A theropod trackway providing evidence of a pathological foot from the exceptional locality of Las Hoyas (upper Barremian, Serranía de Cuenca, Spain).

We describe a trackway (LH-Mg-10-16) occurring in laminated carbonated limestones of the Las Hoyas locality, Serranía de Cuenca, Spain. It is unmistakably a large theropod dinosaur trackway encompassing two unusual aspects, namely, wide-steps, and a set of equally deformed left footprints (with a dislocated digit). The layer also preserves other vertebrate trails (fish Undichna) and different impressions in the sediment. To address these complex settings, we devised a multidisciplinary approach, including the ichnological and taphonomical descriptions, characterisation of the rock lithofacies using thin-sections, 3D structured-light digitalisation with a high precision of 200-400 µm, and a geometric morphometric comparison with a large sample of bipedal dinosaur trackways. Sedimentary analyses showed that the trackway was produced in a humid, benthonic microbial mat, the consistency and plasticity of which enabled the preservation of the details of the movement of the animal. The results of the geometric analysis indicate that the "wide-steps" of the trackway is not unusual compared to other trackways, providing evidence that it was made by a single individual with an estimated hip height approximately 2 m. Analogous pathologies in extant archosaurs that yield the combination of wide steps and deformed digits in the same trackway were considered. All results mutually support the hypothesis that a large theropod dinosaur, with a pathological foot, generated the trackway as it crossed an area of shallow water while slowly walking towards the main water source, thus stepping steadily over the benthonic mat over which multiple fish were swimming.

Helminth eggs from early cretaceous faeces.

The exceptional fossil site of Las Hoyas (upper Barremian, Cuenca, Spain) yields abundant small to medium vertebrate coprolites, hindering the search for parasites. We studied the contents of 29 coprolites that were previously classified into distinct morphotypes. Several parasitic eggs were retrieved from two of these coprolites, confirming the second record of digenea trematode eggs and nematode (ascaridid) eggs from an Early Cretaceous locality. The cylindrical coprolite containing anisakid eggs was likely produced by a crocodylomorph as the parasite host, whereas the bump-headed lace coprolite indicates the role of a fish as an intermediary or definitive host of the trematodes and ascaridids. These trace and body fossils show that the Las Hoyas 126-129 Ma lacustrine ecosystem documents the early connection between basal Gonorynchiformes fish and digenetic trematodes.

A new goniopholidid crocodyliform, Hulkepholis rori sp. nov. from the Camarillas Formation (early Barremian) in Galve, Spain).

BACKGROUND: The neosuchian crocodyliform genus Hulkepholis constitutes the longirostral lineage of the European Goniopholididae. It comprises two species ranging from the Valanginian of southern England to the lower Albian of the northern Teruel (Spain). A new species of Hulkepholis is described based on a partially complete skull from the lower Barremian Camarillas Formation. We investigate its phylogenetic position and the palatal patterns among members of Goniopholididae and the closely related Thalattosuchia and Tethysuchia. METHODS: Phylogenetic relationships were investigated with two matrices using a previously published dataset as the basis: the first differed only by the addition of the new species, the second had newly discovered states for 11 characters, the new species plus several additional specimens of Hulkepholis and Anteophthalmosuchus. Both matrices were processed using TNT v. 1.1, in a heuristic analysis of maximum parsimony, with tree bisection and reconnection 1,000 random addition replicates and saving the 10 most parsimonious trees per replicate, and up to 10 suboptimal trees to calculate Bremer supports. The skull geometry of nine species from Thalattosuchia, Tethysuchia and Goniopholididae was explored to test shape variation between the rostral and postrostral modules, and to visualize the differences on the secondary palate. A set of 18 landmarks was used to delimit significant anatomical features, and the skulls were isotropically scaled using Adobe Illustrator, with the longest skull (Sarcosuchus imperator) as the baseline for comparison. RESULTS: The European lineages of goniopholidids are two clades (Nannosuchus + Goniopholis) plus (Hulkepholis + Anteophthalmosuchus). The new species, Hulkepholis rori sp. nov, shares with the latter clade the following apormorphies: a long anterolateral postorbital process, postorbital process almost reaching the anterior jugal ramus, and basioccipital tubera with lateral edges turned posteriorly. Anteophthalmosuchus was found to be monophyletic, and Hulkepholis paraphyletic due to the poor preservation of H. willetti. Hulkepholis rori is distinguished by having vascular fossae and a mid-protuberance on the ventral surface of the basioccipital, and wide internal fossae in the quadrate. Among Goniopholididae differences on the secondary palate are the presence of a palatal cleft, the narrowness of the secondary choana, and a wide foramen of the median pharyngeal tube. CONCLUSIONS: The new species is the earliest Hulkepholis from the Iberian Peninsula. New characters have been recognized in the organization of the palate and in the occipital region raising unexpected questions on the evolution of Goniopholididae. The set of palatal characters is discussed as part of a singular palatogenesis in Goniopholididae. The protruding occipital areas suggest that the longirostral Hulkepholis would have had an aquatic lifestyle with particular neck and skull movements.

A novel approach for the metric analysis of fern fronds: Growth and architecture of the Mesozoic fern Weichselia reticulata in the light of modern ferns.

The architecture of primary and secondary pinnae of the fossil fern Weichselia reticulata (C. Stokes et Webb) Fontaine is studied based on 28 large specimens from the upper Barremian La Huérguina Formation of the fossil locality Las Hoyas, Cuenca, Spain. The study of the specimens is performed through a morphometric analysis consisting in a reformulation of the Branching Algorithms method of shape description including measurements (insertion angle, distance between pinnae, first segment length and rachis width) and ratios (interval ratio, branching ratio and tapering ratio). A protocol to relocate isolated fragments of fossil pinnae is also established using the interval ratio (distance between pinnae/ previous distance between pinnae) and insertion angle of the pinnae. All specimens show a similar architecture, having elliptic primary pinnae with a sinuous apically tapering primary rachis and triangular secondary pinnae with pinnules of different morphologies. The analysis of the architecture allows to propose that the position of the frond was plagiotropic and that the frond growth was basiplastic for the petiole head and acroplastic for the primary pinnae. The metric method is applied to explore the architecture of four living fern species (Angiopteris evecta (Forst.) Hoffm., Matonia pectinata R.Br., Sphaeropteris cooperi (F.Muell.) R.M.Tryon, and Woodwardia unigemmata (Makino) Nakai). Weichselia architecture results extremely ordered and regular in comparison with the primary pinnae variation of the living species.

Fully fledged enantiornithine hatchling revealed by Laser-Stimulated Fluorescence supports precocial nesting behavior.

Laser-Stimulated Fluorescence (LSF) is used to identify fully fledged feathering in the hatchling enantiornithine bird specimen MPCM-LH-26189, supporting precocial nesting behavior in this extinct group. The LSF results include the detection of a long pennaceous wing feather as well as cover feathers around the body. The LSF technique showed improved detection limits over and above synchrotron and UV imaging which had both been performed on this specimen. The findings underscore the value of using a wide range of analytical techniques.

Exceptional coprolite association from the Early Cretaceous continental Lagerstätte of Las Hoyas, Cuenca, Spain.

Coprolites are some of the most abundant fossils at the Las Hoyas site, a well-known Early Cretaceous Konservat-Lagerstätte located in Cuenca, central Spain. The coprolite association is described, introducing taphonomic features and sedimentological properties. This study is based on a subsample of 433 fossils selected from some 2000 specimens collected. The taphonomic features of the coprolites show that their integrity, absence of desiccation marks, and volume are congruent with faeces produced and deposited in an aquatic ecosystem, which were immediately covered by microbial mats. The highest abundance of coprolites, 96%, occurs in layers linked to the presence of microbial mats. Consequently, it is likely that coprolites are taphonomically autochthonous. A dichotomous key has been made in order to delimit the morphotypes. The key is based on (1) presence/absence of spiral marks, (2) morphology of coprolite ends, including polarity, expansion, and surface, and (3) overall shape, outline, diameter, and constrictions. Twelve different morphotypes are distinguished: spiral, circular, irregular, elongated, rosary, ellipsoidal, cylinder, bump-headed lace, fir-tree, cone, straight lace, and thin lace. The association is dominated by thin-lace and cylinder morphotypes. The sizes, inclusions, and EDX analyses indicate that the Las Hoyas coprolites correspond mostly to carnivorous producers with ichthyophagous diets, as crocodiles, urodelans and different kind of fishes.

Involvement of microbial mats in early fossilization by decay delay and formation of impressions and replicas of vertebrates and invertebrates.

Microbial mats have been hypothesized to improve the persistence and the preservation of organic remains during fossilization processes. We test this hypothesis with long-term experiments (up to 5.5 years) using invertebrate and vertebrate corpses. Once placed on mats, the microbial community coats the corpses and forms a three-dimensional sarcophagus composed of microbial cells and exopolymeric substances (EPS). This coverage provides a template for i) moulding superficial features, resulting in negative impressions, and ii) generating replicas. The impressions of fly setulae, fish scales and frog skin verrucae are shaped mainly by small cells in an EPS matrix. Microbes also replicate delicate structures such as the three successive layers that compose a fish eye. The sarcophagus protects the body integrity, allowing the persistence of inner organs such as the ovaries and digestive apparatus in flies, the swim bladder and muscles in fish, and the bone marrow in frog legs. This study brings strong experimental evidence to the idea that mats favour metazoan fossilization by moulding, replicating and delaying decay. Rapid burial has classically been invoked as a mechanism to explain exceptional preservation. However, mats may play a similar role during early fossilization as they can preserve complex features for a long time.

A Cretaceous eutriconodont and integument evolution in early mammals.

The Mesozoic era (252-66 million years ago), known as the domain of dinosaurs, witnessed a remarkable ecomorphological diversity of early mammals. The key mammalian characteristics originated during this period and were prerequisite for their evolutionary success after extinction of the non-avian dinosaurs 66 million years ago. Many ecomorphotypes familiar to modern mammal fauna evolved independently early in mammalian evolutionary history. Here we report a 125-million-year-old eutriconodontan mammal from Spain with extraordinary preservation of skin and pelage that extends the record of key mammalian integumentary features into the Mesozoic era. The new mammalian specimen exhibits such typical mammalian features as pelage, mane, pinna, and a variety of skin structures: keratinous dermal scutes, protospines composed of hair-like tubules, and compound follicles with primary and secondary hairs. The skin structures of this new Mesozoic mammal encompass the same combination of integumentary features as those evolved independently in other crown Mammalia, with similarly broad structural variations as in extant mammals. Soft tissues in the thorax and abdomen (alveolar lungs and liver) suggest the presence of a muscular diaphragm. The eutriconodont has molariform tooth replacement, ossified Meckel's cartilage of the middle ear, and specialized xenarthrous articulations of posterior dorsal vertebrae, convergent with extant xenarthran mammals, which strengthened the vertebral column for locomotion.

Soft-tissue and dermal arrangement in the wing of an Early Cretaceous bird: Implications for the evolution of avian flight.

Despite a wealth of fossils of Mesozoic birds revealing evidence of plumage and other soft-tissue structures, the epidermal and dermal anatomy of their wing's patagia remain largely unknown. We describe a distal forelimb of an enantiornithine bird from the Lower Cretaceous limestones of Las Hoyas, Spain, which reveals the overall morphology of the integument of the wing and other connective structures associated with the insertion of flight feathers. The integumentary anatomy, and myological and arthrological organization of the new fossil is remarkably similar to that of modern birds, in which a system of small muscles, tendons and ligaments attaches to the follicles of the remigial feathers and maintains the functional integrity of the wing during flight. The new fossil documents the oldest known occurrence of connective tissues in association with the flight feathers of birds. Furthermore, the presence of an essentially modern connective arrangement in the wing of enantiornithines supports the interpretation of these primitive birds as competent fliers.

Morphological integration and functional modularity in the crocodilian skull.

We explored the morphological organization of the skull within Crocodylidae, analyzing functional and phylogenetic interactions between its 2 constituent functional modules: the rostrum and the postrostrum. We used geometric morphometrics to identify localized shape changes, focusing on the differences between the major clades of the crown-group Crocodylia: Alligatoridae and Crocodylidae. We used published bite performance data to correlate rostral function with postrostral morphology. The skull modules appear more integrated within Alligatoridae than within Crocodyliade. Phylogenetic effects on shape variation are more evident in Alligatoridae than in Crocodylidae, where functional parameters concerning the rostral morphology are proportionally more important than phylogeny. Long-snouted species are characterized by low structural performance, which is significantly associated with a reduction of the pterygoid-quadrate cranial nipper, suggesting that the nipper is important for the ingestion of large food items in generalist species. This functional association is coupled with a significant evolutionary allometry at the clade level, while Alligatoridae and Crocodylidae show different degrees of evolutionary allometry for their entire shape and rostrum. The postrostrum is more conservative than the rostrum in terms of morphospace occupation, evolutionary allometry and phylogenetic signal.

3D geometry and quantitative variation of the cervico-thoracic region in Crocodylia.

This study aims to interpret the axial patterning of the crocodylian neck, and to find a potential taxonomic signal that corresponds to vertebral position. Morphological variation in the cervico-thoracic vertebrae is compared in fifteen different crocodylian species using 3D geometric morphometric methods. Multivariate analysis indicated that the pattern of intracolumnar variation was a gradual change in shape of the vertebral series (at the parapophyses, diapophyses, prezygapohyses, and postzygapohyses), in the cervical (C3 to C9) and dorsal (D1-D2) regions which was quite conservative among the crocodylians studied. In spite of this, we also found that intracolumnar shape variation allowed differentiation between two sub regions of the crocodylian neck. Growth is subtly correlated with vertebral shape variation, predicting changes in both the vertebral centrum and the neural spine. Interestingly, the allometric scaling for the pooled sample is equivalently shared by each vertebra studied. However, there were significant taxonomic differences, both in the average shape of the entire neck configuration (regional variation) and by shape variation at each vertebral position (positional variation) among the necks. The average neck vertebra of crocodylids is characterized by a relatively cranio-caudally short neural arch, whereby the spine is relatively longer and pointed orthogonal to the frontal plane. Conversely, the average vertebra in alligatorids has cranio-caudally longer neural spine and arch, with a relatively (dorso-ventrally) shorter spine. At each vertebral position there are significant differences between alligatorids and crocodylids. We discuss that the delayed timing of neurocentral fusion in Alligatoridae possibly explains the observed taxonomic differences.

Shape Variation and Allometry in the Precloacal Vertebral Series of the Snake Daboia russelli (Viperidae) / Variación Morfológica y Alometría de las Vértebras Precloacales en el Ofidio Daboia russelli (Viperidae)

Understanding the variation of the ophidian vertebral morphology is an essential tool in snake paleobiology, but so far this field remains hardly investigated. A major problematic is the still scarce knowledge about the basis of homogeneity of intracolumnar shape variation along the vertebrae of the precloacal region in these animals. For instance, this variation can be overwhelmingly low in cases such as in vipers, for which it seems almost impossible to describe a concrete regionalization of the precloacal region without ambiguity. This study has applied geometric morphometrics to analyze if the shape variation of the vertebrae of the precloacal vertebrae of an adult specimen of Daboia russelli allows differentiating any sort of parcellation within the column of this organism. We have also explored if size is associated with the organization of vertebral shape along the axial skeleton. The multivariate analyses showed that the main pattern of vertebral shape variation in D. russelli concerns the neural spine and the hypapophysis, whereas the shape of the vertebral centrum appears to be nearly invariant along the series. Our analysis also showed that the precloacal region can be sudivided into two portions that merge in a transitional boundary of largest vertebrae in the middle of the column. From this middle region towards the distal ends of the column vertebrae become smaller changing their shapes in two antithetical ways.

Entender la variación en la morfología vertebral de los ofidios es crucial para la paleobiología del grupo pero, hasta ahora, este campo está poco investigado. Uno de los principales problemas es el escaso conocimiento sobre las bases de la homogeneidad en la variación de la forma a lo largo de la región precloacal en estos animales. Por ejemplo, en el caso de las víboras, dicha variación puede ser muy pequeña lo cual hace casi imposible la descripción de una regionalización precisa sin ambigüedad. En este estudio se ha aplicado morfometría geométrica para analizar si la variación de la forma vertebral de un individuo adulto de la especie Daboia russelli permite subdividir la región precloacal. Además, hemos explorado si el tamaño está asociado con la organización vertebral a lo largo del esqueleto axial. Los análisis multivariantes han demostrado que el patrón principal de la variación de la forma vertebral está determinado por la espina neural y la hipapofisis, mientras que el centro vertebral varía poco a lo largo de la serie. Nuestro análisis ha mostrado que la región precloacal puede ser dividida en dos series cuya separación está marcada por las vértebras más grandes, posicionadas aproximadamente en la mitad de la columna. Tomando como referencia la mitad de la columna, hacia los extremos distales, las vértebras tienden a ser más pequeñas cambiando su forma de modo antitético.

A new crested pterosaur from the Early Cretaceous of Spain: the first European tapejarid (Pterodactyloidea: Azhdarchoidea).

BACKGROUND: The Tapejaridae is a group of unusual toothless pterosaurs characterized by bizarre cranial crests. From a paleoecological point of view, frugivorous feeding habits have often been suggested for one of its included clades, the Tapejarinae. So far, the presence of these intriguing flying reptiles has been unambiguously documented from Early Cretaceous sites in China and Brazil, where pterosaur fossils are less rare and fragmentary than in similarly-aged European strata. METHODOLOGY/PRINCIPAL FINDINGS: Europejara olcadesorum gen. et sp. nov. is diagnosed by a unique combination of characters including an unusual caudally recurved dentary crest. It represents the oldest known member of Tapejaridae and the oldest known toothless pterosaur. The new taxon documents the earliest stage of the acquisition of this anatomical feature during the evolutionary history of the Pterodactyloidea. This innovation may have been linked to the development of new feeding strategies. CONCLUSION/SIGNIFICANCE: The discovery of Europejara in the Barremian of the Iberian Peninsula reveals an earlier and broader global distribution of tapejarids, suggesting a Eurasian origin of this group. It adds to the poorly known pterosaur fauna of the Las Hoyas locality and contributes to a better understanding of the paleoecology of this Konservat-Lagerstätte. Finally, the significance of a probable contribution of tapejarine tapejarids to the early angiosperm dispersal is discussed.

New insight on the anatomy and architecture of the avian neurocranium.

This study aims to disentangle the main features of the avian neurocranium at high taxonomic scales using geometric morphometric tools. When surveying the variation across 60% of avian orders (sampled among 72 individuals), our results verify that the central nervous system has an important influence upon the architecture of the avian neurocranium, as in other very encephalized vertebrates such as mammals. When the avian brain expands relative to the cranial base it causes more "reptilian-like" neurocranial configurations to shape into rounder ones. This rounder appearance is achieved because the cranial base becomes relatively shorter and turns its flexure from concave to convex, at the same time forcing the foramen magnum to reorient ventrally instead of caudally. However, our analyses have also revealed that an important morphological difference between birds resides between the occiput and the cranial roof. This variation was unexpected since it had not been reported thus far, and entertains two plausible interpretations. Although it could be due to a trade-off between the relative sizes of the supraoccipital and the parietal bones, the presence of an additional bone (the intra- or post-parietal) between the latter two bones could also explain the variation congruently. This descriptive insight stresses the need for further developmental studies focused in understanding the evolutionary disparity of the avian neurocranium.

Avian skull morphological evolution: exploring exo- and endocranial covariation with two-block partial least squares.

While rostral variation has been the subject of detailed avian evolutionary research, avian skull organization, characterized by a flexed or extended appearance of the skull, has eventually become neglected by mainstream evolutionary inquiries. This study aims to recapture its significance, evaluating possible functional, phylogenetic and developmental factors that may be underlying it. In order to estimate which, and how, elements of the skull intervene in patterning the skull we tested the statistical interplay between a series of old mid-sagittal angular measurements (mostly endocranial) in combination with newly obtained skull metrics based on landmark superimposition methods (exclusively exocranial shape), by means of the statistic-morphometric technique of two-block partial least squares. As classic literature anticipated, we found that the external appearance of the skull corresponds to the way in which the plane of the caudal cranial base is oriented, in connection with the orientations of the plane of the foramen magnum and of the lateral semicircular canal. The pattern of covariation found between metrics conveys flexed or extended appearances of the skull implicitly within a single and statistically significant dimension of covariation. Marked shape changes with which angles covary concentrate at the supraoccipital bone, the cranial base and the antorbital window, whereas the plane measuring the orientation of the anterior portion of the rostrum does not intervene. Statistical covariance between elements of the caudal cranial base and the occiput inplies that morphological integration underlies avian skull macroevolutionary organization as a by-product of the regional concordance of such correlated elements within the early embryonic chordal domain of mesodermic origin.