinct crocodylian lineages.

Hindlimb and pelvic anatomy of Caiman yacare (Archosauria, Pseudosuchia): Myology and osteological correlates with emphasis on lower leg and autopodial musculature.

The anatomy of the archosaurian pelvis and hindlimb has adopted a diversity of successful configurations allowing a wide range of postures during the evolution of the group (e.g., erect, sprawling). For this reason, thorough studies of the structure and function of the pelvic and hindlimb musculature of crocodylians are required and provide the possibility to expand their implications for the evolution of archosaurian locomotion, as well as to identify potential new characters based on muscles and their bony correlates. In this study, we give a detailed description of the pelvic and hindlimb musculature of the South American alligator Caiman yacare, providing comprehensive novel information regarding lower limb and autopodial muscles. Particularly for the pedal muscles, we propose a new classification for the dorsal and ventral muscles of the autopodium based on the organisation of these muscles in successive layers. We have studied the myology in a global background in which we have compared the Caiman yacare musculature with other crocodylians. In this sense, differences in the arrangement of m. flexor tibialis internus 1, m. flexor tibialis externus, m. iliofibularis, mm. puboischiofemorales internii 1 and 2, between Ca. yacare and other crocodylians were found. We also discuss the muscle attachments that have different bony correlates among the crocodylian species and their morphological variation. Most of the correlates did not exhibit great variation among the species compared. The majority of the recognised correlates were identified in the pelvic girdle; additionally, some bony correlates associated with the pedal muscles are highlighted here for the first time. This research provides a wide framework for future studies on comparative anatomy and functional morphology, which could contribute to improving the character definition used in phylogenetic analyses and to understand the patterns of musculoskeletal hindlimb evolution.

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.

Unexpected morphological variability in the eggshells of the South American caimans Caiman latirostris and Caiman yacare.

Eggshell morphology is a valuable indicator of the local conditions within the nests of modern crocodilians and birds. In contrast to these latter, the anatomical structure of the eggshells of most crocodilian species is practically unknown. Here, we provide the first characterization of crocodilian eggshells, using x-ray micro-CT scans. We studied eggshells of Caiman latirostris and Caiman yacare from various developmental stages that coincide with the beginning of embryonic ossification. The new 3D renderings revealed complex ornamentation, unique among crocodilians, and amphora-shaped pore canals, some of which converge in single pore openings. We also documented a high density of pore canals with a gas diffusion capacity 45 times higher than the average predicted for modern avian eggshells. The external ornamentation and the thickness of the compact layer of the eggshells (i.e. excluding ornamentation) showed ontogenetic and interspecific differences that could be related to nesting materials and nesting areas selected by each species. The shell features described here evidence a greater structural complexity than previously recognized in phylogenetically close, sympatric crocodilian species. Further comprehensive morphological analyses on other modern and fossil crocodilian eggshells using micro-CT technology will shed new light on the evolution of reproductive strategies in this intriguing archosaur clade.

Late Campanian-Early Maastrichtian Vertebrates From The James Ross Basin, West Antarctica: Updated Synthesis, Biostratigraphy, And Paleobiogeography.

The Snow Hill Island Formation (SHIF; late Campanian - early Maastrichtian) crops out in the northeast of the Antarctic Peninsula and constitutes the basal part of the late Campanian-early Maastrichtian sedimentary succession of the James Ross Basin (NG Sequence). Its major exposures occur at the James Ross and Vega islands. Several fossil-bearing localities have been identified in the SHIF providing a valuable fauna of invertebrates and vertebrates, and flora. Our study focuses on the vertebrate fauna recovered at Gamma and Cape Lamb members of the SHIF. The marine vertebrate assemblages include chondrichthyans, actinopterygians, and marine reptiles (elasmosaurid plesiosaurs and mosasaurs). A diverse terrestrial vertebrate assemblage has been reported being characterized by dinosaurs (sauropod, elasmarian ornithopods, nodosaurid ankylosaur, and a paravian theropod), pterosaurs and birds. Most SHIF dinosaurs share close affinities with penecontemporaneous taxa from southern South America, indicating that at least some continental vertebrates could disperse between southern South America and Antarctica during the Late Cretaceous. The Snow Hill Island Formation provides the most diverse Late Cretaceous marine and continental faunas from Antarctica. The present study summarizes previous and new vertebrate findings with the best actualized stratigraphical framework, providing a more complete fauna association and analyzing further perspectives.

On the homology of crocodylian post-dentary bones and their macroevolution throughout Pseudosuchia.

The lower jaw of early tetrapods is composed of several intramembranous ossifications. However, a tendency toward the independent reduction of the number of bones has been observed in the mandible of mammals, lepidosaurs, turtles, crocodiles, and birds. Regarding archosaurs, the coronoid and prearticular bones are interpreted to be lost during the evolution of stem-birds and stem-crocodiles, respectively, but the homology of the post-dentary bones retained in living pseudosuchians remains unclear. Here, we combine paleontological and embryological evidence to explore in detail the homology of the crocodylian post-dentary bones. We study the mandible embryogenesis on a sample of 71 embryos of Caiman and compare this pattern with the mandibular transformations observed across pseudosuchian evolution. In the pre-hatching ontogeny of caimans, at least five intramembranous ossification centers are formed along the margins of the internal mandibular fenestra (perifenestral centers) and, subsequently, merge to form the coronoid (three intramembranous centers), angular (one intramembranous center), and articular (one intramembranous and one chondral center). In the fossil record, an independent prearticular is lost around the base of Mesoeucrocodylia (optimized as reappearing in Thalattosuchia if they are placed within Neosuchia), and the coronoid is apomorphically lost in notosuchians. The integration of embryological and paleontological data indicates that most perifenestral centers are involved in the origin of the prearticular of non-mesoeucrocodylian pseudosuchians. These centers are rearranged during the evolution to contribute to different post-dentary bones in mesoeucrocodylians bolstering the idea that the coronoid and the articular of Crocodylia are not completely homologous to those of other diapsids.

New digital braincase endocasts of two species of Desmatosuchus and neurocranial diversity within Aetosauria (Archosauria: Pseudosuchia).

In the present contribution we revise, figure, and redescribe several isolated braincases of the iconic aetosaur Desmatosuchus from the Placerias Quarry locality, Chinle Formation, Arizona, United States. The detailed study of the isolated braincases from the UCMP collection allowed us to assign them at the species-level and recognize two species of Desmatosuchus for the Placerias Quarry: D. spurensis and D. smalli. The former can be distinguished from the latter by the presence of a transverse sulcus on the parietals, deep median pharyngeal recess on the basisphenoid, almost no gap between the basal tubera and the basipterygoid processes, and the exoccipitals meeting at the midline. The presence of D. smalli at the Placerias Quarry has not been previously reported. Based on the braincases UCMP 27408, 27410, 27407, three new brain endocasts were developed through CT scan images, reconstructing the most complete endocranial casts known for an aetosaur, including the encephalon, cranial nerves, inner ear, and endocranial vasculature. The cranial endocasts also exhibited some differences between both species of Desmatosuchus, with D. spurensis having a distinguishable dural expansion and markedly asymmetric anterior and posterior semicircular canals of the labyrinth. Additionally, the combination of osteological features and the endocranial casts allowed us to identify and discuss the presence of an ossified orbitosphenoid on the anteriormost region of the braincase among aetosaurs. Furthermore, we were able to reinterpret some of the observations made by previous authors on the endocast of the holotype of Desmatosuchus spurensis (UMMP VP 7476) and provide some insight into their neurosensory capabilities.

Developmental evolution of the distal ankle in the dinosaur-bird transition.

The adult ankle of early reptiles had five distal tarsal (dt) bones, but in Dinosauria, these were reduced to only two: dt3 and dt4, articulated to metatarsals (mt) mt3 and mt4. Birds have a single distal tarsal ossification center that fuses to the proximal metatarsals to form a new adult skeletal structure: the composite tarsometatarsus. This ossification center develops within a single large embryonic cartilage, but it is unclear if this cartilage results from fusion of earlier cartilages. We studied embryos in species from four different bird orders, an alligatorid, and an iguanid. In all embryos, cartilages dt2, dt3, and dt4 are formed. In the alligatorid and the iguanid, dt2 failed to ossify: only dt3 and dt4 develop into adult bones. In birds, dt2, dt3, and dt4 fuse to form the large distal tarsal cartilage; the ossification center then develops above mt3, in cartilage presumably derived from dt3. During the entire dinosaur-bird transition, a dt2 embryonic cartilage was always formed, as inferred from the embryology of extant birds and crocodilians. We propose that in the evolution of the avian ankle, fusion of cartilages dt3 and dt2 allowed ossification from dt3 to progress into dt2, which began to contribute bone medially, while fusion of dt3 to dt4 enabled the evolutionary loss of the dt4 ossification center. As a result, a single ossification center expands into a plate-like unit covering the proximal ends of the metatarsals, that is key to the development of an integrated tarsometatarsus.

New embryological and palaeontological evidence sheds light on the evolution of the archosauromorph ankle.

The homology and evolution of the archosaur ankle is a controversial topic that has been deeply studied using evidence from both extinct and extant taxa. In early stem archosaurs, the astragalus and calcaneum form the ancestral proximal tarsus and a single ossification composes the centrale series. In more recent stem archosaurs, the centrale is incorporated to the proximal row of tarsals laterally contacting the astragalus. This bone is subsequently lost as an independent ossification before the last common ancestor of birds and crocodilians, but the evolutionary fate of this element remains mostly unexplored. Here, we integrate embryological and palaeontological data with morphogeometric analyses to test the hypothesis of loss of the centrale or, alternatively, its incorporation into the archosaurian astragalus. Our results support the latter hypothesis, indicating that the astragalus developed ancestrally from two ossification centres in stem archosaurs and that the supposed tibiale of bird embryos represents a centrale. This conclusion agrees with previous embryological studies that concluded that the tibiale never develops in diapsids.

Phylogenetic signal analysis in the basicranium of Ursidae (Carnivora, Mammalia).

Ursidae is a monophyletic group comprised of three subfamilies: Tremarctinae, Ursinae and Ailuropodinae, all of which have a rich geographical distribution. The phylogenetic relationships within the Ursidae group have been underexamined, especially regarding morphological traits such as the basicranium. Importantly, the basicranium is a highly complex region that covers a small portion of the skull, combining both structural and functional aspects that determine its morphology. Phylogenetic hypotheses of the Ursidae (including Tremarctinae) have been made based on morphological characters that considers skull, mandible and teeth features, while specific characters of the auditory region and basicranium have not been taken into account. To do this, we analyse the shape and size macroevolution of the basicranium of Ursidae, testing its morphological disparity in a phylogenetic context, which is quantified by means of the phylogenetic signal. We investigated phylogenetical autocorrelation by shape (depicted by Principal Components Analysis scores from previous published analyses) and basicranium size (depicted by centroid size, CS) using an orthonormal decomposition analysis and Abouheif C mean. The main advantages of these methods are that they rely exclusively on cladogram topology and do not require branch-length estimates. Also, an optimisation of the ancestral nodes was performed using TNT 1.5 software. In relation to the phylogenetic signal, both methods showed similar results: the presence of autocorrelation was detected in PC1 and PC2, while in PC3, PC4 and PC5 and in the size of the basicranium (CS), the absence of autocorrelation occurred. The most significant nodes (where there is autocorrelation) are the basal nodes 'Ursidae' and 'Ursinae-Tremarctinae'. Within this last group, distinctive basicranium morphology is observed, being more conservative in Tremarctinae than in Ursinae. The differences between these subfamilies could be related to historical events involving varying food and environmental preferences. The high phylogenetic signal in the node Tremarctinae probably indicates that the basicranium configuration of these bears was obtained early in their evolutionary history. Finally, our results of the basicranium and skull length ratios indicate that in Tremarctinae, the basicranium size was not determined by phylogeny but instead by other factors, such as adaptive responses to climatic changes and competition with other carnivores.

Osteohistological correlates of muscular attachment in terrestrial and freshwater Testudines.

Sharpey's fibers are considered the anatomical structures integrated to the muscles. Since these fibers leave marks at the microscopic level, their presence and distribution are used as evidence of muscle attachment in extinct and extant forms. In recent years, studies have been focusing on muscle-bone and tendon-bone interaction mostly on mammals. The main objective of this work is to contribute to the morphological and histological knowledge of muscle attachment in other amniotes, such as reptiles, and their variation related to different locomotor habits. In this way, a study was performed on terrestrial and aquatic turtles. The musculature related to the movement of the humerus, and pectoral girdle in Chelonoidis chilensis, Phrynops hilarii and Hydromedusa tectifera was analyzed. Dissections were performed mapping the origins and insertions of each muscle and undecalcified thin sections were performed in specific muscular attachment sites. We found some differences which were not previously reported, related to the insertion of the m. pectoralis, the m. coracobrachialis magnus and the origin of the m. tractor radii. The osteohistology revealed the presence of Sharpey's fibers in the cortex of all the bone elements analyzed. Patterns were established in relation to the orientation and density of Sharpey's fibers, which were used for the categorization of each muscle attachment site. The comparative micro-anatomical study of these areas did not reveal any important differences between terrestrial and freshwater turtles in muscles involved with the rotation, abduction and adduction of the humerus. In this way, the preliminary results suggest an absence of correlation between the distribution and density of Sharpey's fibers between different habitat forms, at least in the bones and species analyzed.

Skull ontogeny of extant caimans: a three-dimensional geometric morphometric approach.

Ontogenetic variation of cranial characters used in crocodylian phylogenetic systematics has never been studied. Furthermore, the relationship between diet and skull morphological transformation during ontogeny has not been properly explored yet. We quantify the inter- and intraspecific skull morphological variation in extant caiman species focusing on those areas relevant to systematics and, also investigate the relation between diet and morphological changes during ontogeny. We applied a three-dimensional approach of geometric morphometrics on post-hatching ontogenetic cranial series of Caiman latirostris and C. yacare. In order to incorporate incomplete material, we additionally tested four different methods of missing landmark estimation and apply the thin-plate spline interpolation. We detected morphological changes between species and during ontogeny (snout and pterygoid flanges increase their proportions and, orbits, temporal fenestrae, skull roof and foramen magnum decrease their relative size) that constitutes part of a general morphological change in the cranial ontogeny of crocodylians. Moreover, the negative allometry of the fenestrae and neurocranium and the positive allometry of the splanchnocranium in both caiman species are the plesiomorphic condition, at least, for tetrapods. Shape changes during growth were found to be related to ontogenetic changes in the diet. Dissimilarities between species seem to be related to different mechanical requirements and different use of the habitat. We found inter- and intraspecific variation in some morphological characters with systematic implications (the contact of nasals with naris, the contact of prefrontals in the midline, and the bones that border the suborbital fenestra and the proportion in which one of them participates) that are not currently considered in phylogenetic analyses.

A new Palaeocene crocodylian from southern Argentina sheds light on the early history of caimanines.

Caimanines are crocodylians currently restricted to South and Central America and the oldest members are from lower Palaeocene localities of the Salamanca Formation (Chubut Province, Argentina). We report here a new caimanine from this same unit represented by a skull roof and partial braincase. Its phylogenetic relationships were explored in a cladistic analysis using standard characters and a morphogeometric two-dimensional configuration of the skull roof. The phylogenetic results were used for an event-based supermodel quantitative palaeobiogeographic analysis. The new species is recovered as the most basal member of the South American caimanines, and the Cretaceous North American lineage 'Brachychampsa and related forms' as the most basal Caimaninae. The biogeographic results estimated north-central North America as the ancestral area of Caimaninae, showing that the Cretaceous and Palaeocene species of the group were more widespread than thought and became regionally extinct in North America around the Cretaceous-Palaeocene boundary. A dispersal event from north-central North America during the middle Late Cretaceous explains the arrival of the group to South America. The Palaeogene assemblage of Patagonian crocodylians is composed of three lineages of caimanines as a consequence of independent dispersal events that occurred between North and South America and within South America around the Cretaceous-Palaeogene boundary.

Cranial shape variation in jacarean caimanines (Crocodylia, Alligatoroidea) and its implications in the taxonomic status of extinct species: The case of Melanosuchus fisheri.

Melanosuchus niger (Crocodylia, Alligatoroidea) is one of the six living caimanine species widely distributed throughout the Amazon River basin today. Although there is only one extant species of Melanosuchus, fossil material assigned to this genus, represented by M. fisheri, has been reported from the late Miocene in South America. However, the validity of this taxon has been questioned and a recent investigation indicates that the referred specimen of M. fisheri (MCZ 4336) actually belongs to Globidentosuchus brachyrostris, while those diagnostic characters present in the holotype (MCNC 243) fall into the spectrum of intraspecific variation of M. niger. Here, we compare the skull shape of the holotype of M. fisheri with the ontogenetic series of the four jacarean species (M. niger, Caiman yacare, Caiman crocodilus, and Caiman latirostris) using 2D-geometric morphometric analyses in two different views. The analyses indicate that MCNC 243 falls into the morphospace of M. niger and C. latirostris. Despite strong shape similarities between juveniles of C. latirostris and MCNC 243, further anatomical comparisons reveal notable differences between them. In contrast, no concrete anatomical differences can be found between MCNC 243 and M. niger, although shape analyses indicate that MCNC 243 is relatively robust for its size. Thus, this study is able to confirm that the genus Melanosuchus was present in the late Miocene, but it still remains unclear if MCNC 243 should be treated as a junior synonym or probably a sister species of M. niger. Its Miocene age favors the second option, but as the shape analyses were also not able to extract any diagnostic characters, it should be retained as Melanosuchus cf. niger.

Anatomical study of the auditory region of Arctotherium tarijense (Ursidae, Tremarctinae), an extinct short-faced bear from the Pleistocene of South America.

Here we present the most detailed morphological study of the auditory region of a tremarctinae bear, Arctotherium tarijense Ameghino. In addition, we provide new anatomical information of the Tremarctinae inner ear, such as coplanarity and deviation from orthogonality of the semicircular canals, as an approach to infer the head movements which encountered the extinct forms in locomotion. Based on morphological comparisons, A. tarijense exhibits the following particular features: the cavum tympani presents the highest relative volume compared with other ursids; the processus paraoccipitalis has a foramen that is absent in other tremarctines; there is only one (ventral) recess in the anterior region of the cavum tympani; and the recessus epytimpanicus is the smallest for all ursids studied. In relation to the inner ear, A. tarijense shows the lowest values of orthogonality deviation and highest scores of locomotor agility. Based on this, is possible to make a preliminary proposal that this species had a relative high vestibular sensibility and therefore a better ability to explore different kind of habitats. However, this hypothesis might be contrasted among bears taking into account the orientation of each semicircular canal in a phylogenetic framework.

Skull anatomy of the bizarre crocodylian Mourasuchus nativus (Alligatoridae, Caimaninae).

Mourasuchus is a Miocene alligatorid endemic to South America, and is represented by four species. Together with the closely related Purussaurus, it is a peculiar crocodylian taxon of neogene Caimaninae and one of the most bizarre forms among eusuchian crocodiles. The phylogenetic relationships between Mourasuchus species have not been explored, and detailed skull descriptions are scarce. The goal of this study is to provide new data on skull morphology and cranial recesses in Mourasuchus nativus, including a new tomography analysis (3D modeling). We observed that several diagnostic characters of Purussaurus, such as lack of contact between the nasal and lacrimal, separation of the nasal and frontal by the prefrontals, and the posterior dorsal margin of the skull table, are shared with Mourasuchus. M. nativus is characterized by the presence of solid transverse squamosal eminences, large posttemporal fenestrae, and a quadrate laterocaudal bridge separating V(2) -V(3) trigeminal openings. Compared with other crocodylians, the endocast of M. nativus is similar in shape but quite sigmoid in lateral view, the canal of the supraorbital ramus of V(2) is more vertically oriented, the thick tympanic branch canal opens in a large foramen aligned with trigeminal foramen, and the canal of the vagal (X) tympanic ramus is also very wide. Contrary to extant alligatorids, the median pharyngeal recess remains paired throughout its course and only connects its opposite fellow near the external ventral opening. The knowledge of the internal skull anatomy of Mourasuchus contributes to the understanding of the general morphology of alligatorids, Caimaninae, and their variation.

Osteology and cranial musculature of Caiman latirostris (crocodylia: Alligatoridae).

Caiman latirostris Daudin is one of the extant species of Caimaninae alligatorids characterized taxonomically only by external morphological features. In the present contribution, we describe the cranial osteology and myology of this species and its morphological variation. Several skull dissections and comparisons with other caimans were made. Although jaw muscles of living crocodiles show the same general "Bauplan" and alligatorids seem to have a similar cranial musculature pattern, we describe some morphological variations (e.g., in C. latirostris the superficial portion of the M. adductor mandibulae externus did not reach the postorbital; the M. adductor mandibulae internus pars pterygoideus dorsalis did not reach the pterygoid and lacrimal and contrary to the case of C. crocodilus the M. adductor mandibulae internus pars pterygoideus ventralis attaches to the posterodorsal surface of the pterygoid and the pterygoid aponeurosis, without contacting the dorsal and ventral surface of the pterygoid margin; the M. intermandibularis is attached to the anterior half of the splenial and posteriorly inserts medially by a medial raphe that serves as attachment zone for M. constrictor colli, and the M. constrictor colli profundus presents a medial notch in its anterior margin). In addition, the skull of C. latirostris differs from that of other caimans and possesses several characters that are potential diagnostic features of this species (e.g., outline of glenoid cavity in dorsal view, extension of the rostral ridges, and occlusion of the first dentary tooth). Nevertheless, these characters should be analyzed within the phylogenetic context of the Caimaninae to evaluate its evolutionary implications for the history of the group.