Emerging biological insights enabled by high-resolution 3D motion data: promises, perspectives and pitfalls.

Deconstructing motion to better understand it is a key prerequisite in the field of comparative biomechanics. Since Marey and Muybridge's work, technical constraints have been the largest limitation to motion capture and analysis, which, in turn, limited what kinds of questions biologists could ask or answer. Throughout the history of our field, conceptual leaps and significant technical advances have generally worked hand in hand. Recently, high-resolution, three-dimensional (3D) motion data have become easier to acquire, providing new opportunities for comparative biomechanics. We describe how adding a third dimension of information has fuelled major paradigm shifts, not only leading to a reinterpretation of long-standing scientific questions but also allowing new questions to be asked. In this paper, we highlight recent work published in Journal of Experimental Biology and influenced by these studies, demonstrating the biological breakthroughs made with 3D data. Although amazing opportunities emerge from these technical and conceptual advances, high-resolution data often come with a price. Here, we discuss challenges of 3D data, including low-throughput methodology, costly equipment, low sample sizes, and complex analyses and presentation. Therefore, we propose guidelines for how and when to pursue 3D high-resolution data. We also suggest research areas that are poised for major new biological advances through emerging 3D data collection.

Realistic three-dimensional avian vocal tract model demonstrates how shape affects sound filtering (Passer domesticus).

Despite the complex geometry of songbird's vocal system, it was typically modelled as a tube or with simple mathematical parameters to investigate sound filtering. Here, we developed an adjustable computational acoustic model of a sparrow's upper vocal tract (Passer domesticus), derived from micro-CT scans. We discovered that a 20% tracheal shortening or a 20° beak gape increase caused the vocal tract harmonic resonance to shift toward higher pitch (11.7% or 8.8%, respectively), predominantly in the mid-range frequencies (3-6 kHz). The oropharyngeal-oesophageal cavity (OEC), known for its role in sound filtering, was modelled as an adjustable three-dimensional cylinder. For a constant OEC volume, an elongated cylinder induced a higher frequency shift than a wide cylinder (70% versus 37%). We found that the OEC volume adjustments can modify the OEC first harmonic resonance at low frequencies (1.5-3 kHz) and the OEC third harmonic resonance at higher frequencies (6-8 kHz). This work demonstrates the need to consider the realistic geometry of the vocal system to accurately quantify its effect on sound filtering and show that sparrows can tune the entire range of produced sound frequencies to their vocal system resonances, by controlling the vocal tract shape, especially through complex OEC volume adjustments.

Foot adaptation to climbing in ovenbirds and woodcreepers (Furnariida).

Furnariida (i.e. ovenbirds, woodcreepers and antbirds) cover diverse ecologies and locomotor habits, ranging from strictly terrestrial to climbing birds, with different degrees of acrobatic performances. We know that this variety of locomotor modes is linked to different limb morpho-functional adaptations in other climbing clades of birds, such as woodpeckers and nuthatches. Here, we link the morphological variations to ecological categories, such as different locomotor habits and a gradient of acrobatic performances, in a phylogenetically informed analysis. We used a high-density three-dimensional (3D) geometric morphometric approach on foot bones (tarsometatarsus and all toes) of 55 specimens from 39 species of Furnariida. We found a significant correlation between acrobatic performances and foot bone shapes, partly explained by the phylogenetic relationship between species. Dendrocolaptidae show specific anatomical features, linked to their acrobatic locomotor habits. More specifically, we found that: (1) foot bones are more robust amongst climbing Furnariida, (2) the spread between toes is wider amongst highly acrobatic Furnariida, (3) dermal syndactyly between digits II and III is linked to special osteological features interpreted as functional osteological syndactyly in woodcreepers (tail-assisted climbers) and (4) the hallux claw is straighter than other claws in climbing Furnariida. Our study demonstrates that climbing Furnariida evolved common foot adaptations with subtle phenotypic variations depending on their climbing performances, refining our understanding of how evolution shapes interactions amongst structure, function and ecological traits.

In vivo intraoral waterflow quantification reveals hidden mechanisms of suction feeding in fish.

Virtually all fishes rely on flows of water to transport food to the back of their pharynx. While external flows that draw food into the mouth are well described, how intraoral waterflows manage to deposit food at the esophagus entrance remains unknown. In theory, the posteriorly moving water must, at some point, curve laterally and/or ventrally to exit through the gill slits. Such flows would eventually carry food away from the esophagus instead of toward it. This apparent paradox calls for a filtration mechanism to deviate food from the suction-feeding streamlines. To study this gap in our fundamental understanding of how fishes feed, we developed and applied a new technique to quantify three-dimensional (3D) patterns of intraoral waterflows in vivo. We combined stereoscopic high-speed X-ray videos to quantify skeletal motion (XROMM) with 3D X-ray particle tracking (XPT) of neutrally buoyant spheres of 1.4 mm in diameter. We show, for carp (Cyprinus carpio) and tilapia (Oreochromis niloticus), that water tracers displayed higher curvatures than food tracers, indicating an inertia-driven filtration. In addition, tilapia also exhibited a 'central jet' flow pattern, which aids in quickly carrying food to the pharyngeal jaw region. When the food was trapped at the branchial basket, it was resuspended and carried more centrally by periodical bidirectional waterflows, synchronized with head-bone motions. By providing a complete picture of the suction-feeding process and revealing fundamental differences in food transport mechanisms among species, this novel technique opens a new area of investigation to fully understand how most aquatic vertebrates feed.

To Hop or Not to Hop? The Answer Is in the Bird Trees.

Birds can use different types of gaits to move on the ground: they either walk, hop, or run. Although velocity can easily explain a preference for running, it remains unclear what drives a bird species to favor hopping over walking. As many hopping birds are relatively small and arboreal, we wanted to test the link between size, arboreality, and hopping ability. First, we carried out ancestral character state reconstructions of size range, hopping ability, and habitat traits on over 1000 species of birds. We found that both hopping ability and arboreality were derived and significantly correlated traits in avian evolution. Second, we tested the influence of hopping ability on the morphology of the lower appendicular skeleton by quantifying the shape differences of the pelvis and the three long bones of the hind limbs in 47 avian species with different habitats and gait preferences. We used geometric morphometrics on 3D landmarks, digitized on micro-computed tomography (micro-CT) and surface scans of the pelvis, femur, tibiotarsus, and tarsometatarsus. Locomotion habits significantly influence the conformation of the pelvis, especially at the origin of hip and knee muscle extensors. Interestingly, habitat, more than locomotion habits, significantly changed tarsometatarsus conformation. The morphology of the distal part of the tarsometatarsus constrains digit orientation, which leads to a greater ability to perch, an advantageous trait in arboreality. The results of this work suggest an arboreal origin of hopping and illuminate the evolution of avian terrestrial locomotion.[Anatomy; avian; gait; leg; lifestyle; pelvis; tree-dwelling.].

How to walk carrying a huge egg? Trade-offs between locomotion and reproduction explain the special pelvis and leg anatomy in kiwi (Aves; Apteryx spp.).

Kiwi (Aves; genus Apteryx) are famous for laying an enormous egg in comparison with their relatively small body size. Considering the peculiar gait of this flightless bird, we suspected the existence of morpho-functional trade-offs between reproduction and locomotion. To understand how structural constraints, imposed by a large egg size, might influence the terrestrial locomotion of Apteryx, we analysed the anatomy of the limb osteomuscular system in two species of kiwi (Apteryx mantelli and Apteryx owenii). We performed detailed dissections and brought to light specific anatomical features of kiwi, in comparison with other ratites and neognathous birds. Our osteological study revealed a strongly curved pelvis, a rigid tail, and enlarged ribs. Our myology study showed an unusual location of the caudofemoralis muscle origin and insertion. The insertion of the pars pelvica along the entire caudal face of the femur, contrasts with the proximal insertion usually seen in other birds. Additionally, the pars caudalis originates along the entire tail, whereas it only inserts on the uropygium in the other birds. To interpret these specificities from a functional point of view, we built three-dimensional osteomuscular models based on computed tomography scans, radiographies and our dissections. We chose three postures associated with reproductive constraints: the standing position of a gravid compared with a non-gravid bird, as well as the brooding position. The 3D model of the brooding position suggested that the enlarged ribs could support the bodyweight when leaning on the huge egg in both males and females. Moreover, we found that in gravid females, the unusual shape of the pelvis and tail allowed the huge egg to sit ventrally below the pelvis, whereas it is held closer to the rachis in other birds. The specific conformation of the limb and the insertions of the two parses of the caudofemoralis help to maintain the tail flexed, and to keep the legs adducted when carrying the egg. The caudal location of the hip and its flexed position explains the long stance phase during the strange gait of kiwi, revealing the functional trade-off between reproduction and locomotion in this emblematic New Zealand bird.

Whole-body 3D kinematics of bird take-off: key role of the legs to propel the trunk.

Previous studies showed that birds primarily use their hindlimbs to propel themselves into the air in order to take-off. Yet, it remains unclear how the different parts of their musculoskeletal system move to produce the necessary acceleration. To quantify the relative motions of the bones during the terrestrial phase of take-off, we used biplanar fluoroscopy in two species of birds, diamond dove (Geopelia cuneata) and zebra finch (Taeniopygia guttata). We obtained a detailed 3D kinematics analysis of the head, the trunk and the three long bones of the left leg. We found that the entire body assisted the production of the needed forces to take-off, during two distinct but complementary phases. The first one, a relatively slow preparatory phase, started with a movement of the head and an alignment of the different groups of bones with the future take-off direction. It was associated with a pitch down of the trunk and a flexion of the ankle, of the hip and, to a lesser extent, of the knee. This crouching movement could contribute to the loading of the leg muscles and store elastic energy that could be released in the propulsive phase of take-off, during the extension of the leg joints. Combined with the fact that the head, together with the trunk, produced a forward momentum, the entire body assisted the production of the needed forces to take-off. The second phase was faster with mostly horizontal forward and vertical upward translation motions, synchronous to an extension of the entire lower articulated musculoskeletal system. It led to the propulsion of the bird in the air with a fundamental role of the hip and ankle joints to move the trunk upward and forward. Take-off kinematics were similar in both studied species, with a more pronounced crouching movement in diamond dove, which can be related to a large body mass compared to zebra finch.

Hydrodynamic performance of suction feeding is virtually unaffected by variation in the shape of the posterior region of the pharynx in fish.

To capture prey by suction, fish generate a flow of water that enters the mouth and exits at the back of the head. It was previously hypothesized that prey-capture performance is improved by a streamlined shape of the posterior region of the pharynx, which enables an unobstructed outflow with minimal hydrodynamic resistance. However, this hypothesis remained untested for several decades. Using computational fluid dynamics simulations, we now managed to quantify the effects of different shapes of the posterior pharynx on the dynamics of suction feeding, based on a feeding act of a sunfish (Lepomis gibbosus). In contrast to what was hypothesized, the effects of the imposed variation in shape were negligible: flow velocity patterns remained essentially identical, and the effects on feeding dynamics were negligibly small. This remarkable hydrodynamic insensitivity implies that, in the course of evolution, the observed wedge-like protrusions of the pectoral surfaces of the pharynx probably resulted from spatial constraints and/or mechanical demands on the musculoskeletal linkages, rather than constraints imposed by hydrodynamics. Our study, therefore, exceptionally shows that a streamlined biological shape subjected to fluid flows is not always the result of selection for hydrodynamic improvement.

Transition from wing to leg forces during landing in birds.

Transitions to and from the air are critical for aerial locomotion and likely shaped the evolution of flying animals. Research on take-off demonstrates that legs generate greater body accelerations compared with wings, and thereby contribute more to initial flight velocity. Here, we explored coordination between wings and legs in two species with different wingbeat styles, and quantified force production of these modules during the final phase of landing. We used the same birds that we had previously studied during take-off: zebra finch (Taeniopygia guttata, N=4) and diamond dove (Geopelia cuneata, N=3). We measured kinematics using high-speed video, aerodynamics using particle image velocimetry, and ground-reaction forces using a perch mounted on a force plate. In contrast with the first three wingbeats of take-off, the final four wingbeats during landing featured ~2 times greater force production. Thus, wings contribute proportionally more to changes in velocity during the last phase of landing compared with the initial phase of take-off. The two species touched down at the same velocity (~1 m s(-1)), but they exhibited significant differences in the timing of their final wingbeat relative to touchdown. The ratio of average wing force to peak leg force was greater in diamond doves than in zebra finches. Peak ground reaction forces during landing were ~50% of those during take-off, consistent with the birds being motivated to control landing. Likewise, estimations of mechanical energy flux for both species indicate that wings produce 3-10 times more mechanical work within the final wingbeats of flight compared with the kinetic energy of the body absorbed by legs during ground contact.

Transition from leg to wing forces during take-off in birds.

Take-off mechanics are fundamental to the ecology and evolution of flying animals. Recent research has revealed that initial take-off velocity in birds is driven mostly by hindlimb forces. However, the contribution of the wings during the transition to air is unknown. To investigate this transition, we integrated measurements of both leg and wing forces during take-off and the first three wingbeats in zebra finch (Taeniopygia guttata, body mass 15 g, N=7) and diamond dove (Geopelia cuneata, body mass 50 g, N=3). We measured ground reaction forces produced by the hindlimbs using a perch mounted on a force plate, whole-body and wing kinematics using high-speed video, and aerodynamic forces using particle image velocimetry (PIV). Take-off performance was generally similar between species. When birds were perched, an acceleration peak produced by the legs contributed to 85±1% of the whole-body resultant acceleration in finch and 77±6% in dove. At lift-off, coincident with the start of the first downstroke, the percentage of hindlimb contribution to initial flight velocity was 93.6±0.6% in finch and 95.2±0.4% in dove. In finch, the first wingbeat produced 57.9±3.4% of the lift created during subsequent wingbeats compared with 62.5±2.2% in dove. Advance ratios were <0.5 in both species, even when taking self-convection of shed vortices into account, so it was likely that wing-wake interactions dominated aerodynamics during wingbeats 2 and 3. These results underscore the relatively low contribution of the wings to initial take-off, and reveal a novel transitional role for the first wingbeat in terms of force production.

Walking, paddling, waddling: 3D kinematics anatidae locomotion (Callonetta leucophrys).

Walking and paddling motions were studied in a semiaquatic bird, the ringed teal (Callonetta leucophrys), to investigate the motions associated with movements in two environments with radically divergent physical properties. A three-dimensional (3D) kinematic reconstruction based on nonsynchronous biplanar cineradiographic data was used to quantify the 3D trajectories of the body and hind limb segments. Our study revealed that two subsystems interact to provide propulsion in water and on land. During paddling, the trunk, the femur, and the tibiotarsus are in a stable position and play the role of the hull. The femur and tibiotarsus are positioned laterally and parasagittaly and the intertarsal joint is fixed and positioned caudally allowing large amplitude movements of the "paddle" (tarsometatarsus and palmate foot). During walking, the center of mass is held above the medially oriented foot, providing stability during the single support phase. During stance, the foot is medially oriented because of the lateral and parasagittal positions of the tibiotarsus and tarsometatarsus during both walking and paddling. This position of the foot during walking imposes trunk translation and results in the typical waddling motion of Anatidae. This study provides new insights into how waddling motion relates to semiaquatic birds' ability to move in both terrestrial and aquatic environments.

Bird terrestrial locomotion as revealed by 3D kinematics.

Most birds use at least two modes of locomotion: flying and walking (terrestrial locomotion). Whereas the wings and tail are used for flying, the legs are mainly used for walking. The role of other body segments remains, however, poorly understood. In this study, we examine the kinematics of the head, the trunk, and the legs during terrestrial locomotion in the quail (Coturnix coturnix). Despite the trunk representing about 70% of the total body mass, its function in locomotion has received little scientific interest to date. This prompted us to focus on its role in terrestrial locomotion. We used high-speed video fluoroscopic recordings of quails walking at voluntary speeds on a trackway. Dorso-ventral and lateral views of the motion of the skeletal elements were recorded successively and reconstructed in three dimensions using a novel method based on the temporal synchronisation of both views. An analysis of the trajectories of the body parts and their coordination showed that the trunk plays an important role during walking. Moreover, two sub-systems participate in the gait kinematics: (i) the integrated 3D motion of the trunk and thighs allows for the adjustment of the path of the centre of mass; (ii) the motion of distal limbs transforms the alternating forward motion of the feet into a continuous forward motion at the knee and thus assures propulsion. Finally, head bobbing appears qualitatively synchronised to the movements of the trunk. An important role for the thigh muscles in generating the 3D motion of the trunk is suggested by an analysis of the pelvic anatomy.