Symbiosis between Cretaceous dinosaurs and feather-feeding beetles.

Extant terrestrial vertebrates, including birds, have a panoply of symbiotic relationships with many insects and arachnids, such as parasitism or mutualism. Yet, identifying arthropod-vertebrate symbioses in the fossil record has been based largely on indirect evidence; findings of direct association between arthropod guests and dinosaur host remains are exceedingly scarce. Here, we present direct and indirect evidence demonstrating that beetle larvae fed on feathers from an undetermined theropod host (avian or nonavian) 105 million y ago. An exceptional amber assemblage is reported of larval molts (exuviae) intimately associated with plumulaceous feather and other remains, as well as three additional amber pieces preserving isolated conspecific exuviae. Samples were found in the roughly coeval Spanish amber deposits of El Soplao, San Just, and Peñacerrada I. Integration of the morphological, systematic, and taphonomic data shows that the beetle larval exuviae, belonging to three developmental stages, are most consistent with skin/hide beetles (family Dermestidae), an ecologically important group with extant keratophagous species that commonly inhabit bird and mammal nests. These findings show that a symbiotic relationship involving keratophagy comparable to that of beetles and birds in current ecosystems existed between their Early Cretaceous relatives.

Insect pollination in deep time.

Inferring insect pollination from compression fossils and amber inclusions is difficult because of a lack of consensus on defining an insect pollinator and the challenge of recognizing this ecological relationship in deep time. We propose a conceptual definition for such insects and an operational classification into pollinator or presumed pollinator. Using this approach, we identified 15 insect families that include fossil pollinators and show that pollination relationships have existed since at least the Upper Jurassic (~163 Ma). Insects prior to this can only be classified as presumed pollinators. This gives a more nuanced insight into the origin and evolution of an ecological relationship that is vital to the establishment, composition and conservation of modern terrestrial ecosystems.

Necrophagy by insects in Oculudentavis and other lizard body fossils preserved in Cretaceous amber.

When a vertebrate carcass begins its decay in terrestrial environments, a succession of different necrophagous arthropod species, mainly insects, are attracted. Trophic aspects of the Mesozoic environments are of great comparative interest, to understand similarities and differences with extant counterparts. Here, we comprehensively study several exceptional Cretaceous amber pieces, in order to determine the early necrophagy by insects (flies in our case) on lizard specimens, ca. 99 Ma old. To obtain well-supported palaeoecological data from our amber assemblages, special attention has been paid in the analysis of the taphonomy, succession (stratigraphy), and content of the different amber layers, originally resin flows. In this respect, we revisited the concept of syninclusion, establishing two categories to make the palaeoecological inferences more accurate: eusyninclusions and parasyninclusions. We observe that resin acted as a "necrophagous trap". The lack of dipteran larvae and the presence of phorid flies indicates decay was in an early stage when the process was recorded. Similar patterns to those in our Cretaceous cases have been observed in Miocene ambers and actualistic experiments using sticky traps, which also act as "necrophagous traps"; for example, we observed that flies were indicative of the early necrophagous stage, but also ants. In contrast, the absence of ants in our Late Cretaceous cases confirms the rareness of ants during the Cretaceous and suggests that early ants lacked this trophic strategy, possibly related to their sociability and recruitment foraging strategies, which developed later in the dimensions we know them today. This situation potentially made necrophagy by insects less efficient in the Mesozoic.

The First Representative of the Roachoid Family Spiloblattinidae (Insecta, Dictyoptera) from the Late Pennsylvanian of the Iberian Peninsula.

Sysciophlebia 'sp. form Villablino', the first Iberian representative of the Palaeozoic-Early Mesozoic family Spiloblattinidae, is described and illustrated. Its forewing colour pattern is strongly similar to those of the Gzhelian-early-middle Asselian species Sysciophlebia euglyptica, Sysciophlebia ilfeldensis, Sysciophlebia rubida, and 'Sysciophlebia sp. form KBQ', supporting the currently proposed Gzhelian age for its type locality. It supports the use of the representatives of the Spiloblattinidae for stratigraphic purposes. The diagnoses and limits of the families Subioblattidae, Phyloblattidae, Compsoblattidae, Spiloblattinidae, and of the spiloblattinid genera are discussed.

A braconid wasp (Hymenoptera, Braconidae) from the Lower Cretaceous amber of San Just, eastern Iberian Peninsula.

Braconid parasitoid wasps are a widely diversified group today, while their fossil record from the Mesozoic is currently poorly known. Here, we describe Utrillabraconelectropteron Álvarez-Parra & Engel, gen. et sp. nov., from the upper Albian (Lower Cretaceous) amber of San Just in the eastern Iberian Peninsula. The holotype specimen is incomplete, although the forewing and hind wing venation are well preserved. The new taxon is assigned to the subfamily †Protorhyssalinae (Braconidae) and, based on characteristics of the wing venation, seems to be closely related to Protorhyssalusgoldmani Basibuyuk & Quicke, 1999 and Diorhyssalusallani (Brues, 1937), both from Upper Cretaceous ambers of North America. We discuss the taxonomy of the Cretaceous braconids, considering †Seneciobraconinae as a valid subfamily. We also comment on possible relationships within †Protorhyssalinae, although a phylogenetic analysis is necessary. Additionally, a checklist is included of braconids known from Cretaceous ambers.

Dinosaur bonebed amber from an original swamp forest soil.

Dinosaur bonebeds with amber content, yet scarce, offer a superior wealth and quality of data on ancient terrestrial ecosystems. However, the preserved palaeodiversity and/or taphonomic characteristics of these exceptional localities had hitherto limited their palaeobiological potential. Here, we describe the amber from the Lower Cretaceous dinosaur bonebed of Ariño (Teruel, Spain) using a multidisciplinary approach. Amber is found in both a root layer with amber strictly in situ and a litter layer mainly composed of aerial pieces unusually rich in bioinclusions, encompassing 11 insect orders, arachnids, and a few plant and vertebrate remains, including a feather. Additional palaeontological data-charophytes, palynomorphs, ostracods- are provided. Ariño arguably represents the most prolific and palaeobiologically diverse locality in which fossiliferous amber and a dinosaur bonebed have been found in association, and the only one known where the vast majority of the palaeontological assemblage suffered no or low-grade pre-burial transport. This has unlocked unprecedentedly complete and reliable palaeoecological data out of two complementary windows of preservation-the bonebed and the amber-from the same site.

Cretaceous amniote integuments recorded through a taphonomic process unique to resins.

Fossil records of vertebrate integuments are relatively common in both rocks, as compressions, and amber, as inclusions. The integument remains, mainly the Mesozoic ones, are of great interest due to the panoply of palaeobiological information they can provide. We describe two Spanish Cretaceous amber pieces that are of taphonomic importance, one bearing avian dinosaur feather remains and the other, mammalian hair. The preserved feather remains originated from an avian dinosaur resting in contact with a stalactite-shaped resin emission for the time it took for the fresh resin to harden. The second piece shows three hair strands recorded on a surface of desiccation, with the characteristic scale pattern exceptionally well preserved and the strands aligned together, which can be considered the record of a tuft. These assemblages were recorded through a rare biostratinomic process we call "pull off vestiture" that is different from the typical resin entrapment and embedding of organisms and biological remains, and unique to resins. The peculiarity of this process is supported by actualistic observations using sticky traps in Madagascar. Lastly, we reinterpret some exceptional records from the literature in the light of that process, thus bringing new insight to the taphonomic and palaeoecological understanding of the circumstances of their origins.

A revised definition for copal and its significance for palaeontological and Anthropocene biodiversity-loss studies.

The early fossilization steps of natural resins and associated terminology are a subject of constant debate. Copal and resin are archives of palaeontological and historical information, and their study is critical to the discovery of new and/or recently extinct species and to trace changes in forests during the Holocene. For such studies, a clear, suitable definition for copal is vital and is herein established. We propose an age range for copal (2.58 Ma-1760 AD), including Pleistocene and Holocene copals, and the novel term "Defaunation resin", defined as resin produced after the commencement of the Industrial Revolution. Defaunation resin is differentiated from Holocene copal as it was produced during a period of intense human transformative activities. Additionally, the "Latest Amber Bioinclusions Gap" (LABG) since the late Miocene to the end of the Pleistocene is hereby newly defined, and is characterized by its virtual absence of bioinclusions and the consequent lack of palaeontological information, which in part explains the historical differentiation between amber and copal. Crucial time intervals in the study of resin production, and of the biodiversity that could be contained, are now clarified, providing a framework for and focusing future research on bioinclusions preserved in copal and resin.

DNA from resin-embedded organisms: Past, present and future.

Past claims have been made for fossil DNA recovery from various organisms (bacteria, plants, insects and mammals, including humans) dating back in time from thousands to several million years BP. However, many of these recoveries, especially those described from million-year-old amber (fossil resin), have faced criticism as being the result of modern environmental contamination and for lack of reproducibility. Using modern genomic techniques, DNA can be obtained with confidence from a variety of substrates (e.g. bones, teeth, gum, museum specimens and fossil insects) of different ages, albeit always less than one million years BP, and results can also be obtained from much older materials using palaeoproteomics. Nevertheless, new attempts to determine if ancient DNA (aDNA) is present in insects preserved in 40 000-year old sub-fossilised resin, the precursor of amber, have been unsuccessful or not well documented. Resin-embedded specimens are therefore regarded as unsuitable for genetic studies. However, we demonstrate here, for the first time, that although a labile molecule, DNA is still present in platypodine beetles (Coleoptera: Curculionidae) embedded in six-year-old and two-year-old resin pieces from Hymenaea verrucosa (Angiospermae: Fabaceae) collected in Madagascar. We describe an optimised method which meets all the requirements and precautions for aDNA experiments for our purpose: to explore the DNA preservation limits in resin. Our objective is far from starting an uncontrolled search for aDNA in amber as it was in the past, but to start resolving basic aspects from the DNA preservation in resin and search from the most modern samples to the ancient ones, step by step. We conclude that it is therefore possible to study genomics from resin-embedded organisms, although the time limits remain to be determined.

Phloem sap in Cretaceous ambers as abundant double emulsions preserving organic and inorganic residues.

Fossilized remains preserved in amber provide abundant data on the paleobiota surrounding the resin-producing plants, but relatively scarcer information about the resinous sources themselves. Here, dark pseudoinclusions in kidney-shaped amber pieces from the Early Cretaceous (Albian) amber from Spain are studied. This type of fossilized remain, abundant in Cretaceous ambers, was first interpreted as fossilized vacuole-bearing microorganisms, but later regarded as artifactual and probably secreted by the resinous trees, although their origin remained unclear. Using complementary microscopy (light, electron, confocal), spectroscopy (infrared, micro-Raman), mass spectrometry and elemental analysis techniques, we demonstrate that the pseudoinclusions correspond to droplets of phloem sap containing amber spheroids and preserving both organic and inorganic residues consistent with degraded components from the original sap. The amber pieces containing pseudoinclusions are fossilized, resin-in-sap-in-resin double emulsions, showing banding patterns with differential content of resin-in-sap emulsion droplets. Our findings represent the first time fossilized phloem sap, 105 million years old, has been recognized and characterized, and open new lines of paleontological research with taxonomic, taphonomic, physiological and ecological implications.

Unravelling the mystery of "Madagascar copal": Age, origin and preservation of a Recent resin.

The loss of biodiversity during the Anthropocene is a constant topic of discussion, especially in the top biodiversity hotspots, such as Madagascar. In this regard, the study of preserved organisms through time, like those included in "Madagascar copal", is of relevance. "Madagascar copal" originated from the leguminous tree Hymenaea verrucosa, which produced and produces resin abundantly. In the last 20 years, interest has focused on the scientific study of its biological inclusions, mainly arthropods, described in dozens of publications. The age and origin of the deposits of "Madagascar copal" have not yet been resolved. Our objectives are to determine its age and geographical origin, and thus increase its scientific value as a source of biological/palaeobiological information. Although Hymenaea was established in Madagascar during the Miocene, we did not find geological deposits of copal or amber in the island. It is plausible that the evolution of those deposits was negatively conditioned by the type of soil, by the climate, and by the development of soil/litter microorganisms, which inhibit preservation of the resin pieces in the litter and subsoil over 300 years. Our results indicate that "Madagascar copal" is a Recent resin, up to a few hundred years old, that originated from Hymenaea trees growing in the lowland coastal forests, one of the most endangered ecosystems in the world. The included and preserved biota is representative of that ecosystem today and during historical times. Inclusions in this Recent resin do not have the palaeontological significance that has been mistakenly attributed to them, but they do have relevant implications for studies regarding Anthropocene biodiversity loss in this hottest hotspot.

Amber from the Triassic to Paleogene of Australia and New Zealand as exceptional preservation of poorly known terrestrial ecosystems.

The Northern Hemisphere dominates our knowledge of Mesozoic and Cenozoic fossilized tree resin (amber) with few findings from the high southern paleolatitudes of Southern Pangea and Southern Gondwana. Here we report new Pangean and Gondwana amber occurrences dating from ~230 to 40 Ma from Australia (Late Triassic and Paleogene of Tasmania; Late Cretaceous Gippsland Basin in Victoria; Paleocene and late middle Eocene of Victoria) and New Zealand (Late Cretaceous Chatham Islands). The Paleogene, richly fossiliferous deposits contain significant and diverse inclusions of arthropods, plants and fungi. These austral discoveries open six new windows to different but crucial intervals of the Mesozoic and early Cenozoic, providing the earliest occurrence(s) of some taxa in the modern fauna and flora giving new insights into the ecology and evolution of polar and subpolar terrestrial ecosystems.

Direct evidence for eudicot pollen-feeding in a Cretaceous stinging wasp (Angiospermae; Hymenoptera, Aculeata) preserved in Burmese amber.

Angiosperms and their insect pollinators form a foundational symbiosis, evidence for which from the Cretaceous is mostly indirect, based on fossils of insect taxa that today are anthophilous, and of fossil insects and flowers that have apparent anthophilous and entomophilous specializations, respectively. We present exceptional direct evidence preserved in mid-Cretaceous Burmese amber, 100 mya, for feeding on pollen in the eudicot genus Tricolporoidites by a basal new aculeate wasp, Prosphex anthophilos, gen. et sp. nov., in the lineage that contains the ants, bees, and other stinging wasps. Plume of hundreds of pollen grains wafts from its mouth and an apparent pollen mass was detected by micro-CT in the buccal cavity: clear evidence that the wasp was foraging on the pollen. Eudicots today comprise nearly three-quarters of all angiosperm species. Prosphex feeding on Tricolporoidites supports the hypothesis that relatively small, generalized insect anthophiles were important pollinators of early angiosperms.

A mantidfly in Cretaceous Spanish amber provides insights into the evolution of integumentary specialisations on the raptorial foreleg.

Multiple predatory insect lineages have developed a raptorial lifestyle by which they strike and hold prey using modified forelegs armed with spine-like structures and other integumentary specialisations. However, how structures enabling the raptorial function evolved in insects remains largely hypothetical or inferred through phylogeny due to the rarity of meaningful fossils. This is particularly true for mantidflies (Neuroptera: Mantispidae), which have a scarce fossil record mostly based on rock compressions, namely isolated wings. Here, Aragomantispa lacerata gen. et sp. nov. is described from ca. 105-million-year-old San Just amber (Spain), representing the oldest and one of the few mantidflies hitherto described from amber. The fossil shows exquisitely preserved forefemoral spine-like structures composed of integumentary processes each bearing a modified seta, and prostrate setae on foretibiae and foretarsi. The fine morphology of these structures was unknown in fossil mantidflies. An assessment of integumentary specialisations from raptorial forelegs across mantispoid lacewings is provided. The present finding reveals how the specialised foreleg armature associated to the raptorial lifestyle in extant mantidflies was present yet not fully established by the Early Cretaceous, at least in some lineages, and provides palaeontological evidence supporting certain evolutionary patterns of acquisition of integumentary specialisations related to the raptorial function in the group.

Ancient amino acids from fossil feathers in amber.

Ancient protein analysis is a rapidly developing field of research. Proteins ranging in age from the Quaternary to Jurassic are being used to answer questions about phylogeny, evolution, and extinction. However, these analyses are sometimes contentious, and focus primarily on large vertebrates in sedimentary fossilisation environments; there are few studies of protein preservation in fossils in amber. Here we show exceptionally slow racemisation rates during thermal degradation experiments of resin enclosed feathers, relative to previous thermal degradation experiments of ostrich eggshell, coral skeleton, and limpet shell. We also recover amino acids from two specimens of fossil feathers in amber. The amino acid compositions are broadly similar to those of degraded feathers, but concentrations are very low, suggesting that much of the original protein has been degraded and lost. High levels of racemisation in more apolar, slowly racemising amino acids suggest that some of the amino acids were ancient and therefore original. Our findings indicate that the unique fossilisation environment inside amber shows potential for the recovery of ancient amino acids and proteins.

A soil-carrying lacewing larva in Early Cretaceous Lebanese amber.

Diverse organisms protect and camouflage themselves using varied materials from their environment. This adaptation and associated behaviours (debris-carrying) are well known in modern green lacewing larvae (Neuroptera: Chrysopidae), mostly due to the widespread use of these immature insects in pest control. However, the evolutionary history of this successful strategy and related morphological adaptations in the lineage are still far from being understood. Here we describe a novel green lacewing larva, Tyruschrysa melqart gen. et sp. nov., from Early Cretaceous Lebanese amber, carrying a preserved debris packet composed by soil particles entangled among specialised setae of extremely elongate tubular tubercles. The new morphotype has features related to the debris-carrying habit that are unknown from extant or extinct green lacewings, namely a high number of tubular tubercle pairs on the abdomen and tubular tubercle setae with mushroom-shaped endings that acted as anchoring points for debris. The current finding expands the diversity of exogenous materials used by green lacewing larvae in deep time, and represents the earliest direct evidence of debris-carrying in the lineage described to date. The debris-carrying larval habit likely played a significant role during the initial phases of diversification of green lacewings.

Amber in prehistoric Iberia: New data and a review.

Provenancing exotic raw materials and reconstructing the nature and routes of exchange is a major concern of prehistoric archaeology. Amber has long been recognised as a key commodity of prehistoric exchange networks in Europe. However, most science-based studies so far have been localised and based on few samples, hence making it difficult to observe broad geographic and chronological trends. This paper concentrates on the nature, distribution and circulation of amber in prehistoric Iberia. We present new standardised FTIR analyses of 22 archaeological and geological samples from a large number of contexts across Iberia, as well as a wide scale review of all the legacy data available. On the basis of a considerable body of data, we can confirm the use of local amber resources in the Northern area of the Iberian Peninsula from the Palaeolithic to the Bronze Age; we push back the arrival of Sicilian amber to at least the 4th Millennium BC, and we trace the appearance of Baltic amber since the last quarter of the 2nd Millennium BC, progressively replacing Sicilian simetite. Integrating these data with other bodies of archaeological information, we suggest that the arrival of Baltic amber was part of broader Mediterranean exchange networks, and not necessarily the result of direct trade with the North. From a methodological perspective, thanks to the analyses carried out on both the vitreous core and the weathered surfaces of objects made of Sicilian simetite, we define the characteristic FTIR bands that allow the identification of Sicilian amber even in highly deteriorated archaeological samples.

Arthropods in modern resins reveal if amber accurately recorded forest arthropod communities.

Amber is an organic multicompound derivative from the polymerization of resin of diverse higher plants. Compared with other modes of fossil preservation, amber records the anatomy of and ecological interactions between ancient soft-bodied organisms with exceptional fidelity. However, it is currently suggested that ambers do not accurately record the composition of arthropod forest paleocommunities, due to crucial taphonomic biases. We evaluated the effects of taphonomic processes on arthropod entrapment by resin from the plant Hymenaea, one of the most important resin-producing trees and a producer of tropical Cenozoic ambers and Anthropocene (or subfossil) resins. We statistically compared natural entrapment by Hymenaea verrucosa tree resin with the ensemble of arthropods trapped by standardized entomological traps around the same tree species. Our results demonstrate that assemblages in resin are more similar to those from sticky traps than from malaise traps, providing an accurate representation of the arthropod fauna living in or near the resiniferous tree, but not of entire arthropod forest communities. Particularly, arthropod groups such as Lepidoptera, Collembola, and some Diptera are underrepresented in resins. However, resin assemblages differed slightly from sticky traps, perhaps because chemical compounds in the resins attract or repel specific insect groups. Ground-dwelling or flying arthropods that use the tree-trunk habitat for feeding or reproduction are also well represented in the resin assemblages, implying that fossil inclusions in amber can reveal fundamental information about biology of the past. These biases have implications for the paleoecological interpretation of the fossil record, principally of Cenozoic amber with angiosperm origin.

Mating and aggregative behaviors among basal hexapods in the Early Cretaceous.

Among the many challenges in paleobiology is the inference and reconstruction of behaviors that rarely, if ever, leave a physical trace on the environment that is suitable for fossilization. Of particular significance are those behaviors tied to mating and courtship, individual interactions critical for species integrity and continuance, as well as those for dispersal, permitting the taxon to expand its distribution as well as access new habitats in the face of local or long-term environmental change. In this context, two recently discovered fossils from the Early Cretaceous amber of Spain (ca. 105 mya) give a detailed view of otherwise fleeting ethologies in Collembola. These occurrences are phylogenetically spaced across the class, and from species representing the two major clades of springtails-Symphypleona and Entomobryomorpha. Specifically, we report unique evidence from a symphypleonan male (Pseudosminthurides stoechus Sánchez-García & Engel, 2016) with modified antennae that may have functioned as a clasping organ for securing females during mating on water's surface, and from an aggregation of entomobryomorphan individuals (Proisotoma communis Sánchez-García & Engel, 2016) purportedly representing a swarming episode on the forest floor. We demonstrate that the mating behavioral repertoire in P. stoechus, which is associated with considerable morphological adaptations, likely implied elaborate courtship and maneuvering for guarantee sperm transfer in an epineustic species. These discoveries reveal significant behaviors consistent with modern counterparts and a generalized stasis for some ancient hexapod ethologies associated with complex mating and courtship and social or pre-social aggregations, so critical to specific constancy and dispersal.

Publisher Correction: Ticks parasitised feathered dinosaurs as revealed by Cretaceous amber assemblages.

The originally published version of this Article was updated shortly after publication to add the word 'Ticks' to the title, following its inadvertent removal during the production process. This has now been corrected in both the PDF and HTML versions of the Article.