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AIMS: To describe the gross and light microscopic characteristics of skin lesions observed on the ventral skin of captive Archey’s frogs (Leiopelma archeyi) between 2000 and 2012, and to investigate their occurrence, possible aetiology and association with survival. METHODS: Postmortem skin samples were obtained for histological evaluation from 37 frogs, with and without skin lesions, that died while in captivity at Auckland Zoo between 2000 and 2012. Four frogs with skin lesions were biopsied under general anaesthesia and samples used for both light and transmission electron microscopy. The records of 94 frogs held at the University of Otago and Auckland Zoo between 2000–2012 were reviewed, which included some frogs recently collected from the wild. Information about the occurrence of skin lesions, and mortality associated with skin lesions was collated. RESULTS: Grossly the skin lesions varied in appearance; most were circular, pale grey papules, which measured from <0.5–1.5 mm in diameter with no umbilication. The overlying epidermis was not fragile and there was no associated inflammation. Contents often appeared clear or semi-transparent. Lesions were located predominantly on ventral surfaces including trunk, thighs, lower legs and forearms, and gular region, but not on digits. The number ranged from single to multiple, often confluent lesions covering the entire ventral surface of the frog. Histologically the lesions consisted of enlarged proliferating mucous glands that expanded the dermis and elevated the epidermis. They were semi-organised, solid or occasionally cavitated acinar structures with central lumina which sometimes contained mucus. Nuclei showed moderate anisokaryosis and mitotic figures were uncommon. Transmission electron microscopy did not show any infectious agents. Between 2000 and 2012, skin lesions were recorded in 35/94 (37%) frogs. The size and location of skin lesions varied over time, with some resolving and sometimes reappearing. Skin lesions were not associated with an increased risk of death. CONCLUSIONS: The skin lesions had the gross and microscopic characteristics of adenomatous hyperplasia of the dermal mucous glands. CLINICAL RELEVANCE: The aetiology of this adenomatous hyperplasia is unknown, but factors associated with the captive environment are the most likely cause. This is the first description of adenomatous hyperplasia of the cutaneous mucous glands in amphibians. [ABSTRACT FROM AUTHOR]

Surveys were distributed to New Zealand land users in 1998 and 2008 to acquire information about New Zealand frogs with the aim of compiling and mapping their distribution and inferred population trends without costly and time-consuming field surveys. The overall frog population trend was reported as declining, with possible causes reported as an increase in agriculture, an increase in the distribution of predatory fish and disease. The resultant maps could be used for four main purposes: 1) to identify regions where Litoria populations are known to occur, which can be eliminated when considering suitable regions for translocation of Leiopelma; 2) to identify growing or stable populations of Litoria species, which may assist future disease surveys, population monitoring and to identify sources of genetic material that may serve as an Ark for declining Australian populations; 3) to highlight populations that are in decline to enable effective targeting of detailed disease studies; and 4) to approximate the stability of amphibian populations in the absence of more accurate, but costly, scientific monitoring. [ABSTRACT FROM AUTHOR]

The secretion of defensive chemicals onto the skin is a widely used mechanism for predator defence in anurans. Secretions often consist of a mixture of bioactive peptides with cytotoxic or neurotoxic effects. In New Zealand, introduced rodents have been suggested as main drivers for declines and extinctions of endemic frogs. We demonstrate the efficacy of Leiopelma pakeka secretions in deterring rats (Rattus norvegicus Long-Evans) from ingesting secretion-covered food and showed that they can successfully lyse rat erythrocytes. When offered a choice, rats displayed a significant preference for food pellets coated with water over those covered in frog secretions. Direct oral exposure to the secretions has no significant effects on water or food intake of rats. Video analysis showed no significant difference in the proportion of time rats spent grooming, rising on hind legs, motionless or investigating associated with exposure to the secretions. This study provides new insight into the defensive function of leiopelmatid skin secretions. [ABSTRACT FROM PUBLISHER]

New Zealand's endemic leiopelmatid frogs are all threatened with extinction. There is no obvious physical or behavioural difference between males and females of three of the four species and this lack of a sex identification technique is hindering conservation management. Twenty-one morphological features were measured for 67 live Leiopelma pakeka of known sex to determine whether a slight morphological difference existed that could be used for sex identification. Females were significantly longer in snout-to-vent length (SVL) than males, but as a great deal of overlap exists between sexes, this is not an ideal sex identification trait. No other physical characteristic was significantly different between sexes when adjusted for SVL. Discriminant analysis using a suite of traits was also unable to discriminate between sexes for frogs in the male-female overlap size range. The measurement of physical characteristics is inadequate for identifying sex in L.pakeka and other methods for sex identification must be developed. The reasons for the lack of sexual difference are unknown. [ABSTRACT FROM AUTHOR]

Axial compressive loading, the principal cause of spinal cord quadriplegia in American football, is produced when a player is forcibly struck on the crown of the helmet. This impact subjects the small cervical vertebrae to a large compressive force that often produces stress that exceeds the failure limit of the spine. Several factors influence the outcome in axial collisions, including the available kinetic energy, the displacement needed to dissipate the energy, and the end conditions of the collision (i.e., the position of the head). Effective intervention of this catastrophic injury requires the melding of information from the fields of biomechanics and epidemiology. From a biomechanical perspective, neck loading should be kept at a level that is below the failure limit of the cervical spine. The epidemiologic rate at which these injuries develop among football players suggests that cervical quadriplegia is rare. Thus, protective devices intended to lower the forces on the cervical spine may not succeed in dramatically reducing the incidence of this injury. Because this injury is rare, it is important to consider that introducing new protective equipment, intended for intervention of one problem (i.e., cervical quadriplegia), may lead to other injuries. [ABSTRACT FROM AUTHOR]

This paper is the first in a two-part series that charts the evolution of appendicular musculature along the mammalian stem lineage, drawing upon the exceptional fossil record of extinct synapsids. Here, attention is focused on muscles of the forelimb. Understanding forelimb muscular anatomy in extinct synapsids, and how this changed on the line to mammals, can provide important perspective for interpreting skeletal and functional evolution in this lineage, and how the diversity of forelimb functions in extant mammals arose. This study surveyed the osteological evidence for muscular attachments in extinct mammalian and nonmammalian synapsids, two extinct amniote outgroups, and a large selection of extant mammals, saurians, and salamanders. Observations were integrated into an explicit phylogenetic framework, comprising 73 character-state complexes covering all muscles crossing the shoulder, elbow, and wrist joints. These were coded for 33 operational taxonomic units spanning >330 Ma of tetrapod evolution, and ancestral state reconstruction was used to evaluate the sequence of muscular evolution along the stem lineage from Amniota to Theria. In addition to producing a comprehensive documentation of osteological evidence for muscle attachments in extinct synapsids, this work has clarified homology hypotheses across disparate taxa and helped resolve competing hypotheses of muscular anatomy in extinct species. The evolutionary history of mammalian forelimb musculature was a complex and nonlinear narrative, punctuated by multiple instances of convergence and concentrated phases of anatomical transformation. More broadly, this study highlights the great insight that a fossil-based perspective can provide for understanding the assembly of novel body plans., (© 2023 American Association for Anatomy.)

This paper is the second in a two-part series that charts the evolution of appendicular musculature along the mammalian stem lineage, drawing upon the exceptional fossil record of extinct synapsids. Here, attention is focused on muscles of the hindlimb. Although the hindlimb skeleton did not undergo as marked a transformation on the line to mammals as did the forelimb skeleton, the anatomy of extant tetrapods indicates that major changes to musculature have nonetheless occurred. To better understand these changes, this study surveyed the osteological evidence for muscular attachments in extinct mammalian and nonmammalian synapsids, two extinct amniote outgroups, and a large selection of extant mammals, saurians, and salamanders. Observations were integrated into an explicit phylogenetic framework, comprising 80 character-state complexes covering all muscles crossing the hip, knee, and ankle joints. These were coded for 33 operational taxonomic units spanning >330 Ma of tetrapod evolution, and ancestral state reconstruction was used to evaluate the sequence of muscular evolution along the stem lineage from Amniota to Theria. The evolutionary history of mammalian hindlimb musculature was complex, nonlinear, and protracted, with several instances of convergence and pulses of anatomical transformation that continued well into the crown group. Numerous traits typically regarded as characteristically "mammalian" have much greater antiquity than previously recognized, and for some traits, most synapsids are probably more reflective of the ancestral amniote condition than are extant saurians. More broadly, this study highlights the utility of the fossil record in interpreting the evolutionary appearance of distinctive anatomies., (© 2023 American Association for Anatomy.)

Theropods are obligate bipedal dinosaurs that appeared 230 million years ago and are still extant as birds. Their history is characterized by extreme variations in body mass, with gigantism evolving convergently between many lineages. However, no quantification of hindlimb functional morphology has shown if these body mass increases led to similar specializations between distinct lineages. Here we studied femoral shape variation across 41 species of theropods (n= 68 specimens) using a high-density 3D geometric morphometric approach. We demonstrated that the heaviest theropods evolved wider epiphyses and a more distally located fourth trochanter, as previously demonstrated in early archosaurs, along with an upturned femoral head and a mediodistal crest that extended proximally along the shaft. Phylogenetically informed analyses highlighted that these traits evolved convergently within six major theropod lineages, regardless of their maximum body mass. Conversely, the most gracile femora were distinct from the rest of the dataset, which we interpret as a femoral specialization to "miniaturization" evolving close to Avialae (bird lineage). Our results support a gradual evolution of known "avian" features, such as the fusion between lesser and greater trochanters and a reduction of the epiphyses' offset, independently from body mass variations, which may relate to a more "avian" type of locomotion (more knee-than hip-driven). The distinction between body mass variations and a more "avian" locomotion is represented by a decoupling in the mediodistal crest morphology, whose biomechanical nature should be studied to better understand the importance of its functional role in gigantism, miniaturization and higher parasagittal abilities., Competing Interests: Competing interests. The authors declare no competing interest.

Animal performance fundamentally influences behaviour, ecology, and evolution. It typically varies monotonously with size. A notable exception is maximum running speed; the fastest animals are of intermediate size. Here we show that this peculiar allometry results from the competition between two musculoskeletal constraints: the kinetic energy capacity, which dominates in small animals, and the work capacity, which reigns supreme in large animals. The ratio of both capacities defines the physiological similarity index Γ, a dimensionless number akin to the Reynolds number in fluid mechanics. The scaling of Γ indicates a transition from a dominance of muscle forces to a dominance of inertial forces as animals grow in size; its magnitude defines conditions of "dynamic similarity" that enable comparison and estimates of locomotor performance across extant and extinct animals; and the physical parameters that define it highlight opportunities for adaptations in musculoskeletal "design" that depart from the eternal null hypothesis of geometric similarity. The physiological similarity index challenges the Froude number as prevailing dynamic similarity condition, reveals that the differential growth of muscle and weight forces central to classic scaling theory is of secondary importance for the majority of terrestrial animals, and suggests avenues for comparative analyses of locomotor systems., (© 2024. The Author(s).)

In vertebrates, active movement is driven by muscle forces acting on bones, either directly or through tendinous insertions. There has been much debate over how muscle size and force are reflected by the muscular attachment areas (AAs). Here we investigate the relationship between the physiological cross-sectional area (PCSA), a proxy for the force production of the muscle, and the AA of hindlimb muscles in Nile crocodiles and five bird species. The limbs were held in a fixed position whilst blunt dissection was carried out to isolate the individual muscles. AAs were digitised using a point digitiser, before the muscle was removed from the bone. Muscles were then further dissected and fibre architecture was measured, and PCSA calculated. The raw measures, as well as the ratio of PCSA to AA, were studied and compared for intra-observer error as well as intra- and interspecies differences. We found large variations in the ratio between AAs and PCSA both within and across species, but muscle fascicle lengths are conserved within individual species, whether this was Nile crocodiles or tinamou. Whilst a discriminant analysis was able to separate crocodylian and avian muscle data, the ratios for AA to cross-sectional area for all species and most muscles can be represented by a single equation. The remaining muscles have specific equations to represent their scaling, but equations often have a relatively high success at predicting the ratio of muscle AA to PCSA. We then digitised the muscle AAs of Coelophysis bauri, a dinosaur, to estimate the PCSAs and therefore maximal isometric muscle forces. The results are somewhat consistent with other methods for estimating force production, and suggest that, at least for some archosaurian muscles, that it is possible to use muscle AA to estimate muscle sizes. This method is complementary to other methods such as digital volumetric modelling., (© 2022 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.)

Significant evolutionary shifts in locomotor behaviour often involve comparatively subtle anatomical transitions. For dinosaurian and avian evolution, medial overhang of the proximal femur has been central to discussions. However, there is an apparent conflict with regard to the evolutionary origin of the dinosaurian femoral head, with neontological and palaeontological data suggesting seemingly incongruent hypotheses. To reconcile this, we reconstructed the evolutionary history of morphogenesis of the proximal end of the femur from early archosaurs to crown birds. Embryological comparison of living archosaurs (crocodylians and birds) suggests the acquisition of the greater overhang of the femoral head in dinosaurs results from additional growth of the proximal end in the medial-ward direction. On the other hand, the fossil record suggests that this overhang was acquired by torsion of the proximal end, which projected in a more rostral direction ancestrally. We reconcile this apparent conflict by inferring that the medial overhang of the dinosaur femoral head was initially acquired by torsion, which was then superseded by mediad growth. Details of anatomical shifts in fossil forms support this hypothesis, and their biomechanical implications are congruent with the general consensus regarding broader morpho-functional evolution on the avian stem.

The last common ancestor of birds and crocodylians plus all of its descendants (clade Archosauria) dominated terrestrial Mesozoic ecosystems, giving rise to disparate body plans, sizes, and modes of locomotion. As in the fields of vertebrate morphology and paleontology more generally, studies of archosaur skeletal structure have come to depend on tools for acquiring, measuring, and exploring three-dimensional (3-D) digital models. Such models, in turn, form the basis for many analyses of musculoskeletal function. A set of shared conventions for describing 3-D pose (joint or limb configuration) and 3-D kinematics (change in pose through time) is essential for fostering comparison of posture/movement among such varied species, as well as for maximizing communication among scientists. Following researchers in human biomechanics, we propose a standard methodological approach for measuring the relative position and orientation of the major segments of the archosaur pelvis and hindlimb in 3-D. We describe the construction of anatomical and joint coordinate systems using the extant guineafowl and alligator as examples. Our new standards are then applied to three extinct taxa sampled from the wider range of morphological, postural, and kinematic variation that has arisen across >250 million years of archosaur evolution. These proposed conventions, and the founding principles upon which they are based, can also serve as starting points for measuring poses between elements within a hindlimb segment, for establishing coordinate systems in the forelimb and axial skeleton, or for applying our archosaurian system more broadly to different vertebrate clades., (© 2022 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.)

The global reach of zoos has enormous potential to raise awareness of conservation issues such as the amphibian extinction crisis. One mechanism of achieving this is through targeted, time-limited campaigns. However, the longer-term impact of such campaigns on conservation outcomes is rarely evaluated. The goal of this study is to evaluate how zoos view "The Year of The Frog," a campaign launched by zoos in 2008, with the aim of raising awareness of the global amphibian crisis and generating funds to support amphibian conservation initiatives. A questionnaire-based survey of 43 zoos from fourteen countries was used to evaluate how zoo practitioners believed the campaign influenced amphibian conservation some 10 years after its launch. Survey results showed that the campaign not only contributed to the improvement of research into amphibian husbandry in zoos but also increased support of in situ conservation of amphibians. Zoos focused on exotic species, but developed countries led most of the actions of the campaign. The campaign has inspired and encouraged some zoos and other ex situ conservation organizations to embrace amphibian conservation. The next step is to increase collaboration with more organizations through, for example, AArk and IUCN SSC/ASG, particularly in countries that have high amphibian biodiversity., (© 2021 Wiley Periodicals LLC.)

Bats are the most diverse mammalian order second to rodents, with 1400+ species globally. In the tropics, it is possible to find more than 60 bat species at a single site. However, monitoring bats is challenging due to their small size, ability to fly, cryptic nature, and nocturnal activity. Recently, bioacoustic techniques have been incorporated into survey methods, either through passive acoustic monitoring or acoustic bat lures. Lures have been developed on the premise that broadcasting acoustic stimuli increases the number of captures in harp traps or mist nets. However, this is a relatively new, niche method. This study tested the efficacy of two commonly used acoustic bat lure devices, broadcasting two different acoustic stimuli, to increase forest understory bat captures in the tropics. This is the first time an acoustic bat lure has been systematically tested in a tropical rainforest, and the first study to compare two lure devices (Sussex AutoBat and Apodemus BatLure). Using a paired experimental design, two synthesized acoustic stimuli were broadcasted, a feeding call and a social call, to understand the importance of the call type used on capture rates and genus-specific responses. Using an acoustic lure significantly increased capture rates, while the type of device did not impact capture rates. The two acoustic stimuli had an almost even distribution of captures, suggesting that the type of call may be less important than previously thought. Results indicate a possible deterrent effect on Rhinolophous sp., while being particularly effective for attracting bats in the genera Murina and Kerivoula . This study highlights the effectiveness of lures, however, also indicates that lure effects can vary across genera. Therefore, lures may bias survey results by altering the species composition of bats caught. Future research should focus on a single species or genus, using synthesized calls of conspecifics, to fully understand the effect of lures., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Amidst a global amphibian decline, captive breeding has become an important component of many conservation management programs. Some species fail to readily reproduce in captivity, including leiopelmatid frogs, an archaic lineage endemic to New Zealand. Assisted reproductive technologies can improve the reproductive potential and genetic management of captive programs. Herein we describe efforts to induce spermiation and mating in the threatened Hamilton's frog Leiopelma hamiltoni using a gonadotrophin-releasing hormone agonist and human chorionic gonadotrophin. The spermiation response was poor, with few individuals releasing spermatozoa and a low sperm concentration (mean (±s.e.) 27.4±8.5×103 spermatozoa mL-1 ) when spermiation was stimulated. The sperm concentration peaked 12-24h after hormone administration, suggesting this species may be slower to react than other anurans. Spermic urine was present in five males before hormone treatment. The sperm concentration was higher in this group (128.7±54.9×103 spermatozoa mL-1 ) than in the hormone-induced males. All spermatozoa observed exhibited no motility or, at most, a limited swaying movement with no forward progression. In separate hormone induction trials with 60 individuals (two males to one female per trial), no mating was exhibited during the 12h after treatment. Further studies to hormonally stimulate gamete release and mating are needed in leiopelmatids.

Skeletal muscle mass, architecture and force-generating capacity are well known to scale with body size in animals, both throughout ontogeny and across species. Investigations of limb muscle scaling in terrestrial amniotes typically focus on individual muscles within select clades, but here this question was examined at the level of the whole limb across amniotes generally. In particular, the present study explored how muscle mass, force-generating capacity (measured by physiological cross-sectional area) and internal architecture (fascicle length) scales in the fore- and hindlimbs of extant mammals, non-avian saurians ('reptiles') and bipeds (birds and humans). Sixty species spanning almost five orders of magnitude in body mass were investigated, comprising previously published architectural data and new data obtained via dissections of the opossum Didelphis virginiana and the tegu lizard Salvator merianae . Phylogenetic generalized least squares was used to determine allometric scaling slopes (exponents) and intercepts, to assess whether patterns previously reported for individual muscles or functional groups were retained at the level of the whole limb, and to test whether mammals, reptiles and bipeds followed different allometric trajectories. In general, patterns of scaling observed in individual muscles were also observed in the whole limb. Reptiles generally have proportionately lower muscle mass and force-generating capacity compared to mammals, especially at larger body size, and bipeds exhibit strong to extreme positive allometry in the distal hindlimb. Remarkably, when muscle mass was accounted for in analyses of muscle force-generating capacity, reptiles, mammals and bipeds almost ubiquitously followed a single common scaling pattern, implying that differences in whole-limb force-generating capacity are principally driven by differences in muscle mass, not internal architecture. In addition to providing a novel perspective on skeletal muscle allometry in animals, the new dataset assembled was used to generate pan-amniote statistical relationships that can be used to predict muscle mass or force-generating capacity in extinct amniotes, helping to inform future reconstructions of musculoskeletal function in the fossil record., Competing Interests: Stephanie E. Pierce is an Academic Editor for PeerJ., (© 2021 Bishop et al.)

Locomotion has influenced the ecology, evolution, and extinction of species throughout history, yet studying locomotion in the fossil record is challenging. Computational biomechanics can provide novel insight by mechanistically relating observed anatomy to whole-animal function and behavior. Here, we leverage optimal control methods to generate the first fully predictive, three-dimensional, muscle-driven simulations of locomotion in an extinct terrestrial vertebrate, the bipedal non-avian theropod dinosaur Coelophysis . Unexpectedly, our simulations involved pronounced lateroflexion movements of the tail. Rather than just being a static counterbalance, simulations indicate that the tail played a crucial dynamic role, with lateroflexion acting as a passive, physics-based mechanism for regulating angular momentum and improving locomotor economy, analogous to the swinging arms of humans. We infer this mechanism to have existed in many other bipedal non-avian dinosaurs as well, and our methodology provides new avenues for exploring the functional diversity of dinosaur tails in the future.

We developed a three-dimensional, computational biomechanical model of a juvenile Nile crocodile (Crocodylus niloticus) pelvis and hindlimb, composed of 47 pelvic limb muscles, to investigate muscle function. We tested whether crocodiles, which are known to use a variety of limb postures during movement, use limb orientations (joint angles) that optimise the moment arms (leverages) or moment-generating capacities of their muscles during different limb postures ranging from a high walk to a sprawling motion. We also describe the three-dimensional (3D) kinematics of the crocodylian hindlimb during terrestrial locomotion across an instrumented walkway and a treadmill captured via X-ray Reconstruction of Moving Morphology (biplanar fluoroscopy; 'XROMM'). We reconstructed the 3D positions and orientations of each of the hindlimb bones and used dissection data for muscle lines of action to reconstruct a focal, subject-specific 3D musculoskeletal model. Motion data for different styles of walking (a high, crouched, bended and two types of sprawling motion) were fed into the 3D model to identify whether any joints adopted near-optimal poses for leverage across each of the behaviours. We found that (1) the hip adductors and knee extensors had their largest leverages during sprawling postures and (2) more erect postures typically involved greater peak moment arms about the hip (flexion-extension), knee (flexion) and metatarsophalangeal (flexion) joints. The results did not fully support the hypothesis that optimal poses are present during different locomotory behaviours because the peak capacities were not always reached around mid-stance phase. Furthermore, we obtained few clear trends for isometric moment-generating capacities. Therefore, perhaps peak muscular leverage in Nile crocodiles is instead reached either in early/late stance or possibly during swing phase or other locomotory behaviours that were not studied here, such as non-terrestrial movement. Alternatively, our findings could reflect a trade-off between having to execute different postures, meaning that hindlimb muscle leverage is not optimised for any singular posture or behaviour. Our model, however, provides a comprehensive set of 3D estimates of muscle actions in extant crocodiles which can form a basis for investigating muscle function in extinct archosaurs., (© 2021 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.)

Joint mobility is a key factor in determining the functional capacity of tetrapod limbs, and is important in palaeobiological reconstructions of extinct animals. Recent advances have been made in quantifying osteological joint mobility using virtual computational methods; however, these approaches generally focus on the proximal limb joints and have seldom been applied to fossil mammals. Palorchestes azael is an enigmatic, extinct ~1000 kg marsupial with no close living relatives, whose functional ecology within Australian Pleistocene environments is poorly understood. Most intriguing is its flattened elbow morphology, which has long been assumed to indicate very low mobility at this important joint. Here, we tested elbow mobility via virtual range of motion (ROM) mapping and helical axis analysis, to quantitatively explore the limits of Palorchestes' elbow movement and compare this with their living and extinct relatives, as well as extant mammals that may represent functional analogues. We find that Palorchestes had the lowest elbow mobility among mammals sampled, even when afforded joint translations in addition to rotational degrees of freedom. This indicates that Palorchestes was limited to crouched forelimb postures, something highly unusual for mammals of this size. Coupled flexion and abduction created a skewed primary axis of movement at the elbow, suggesting an abducted forelimb posture and humeral rotation gait that is not found among marsupials and unlike that seen in any large mammals alive today. This work introduces new quantitative methods and demonstrates the utility of comparative ROM mapping approaches, highlighting that Palorchestes' forelimb function was unlike its contemporaneous relatives and appears to lack clear functional analogues among living mammals., (© 2021 Anatomical Society.)

Jumping is a common, but demanding, behavior that many animals employ during everyday activity. In contrast to jump-specialists such as anurans and some primates, jumping biomechanics and the factors that influence performance remains little studied for generalized species that lack marked adaptations for jumping. Computational biomechanical modeling approaches offer a way of addressing this in a rigorous, mechanistic fashion. Here, optimal control theory and musculoskeletal modeling are integrated to generate predictive simulations of maximal height jumping in a small ground-dwelling bird, a tinamou. A three-dimensional musculoskeletal model with 36 actuators per leg is used, and direct collocation is employed to formulate a rapidly solvable optimal control problem involving both liftoff and landing phases. The resulting simulation raises the whole-body center of mass to over double its standing height, and key aspects of the simulated behavior qualitatively replicate empirical observations for other jumping birds. However, quantitative performance is lower, with reduced ground forces, jump heights, and muscle-tendon power. A pronounced countermovement maneuver is used during launch. The use of a countermovement is demonstrated to be critical to the achievement of greater jump heights, and this phenomenon may only need to exploit physical principles alone to be successful; amplification of muscle performance may not necessarily be a proximate reason for the use of this maneuver. Increasing muscle strength or contractile velocity above nominal values greatly improves jump performance, and interestingly has the greatest effect on more distal limb extensor muscles (i.e., those of the ankle), suggesting that the distal limb may be a critical link for jumping behavior. These results warrant a re-evaluation of previous inferences of jumping ability in some extinct species with foreshortened distal limb segments, such as dromaeosaurid dinosaurs. Simulations prédictives de la fonction musculo-squelettique et des performances de saut chez un oiseau généralisé Sauter est un comportement commun, mais exigeant, que de nombreux animaux utilisent au cours de leurs activités quotidiennes. Contrairement aux spécialistes du saut tels que les anoures et certains primates, la biomécanique du saut et les facteurs qui influencent la performance restent peu étudiés pour les espèces généralisées qui n'ont pas d'adaptations marquées pour le saut. Les approches de modélisation biomécanique computationnelle offrent un moyen d'aborder cette question de manière rigoureuse et mécaniste. Ici, la théorie du contrôle optimal et la modélisation musculo-squelettique sont intégrées pour générer des simulations prédictives du saut en hauteur maximal chez un petit oiseau terrestre, le tinamou. Un modèle musculo-squelettique tridimensionnel avec 36 actionneurs par patte est utilisé, et une méthode numérique nommée "direct collocation" est employée pour formuler un problème de contrôle optimal rapidement résoluble impliquant les phases de décollage et d'atterrissage. La simulation qui en résulte élève le centre de masse du corps entier à plus du double de sa hauteur debout, et les aspects clés du comportement simulé reproduisent qualitativement les observations empiriques d'autres oiseaux sauteurs. Cependant, les performances quantitatives sont moindres, avec une réduction des forces au sol, des hauteurs de saut et de la puissance musculo-tendineuse. Une manœuvre de contre-mouvement prononcée est utilisée pendant le lancement. Il a été démontré que l'utilisation d'un contre-mouvement est essentielle à l'obtention de hauteurs de saut plus importantes, et il se peut que ce phénomène doive exploiter uniquement des principes physiques pour réussir; l'amplification de la performance musculaire n'est pas nécessairement une raison immédiate de l'utilisation de cette manœuvre. L'augmentation de la force musculaire ou de la vitesse de contraction au-dessus des valeurs nominales améliore grandement la performance de saut et, fait intéressant, a le plus grand effet sur les muscles extenseurs des membres plus distaux (c'est-à-dire ceux de la cheville), ce qui suggère que le membre distal peut être un lien critique pour le comportement de saut. Ces résultats justifient une réévaluation des déductions précédentes de la capacité de sauter chez certaines espèces éteintes avec des segments de membres distaux raccourcis, comme les dinosaures droméosauridés. Voorspellende simulaties van musculoskeletale functie en springprestaties bij een gegeneraliseerde vogel Springen is een veel voorkomend, maar veeleisend, gedrag dat veel dieren toepassen tijdens hun dagelijkse bezigheden. In tegenstelling tot de springspecialisten zoals de anura en sommige primaten, is de biomechanica van het springen en de factoren die de prestaties beïnvloeden nog weinig bestudeerd voor algemene soorten die geen uitgesproken adaptaties voor het springen hebben. Computationele biomechanische modelbenaderingen bieden een manier om dit op een rigoureuze, mechanistische manier aan te pakken. Hier worden optimale controle theorie en musculoskeletale modellering geïntegreerd om voorspellende simulaties te genereren van maximale hoogtesprong bij een kleine grondbewonende vogel, een tinamou. Een driedimensionaal musculoskeletaal model met 36 actuatoren per poot wordt gebruikt, en directe collocatie wordt toegepast om een snel oplosbaar optimaal controleprobleem te formuleren dat zowel de opstijg-als de landingsfase omvat. De resulterende simulatie verhoogt het lichaamszwaartepunt tot meer dan het dubbele van de stahoogte, en belangrijke aspecten van het gesimuleerde gedrag komen kwalitatief overeen met empirische waarnemingen voor andere springende vogels. De kwantitatieve prestaties zijn echter minder, met verminderde grondkrachten, spronghoogtes en spierpeeskracht. Tijdens de lancering wordt een uitgesproken tegenbewegingsmanoeuvre gebruikt. Aangetoond is dat het gebruik van een tegenbeweging van cruciaal belang is voor het bereiken van grotere spronghoogten, en dit fenomeen hoeft alleen op fysische principes te berusten om succesvol te zijn; versterking van de spierprestaties hoeft niet noodzakelijk een proximate reden te zijn voor het gebruik van deze manoeuvre. Het verhogen van de spierkracht of van de contractiesnelheid boven de nominale waarden verbetert de sprongprestatie aanzienlijk, en heeft interessant genoeg het grootste effect op de meer distale extensoren van de ledematen (d.w.z. die van de enkel), wat suggereert dat de distale ledematen een kritieke schakel kunnen zijn voor het springgedrag. Deze resultaten rechtvaardigen een herevaluatie van eerdere conclusies over springvermogen bij sommige uitgestorven soorten met voorgekorte distale ledematen, zoals dromaeosauride dinosauriërs. Prädiktive Simulationen der muskuloskelettalen Funktion und Sprungleistung bei einem generalisierten Vogel Springen ist ein übliches jedoch anstrengendes Verhalten, das viele Tiere bei ihren täglichen Aktivitäten einsetzen. Im Gegensatz zu Springspezialisten, wie Fröschen und einigen Primaten, sind bei allgemeinen Arten, welche keine ausgeprägten Anpassung für Sprungverhalten aufweisen, die Biomechanik beim Springen und die Faktoren, welche die Leistungsfähigkeit beeinflussen, noch wenig untersucht. Computergestützte biomechanische Modellierungsverfahren bieten hier eine Möglichkeit, dies in einer gründlichen, mechanistischen Weise anzugehen. In dieser Arbeit werden die optimale Steuerungstheorie und Muskel-Skelett-Modellierung zusammen eingesetzt, um die maximale Sprunghöhe eines kleinen bodenlebenden Vogels, eines Perlsteisshuhns, zu simulieren und zu prognostizieren. Es wird ein dreidimensionales Muskel-Skelett-Modell mit 36 Aktuatoren pro Bein verwendet, und durch direkte Kollokation wird ein schnell lösbares optimales Steuerungsproblem formuliert, das sowohl die Abstoss- als auch die Landephase umfasst. Die daraus folgende Simulation bringt den Ganzkörperschwerpunkt auf mehr als das Doppelte seiner Standhöhe und entscheidende Aspekte des simulierten Verhaltens entsprechen qualitativ empirischen Beobachtungen für andere springende Vögel. Allerdings ist die quantitative Leistungsfähigkeit geringer, mit reduzierten Bodenkräften, Sprunghöhen und Muskel-Sehnen-Kräften. Beim Abstossen wird ein ausgeprägtes Gegenbewegungsmanöver durchgeführt. Die Durchführung einer Gegenbewegung ist nachweislich entscheidend für das Erreichen grösserer Sprunghöhen, wobei dieses Phänomen möglicherweise nur physikalische Prinzipien auszuschöpfen braucht, um erfolgreich zu sein. Die Verstärkung der Muskelleistung ist daher möglicherweise nicht zwingend ein unmittelbarer Grund für die Verwendung dieses Manövers. Eine Erhöhung der Muskelkraft oder der Kontraktionsgeschwindigkeit über die Nominalwerte hinaus führt zu einer erheblichen Zunahme der Sprungleistung und hat interessanterweise den grössten Effekt bei den weiter distal gelegenen Streckmuskeln der Beine (d.h. bei denjenigen des Sprunggelenks), was darauf hindeutet, dass die distale Gliedmasse ein entscheidendes Element für das Sprungverhalten sein könnte. Diese Ergebnisse geben Anlass zur Überprüfung früherer Schlussfolgerungen hinsichtlich der Sprungfähigkeit einiger ausgestorbener Arten mit verkürzten distalen Gliedmassen, wie beispielsweise bei dromaeosauriden Dinosauriern., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

The arrangement and physiology of muscle fibres can strongly influence musculoskeletal function and whole-organismal performance. However, experimental investigation of muscle function during in vivo activity is typically limited to relatively few muscles in a given system. Computational models and simulations of the musculoskeletal system can partly overcome these limitations, by exploring the dynamics of muscles, tendons and other tissues in a robust and quantitative fashion. Here, a high-fidelity, 26-degree-of-freedom musculoskeletal model was developed of the hindlimb of a small ground bird, the elegant-crested tinamou (Eudromia elegans, ~550 g), including all the major muscles of the limb (36 actuators per leg). The model was integrated with biplanar fluoroscopy (XROMM) and forceplate data for walking and running, where dynamic optimization was used to estimate muscle excitations and fibre length changes throughout both gaits. Following this, a series of static simulations over the total range of physiological limb postures were performed, to circumscribe the bounds of possible variation in fibre length. During gait, fibre lengths for all muscles remained between 0.5 to 1.21 times optimal fibre length, but operated mostly on the ascending limb and plateau of the active force-length curve, a result that parallels previous experimental findings for birds, humans and other species. However, the ranges of fibre length varied considerably among individual muscles, especially when considered across the total possible range of joint excursion. Net length change of muscle-tendon units was mostly less than optimal fibre length, sometimes markedly so, suggesting that approaches that use muscle-tendon length change to estimate optimal fibre length in extinct species are likely underestimating this important parameter for many muscles. The results of this study clarify and broaden understanding of muscle function in extant animals, and can help refine approaches used to study extinct species., Competing Interests: The authors have declared that no competing interests exist.

Emerging infectious diseases are on the rise in many different taxa, including, among others, the amphibian batrachochytrids, the snake fungal disease and the Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) virus, responsible for Coronavirus disease 2019 (COVID-19) in mammals. Following the onset of the pandemic linked to COVID-19, eastern Asia has shown strong leadership, taking actions to regulate the trade of potential vector species in several regions. These actions were taken in response to an increase in public awareness, and the need for a quick reaction to mitigate against further pandemics. However, trade restrictions rarely affect amphibians, despite the risk of pathogen transmission, directly, or indirectly through habitat destruction and the loss of vector consumption. Thus, species that help alleviate the risk of zoonoses or provide biological control are not protected. Hence, in view of the global amphibian decline and the risk of zoonoses, we support the current wildlife trade regulations and support measures to safeguard wildlife from overexploitation. The current period of regulation overhaul should be used as a springboard for amphibian conservation. To mitigate risks, we suggest the following stipulations specifically for amphibians. I) Restrictions to amphibian farming in eastern Asia, in relation to pathogen transmission and the establishment of invasive species. II) Regulation of the amphibian pet trade, with a focus on potential vector species. III) Expansion of the wildlife trade ban, to limit the wildlife-human-pet interface. The resulting actions will benefit both human and wildlife populations, as they will lead to a decrease in the risk of zoonoses and better protection of the environment., Significance Statement: There is an increasing number of emerging infectious diseases impacting all species, including amphibians, reptiles and mammals. The latest threat to humans is the virus responsible for COVID-19, and the resulting pandemic. Countries in eastern Asia have taken steps to regulate wildlife trade and prevent further zoonoses thereby decreasing the risk of pathogens arising from wild species. However, as amphibians are generally excluded from regulations we support specific trade restrictions: I) Restrictions to amphibian farming; II) regulation of the amphibian pet trade; III) expansion of the wildlife trade ban. These restrictions will benefit both human and wildlife populations by decreasing the risks of zoonoses and better protecting the environment., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 Elsevier Ltd. All rights reserved.)

Differentiation of cultured skeletal myoblasts is induced by extrinsic signals that include reduction in ambient mitogen concentration and increased cell density. Using an established murine myoblast cell line (C2C12), we have found that experimental reduction of the nucleoporin p62 (Nup62) content of myoblasts enhances differentiation in high-mitogen medium, while forced expression of Nup62 inhibits density-induced differentiation. In contrast, differentiation of myoblasts induced by low-mitogen medium was unaffected by ectopic Nup62 expression. Further analyses suggested that Nup62 content affects density-induced myoblast differentiation through a mechanism involving activation of p38 MAP kinase. Nuclear pore complex (NPC) composition, in particular changes in NUP62 content, may be altered during viral infection, differentiation, and in neoplastic growth. The results support a functional role for changes in Nup62 composition in NPCs and density-induced myogenic differentiation, and suggest a link between loss of Nup62 content and induction of an intracellular stress signaling pathways., (Copyright © 2020. Published by Elsevier B.V.)

The locomotion strategies of fossil invertebrates are typically interpreted on the basis of morphological descriptions. However, it has been shown that homologous structures with disparate morphologies in extant invertebrates do not necessarily correlate with differences in their locomotory capability. Here, we present a new methodology for analysing locomotion in fossil invertebrates with a rigid skeleton through an investigation of a cornute stylophoran, an extinct fossil echinoderm with enigmatic morphology that has made its mode of locomotion difficult to reconstruct. We determined the range of motion of a stylophoran arm based on digitized three-dimensional morphology of an early Ordovician form, Phyllocystis crassimarginata . Our analysis showed that efficient arm-forward epifaunal locomotion based on dorsoventral movements, as previously hypothesized for cornute stylophorans, was not possible for this taxon; locomotion driven primarily by lateral movement of the proximal aulacophore was more likely. Three-dimensional digital modelling provides an objective and rigorous methodology for illuminating the movement capabilities and locomotion strategies of fossil invertebrates., Competing Interests: We declare we have no competing interests., (© 2020 The Authors.)

Despite the global declines in the rate of amphibians, evaluation of public understanding of the crisis has not yet been carried out. We surveyed visitors (n = 1,293) at 15 zoos in Brazil, New Zealand, and the United Kingdom, using a certainty-based assessment method to compare visitor knowledge of the global amphibian crisis. We further analyzed zoo educational material about amphibians to explore its potential to raise awareness through amphibian-focused environmental education. Visitors in the three countries had relatively little understanding of amphibians and the global amphibian crisis. When the degree of confidence in answering the questions (high, medium, and low) is accounted for, correct answers varied between 28% and 39%. This compared to scores of between 58% and 73% when the degree of confidence in responding was not accounted for. However, specific areas of knowledge (e.g., biology, conservation, biogeography, and conceptual ideas) varied significantly across the countries. Visitors had a weaker grasp of biogeographical and conservation issues than general amphibian biology. Zoo visitors in Brazil knew less about amphibian conservation than those in New Zealand or the United Kingdom. There was less amphibian-focused content in educational materials in zoos in Brazil than there was in the United Kingdom. Improving information about the global amphibian crisis may increase support for future conservation actions. Outreach education is one of the most important approaches in any strategic planning for conservation of species. Amphibian-focused environmental education at institutions such as zoos and aquaria can be a crucial intervention to support amphibian conservation worldwide., (© 2019 Wiley Periodicals, Inc.)

Dromaeosaurids were a clade of bird-like, carnivorous dinosaurs that are well known for their characteristic morphology of pedal digit II, which bore an enlarged, sickle-shaped claw and permitted an extreme range of flexion-extension. Proposed functions for the claw often revolve around predation, but the exact manner of use varies widely. Musculoskeletal modelling provides an avenue to quantitatively investigate the biomechanics of this enigmatic system, and thereby test different behavioural hypotheses. Here, a musculoskeletal model of the hindlimb and pes of Deinonychus was developed, and mathematical optimization was used to assess the factors that maximize production of force at the claw tip. Optimization revealed that more crouched hindlimb postures (i.e., more flexed knees and ankles) and larger flexor muscle volumes consistently increased claw forces, although the optimal degree of digit flexion or extension depended on assumptions of muscle activity and fibre operating range. Interestingly, the magnitude of force capable of being produced at the claw tip was relatively small, arguing against regular transmission of a large proportion of body weight into a substrate principally via the claw tip. Such transmission would therefore likely have needed to occur via more proximal parts of the foot. Collectively, the results best support a grasping function for digit II (e.g., restraint of prey smaller than the dromaeosaurid's own body size), although other behaviours involving flexed hindlimbs cannot be excluded., Competing Interests: The author declares there are no competing interests.

The decline of amphibians has been of international concern for more than two decades, and the global spread of introduced fauna is a major factor in this decline. Conservation management decisions to implement control of introduced fauna are often based on diet studies. One of the most common metrics to report in diet studies is Frequency of Occurrence (FO), but this can be difficult to interpret, as it does not include a temporal perspective. Here, we examine the potential for FO data derived from molecular diet analysis to inform invasive species management, using invasive ship rats ( Rattus rattus ) and endemic frogs ( Leiopelma spp.) in New Zealand as a case study. Only two endemic frog species persist on the mainland. One of these, Leiopelma archeyi , is Critically Endangered (IUCN 2017) and ranked as the world's most evolutionarily distinct and globally endangered amphibian (EDGE, 2018). Ship rat stomach contents were collected by kill-trapping and subjected to three methods of diet analysis (one morphological and two DNA-based). A new primer pair was developed targeting all anuran species that exhibits good coverage, high taxonomic resolution, and reasonable specificity. Incorporating a temporal parameter allowed us to calculate the minimum number of ingestion events per rat per night, providing a more intuitive metric than the more commonly reported FO. We are not aware of other DNA-based diet studies that have incorporated a temporal parameter into FO data. The usefulness of such a metric will depend on the study system, in particular the feeding ecology of the predator. Ship rats are consuming both species of native frogs present on mainland New Zealand, and this study provides the first detections of remains of these species in mammalian stomach contents.

Amphibian skin is highly variable in structure and function across anurans, and plays an important role in physiological homeostasis and immune defence. For example, skin sloughing has been shown to reduce pathogen loads on the skin, such as the lethal fungus Batrachochytrium dendrobatidis ( Bd), but interspecific variation in sloughing frequency is largely unknown. Using phylogenetic linear mixed models, we assessed the relationship between skin turnover rate, skin morphology, ecological traits and overall evidence of Bd-driven declines. We examined skin sloughing rates in 21 frog species from three continents, as well as structural skin characteristics measured from preserved specimens. We found that sloughing rate varies significantly with phylogenetic group, but was not associated with evidence of Bd-driven declines, or other skin characteristics examined. This is the first comparison of sloughing rate across a wide range of amphibian species, and creates the first database of amphibian sloughing behaviour. Given the strong phylogenetic signal observed in sloughing rate, approximate sloughing rates of related species may be predicted based on phylogenetic position. While not related to available evidence of declines, understanding variation in sloughing rate may help explain differences in the severity of infection in genera with relatively slow skin turnover rates (e.g. Atelopus).

Musculoskeletal injuries (MSI) in the military reduce soldier capability and impose substantial costs. Characterizing biomechanical surrogates of MSI during commonly performed military tasks (e.g., load carriage) is necessary for evaluating the effectiveness of possible interventions to reduce MSI risk. This study determined the effects of body-borne load distribution, load magnitude, and walking speed on tibiofemoral contact forces. Twenty-one Australian Army Reserve soldiers completed a treadmill walking protocol in an unloaded condition and wearing four armor types (standard-issue and three prototypes) with two load configurations (15 and 30 kg) for a total of 8 armor x load ensembles. In each ensemble, participants completed a 5-minute warm-up, and then walked for 10 minutes at both moderate (1.53 m⋅s-1) and fast (1.81 m⋅s-1) speeds. During treadmill walking, three-dimensional kinematics, ground reaction forces, and muscle activity from nine lower-limb muscles were collected in the final minute of each speed. These data were used as inputs into a neuromusculoskeletal model, which estimated medial, lateral and total tibiofemoral contact forces. Repeated measures analyses of variance revealed no differences for any variables between armor types, but peak medial compartment contact forces increased when progressing from moderate to fast walking and with increased load (p<0.001). Acute exposure to load carriage increased estimated tibiofemoral contact forces 10.1 and 19.9% with 15 and 30kg of carried load, respectively, compared to unloaded walking. These results suggest that soldiers carrying loads in excess of 15 kg for prolonged periods could be at greater risk of knee MSI than those with less exposure., Competing Interests: The authors have declared that no competing interests exist.