Reinterpretation of tuberculate cervical vertebrae of Eocene birds as an exceptional anti-predator adaptation against the mammalian craniocervical killing bite.

We report avian cervical vertebrae from the Quercy fissure fillings in France, which are densely covered with villi-like tubercles. Two of these vertebrae stem from a late Eocene site, another lacks exact stratigraphic data. Similar cervical vertebrae occur in avian species from Eocene fossils sites in Germany and the United Kingdom, but the new fossils are the only three-dimensionally preserved vertebrae with pronounced surface sculpturing. So far, the evolutionary significance of this highly bizarre morphology, which is unknown from extant birds, remained elusive, and even a pathological origin was considered. We note the occurrence of similar structures on the skull of the extant African rodent Lophiomys and detail that the tubercles represent true osteological features and characterize a distinctive clade of Eocene birds (Perplexicervicidae). Micro-computed tomography (µCT) shows the tubercles to be associated with osteosclerosis of the cervical vertebrae, which have a very thick cortex and much fewer trabecles and pneumatic spaces than the cervicals of most extant birds aside from some specialized divers. This unusual morphology is likely to have served for strengthening the vertebral spine in the neck region, and we hypothesize that it represents an anti-predator adaptation against the craniocervical killing bite ("neck bite") that evolved in some groups of mammalian predators. Tuberculate vertebrae are only known from the Eocene of Central Europe, which featured a low predation pressure on birds during that geological epoch, as is evidenced by high numbers of flightless avian species. Strengthening of the cranialmost neck vertebrae would have mitigated attacks by smaller predators with weak bite forces, and we interpret these vertebral specializations as the first evidence of "internal bony armor" in birds.

An Unbiased Molecular Approach Using 3'-UTRs Resolves the Avian Family-Level Tree of Life.

Presumably, due to a rapid early diversification, major parts of the higher-level phylogeny of birds are still resolved controversially in different analyses or are considered unresolvable. To address this problem, we produced an avian tree of life, which includes molecular sequences of one or several species of ∼90% of the currently recognized family-level taxa (429 species, 379 genera) including all 106 family-level taxa of the nonpasserines and 115 of the passerines (Passeriformes). The unconstrained analyses of noncoding 3-prime untranslated region (3'-UTR) sequences and those of coding sequences yielded different trees. In contrast to the coding sequences, the 3'-UTR sequences resulted in a well-resolved and stable tree topology. The 3'-UTR contained, unexpectedly, transcription factor binding motifs that were specific for different higher-level taxa. In this tree, grebes and flamingos are the sister clade of all other Neoaves, which are subdivided into five major clades. All nonpasserine taxa were placed with robust statistical support including the long-time enigmatic hoatzin (Opisthocomiformes), which was found being the sister taxon of the Caprimulgiformes. The comparatively late radiation of family-level clades of the songbirds (oscine Passeriformes) contrasts with the attenuated diversification of nonpasseriform taxa since the early Miocene. This correlates with the evolution of vocal production learning, an important speciation factor, which is ancestral for songbirds and evolved convergent only in hummingbirds and parrots. As 3'-UTR-based phylotranscriptomics resolved the avian family-level tree of life, we suggest that this procedure will also resolve the all-species avian tree of life.

A tiny, long-legged raptor from the early Oligocene of Poland may be the earliest bird-eating diurnal bird of prey.

We report a small hawk-like diurnal bird from the early Oligocene (30-31 million years ago) of Poland. Aviraptor longicrus, n. gen. et sp. is of a size comparable with the smallest extant Accipitridae. The new species is characterized by very long legs, which, together with the small size, suggest an avivorous (bird-eating) feeding behavior. Overall, the new species resembles extant sparrowhawks (Accipiter spp.) in the length proportions of the major limb bones, even though some features indicate that it convergently acquired an Accipiter-like morphology. Most specialized avivores amongst extant accipitrids belong to the taxon Accipiter and predominantly predate small forest passerines; the smallest Accipiter species also hunts hummingbirds. Occurrence of a possibly avivorous raptor in the early Oligocene of Europe is particularly notable because A. longicrus coexisted with the earliest Northern Hemispheric passerines and modern-type hummingbirds. We therefore hypothesize that the diversification of these birds towards the early Oligocene may have triggered the evolution of small-sized avivorous raptors, and the new fossil may exemplify one of the earliest examples of avian predator/prey coevolution.

Pelvis morphology suggests that early Mesozoic birds were too heavy to contact incubate their eggs.

Numerous new fossils have driven an interest in reproduction of early birds, but direct evidence remains elusive. No Mesozoic avian eggs can be unambiguously assigned to a species, which hampers our understanding of the evolution of contact incubation, which is a defining feature of extant birds. Compared to living species, eggs of Mesozoic birds are relatively small, but whether the eggs of Mesozoic birds could actually have borne the weight of a breeding adult has not yet been investigated. We estimated maximal egg breadth for a range of Mesozoic avian taxa from the width of the pelvic canal defined by the pubic symphysis. Known elongation ratios of Mesozoic bird eggs allowed us to predict egg mass and hence the load mass an egg could endure before cracking. These values were compared to the predicted body masses of the adult birds based on skeletal remains. Based on 21 fossil species, we show that for nonornithothoracine birds body mass was 187% of the load mass of the eggs. For Enantiornithes, body mass was 127% greater than the egg load mass, but some early Cretaceous ornithuromorphs were 179% heavier than their eggs could support. Our indirect approach provides the best evidence yet that early birds could not have sat on their eggs without running the risk of causing damage. We suggest that contact incubation evolved comparatively late in birds.

Preservation of uropygial gland lipids in a 48-million-year-old bird.

Although various kinds of organic molecules are known to occur in fossils and rocks, most soft tissue preservation in animals is attributed to melanin or porphyrins. Lipids are particularly stable over time-as diagenetically altered 'geolipids' or as major molecular constituents of kerogen or fossil 'geopolymers'-and may be expected to be preserved in certain vertebrate tissues. Here we analysed lipid residues from the uropygial gland of an early Eocene bird using pyrolysis gas chromatography mass spectroscopy. We found a pattern of aliphatic molecules in the fossil gland that was distinct from the host oil shale sediment matrix and from feathers of the same fossil. The fossil gland contained abundant n-alkenes, n-alkanes and alkylbenzenes with chain lengths greater than 20, as well as functionalized long-chain aldehydes, ketones, alkylnitriles and alkylthiophenes that were not detected in host sediment or fossil feathers. By comparison with modern bird uropygial gland wax esters, we show that these molecular fossils are likely derived from endogenous wax ester fatty alcohols and fatty acids that survived initial decay and underwent early diagenetic geopolymerization. These data demonstrate the high fidelity preservation of the uropygial gland waxes and showcase the resilience of lipids over geologic time and their potential role in the exceptional preservation of lipid-rich tissues of macrofossils.

A new fossil from the mid-Paleocene of New Zealand reveals an unexpected diversity of world's oldest penguins.

We describe leg bones of a giant penguin from the mid-Paleocene Waipara Greensand of New Zealand. The specimens were found at the type locality of Waimanu manneringi and together with this species they constitute the oldest penguin fossils known to date. Tarsometatarsus dimensions indicate a species that reached the size of Anthropornis nordenskjoeldi, one of the largest known penguin species. Stem group penguins therefore attained a giant size very early in their evolution, with this gigantism existing for more than 30 million years. The new fossils are from a species that is phylogenetically more derived than Waimanu, and the unexpected coexistence of Waimanu with more derived stem group Sphenisciformes documents a previously unknown diversity amongst the world's oldest penguins. The characteristic tarsometatarsus shape of penguins evolved early on, and the significant morphological disparity between Waimanu and the new fossil conflicts with recent Paleocene divergence estimates for penguins, suggesting an older, Late Cretaceous, origin.

Eocene fossil is earliest evidence of flower-visiting by birds.

Birds are important pollinators, but the evolutionary history of ornithophily (bird pollination) is poorly known. Here, we report a skeleton of the avian taxon Pumiliornis from the middle Eocene of Messel in Germany with preserved stomach contents containing numerous pollen grains of an eudicotyledonous angiosperm. The skeletal morphology of Pumiliornis is in agreement with this bird having been a, presumably nectarivorous, flower-visitor. It represents the earliest and first direct fossil evidence of flower-visiting by birds and indicates a minimum age of 47 million years for the origin of bird-flower interactions. As Pumiliornis does not belong to any of the modern groups of flower-visiting birds, the origin of ornithophily in some angiosperm lineages may have predated that of their extant avian pollinators.

Earliest and first Northern Hemispheric hoatzin fossils substantiate Old World origin of a "Neotropic endemic".

The recent identification of hoatzins (Opisthocomiformes) in the Miocene of Africa showed part of the evolution of these birds, which are now only found in South America, to have taken place outside the Neotropic region. Here, we describe a new fossil species from the late Eocene of France, which constitutes the earliest fossil record of hoatzins and the first one from the Northern Hemisphere. Protoazin parisiensis gen. et sp. nov. is more closely related to South American Opisthocomiformes than the African taxon Namibiavis and substantiates an Old World origin of hoatzins, as well as a relictual distribution of the single extant species. Although recognition of hoatzins in Europe may challenge their presumed transatlantic dispersal, there are still no North American fossils in support of an alternative, Northern Hemispheric, dispersal route. In addition to Opisthocomiformes, other avian taxa are known from the Cenozoic of Europe, the extant representatives of which are only found in South America. Recognition of hoatzins in the early Cenozoic of Europe is of particular significance because Opisthocomiformes have a fossil record in sub-Saharan Africa, which supports the hypothesis that extinction of at least some of these "South American" groups outside the Neotropic region was not primarily due to climatic factors.

Comparative morphology of the avian bony columella.

In birds, the columella is the only bony element of the sound conducting apparatus, conveying vibrations of the cartilaginous extracolumella to the fluid of the inner ear. Although avian columellar morphology has attracted some attention over the past century, it nonetheless remains poorly described in the literature. The few existing studies mostly focus on morphological descriptions in relatively few taxa, with no taxonomically broad surveys yet published. Here we use observations of columellae from 401 extant bird species to provide a comprehensive survey of columellar morphology in a phylogenetic context. We describe the columellae of several taxa for the first time and identify derived morphologies characterizing higher-level clades based on current phylogenies. In particular, we identify a derived columellar morphology diagnosing a major subclade of Accipitridae. Within Suliformes, we find that Fregatidae, Sulidae, and Phalacrocoracidae share a derived morphology that is absent in Anhingidae, suggesting a secondary reversal. Phylogenetically informed comparisons allow recognition of instances of homoplasy, including the distinctive bulbous columellae in suboscine passerines and taxa belonging to Eucavitaves, and bulging footplates that appear to have evolved at least twice independently in Strigiformes. We consider phylogenetic and functional factors influencing avian columellar morphology, finding that aquatic birds possess small footplates relative to columellar length, possibly related to hearing function in aquatic habitats. By contrast, the functional significance of the distinctive bulbous basal ends of the columellae of certain arboreal landbird taxa remains elusive.

A survey of the uncinate bone and other poorly known ossicles associated with the lacrimal/ectethmoid complex of the avian skull.

In several taxa of Neornithes (crown group birds), the lacrimal/ectethmoid complex exhibits small bones, the comparative osteology of which is poorly studied. Some of these ossicles-which are commonly known as uncinate bones (ossa uncinata or ossa lacrimopalatina)-were already described two centuries ago, but knowledge of their distribution and morphological variability in higher level clades is incomplete. In the present study, a detailed survey of the occurrence of uncinate bones is given, and these ossicles are for the first time reported in the gruiform Psophiidae, some Rallidae, and in the Otidiformes. Their presence in the latter taxon is of particular interest, because in current molecular analyses, the Otidiformes result as close relatives of the Musophagiformes, in which the uncinate bone is particularly large. The uncinate bones of most other neornithine clades, however, appear to have evolved multiple times independently through parallel evolution from the same ligamentous structures. A few earlier authors assumed that the uncinate bone is homologous to the ectopterygoid of non-avian theropods. Although this remains a viable hypothesis, more data on the occurrence of the ectopterygoid in Mesozoic birds are needed for well-supported conclusions. Here, it is noted that the ontogenetic development of the uncinate bone appears to be correlated with that of the ectethmoid, which is another bone in the skull of neornithine birds that is of unknown origin.

The exquisitely preserved integument of Psittacosaurus and the scaly skin of ceratopsian dinosaurs.

The Frankfurt specimen of the early-branching ceratopsian dinosaur Psittacosaurus is remarkable for the exquisite preservation of squamous (scaly) skin and other soft tissues that cover almost its entire body. New observations under Laser-Stimulated Fluorescence (LSF) reveal the complexity of the squamous skin of Psittacosaurus, including several unique features and details of newly detected and previously-described integumentary structures. Variations in the scaly skin are found to be strongly regionalized in Psittacosaurus. For example, feature scales consist of truncated cone-shaped scales on the shoulder, but form a longitudinal row of quadrangular scales on the tail. Re-examined through LSF, the cloaca of Psittacosaurus has a longitudinal opening, or vent; a condition that it shares only with crocodylians. This implies that the cloaca may have had crocodylian-like internal anatomy, including a single, ventrally-positioned copulatory organ. Combined with these new integumentary data, a comprehensive review of integument in ceratopsian dinosaurs reveals that scalation was generally conservative in ceratopsians and typically consisted of large subcircular-to-polygonal feature scales surrounded by a network of smaller non-overlapping polygonal basement scales. This study highlights the importance of combining exceptional specimens with modern imaging techniques, which are helping to redefine the perceived complexity of squamation in ceratopsians and other dinosaurs.

Out of Africa: Fossils shed light on the origin of the hoatzin, an iconic Neotropic bird.

We describe the earliest fossils of the enigmatic avian taxon Opisthocomiformes (hoatzins) from the Oligo-Miocene (22-24 mya) of Brazil. The bones, a humerus, scapula and coracoid, closely resemble those of the extant hoatzin, Opisthocomus hoazin. The very similar osteology of the pectoral girdle in the new Brazilian fossil compared to the extant O. hoazin, in which it reflects peculiar feeding adaptations, may indicate that hoatzins had already evolved their highly specialized feeding behavior by the mid-Cenozoic. We further show that Namibiavis senutae from the early Miocene of Namibia is another, previously misclassified representative of Opisthocomiformes, which documents that the extant Neotropic distribution of hoatzins is relictual. Because of the weak flight capabilities of hoatzins, their occurrence on both sides of the South Atlantic is of particular biogeographic interest. We detail that this distribution pattern is best explained by dispersal from Africa to South America, and that Opisthocomiformes provide the first example of transatlantic rafting among birds.

On the comparative morphology of the juvenile avian skull: An assessment of squamosal shape across avian higher-level taxa.

The comparative morphology of juvenile avian skulls is poorly known. Here, we survey the shape of the squamosal (os squamosum) across juvenile skulls of avian higher-level clades. In all palaeognathous birds, the rostral end of the squamosal does not surpass the parietal and does not reach the frontal. This morphology is likely to be plesiomorphic for neornithine birds. A short squamosal also occurs in some Neognathae, but in most neognathous birds the squamosal contacts the frontal, and in some taxa the bone is strongly elongated and distinctly surpasses the parietal. Some clades show a notable variation in squamosal morphology. This is, for example, true for Strigiformes, where the taxon Athene differs from the other examined owls in squamosal size, and for the Passeriformes, where Old World Suboscines are characterized by a distinctive squamosal morphology. A unique derived squamosal morphology is for the first time reported for the Apodidae and Hemiprocnidae, in which the bone forms a elongated rostral process that runs along most of the orbital rim. In non-avian theropods, the squamosal articulates with the postorbital and delimits the upper temporal opening. Extant birds lack a postorbital, but a topological correlation between the squamosal and the postorbital process is maintained in most taxa of the Neognathae. The phylogenetic significance of squamosal morphology is diminished by the fact that closely related taxa often show very disparate shapes of the bone, and squamosal morphology appears to be determined by multiple functional constraints including skull geometry, brain morphology and, possibly, nostril type.

Structural coloration in a fossil feather.

Investigation of feathers from the famous Middle Eocene Messel Oil Shale near Darmstadt, Germany shows that they are preserved as arrays of fossilized melanosomes, the surrounding beta-keratin having degraded. The majority of feathers are preserved as aligned rod-shaped eumelanosomes. In some, however, the barbules of the open pennaceous, distal portion of the feather vane are preserved as a continuous external layer of closely packed melanosomes enclosing loosely aligned melanosomes. This arrangement is similar to the single thin-film nanostructure that generates an iridescent, structurally coloured sheen on the surface of black feathers in many lineages of living birds. This is, to our knowledge, the first evidence of preservation of a colour-producing nanostructure in a fossil feather and confirms the potential for determining colour differences in ancient birds and other dinosaurs.

The large-sized darter Anhinga pannonica (Aves, Anhingidae) from the late Miocene hominid Hammerschmiede locality in Southern Germany.

We report fossils of the darter Anhinga pannonica Lambrecht, 1916 from two late Miocene (Tortonian, 11.62 and 11.44 Ma) avifaunas in Southern Germany. The material from the hominid locality Hammerschmiede near Pforzen represents the most comprehensive record of this species and includes most major postcranial elements except for the tarsometatarsus. We furthermore show that the putative cormorant Phalacrocorax brunhuberi (von Ammon, 1918) from the middle Miocene of Regensburg-Dechbetten is another, previously misclassified, record of A. pannonica, and this may also be true for early Miocene fossils described as P. intermedius Milne-Edwards, 1867. A. pannonica was distinctly larger than extant darters and reached the size of A. grandis from the late Miocene of North America. We detail that only fossils from the Miocene of Europe and Africa can be referred to A. pannonica, whereas putative records from Asia fall within the size range of extant darters. A. pannonica appears to have been a long-living species (16 to 6 Ma) with an extensive distribution from the equator to the northern mid-latitudes. The extinction of large-sized darters in Europe is likely to have been due to climatic cooling in the late Neogene, but the reasons for their disappearance in Africa and South America remain elusive.

Reanalysis of putative ovarian follicles suggests that Early Cretaceous birds were feeding not breeding.

We address the identity of putative ovarian follicles in Early Cretaceous bird fossils from the Jehol Biota (China), whose identification has previously been challenged. For the first time, we present a link to the botanical fossil record, showing that the "follicles" of some enantiornithine fossils resemble plant propagules from the Jehol Biota, which belong to Carpolithes multiseminalis. The botanical affinities of this "form-taxon" are currently unresolved, but we note that C. multiseminalis propagules resemble propagules associated with cone-like organs described as Strobilites taxusoides, which in turn are possibly associated with sterile foliage allocated to Liaoningcladus. Laser-Stimulated Fluorescence imaging furthermore reveals different intensities of fluorescence of "follicles" associated with a skeleton of the confuciusornithid Eoconfuciusornis zhengi, with a non-fluorescent circular micro-pattern indicating carbonaceous (or originally carbonaceous) matter. This is inconsistent with the interpretation of these structures as ovarian follicles. We therefore reaffirm that the "follicles" represent ingested food items, and even though the exact nature of the Eoconfuciusornis stomach contents remains elusive, at least some enantiornithines ingested plant propagules.

Oldest Finch-Beaked Birds Reveal Parallel Ecological Radiations in the Earliest Evolution of Passerines.

Beak shape plays a key role in avian radiations and is one of the most intensely studied aspects of avian evolution and ecology [1-4]. Perhaps no other group is more closely associated with the study of beak shape than Passeriformes (passerines or perching birds), the most species-rich ordinal clade of modern birds. However, despite their extraordinary present-day diversity, our understanding of early passerine evolution has been hindered by their sparse fossil record [5, 6]. Here, we describe two new species of early Eocene stem passerines from the Green River Formation of the United States and the Messel Formation of Germany. These species are the oldest fossil birds to exhibit a finch-like beak and provide the earliest evidence for a diet focused on small, hard seeds in crown birds. Given that granivory is a key adaptation that allows passerines to exploit open temperate environments, it is notable that both species occurred in subtropical environments [7, 8]. Phylogenetic analyses place both species within the Psittacopedidae, an extinct Eocene clade of zygodactyl stem passeriforms that also includes the slender-beaked nectarivorous Pumiliornis, the short-beaked Psittacopes, and the thrush-beaked Morsoravis. Our results reveal that stem passerines attained a diversity of beak shapes paralleling many of the morphotypes present in extant passerine finches, thrushes, and sunbirds, more than 35 million years before these morphotypes arose in the crown group. Extinction of these ecologically diverse fossil taxa may be linked to more sophisticated nest construction in anisodactyl crown passerines versus cavity-nesting in Eocene zygodactyl stem passerines [9].

Detection of lost calamus challenges identity of isolated Archaeopteryx feather.

In 1862, a fossil feather from the Solnhofen quarries was described as the holotype of the iconic Archaeopteryx lithographica. The isolated feather's identification has been problematic, and the fossil was considered either a primary, secondary or, most recently, a primary covert. The specimen is surrounded by the 'mystery of the missing quill'. The calamus described in the original paper is unseen today, even under x-ray fluorescence and UV imaging, challenging its original existence. We answer this question using Laser-Stimulated Fluorescence (LSF) through the recovery of the geochemical halo from the original calamus matching the published description. Our study therefore shows that new techniques applied to well-studied iconic fossils can still provide valuable insights. The morphology of the complete feather excludes it as a primary, secondary or tail feather of Archaeopteryx. However, it could be a covert or a contour feather, especially since the latter are not well known in Archaeopteryx. The possibility remains that it stems from a different feathered dinosaur that lived in the Solnhofen Archipelago. The most recent analysis of the isolated feather considers it to be a primary covert. If this is the case, it lacks a distinct s-shaped centerline found in modern primary coverts that appears to be documented here for the first time.

Characterization of melanosomes involved in the production of non-iridescent structural feather colours and their detection in the fossil record.

Non-iridescent structural colour in avian feathers is produced by coherent light scattering through quasi-ordered nanocavities in the keratin cortex of the barbs. To absorb unscattered light, melanosomes form a basal layer underneath the nanocavities. It has been shown that throughout Aves, melanosome morphology reflects broad categories of melanin-based coloration, as well as iridescence, allowing identification of palaeocolours in exceptionally preserved fossils. However, no studies have yet investigated the morphology of melanosomes in non-iridescent structural colour. Here, we analyse a wide sample of melanosomes from feathers that express non-iridescent structural colour from a phylogenetically broad range of extant avians to describe their morphology and compare them with other avian melanosome categories. We find that investigated melanosomes are typically wide (approx. 300 nm) and long (approx. 1400 nm), distinct from melanosomes found in black, brown and iridescent feathers, but overlapping significantly with melanosomes from grey feathers. This may suggest a developmental, and perhaps evolutionary, relationship between grey coloration and non-iridescent structural colours. We show that through analyses of fossil melanosomes, melanosomes indicative of non-iridescent structural colour can be predicted in an Eocene stem group roller ( Eocoracias: Coraciiformes) and with phylogenetic comparative methods the likely hue can be surmised. The overlap between melanosomes from grey and non-iridescent structurally coloured feathers complicates their distinction in fossil samples where keratin does not preserve. However, the abundance of grey coloration relative to non-iridescent structural coloration makes the former a more likely occurrence except in phylogenetically bracketed specimens like the specimen of Eocoracias studied here.