Parallel-fibred Bone Research Articles

The osteohistology of Orthosuchus stormbergi using synchrotron radiation microcomputed tomography.

Orthosuchus stormbergi was a small-bodied crocodyliform, representative of a diverse assemblage of Early Jurassic, early branching crocodylomorph taxa from the upper Elliot Formation of South Africa. The life history of these early branching taxa remains poorly understood, with only sparse investigations into their osteohistology, yet species like Orthosuchus have potential to inform about the macroevolution of growth strategies on the stem leading to crown crocodilians. In order to elucidate the growth patterns of Orthosuchus, we used propagation phase contrast X-ray synchrotron micro-computed tomography to virtually image the osteohistology of the postcrania of two specimens, including multiple elements from the type (SAM-PK-K409), and the femur of a referred specimen (BP/1/4242). In total, we scanned nine mid-diaphyseal sections of the humerus, radius, ulna, radiale, femur, tibia, fibula, and a rib. We then compared our results to osteohistological sections of crocodylomorph taxa from the published literature. Our results show that the most predominant bone tissue type in Orthosuchus is lamellar, with a few patches of woven and parallel-fibred bone. The type specimen contains four to five lines of arrested growth and the hindlimb elements present outer circumferential lamellae, whereas the referred specimen contains six to seven. Both specimens grew at similar rates, reaching adult skeletal body size at year four or five. The sectioned bones, most notably the radius and ulna, are comparatively thick walled and compact. Our virtual osteohistological sections are one of the first for an early branching crocodyliform, and the broad sample of skeletal elements makes Orthosuchus a key anchor point for understanding the plesiomorphic life history traits of the clade.

Comparative bone histology of two thalattosaurians (Diapsida: Thalattosauria): Askeptosaurus italicus from the Alpine Triassic (Middle Triassic) and a Thalattosauroidea indet. from the Carnian of Oregon (Late Triassic)

Here, we present the first bone histological and microanatomical study of thalattosaurians, an enigmatic group among Triassic marine reptiles. Two taxa of thalattosaurians, the askeptosauroid Askeptosaurus italicus and one as yet undescribed thalattosauroid, are examined. Both taxa have a rather different microanatomy, tissue type, and growth pattern. Askeptosaurus italicus from the late Anisian middle Besano Formation of the southern Alpine Triassic shows very compact tissue in vertebrae, rib, a gastralium, and femora, and all bones are without medullary cavities. The tissue shows moderate to low vascularization, dominated by highly organized and very coarse parallel-fibred bone, resembling interwoven tissue. Vascularization is dominated by simple longitudinal vascular canals, except for the larger femur of Askeptosaurus, where simple vascular canals dominate in a radial arrangement. Growth marks stratify the cortex of femora. The vertebrae and humeri from the undescribed thalattosauroid from the late Carnian of Oregon have primary and secondary cancellous bone, resulting in an overall low bone compactness. Two dorsal vertebral centra show dominantly secondary trabeculae, whereas a caudal vertebral centrum shows much primary trabecular bone, globuli ossei, and cartilage, indicating an earlier ontogenetic stage of the specimens or paedomorphosis. The humeri of the thalattosauroid show large, simple vascular canals that are dominantly radially oriented in a scaffold of woven and loosely organized parallel-fibred tissue. Few of the simple vascular canals are thinly but only incompletely lined by parallel-fibered tissue. In the Oregon material, changes in growth rate are only indicated by changes in vascular organization but no distinct growth marks were identified. The compact bone of Askeptosaurus is best comparable to some pachypleurosaurs, whereas its combination of tissue and vascularity is similar to eosauropterygians in general, except for the coarse nature of its parallel-fibred tissue. The cancellous bone of the Oregon thalattosauroid resembles what is documented in ichthyosaurs and plesiosaurs. However, in contrast to these its tissue does not consist of fibro-lamellar bone type. Tissue types of both thalattosaurian taxa indicate rather different growth rates and growth patterns, associated with different life history strategies. The microanatomy reflects different life styles that fit to the different environments in which they had been found (intraplatform basin vs. open marine). Both thalattosaurian taxa differ from each other but in sum also from all other marine reptile taxa studied so far. Thalattosaurian bone histology documents once more that bone histology provides for certain groups (i.e., Triassic Diapsida) only a poor phylogenetic signal and is more influenced by exogenous factors. Differences in lifestyle, life history traits, and growth rate and pattern enabled all these Triassic marine reptiles to live contemporaneously in the same habitat managing to avoid substantial competition.

Palaeobiological inferences of "rauisuchians" Fasolasuchus tenax (Los Colorados Fm., Argentina) and Prestosuchus chiniquensis (Santa Maria Super sequence, Brazil) from the Middle-Upper Triassic of South America based on microstructural analyses.

"Rauisuchia" is a non-monophyletic group of quadrupedal and carnivorous pseudosuchians that inhabited the entire world during the Middle-Upper Triassic period (Anisian/Ladinian-Rhaetian). In South America, "rauisuchians" reached the largest sizes among continental carnivores. Despite their important ecological role, some aspects of their palaeobiology have been poorly examined. Here, we study appendicular bones, dorsal ribs and osteoderms of two genera, the Argentinean Fasolasuchus tenax (PVL 3850, holotype) and the Brazilian Prestosuchus chiniquensis (SNSB-BSPG AS XXV) respectively. The femur of F. tenax is formed by laminar fibrolamellar bone, which is composed of non-fully monorefringent woven-fibred matrix and primary osteons; the dorsal rib has a Haversian bone composition with an external fundamental system recorded and the osteoderm is formed by well-organised parallel-fibred bone. The femur, humerus and fibula of P. chiniquensis are mostly composed of strongly arranged parallel-fibred bone and a laminar vascularisation. The minimal ages obtained correspond to 9 years for F. tenax (based on the maximum number of growth marks in the osteoderm) and 4 years for P. chiniquensis (obtained from the highest count of growth marks in the femur and in the humerus). F. tenax attained somatic and skeletal maturity, while P. chiniquensis was near to reaching skeletal and sexual maturity, but it was somatically immature. The overall rapid growth rate and the high and uniform vascularisation seems to imply that these features are common in most of "rauisuchians", except in P. chiniquensis.

Bone histology of Neogene angulate tortoises (Testudines: Testudinidae) from South Africa: palaeobiological and skeletochronological implications.

Here we examine the tibial microstructure of modern and fossil angulate tortoises to assess the histology and growth from the late Miocene-early Pliocene, Pleistocene through to modern forms. The cross-sections of all the tibiae sampled revealed highly vascularized, uninterrupted, fibrolamellar bone tissue during early ontogeny, which suggests that early growth was fast. However, later in ontogeny, growth was slower, as indicated by the deposition of parallel-fibred bone tissue in the outer cortex, and even ceased periodically, as indicated by lines of arrested growth. Comparative analyses of the growth rates of the tortoises from different time periods showed that the tortoises from the late Miocene-early Pliocene Langebaanweg locality and from Diepkloof Rock Shelter had relatively slower growth rates under less optimal growth conditions. Additionally, these prehistoric specimens show extensive remodelling, and several generations of secondary osteons further suggest functional and/or metabolic stresses on the skeleton. Palaeoenvironmental reconstructions suggest that it was mostly cooler and drier with seasonal fluctuations in late Miocene-early Pliocene, and it is likely that Chersina responded to these conditions by having a lower rate of growth as compared with their modern counterparts, which thrive in the current prevailing more favourable Mediterranean type of climate.

Comparative bone microstructure of two non-mammaliaform cynodonts from the Late Triassic (Carnian) Chañares formation of Northwestern Argentina

ABSTRACT The aim of the present contribution is to examine the bone histology of two non-mammaliaform cynodonts recovered from the Late Triassic (Carnian) Chañares Formation, at Los Chañares locality, La Rioja province, Northwestern Argentina. The taxa analysed here are the Traversodontidae Massetognathus pascuali and the Probainognathidae Probainognathus jenseni, both forms known from multiple skeletons. Our osteohistological results reveal an uninterrupted growth pattern and high intraspecific histovariability in Massetognathus. On the other hand, Probainognathus presents cyclical growth, a cortex dominated by parallel-fibred bone tissue and scarce vascularisation. In spite of being derived non-mammaliaform eucynodonts, Massetognathus and Probainognathus show high growth plasticity, a feature usually regarded as plesiomorphic for cynodonts. Both taxa show a thick cortex in the long bones, a feature that could be linked to counteract the strong flexion forces during digging.

Interelemental osteohistological variation in Massospondylus carinatus and its implications for locomotion.

Massospondylus carinatus Owen, 1854 is an iconic basal sauropodomorph dinosaur from the Early Jurassic of southern Africa. Over 200 specimens have been referred to this taxon, spanning the entire ontogenetic series from embryo to adult. Consequently, it provides an ideal sample for investigating dinosaur developmental biology, including growth patterns and growth rates, through osteohistological analysis. Massospondylus carinatus was the first early-branching sauropodomorph dinosaur for which a femoral growth series was sampled. Since then, growth series of other non-avian dinosaur taxa have shown that growth plasticity, interelemental variation, and ontogenetic locomotory shifts can complicate our understanding of growth curves and patterns. To investigate these questions further, it is necessary to sample multiple skeletal elements from multiple individuals across a large range of sizes, something that is often hindered by the incompleteness of the fossil record. Here, we conducted a broad, multielement osteohistological study of long bones (excluding metapodials) from 27 specimens of Massospondylus carinatus that span its ontogenetic series. Our study reveals substantial variations in growth history. A cyclical woven-parallel complex is the predominant bone tissue pattern during early and mid-ontogeny, which transitions to slower forming parallel-fibred bone during very late ontogeny. The bone tissue is interrupted by irregularly spaced cyclical growth marks (CGMs) including lines of arrested growth indicating temporary cessations in growth. These CGMs show that the previously recorded femoral growth plasticity is also visible in other long bones, with a poor correlation between body size (measured by midshaft circumference) and CGM numbers. Furthermore, we found that the growth trajectory for an individual can vary depending on which limb element is studied. This makes the establishment of an accurate growth curve and determination of the onset of reproductive maturity difficult for this taxon. Finally, we found no evidence of differential growth rates in forelimb vs hindlimb samples from the same individual, providing further evidence falsifying hypothesised ontogenetic postural shifts in Massospondylus carinatus.

Histological Diversity And Evolution Of Lizard Osteoderms

Many reptiles reinforce the dermis with discrete mineralized organs known as osteoderms. Among lizards, osteoderms demonstrate species‐specific differences in size, shape, and distribution across the body. Whether osteoderms also vary in details of tissue composition, including the organization of the fibrillar matrix, remains unclear. Here, we investigate osteoderm histology in three species‐rich lizard groups: gekkotans (geckos), scincids (skinks), and anguimorphans (anguids, helodermatids, and related taxa). With one possible exception, all lizard osteoderms are dominated by bone tissue. Unexpectedly, representative members of all the major groups develop an enigmatic, collagen‐poor capping tissue. Gekkotan osteoderms are rare (<2% of species) and demonstrate considerable histological heterogeneity between species. This includes variation in bone matrices, and the development of additional (non‐osseous) skeletal tissues. In the gecko genus Geckolepis, putative osteoderms appear to lack bone and instead are composed of dense collagen plates capped by a vitreous collagen‐poor tissue. Unlike geckos, osteoderms are common to virtually all skinks. Most skink osteoderms are compound elements, composed of multiple conjoined smaller plates (referred to as osteodermites). Histologically, skink osteoderms are dominated by lamellar bone with well‐organized Sharpey’s fibres linking adjacent plates together and some woven and parallel‐fibred bone. Most species develop the collagen‐poor capping tissue, particularly where adjacent osteodermites articulate with one another. Osteoderms are also common to many anguimorphans, but the histology varies considerably between taxa. While anguid and helodermatid osteoderms demonstrate multiple bone matrices (woven‐fibred, parallel‐fibred, lamellar bone, and Sharpey‐fibred), Shinisaurus osteoderms are primarily woven‐fibred and Sharpey‐fibred bone, and those of Varanus are mostly parallel‐fibred bone. Expression of the capping tissue also varies, from thick and well‐defined (Heloderma) to discontinuous (e.g., several anguids) or entirely absent (Shinisaurus, Varanus). We propose that the histological diversity observed in lizard osteoderms is driven by factors including phylogeny, ecology, and function. The identity of the capping tissue remains unresolved, but our findings reveal it is far more widespread than previously considered. The results presented here provide important steps towards understanding the diversity and evolution of the skeleton in the skin.

Palaeobiological inferences of the aetosaur Aetosauroides scagliai (Archosauria: Pseudosuchia) based on microstructural analyses of its appendicular bones

ABSTRACT Aetosaurs were a group of armoured pseudosuchians, recorded in most of the Upper Triassic continental deposits worldwide. Several osteohistological contributions of aetosaurs focused on their osteoderms, but rarely on appendicular bones. Here, we analyse the microstructure of the humerus, femur and tibia of Aetosauroides scagliai (specimens PVL 2073 [holotype] and PVL 2052). These exhibit cortical bone formed by highly vascularised fibrolamellar bone present in the inner portion of the cortex, mixed with scarce parallel-fibred bone. Also, they show parallel-fibred bone in the outermost portion of the cortex. A general growth pattern that includes a first rapid stage followed by a slow stage is reported. Nevertheless, the growth rate and the presence of parallel-fibred bone embedded in fibrolamellar bone layers recognise more variation within Aetosauria. The value of appendicular bones and osteoderms as age estimators is variable, the first being useful in early stages, and in late stages the osteoderms are better (based on the particular growth of osteoderms). Through morphological (neurocentral sutures) and histological (EFS absent) information, the holotype (PVL 2073) was recovered between juvenile and subadult stages. Using a statistical model that combines microanatomical and morphological data, a terrestrial lifestyle is inferred for Aetosauroides, which concur with previous analyses.

Osteohistology of non-mammalian therapsids from Puesto Viejo Group, Argentina

ABSTRACT The Puesto Viejo Group is an important unit for the knowledge Triassic faunas especially non-mammalian therapsids. This group crops out in the San Rafael Block, southwest Mendoza, Argentina. It is composed of the basal Quebrada de los Fósiles Formation (QF) which is overlaid by the Río Seco de la Quebrada Formation (RSQ). This report comprises tetrapod remains from the RSQ Formation at the Quebrada del Durazno area that includes, until now, exclusively therapsids. The aim of this study is to analyse the bone microstructure of cynodonts and dicynodonts to elucidate the histological features and growth patterns. Histological thin sections made in ribs and appendicular bones reveal a predominance of fibrolamellar bone tissue suggesting an overall fast bone growth, however, the presence of parallel-fibred bone suggest periods of slowing down of the growth. The existence of growth marks indicates periodic interruptions of growth, which is likely the expression of their ability to rapid respond to environmental changes. This contribution offers new insights into the bone microstructure of tetrapod assemblages from RSQ Formation during the Upper Triassic span.

Mandible histology in Metoposaurus krasiejowensis (Temnospondyli, Stereospondyli) from the Upper Triassic of Poland.

Recent studies that have systematically augmented our knowledge of dermal bones of the Late Triassic temnospondyl amphibian Metoposaurus krasiejowensis have mostly focused on shoulder girdle elements and the skull. So far, histological data on the mandible are still scant. For the present study, two mandibles have been examined, using 50 standard thin sections. Dermal bones of the mandible reveal a uniform diploë structure, with the external cortex consisting of moderately vascularised, parallel-fibred bone, as well as a distinct alternation of thick zones and thinner annuli. Dense bundles of well-mineralised Sharpey’s fibres are seen in the external cortex over the entire length of the mandible. The trabecular middle region is highly porous and well vascularised, showing small primary vascular canals and more numerous secondary osteons; irregular erosion spaces occur in large numbers as well. The thin and poorly vascular internal cortex consists of parallel-fibred bone. The articular is not a dermal bone in origin, having been formed of a thin layer of avascular cortex and a very extensive, trabecular middle region. In contrast to the dermal bones of the mandible, the articular developed from a cartilaginous precursor, as evidenced by numerous remains of calcified cartilage in the central parts of the bone. Histological variability is extremely high along the mandible, its anterior part being characterised by high compactness and biomechanically good resistance in contrast to the highly porous posterior parts. Distinct variations of bone thickness and degree of bone porosity in specific areas of the mandible, may be due to local differences in biomechanics during feeding. The microstructure of the mandible corroborates a previous study of the active and ambush predation strategy in metoposaurids.

Microanatomy and histology of frontal bones in two species of Albanerpeton sensu lato (Lissamphibia: Albanerpetontidae) from the Upper Cretaceous Oldman Formation in southeastern Alberta, Canada

ABSTRACT Albanerpetontids are an extinct clade of salamander-like lissamphibians characterised by features such as fused frontals and sculptured skull roof bones. Here we provide a histological analysis of the fused and sculptured frontals in two Late Cretaceous species of the type genus Albanerpeton sensu lato that differ in body size: moderate-sized ‘Al.’ gracile and larger-sized ‘Al.’ nexuosum. Despite general similarities in microanatomy/histology (no sutural trace remains between left and right frontals; sculpture formed by appositional growth of bone; predominance of parallel-fibred bone; and lack of growth marks, secondary osteons, and Sharpey’s fibres) and the presence of sub-ventrolateral crest canals (possible albanerpetontid synapomorphy), ‘Al.’ nexuosum differs from ‘Al.’ gracile in exhibiting: a distinct three-layered diploë structure; bone remodelling; a higher degree of vascularisation; and more numerous osteocytic lacunae. We suggest that (1) histological differences between ‘Al.’ gracile and ‘Al.’ nexuosum are related to differences in absolute body sizes and relative development of sculpture and robustness of frontals and (2) the sub-ventrolateral crest canals in albanerpetontids are homologous to the canalis arteriae orbitonasalis in modern anurans. Two features (internal vascularisation and lack of Sharpey’s fibres) support a recent suggestion that albanerpetontids may have relied entirely on cutaneous respiration.

COMPARISON OF BONE HEALING AFTER THE USE OF A THREE DIMENSIONAL MINIPLATE AND A CONVENTIONAL MINIPLATE IN MANDIBULAR BODY OSTEOTOMY (AN EXPERIMENTAL STUDY)

Introduction: Mandibular fractures comprise most of the traumatic injuries treated by an oral and maxillofacial surgeon. The aim of fracture treatment is to restore the architecture and function of bone. Many techniques for treating mandibular fractures have evolved over the years. The design of the three dimensional (3D) plate is conceptually that of 2 linear plates connected by reinforcing vertical struts providing greater resistance against gap opening at the inferior border with biting forces. Objectives: To compare histologically bone healing of mandibular body osteotomy after using either a 3D miniplate or conventional miniplates. Materials and methods: This study was conducted on twelve healthy dogs. An iatrogenic osteotomy defect was performed intra-orally between the mandibular 3rd and 4th premolars using a surgical micro reciprocating saw. The fragments will be reduced bimanually and stabilized: Group A (six dogs): 3D miniplates. Group B (six dogs): conventional miniplates. The dogs were euthanized at 2 and 6 weeks postoperatively and the osteotomy sites were excised, divided into three thirds (Tension Third, TT; Intermediary Third, IT; Compression Third, CT) and prepared for histological analysis. Results:The osteotomy site was completely filled with parallel-fibred bone, with more newly formed Haversian systems in Group A compared with Group B, bone repair did not reach the typical aspects of the parent lamellar bone. The percentages of newly formed bone at 2 weeks in group A according to thirds (TT, IT and CT) that reported have a statistically significant difference in comparison with group B. At 6 weeks, there were no statistically significant differences between the two groups among the TT and CT but there was a statistically significant difference (p4 = 0.039) in the IT. Conclusions: 3-D plates could be a better option than conventional miniplates.

Bone microstructure of the pareiasaur Provelosaurus americanus from the Middle Permian of southern Brazil

ABSTRACT Provelosaurus americanus from the Guadalupian of Brazil, is the only species of pareiasaur known from South America and its studies are limited to anatomical descriptions. Here, we examined the microstructure of limb bones, a rib fragment and osteoderms of P. americanus, aiming to answer questions related to its paleobiology. The bone tissues of this specimen comprise poorly vascularised parallel-fibred bone interrupted by growth marks indicating slow, cyclical growth. This is consistent with the pattern found in other pareisaurs from South Africa. However, the space between the LAGs are irregular and there is no clear decreasing in vascularity toward the periphery of the bone, suggesting that it did not reach skeletal maturity. The highest number of LAGs was found in the rib, suggesting that our sample lived a minimum of fifteen years. The osteoderms present a trilaminar structure consisting of a diploe composed of a cancellous core surrounded by two cortical layers. A diploe is absent from the osteoderms of previously studied South African pareiasaurs. The osteogenesis of the osteoderms is intramembraneous, but a metaplastic ossification during the early developmental stage cannot be entirely discarded, due to extensive remodeling in the central core that could have destroyed potential metaplastic tissue.

Ontogenetic changes in long-bone histology of the cryptobranchid Eoscapherpeton asiaticum (Amphibia: Caudata) from the Late Cretaceous of Uzbekistan

This paper presents histological data on the long bones of different size (age) individuals of the basal cryptobranchid salamander Eoscapherpeton asiaticum from the Upper Cretaceous (Turonian) of Uzbekistan. E. asiaticum is similar to modern members of Cryptobranchidae in being relatively large (estimated body length up to 50–60cm), aquatic, and neotenic. The analysis of growth series of femora demonstrates a significant histological maturation during ontogeny, expressed by the progressive appearance of highly organized parallel-fibred bone in the peripheral part of the periosteal cortex, appearance and increasing number of bone remodeling features, progressive resorption of calcified cartilage in the diaphyseal areas and formation of endochondral bone lining the erosion cavities in the calcified cartilage, progressive thickening of endosteal inner circumferential layer and increasing of vascularity and appearance of vascular network of longitudinal and oblique canals in the cortex. These ontogenetic changes in the long-bone histology of E. asiaticum generally correspond to those of other salamanders, except the appearance of the vascular network in the periosteal cortex — the feature that is characteristic for cryptobranchids and connected with their large body size. According to new data, the large Cenozoic cryptobranchids appear to have attained their larger size by extending the skeletal growth period.

Bone microanatomy of Pepesuchus deiseae (Mesoeucrocodylia, Peirosauridae) reveals a mature individual from the Upper Cretaceous of Brazil

Peirosauridae is placed within Notosuchia with members found in Cretaceous deposits of Africa and South America. These crocodyliforms were considered most terrestrial predators, according to their morphological features. Our data bring contributions to infer about peirosaurids habits from the bone histology and to inform the autoecology of the Pepesuchus deiseae (MN 7466-V). We prepared thin sections of the appendicular bones and an osteoderm from MN 7466-V. Both metacarpals show a broad cortex and a tiny marrow cavity, one has seven cyclical growth marks and Haversian bone in the endosteal region. The ulnare has a highly porous bone with several resorption cavities. The osteoderm is composed of parallel-fibred bone (PFB) forming the basal and external cortex and woven-fibred bone (WFB) constituting the internal core. The tibia shows the presence of external fundamental system (EFS) and the cortex presents WFB and PFB intercalated. The metacarpals exhibit osteosclerosis and the ulnare shows osteoporotic-like bone. In the metacarpals, the occurrence of bone mass increase would have implied rather life in shallow-water environments. The ulnare shows a reduction in bone mass, providing a better maneuverability between its joints with the carpal and ulna. The tibia was the only element indicating the specimen reached the somatic maturity by the presence of EFS. Therefore, this bone was used as a proxy to the maximum growth rate. Currently, the presence of WFB in advanced ontogenetic stage indicates a peculiar feature solely recorded in eusuchian long bones. The resorption cavities in the internal core of the osteoderm probably indicate bone resorption for calcium mobilization during the eggshell formation. In addition, the Haversian bone is an indication that metacarpal A belonged to mature individual, which had accomplished some ovogenetic cycles. In conclusion, our analyses suggest that MN 7466-V was an adult and mature female individual.

Linderochelys rinconensis (Testudines: Pan-Chelidae) from the Upper Cretaceous of northern Patagonia: New insights from shell bone histology, morphology and diagenetic implications

The panchelid turtle Linderochelys rinconensis de la Fuente, Calvo and González Riga, 2007 was described on the basis of several postcranial elements from the Rio Neuquén Subgroup of Loma del Lindero site, near Rincon de los Sauces (Neuquén, Argentina). New preparation of the type material revealed new parts of the animal, including cervical vertebrae, parts of the carapace and a right tibia. The study of the newly recovered material herein improves our knowledge of the macroanatomy of this taxon. The first costal shows an axillary buttress similar to Australasian chelids and the cervical vertebrae morphology suggests a short-necked chelid condition. The re-examination of the type material of Linderochelys and the discovery of new material allowed new studies on microanatomy, osteohistology and diagenesis. Osteohistology suggests some similarities with Patagonian Cretaceous and Palaeocene Yaminuechelys, based on the presence of parallel-fibred bone in the external cortex and a much thinner internal cortex. Paleoecological studies on the proportions of forelimbs and microanatomy point towards aquatic or semi-aquatic habits, closely related to the condition seen in some extant South American chelids, such as Phrynops hilarii, Phrynops geoffroanus, and Mesoclemmys gibba. The fossil-diagenetic processes inferred substitution, fracturing, plastic deformation and permineralization events. Combined analyses through scanning electron microscopy with energy-dispersive X-ray spectrometry and petrographic studies revealed the substitution of hydroxyapatite by francolite. Permineralization stages include infilling of vascular canals, trabeculae and fractures with iron oxides and carbonate minerals during their burial history.

Turtle shell bone and osteoderm histology of Mesozoic and Cenozoic stem-trionychian Adocidae and Nanhsiungchelyidae (Cryptodira: Adocusia) from Central Asia, Mongolia, and North America

Abstract. The record of fossil turtles from the Cretaceous and Cenozoic of Asia and North America is very rich, including several lineages of cryptodiran turtles. Here we survey the shell bone histology of two important closely related groups of stem trionychians, the Adocidae and Nanhsiungchelyidae, which have representatives in both Asia and North America. All studied taxa show shell bones in which the diploe is framed by well-developed cortical compact bone layers. Taxa of both groups also express external regular surface sculpturing of their shell bones, and in the case of the nanhsiungchelyid genus Basilemys also on the osteoderms, which is also reflected in the internal histological bone structures. Besides similarities of the regular ornamentation patterns, both groups share a number of microanatomical and histological characters such as the zonation of external cortex with rather homogeneous fine-fibred interwoven structural fibres (ISF) in the more internal zone and a dominance of vertically oriented fibres in the ISF and the presence of growth marks in the more external zone. On the other hand, growth marks, i.e. lines of arrested growth, which are visible as wavy lines in thin sections, extend subparallel to the external bone surface in adocids, but they are not parallel/subparallel in nanhsiungchelyids. Thickness and structure of bone trabeculae in the cancellous interior regions depends on the shell bone thickness of the individual samples. The internal cortices of all taxa except the North American samples of Adocus usually consist of parallel-fibred bone that locally grades into lamellar bone. Secondary bone remodelling is more frequent in nanhsiungchelyids compared to adocids, and Sharpey's fibres that extend perpendicular to the bone margins extending across subparallel growth marks are more commonly found in adocids. In addition, bone histology served to identify trionychid specimens in the adocid and nanhsiungchelyid samples, especially as bone surface sculpturing patterns were weathered or eroded in those cases. The histological data present thus supplement the numerous previously reported differences in external shell morphology between adocids and nanhsiungchelyids on the one hand and trionychids and carettochelyids on the other.

Histology of sculptured cranial dermal bones of the stem salamander Kokartus honorarius (Amphibia: Caudata) from the Middle Jurassic of Kyrgyzstan

Kokartus honorarius from the Middle Jurassic (Bathonian) of Kyrgyzstan is one of the oldest and basalmost salamanders in the fossil record, and this taxon is useful for documenting a primitive caudate condition and patterns of character state transformations within Caudata. Here, we provide a histological analysis of its dermal bones based on two thin sections of a fragment of a sculptured dermal skull roof bone (squamosal?). The dermal bone histology of Kokartus is similar to that of basal crown-group tetrapods (e.g. temnospondyls, lepospondyls and seymouriamorphs) in the following features: the presence of basic pattern of organisation of dermal bones (diploë structure); the cancellous middle region occupies larger space than the compact cortices; active remodelling with the formation of numerous erosion bays and secondary osteons; high number of vascular canals with the formation a dense vascular network (= ‘rete vasculosum’); and parallel-fibred bone is a dominant type of primary bone matrix. Crown-group salamanders retained the diploë structure, but demonstrate features of simplification (in comparison with Kokartus): absence of pronounced sculpture, lower degree of vascularisation and lower degree of remodelling. The simplification of the histological structure is a derived feature for the Caudata which appeared during the transition from stem-group to crown-group salamanders.

Shell bone histology of the long-necked chelid Yaminuechelys (Testudines: Pleurodira) from the late Cretaceous-early Palaeocene of Patagonia with comments on the histogenesis of bone ornamentation.

Yaminuechelys is a long-necked chelid turtle whose remains have been recovered from outcrops of the Santonian-Maastrichtian and Danian of South America. With the purpose of providing data about shell sculpturing origin and palaeoecology, the bone histology of several shell elements (including neural, costal, peripheral and plastral plates) of Yaminuechelys is described herein. Histological analysis reveals that Yaminuechelys shares with Chelidae the presence of interwoven structural fibre bundles in the external cortex, and parallel-fibred bone of the internal cortex. The presence of resorption lines in several samples indicates that the particular ornamentation of the external surfaces originated, at least in part, by focalized resorption and new bone deposition. This mechanism for ornamentation origin and maintenance is here described for the first time in a turtle. Compactness of the shell bones is consistent with an aquatic habitat, which supports previous hypothesis based on palaeoenvironmental and morphological data.

A new adult specimen of the basalmost ornithuromorph bird Archaeorhynchus spathula (Aves: Ornithuromorpha) and its implications for early avian ontogeny

Most living birds characteristically grow rapidly and reach adult size within a year. Nevertheless, little is known about how such an advanced developmental strategy evolved despite many discoveries of early fossil birds. Here we assess the long-bone histology from a new adult specimen of Archaeorhynchus spathula, the basalmost taxon of Ornithuromorpha. Ornithuromorpha is the most inclusive clade containing extant birds but not the Mesozoic Enantiornithes. Histological analysis reveals that the cortex is composed of parallel-fibred bone with three lines of arrested growth, indicative of slow and annually interrupted growth for this taxon. Such bone histology is significantly different from that of other known basal ornithuromorphs, but resembles that of enantiornithines, which leads us to suggest protracted slow growth in the common ancestor of Ornithuromorpha and Enantiornithes. The fusion sequence of the tarsometatarsus between Enantiornithes and Ornithuromorpha has long been hypothesized to be different and regarded as indicative that enantiornithines are not close relatives of ornithuromorphs. Due to a lack of fossils recording early ontogenetic stages, little is known about the development of the tarsometatarsus in basal ornithuromorphs, making this hypothesis impossible to test. Here we show that the fusion sequence of the tarsometatarsus in Archaeorhynchus is similar to that of enantiornithines, and that the proximal-early fusion in tarsometatarsus represents a plesiomorphic trait for basal birds. Our findings also shed light on ontogenetic variation of sternal morphology, which highlights the importance of ontogeny in the taxonomic and phylogenetic study of early birds.