Anatomically grounded estimation of hindlimb muscle sizes in Archosauria.

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.

A general locomotion control framework for multi-legged locomotors.

Serially connected robots are promising candidates for performing tasks in confined spaces such as search and rescue in large-scale disasters. Such robots are typically limbless, and we hypothesize that the addition of limbs could improve mobility. However, a challenge in designing and controlling such devices lies in the coordination of high-dimensional redundant modules in a way that improves mobility. Here we develop a general framework to discover templates to control serially connected multi-legged robots. Specifically, we combine two approaches to build a general shape control scheme which can provide baseline patterns of self-deformation ('gaits') for effective locomotion in diverse robot morphologies. First, we take inspiration from a dimensionality reduction and a biological gait classification scheme to generate cyclic patterns of body deformation and foot lifting/lowering, which facilitate the generation of arbitrary substrate contact patterns. Second, we extend geometric mechanics, which was originally introduced to study swimming at low Reynolds numbers, to frictional environments, allowing the identification of optimal body-leg coordination in this common terradynamic regime. Our scheme allows the development of effective gaits on flat terrain with diverse numbers of limbs (4, 6, 16, and even 0 limbs) and backbone actuation. By properly coordinating the body undulation and leg placement, our framework combines the advantages of both limbless robots (modularity and narrow profile) and legged robots (mobility). Our framework can provide general control schemes for the rapid deployment of general multi-legged robots, paving the way toward machines that can traverse complex environments. In addition, we show that our framework can also offer insights into body-leg coordination in living systems, such as salamanders and centipedes, from a biomechanical perspective.

Computational modelling of muscle fibre operating ranges in the hindlimb of a small ground bird (Eudromia elegans), with implications for modelling locomotion in extinct species.

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.

Musculoskeletal modelling of the Nile crocodile (Crocodylus niloticus) hindlimb: Effects of limb posture on leverage during terrestrial locomotion.

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.

Anesthesia and Anesthetic-Related Complications of 8 Elegant-Crested Tinamous (Eudromia elegans) Undergoing Experimental Surgery.

The aim of this study was to describe the anesthetic effects of an injectable anesthetic protocol, based on ketamine, midazolam, and medetomidine, followed by inhalational sevoflurane, in 8 elegant-crested tinamous (Eudromia elegans) undergoing experimental surgery. Initial doses for both injectable agents were tested in 1 bird and then refined with an algorithm based on the effects observed in the pilot procedure. Heart and respiratory rates, as well as nociceptive reflexes, were evaluated before anesthesia (baseline) and intraoperatively, at 10 minute intervals. The time from injection to anesthetic induction and surgical anesthesia, as well as the time from atipamezole injection to recovery, was recorded for each bird. The median doses of medetomidine and ketamine were 0.075 mg/kg and 33 mg/kg, respectively. Anesthetic induction was achieved within 10 (range, 4-45) minutes from intramuscular injection, whereas time to surgical anesthesia was 22 ±16 minutes. The baseline heart rate values were significantly higher than those measured intraoperatively at any time point (P = .001). Intraoperatively, 5 of 8 tinamous (63%) developed cardiac arrhythmias. Other encountered complications were regurgitation in 2 birds (25%), cardiac arrest in 1 bird (13%) soon after injection of the anesthetic agents, and prolonged recovery in another bird (13%), which was euthanized. Necropsy of the 2 fatal outcomes (25%) showed evidence of hepatic lipidosis in both (100%) and intramyocardial fat accumulation in 1 bird (50%). This report highlights the challenges of tinamou anesthesia. Cardiac complications are common in this species, and close monitoring of intraoperative cardiovascular variables is recommended for prompt recognition and treatment.

Appendicular Muscle Physiology and Biomechanics in Crocodylus niloticus.

Archosaurian reptiles (including living crocodiles and birds) had an explosive diversification of locomotor form and function since the Triassic approximately 250 million years ago. Their limb muscle physiology and biomechanics are pivotal to our understanding of how their diversity and evolution relate to locomotor function. Muscle contraction velocity, force, and power in extinct archosaurs such as early crocodiles, pterosaurs, or non-avian dinosaurs are not available from fossil material, but are needed for biomechanical modeling and simulation. However, an approximation or range of potential parameter values can be obtained by studying extant representatives of the archosaur lineage. Here, we study the physiological performance of three appendicular muscles in Nile crocodiles (Crocodylus niloticus). Nile crocodile musculature showed high power and velocity values-the flexor tibialis internus 4 muscle, a small "hamstring" hip extensor, and knee flexor actively used for terrestrial locomotion, performed particularly well. Our findings demonstrate some physiological differences between muscles, potentially relating to differences in locomotor function, and muscle fiber type composition. By considering these new data from a previously unstudied archosaurian species in light of existing data (e.g., from birds), we can now better bracket estimates of muscle parameters for extinct species and related extant species. Nonetheless, it will be important to consider the potential specialization and physiological variation among muscles, because some archosaurian muscles (such as those with terrestrial locomotor function) may well have close to double the muscle power and contraction velocity capacities of others.

Les archosaures, le groupe de reptiles incluant les oiseaux et les crocodiles actuels, sont caractérisés par une diversification importante de leurs formes et fonctions locomotrices depuis le Trias il y a environ 250 millions d'années. Des études biomécaniques et musculaires centrées sur les membres appendiculaires sont donc essentielles pour comprendre le lien qui unit les fonctions locomotrices des archosaures avec leur histoire évolutive et leur forte diversité. Les données les plus fréquemment utilisées, telles que la vitesse de contraction et la force musculaire, ne sont pas accessibles pour les archosaures éteints tels que ceux issus de la lignée fossile des crocodiles (pseudosuchiens), les ptérosaures ainsi que les dinosaures non-aviens. Ces données sont pourtant nécessaires à l'établissement de modélisations et de simulations biomécaniques à l'échelle du groupe. Il est cependant possible d'obtenir une estimation de ces paramètres à partir des archosaures actuels. Cette étude présente en détails la physiologie de trois Crocodiles du Nil (Crocodylus niloticus) en détaillant les performances musculaires de leur appareil locomoteur. Les muscles des Crocodiles du Nil présentent des forces et des vitesses de contraction élevées. Les performances du muscle flexeur tibialis internus 4, qui est un petit muscle ischio-jambier entre la hanche et le genou fréquemment sollicité chez les animaux terrestres, s'avèrent être particulièrement élevées. Notre étude met en évidence des différences de physiologie entre les muscles, potentiellement liées aux différences de fonctions locomotrices et à la composition des différents types de fibres musculaires. En couplant ces nouvelles données avec celles déjà connues chez les oiseaux, il est possible de mieux estimer les paramètres musculaires d'espèces éteintes ainsi que d'espèces actuelles phylogénétiquement proches. Il est également essentiel de considérer les différentes spécialisations ainsi que les variations de physiologie musculaire. En effet, les muscles de certains archosaures, en particulier ceux dotés d'un mode de locomotion terrestre, pourrait présenter des forces et vitesses de contractions musculaires bien supérieures à celles d'autres espèces.By Romain Pintore, RVC.

Relating neuromuscular control to functional anatomy of limb muscles in extant archosaurs.

Electromyography (EMG) is used to understand muscle activity patterns in animals. Understanding how much variation exists in muscle activity patterns in homologous muscles across animal clades during similar behaviours is important for evaluating the evolution of muscle functions and neuromuscular control. We compared muscle activity across a range of archosaurian species and appendicular muscles, including how these EMG patterns varied across ontogeny and phylogeny, to reconstruct the evolutionary history of archosaurian muscle activation during locomotion. EMG electrodes were implanted into the muscles of turkeys, pheasants, quail, guineafowl, emus (three age classes), tinamous and juvenile Nile crocodiles across 13 different appendicular muscles. Subjects walked and ran at a range of speeds both overground and on treadmills during EMG recordings. Anatomically similar muscles such as the lateral gastrocnemius exhibited similar EMG patterns at similar relative speeds across all birds. In the crocodiles, the EMG signals closely matched previously published data for alligators. The timing of lateral gastrocnemius activation was relatively later within a stride cycle for crocodiles compared to birds. This difference may relate to the coordinated knee extension and ankle plantarflexion timing across the swing-stance transition in Crocodylia, unlike in birds where there is knee flexion and ankle dorsiflexion across swing-stance. No significant effects were found across the species for ontogeny, or between treadmill and overground locomotion. Our findings strengthen the inference that some muscle EMG patterns remained conservative throughout Archosauria: for example, digital flexors retained similar stance phase activity and M. pectoralis remained an 'anti-gravity' muscle. However, some avian hindlimb muscles evolved divergent activations in tandem with functional changes such as bipedalism and more crouched postures, especially M. iliotrochantericus caudalis switching from swing to stance phase activity and M. iliofibularis adding a novel stance phase burst of activity.

Terrestrial capture of prey by the reedfish, a model species for stem tetrapods.

Due to morphological resemblance, polypterid fishes are used as extant analogues of Late Devonian lobe-finned sarcopterygians to identify the features that allowed the evolution of a terrestrial lifestyle in early tetrapods. Previous studies using polypterids showed how terrestrial locomotion capacity can develop, and how air ventilation for breathing was possible in extinct tetrapodomorphs. Interestingly, one polypterid species, the reedfish Erpetoichthys calabaricus, has been noted being capable of capturing prey on land. We now identified the mechanism of terrestrial prey-capture in reedfish. We showed that this species uses a lifted trunk and downward inclined head to capture ground-based prey, remarkably similar to the mechanism described earlier for eel-catfish. Reedfish similarly use the ground support and flexibility of their elongated body to realize the trunk elevation and dorsoventral flexion of the anterior trunk region, without a role for the pectoral fins. However, curving of the body to lift the trunk may not have been an option for the Devonian tetrapodomorphs as they are significantly less elongated than reedfish and eel-catfish. This would imply that, in contrast to the eel-like extant species, evolution of the capacity to capture prey on land in early tetrapods may be linked to the evolution of the pectoral system to lift the anterior part of the body.

Functional morphology and kinematics of terrestrial feeding in the largescale foureyes (Anableps anableps).

A major challenge for aquatic vertebrates that invade land is feeding in the terrestrial realm. The capacity of the gape to become parallel with the ground has been shown to be a key factor to allow fishes to feed on prey lying on a terrestrial surface. To do so, two strategies have been identified that involve a nose-down tilting of the head: (1) by pivoting on the pectoral fins as observed in mudskippers, and (2) curling of the anterior part of the body supported by a long and flexible eel-like body as shown in eel-catfish. Although Anableps anableps successfully feeds on land, it does not possess an eel-like body or pectoral fins to support or lift the anterior part of the body. We identified the mechanism of terrestrial prey capture in A. anableps by studying kinematics and functional morphology of the cranial structures, using high-speed video and graphical 3D reconstructions from computed tomography scans. In contrast to the previously described mechanisms, A. anableps relies solely on upper and lower jaw movement for re-orientation of the gape towards the ground. The premaxilla is protruded anteroventrally, and the lower jaw is depressed to a right angle with the substrate. Both the lower and upper jaws are selectively positioned onto the prey. Anableps anableps thereby uses the jaw protrusion mechanism previously described for other cyprinodontiforms to allow a continued protrusion of the premaxilla even while closing the jaws. Several structural adaptations appear to allow more controlled movements and increased amplitude of anterior and ventral protrusion of the upper jaw compared with other cyprinodontiforms.

A fish that uses its hydrodynamic tongue to feed on land.

To capture and swallow food on land, a sticky tongue supported by the hyoid and gill arch skeleton has evolved in land vertebrates from aquatic ancestors that used mouth-cavity-expanding actions of the hyoid to suck food into the mouth. However, the evolutionary pathway bridging this drastic shift in feeding mechanism and associated hyoid motions remains unknown. Modern fish that feed on land may help to unravel the physical constraints and biomechanical solutions that led to terrestrialization of fish-feeding systems. Here, we show that the mudskipper emerges onto land with its mouth cavity filled with water, which it uses as a protruding and retracting 'hydrodynamic tongue' during the initial capture and subsequent intra-oral transport of food. Our analyses link this hydrodynamic action of the intra-oral water to a sequence of compressive and expansive cranial motions that diverge from the general pattern known for suction feeding in fishes. However, the hyoid motion pattern showed a remarkable resemblance to newts during tongue prehension. Consequently, although alternative scenarios cannot be excluded, hydrodynamic tongue usage may be a transitional step onto which the evolution of adhesive mucosa and intrinsic lingual muscles can be added to gain further independence from water for terrestrial foraging.

Functional anatomy and kinematics of the oral jaw system during terrestrial feeding in Periophthalmus barbarus.

The Atlantic mudskipper, Periophthalmus barbarus, is an amphibious fish that successfully overcomes the numerous physical challenges of capturing prey in a terrestrial environment. However, it is unclear what changes in the morphology and function of the feeding apparatus contribute to the mudskipper's successful transition from aquatic to terrestrial capture of prey. In particular, how does the mudskipper achieve effective prehension of land-based prey using its percomorph feeding apparatus? To address that question, we performed a morphological analysis of the feeding apparatus of P. barbarus based on microcomputed tomography scanning, histological sectioning, and dissections as well as a kinematic analysis based on high-speed video and X-ray video to quantify the movements of the oral jaw apparatus elements. Our results show that the neurocranium remains in a fixed position relative to the pectoral girdle as the fish pivots over its pectoral fins toward the prey. The premaxilla rotates dorsally and protrudes downward over the prey. The dentary is rotated ventrally over an angle of 120°, which is facilitated by an intramandibular joint. These motions of the neurocranium, premaxilla, and dentary reorient the mouth aperture so it is parallel to the substrate, thereby allowing the jaws to be placed over the prey. The prey is grabbed between the oral teeth or scooped into the mouth primarily via rapid closing motion of the lower jaw. This analysis of P. barbarus clarifies the morphological and kinematic characteristics required by fish to become successful terrestrial feeders at the environmental transition between water and land.