From quadrupedal to bipedal walking 'on the fly': the mechanics of dynamical mode transition in primates.

We investigated how baboons transition from quadrupedal to bipedal walking without any significant interruption in their forward movement (i.e. transition 'on the fly'). Building on basic mechanical principles (momentum only changes when external forces/moments act on the body), insights into possible strategies for such a dynamical mode transition are provided and applied first to the recorded planar kinematics of an example walking sequence (including several continuous quadrupedal, transition and subsequent bipedal steps). Body dynamics are calculated from the kinematics. The strategy used in this worked example boils down to: crouch the hind parts and sprint them underneath the rising body centre of mass. Forward accelerations are not in play. Key characteristics of this transition strategy were extracted: progression speed, hip height, step duration (frequency), foot positioning at touchdown with respect to the hip and the body centre of mass (BCoM), and congruity between the moments of the ground reaction force about the BCoM and the rate of change of the total angular moment. Statistical analyses across the full sample (15 transitions of 10 individuals) confirm this strategy is always used and is shared across individuals. Finally, the costs (in J kg-1 m-1) linked to on the fly transitions were estimated. The costs are approximately double those of both the preceding quadrupedal and subsequent bipedal walking. Given the short duration of the transition as such (<1 s), it is argued that the energetic costs to change walking posture on the fly are negligible when considered in the context of the locomotor repertoire.

Parallel evolution of semicircular canal form and sensitivity in subterranean mammals.

The vertebrate vestibular system is crucial for balance and navigation, and the evolution of its form and function in relation to species' lifestyle and mode of locomotion has been the focus of considerable recent study. Most research, however, has concentrated on aboveground mammals, with much less published on subterranean fauna. Here, we explored variation in anatomy and sensitivity of the semicircular canals among 91 mammal species, including both subterranean and non-subterranean representatives. Quantitative phylogenetically informed analyses showed significant widening of the canals relative to radius of curvature in subterranean species. A relative canal width above 0.166 indicates with 95% certainty that a species is subterranean. Fluid-structure interaction modelling predicted that canal widening leads to a substantial increase in canal sensitivity; a reasonably good estimation of the absolute sensitivity is possible based on the absolute internal canal width alone. In addition, phylogenetic comparative modelling and functional landscape exploration revealed repeated independent evolution of increased relative canal width and anterior canal sensitivity associated with the transition to a subterranean lifestyle, providing evidence of parallel adaptation. Our results suggest that living in dark, subterranean tunnels requires good balance and/or navigation skills which may be facilitated by more sensitive semicircular canals.

Synchrotron radiation X-ray microtomography for the visualization of intra-cochlear anatomy in human temporal bones implanted with a perimodiolar cochlear implant electrode array.

Recently, synchrotron radiation computed microtomography (SRµCT) has emerged as a promising tool for non-destructive, in situ visualization of cochlear implant electrode arrays inserted into a human cochlea. Histological techniques have been the `gold standard' technique for accurate localization of cochlear implant electrodes but are suboptimal for precise three-dimensional measurements. Here, an SRµCT experimental setup is proposed that offers the benefit of a high spatial and contrast resolution (isotropic voxel size = 4.95 µm and propagation-based phase-contrast imaging), while visualizing the soft-tissue structures and electrode array of the cochlear implant simultaneously. In this work, perimodiolar electrode arrays have been tested, which incorporate thick and closely spaced platinum-iridium contacts and wiring. These data can assist cochlear implant and hearing research, can be used to verify electrode segmentation techniques for clinical computed tomography or could be utilized to evaluate cochlear implant electrode array designs.

Habitat use and vestibular system's dimensions in lacertid lizards.

The vestibular system is crucial for movement control during locomotion. As the dimensions of the vestibular system determine the fluid dynamics of the endolymph and, as such, the system's function, we investigate the interaction between vestibular system size, head size and microhabitat use in lizards. We grouped 24 lacertid species in three microhabitat types, we acquired three-dimensional models of the bony vestibular systems using micro-computer tomography scanning, and we performed linear and surface measurements. All vestibular measurements scale with a negative allometry with head size, suggesting that smaller heads house disproportionally large ears. As the sensitivity of the vestibular system is positively related to size, a sufficiently large vestibular system in small-headed animals may meet the sensitivity demands during challenged locomotion. We also found that the microhabitat affects the locomotor dynamics: lizards inhabiting open microhabitats run at higher dimensionless speeds. On the other hand, no statistical relationship exists between dimensionless speed and the vestibular system dimensions. Hence, if the vestibular size would differ between microhabitats, this would be a direct effect (i.e. imposed, for instance, by requirements for manoeuvring, balance control, etc.), rather than depending on the lizards' intrinsic running speed. However, we found no effect of the microhabitat on the allometric relationship between head and vestibular system size. The finding that microhabitat is not reflected in the vestibular system size (hence sensitivity) of the lacertids in this study is possibly due to spatial constraints of the skull.

Compliant legs enable lizards to maintain high running speeds on complex terrains.

Substrate variations are likely to constrain animal performance in natural environments, as running over complex terrains challenges the dynamic stability of the body differently in each step. Yet, being able to negotiate complex terrains at top speed is a strong advantage for animals that have to deal with predators and evasive prey. Little is known on how animals negotiate such terrain variability at high speed. We investigated this in fast-running Acanthodactylus boskianus lizards, by measuring their 3D kinematics using four synchronised high-speed video cameras (325 Hz) on an adaptable racetrack. This racetrack was covered with four different substrates, representing increasing levels of terrain complexity. We found that the lizards deal with this complexity gradient by gradually adopting more erect parasagittal leg postures. Legs in a more-erect position are more compliant and are therefore highly adjustable on complex terrains. Additionally, the lizards stabilise their head, which facilitates vestibular and visual perception. Together, compliant legs and head stabilisation enable the lizards to minimise movements of the body centre of mass, even when running on highly irregular terrains. This suggests that the head and the centre of mass are the priority targets for running on uneven terrains. As a result, running performance (mean forward speed) decreases only slightly, and only on the most challenging substrate under investigation.

Enhanced contrast in X-ray microtomographic images of the membranous labyrinth using different X-ray sources and scanning modes.

The vestibular system, located in the inner ear, plays a crucial role in balance and gaze stabilisation by sensing head movements. The interconnected tubes with membranous walls of the vestibular system are located in the skull bone (the 'membranous labyrinth'). Unfortunately, these membranes are very hard to visualise using three-dimensional (3D) X-ray imaging techniques. This difficulty arises due to the embedment of the membranes in the dense skull bone, the thinness of the membranes, and the small difference in X-ray absorption between the membranes and the surrounding fluid. In this study, we compared the visualisation of very small specimens (lizard heads with vestibular systems smaller than 3 mm) by X-ray computed micro-tomography (µCT) based on synchrotron radiation and conventional sources. A visualisation protocol using conventional X-ray µCT would be very useful thanks to the ease of access and lower cost. Careful optimisation of the acquisition parameters enables detection of the membranes by using µCT scanners based on conventional microfocus sources, but in some cases a low contrast-to-noise ratio (CNR) prevents fast and reliable segmentation of the membranes. Synchrotron radiation µCT proved to be preferable for the visualisation of the small samples with very thin membranes, because of their high demands for spatial and contrast resolution. The best contrast was obtained by using synchrotron radiation µCT working in phase-contrast mode, leading to up to twice as high CNRs than the best conventional µCT results. The CNR of the synchrotron radiation µCT scans was sufficiently high enough to enable the construction of a 3D model by the means of semi-automatic segmentation of the membranous labyrinth. Membrane thickness was found to range between 2.7 and 36.3 µm. Hence, the minimal membrane thickness was found to be much smaller than described previously in the literature (between 10 and 50 µm).

How innate is locomotion in precocial animals? A study on the early development of spatio-temporal gait variables and gait symmetry in piglets.

Locomotion is one of the most important ecological functions in animals. Precocial animals, such as pigs, are capable of independent locomotion shortly after birth. This raises the question whether coordinated movement patterns and the underlying muscular control in these animals is fully innate or whether there still exists a rapid maturation. We addressed this question by studying gait development in neonatal pigs through the analysis of spatio-temporal gait characteristics during locomotion at self-selected speed. To this end, we made video recordings of piglets walking along a corridor at several time points (from 0 h to 96 h). After digitization of the footfalls, we analysed self-selected speed and spatio-temporal characteristics (e.g. stride and step lengths, stride frequency and duty factor) to study dynamic similarity, intralimb coordination and interlimb coordination. To assess the variability of the gait pattern, left-right asymmetry was studied. To distinguish neuromotor maturation from effects caused by growth, both absolute and normalized data (according to the dynamic similarity concept) were included in the analysis. All normalized spatio-temporal variables reached stable values within 4 h of birth, with most of them showing little change after the age of 2 h. Most asymmetry indices showed stable values, hovering around 10%, within 8 h of birth. These results indicate that coordinated movement patterns are not entirely innate, but that a rapid neuromotor maturation, potentially also the result of the rearrangement or recombination of existing motor modules, takes place in these precocial animals.

Jaw morphology and fighting forces in stag beetles.

The jaws of different species of stag beetles show a large variety of shapes and sizes. The male jaws are used as weapons in fights, and they may exert a very forceful bite in some species. We investigated in 16 species whether and how the forcefulness of their bite is reflected in their jaw morphology. We found a large range of maximal muscle forces (1.8-33 N; factor of 18). Species investing in large bite muscles also have disproportionately large jaw volumes. They use this additional jaw volume to elongate their jaws, increasing their chances of winning in battles. The fact that this also decreases the mechanical advantage is largely compensated for by elongated in-levers. As a result, high muscle forces are correlated with elevated bite forces (0.27-7.6 N; factor of 28). Despite the large difference in the forcefulness of their bite, all investigated species experience similar Von Mises stresses in their jaws while biting (29-114 MPa; factor of 4.0; calculated with finite element simulations). Hence, stag beetles have successfully adapted their jaw anatomy according to their bite force in fights.

Biomechanical determinants of bite force dimorphism in Cyclommatus metallifer stag beetles.

In the stag beetle family (Lucanidae), males have diverged from females by sexual selection. The males fight each other for mating opportunities with their enlarged mandibles. It is known that owners of larger fighting apparatuses are favoured to win the male-male fights, but it was unclear whether male stag beetles also need to produce high bite forces while grabbing and lifting opponents in fights. We show that male Cyclommatus metallifer stag beetles bite three times as forcefully as females. This is not entirely unexpected given the spectacular nature of the fights, but all the more impressive given the difficulty of achieving this with their long mandibles (long levers). Our results suggest no increase in male intrinsic muscle strength to accomplish this. However, morphological analyses show that the long mandibular output levers in males are compensated by elongated input levers (and thus a wider anterior side of the head). The surplus of male bite force capability is realized by enlargement of the closer muscles of the mandibles, while overall muscle force direction remained optimal. To enable the forceful bites required to ensure male reproductive success, male head size and shape are adapted for long input levers and large muscles. Therefore, the entire head should be regarded as an integral part of male armature.

Volume shrinkage of bone, brain and muscle tissue in sample preparation for micro-CT and light sheet fluorescence microscopy (LSFM).

Two methods are especially suited for tomographic imaging with histological detail of macroscopic samples that consist of multiple tissue types (bone, muscle, nerve or fat): Light sheet (based) fluorescence microscopy (LSFM) and micro-computed tomography (micro-CT). Micro-CT requires staining with heavy chemical elements (and thus fixation and sometimes dehydration) in order to make soft tissue imageable when measured alongside denser structures. LSMF requires fixation, decalcification, dehydration, clearing and staining with a fluorescent dye. The specimen preparation of both imaging methods is prone to shrinkage, which is often not mentioned, let alone quantified. In this paper the presence and degree of shrinkage are quantitatively identified for the selected preparation methods/stains. LSFM delivers a volume shrinkage of 17% for bone, 56% for muscle and 62% for brain tissue. The three most popular micro-CT stains (phosphotungstic acid, iodine with potassium iodide, and iodine in absolute ethanol) deliver a volume shrinkage ranging from 10 to 56% for muscle and 27-66% for brain, while bone does not shrink in micro-CT preparation.

Keels of boxfish carapaces strongly improve stabilization against roll.

Boxfish (Ostraciidae) have peculiar body shapes, with conspicuous keels formed by their bony carapaces. Previous studies have proposed various hydrodynamic roles for these keels, including reducing drag during swimming, contributing to passive stabilization of the swimming course, or providing resistance against roll rotations. Here, we tested these hypotheses using computational fluid dynamics simulations of five species of Ostraciidae with a range of carapace shapes. The hydrodynamic performance of the original carapace surface models, obtained from laser scanning of museum specimens, was compared with models where the keels had been digitally reduced. The original carapaces showed no reduced drag or increased passive stability against pitch and yaw compared to the reduced-keel carapaces. However, consistently for all studied species, a strong increase in roll drag and roll-added mass was observed for the original carapaces compared to the reduced-keel carapaces, despite the relatively small differences in keel height. In particular, the damping of roll movement by resistive drag torques increased considerably by the presence of keels. Our results suggest that the shape of the boxfish carapace is important in enabling the observed roll-free forward swimming of boxfish and may facilitate the control of manoeuvres.

Living in darkness: Exploring adaptation of Proteus anguinus in 3 dimensions by X-ray imaging.

BACKGROUND: Lightless caves can harbour a wide range of living organisms. Cave animals have evolved a set of morphological, physiological, and behavioural adaptations known as troglomorphisms, enabling their survival in the perpetual darkness, narrow temperature and humidity ranges, and nutrient scarcity of the subterranean environment. In this study, we focused on adaptations of skull shape and sensory systems in the blind cave salamander, Proteus anguinus, also known as olm or simply proteus-the largest cave tetrapod and the only European amphibian living exclusively in subterranean environments. This extraordinary amphibian compensates for the loss of sight by enhanced non-visual sensory systems including mechanoreceptors, electroreceptors, and chemoreceptors. We compared developmental stages of P. anguinus with Ambystoma mexicanum, also known as axolotl, to make an exemplary comparison between cave- and surface-dwelling paedomorphic salamanders. FINDINGS: We used contrast-enhanced X-ray computed microtomography for the 3D segmentation of the soft tissues in the head of P. anguinus and A. mexicanum. Sensory organs were visualized to elucidate how the animal is adapted to living in complete darkness. X-ray microCT datasets were provided along with 3D models for larval, juvenile, and adult specimens, showing the cartilage of the chondrocranium and the position, shape, and size of the brain, eyes, and olfactory epithelium. CONCLUSIONS: P. anguinus still keeps some of its secrets. Our high-resolution X-ray microCT scans together with 3D models of the anatomical structures in the head may help to elucidate the nature and origin of the mechanisms behind its adaptations to the subterranean environment, which led to a series of troglomorphisms.

Modelling shows that stimulation of the semicircular canals depends on the rotation centre.

The vestibular system in the inner ear senses head movements, which is indispensable for balance during locomotion. The semicircular canals of the vestibular system are stimulated during head rotations, due to the interplay between its endolymph fluid and its cupula membrane. Until now, it has been assumed that the stimulation of the canals is independent of the rotation centre. However, Fluid-Structure Interaction investigation now shows that the horizontal canal is more strongly stimulated when the rotation centre lies further from the canal. Such eccentric rotations result in a different stimulation of the left and the right canal, because the distance to the rotation centre differs between both ears. The dissimilar stimulation enables localization of the rotation centre based on information from the canals alone, similar to sound localization based on the interaural time difference for hearing. Additionally, a rotation centre further away from the canal will probably elicit a larger reflexive eye movement. This is exactly what is required to compensate the larger change in viewing angle, and it corresponds to experimental findings. Two fictitious forces (d'Alembert forces), the centrifugal force and the Euler force, cause the rotation centre-dependency of the canal stimulation. Implications for (pre)clinical eccentric rotation experiments are discussed.

High ellipticity reduces semi-circular canal sensitivity in squamates compared to mammals.

The semi-circular canals in the inner ear sense head rotations. It is widely recognised that the anatomy of the semi-circular canals is often adapted to the species-specific agility, in order to provide the necessary sensitivity. Based on research on mammals, the ellipticity of the semi-circular canal was so far considered as a non-important factor herein. A dataset of 125 squamate species and 156 mammalian species, now shows that the posterior semi-circular canal of squamates is much more elliptical (eccentricities ranging between 0.76 and 0.94) than that of mammals (eccentricities ranging between 0 and 0.71). Fluid-Structure Interaction computer models show that the effect of the ellipticity on sensitivity is strongest in small semi-circular canals. This new insight indicates that the high ellipticity in squamates leads to a severe reduction in sensitivity of up to 45%. In mammals, on the other hand, the reduction in sensitivity is limited to 13%, which is consistent with previous literature that found a limited effect of semi-circular canal ellipticity in mammals. Further, there is a strongly negative correlation between semi-circular canal size and eccentricity in squamates, which is absent in mammals. Hence, the smallest squamates have the most elliptical semi-circular canals. In general, the smaller the semi-circular canal, the less sensitive it is. Therefore, the highly elliptical squamate canals are probably the result of fitting the largest possible canal in small and flat head. Miniaturising the canals while maintaining a circular shape would reduce the sensitivity by another 73% compared to the highly elliptical canals.

Small vertebrates running on uneven terrain: a biomechanical study of two differently specialised lacertid lizards.

While running, small animals frequently encounter large terrain variations relative to their body size, therefore, terrain variations impose important functional demands on small animals. Nonetheless, we have previously observed in lizards that running specialists can maintain a surprisingly good running performance on very uneven terrains. The relatively large terrain variations are offset by their capacity for leg adjustability that ensures a 'smooth ride' of the centre of mass (CoM). The question as to how the effect of an uneven terrain on running performance and locomotor costs differs between species exhibiting diverse body build and locomotor specializations remains. We hypothesise that specialized runners with long hind limbs can cross uneven terrain more efficiently than specialized climbers with a dorso-ventrally flattened body and equally short fore and hind limbs. This study reports 3D kinematics using high-speed videos (325 Hz) to investigate leg adjustability and CoM movements in two lacertid lizards (Acanthodactylus boskianus, running specialist; Podarcis muralis, climbing specialist). We investigated these parameters while the animals were running on a level surface and over a custom-made uneven terrain. We analysed the CoM dynamics, we evaluated the fluctuations of the positive and negative mechanical energy, and we estimated the overall cost of transport. Firstly, the results reveal that the climbers ran at lower speeds on flat level terrain but had the same cost of transport as the runners. Secondly, contrary to the running specialists, the speed was lower and the energy expenditure higher in the climbing specialists while running on uneven terrain. While leg movements adjust to the substrates' variations and enhance the stability of the CoM in the running specialist, this is not the case in the climbing specialist. Although their legs are kept more extended, the amplitude of movement does not change, resulting in an increase of the movement of the CoM and a decrease in locomotor efficiency. These results are discussed in light of the respective (micro-)habitat of these species and suggest that energy economy can also be an important factor for small vertebrates.

Head stabilisation in fast running lizards.

The cyclic patterns of terrestrial animal locomotion are frequently perturbed in natural environments. The terrain can be complex or inclined, the substrate can move unexpectedly and animals can misjudge situations. Loosing stability due to perturbations increases the probability of capture by predators and decreases the chance of successful prey capture and winning intraspecific battles. When controlled corrective actions are necessary to negotiate perturbations, animals rely on their exteroceptive and proprioceptive senses to monitor the environment and their own body movements. The vestibular system in the inner ear perceives linear and angular accelerations. This information enables gaze stabilisation and the creation of a stable, world-bound reference frame for the integration of the information of other senses. During locomotion, both functions are known to be facilitated by head stabilisation in several animals with an erect posture. Animals with a sprawled body posture, however, undergo very large body undulations while running. Using high speed video recordings, we tested whether they nevertheless stabilise their head during running, and how this is influenced by perturbations. We found that running Acanthodactylus boskianus lizards strongly stabilise their head yaw rotations when running on a flat, straight runway: the head rotation amplitude is only 4.76±0.99°, while the adjacent trunk part rotates over 27.0±3.8°. Lateral head translations are not stabilised (average amplitude of 7.4±2.0mm). When the lizards are experimentally perturbed by a large and unexpected lateral substrate movement, lateral translations of both the head and the body decrease (on average by 1.52±0.81mm). At the same time, the rotations of the head and trunk also decrease (on average by 1.62°±7.21°). These results show that head stabilisation intensifies because of the perturbation, which emphasises the importance of vestibular perception and balance in these fast and manoeuvrable animals.

Evolutionary morphology of the lizard chemosensory system.

Foraging mode plays a pivotal role in traditional reconstructions of squamate evolution. Transitions between modes are said to spark concerted changes in the morphology, physiology, behaviour, and life history of lizards. With respect to their sensory systems, species that adopt a sit-and-wait strategy are thought to rely on visual cues primarily, while actively hunting species would predominantly use chemical information. The morphology of the tongue and the vomeronasal-organs is believed to mirror this dichotomy. Still, support for this idea of concerted evolution of the morphology of the lizard sensory system merely originates from studies comparing only a few, distantly related taxa that differ in many aspects of their biology besides foraging mode. Hence, we compared vomeronasal-lingual morphology among closely related lizard species (Lacertidae). Our findings show considerable interspecific variation indicating that the chemosensory system of lacertids has undergone substantial change over a short evolutionary time. Although our results imply independent evolution of tongue and vomeronasal-organ form, we find evidence for co-variation between sampler and sensor, hinting towards an 'optimization' for efficient chemoreception. Furthermore, our findings suggest species' degree of investment in chemical signalling, and not foraging behaviour, as a leading factor driving the diversity in vomeronasal-lingual morphology among lacertid species.

Author Correction: Evolutionary morphology of the lizard chemosensory system.

A correction to this article has been published and is linked from the HTML version of this paper. The error has not been fixed in the paper.

Role of stag beetle jaw bending and torsion in grip on rivals.

In aggressive battles, the extremely large male stag beetle jaws have to withstand strongly elevated bite forces. We found several adaptations of the male Cyclommatus metallifer jaw morphology for enhanced robustness that conspecific females lack. As a result, males improve their grip on opponents and they maintain their safety factor (5.2-7.2) at the same level as that of females (6.8), despite their strongly elevated bite muscle force (3.9 times stronger). Males have a higher second moment of area and torsion constant than females, owing to an enhanced cross-sectional area and shape. These parameters also increase faster with increasing bending moment towards the jaw base in males than in females. Male jaws are more bending resistant against the bite reaction force than against perpendicular forces (which remain lower in battles). Because of the triangular cross section of the male jaw base, it twists more easily than it bends. This torsional flexibility creates a safety system against overload that, at the same time, secures a firm grip on rivals. We found no structural mechanical function of the large teeth halfway along the male jaws. Therefore, it appears that the main purpose of these teeth is a further improvement of grip on rivals.

On the connection between the tympanic membrane and the malleus.

BACKGROUND: The tympano-mallear connection (TMC) is the soft-tissue connection between the tympanic membrane (TM) and the manubrium of the malleus. Some studies suggest that its mechanical properties may have a substantial influence on the mechanics and transfer function of the middle ear. However, relatively little is known about the dimensions of the TMC and its variability among individuals. METHOD: Thirteen samples were collected from human temporal bones, consisting of only the malleus and the TM. They were imaged using µCT without contrast enhancing agent. From the µCT images, the TMC dimensions were measured in both anterior-posterior direction (TMC width) and medial-lateral direction (TMC thickness). Three selected samples were examined using histological microscopy. RESULTS: Both TMC width and thickness featured a large variability among individuals. The minimal TMC width along the manubrium for different individuals covered a range between 83 and 840 µm. The minimal thickness ranged from 48 to 249 µm and the maximal thickness from 236 to 691 µm. Histological sections showed that the TMC consists of a narrow core of dense regular connective tissue, surrounded by loose connective tissue. In some samples, either of these two components was absent in the TMC at some manubrium locations. The configuration of these components varied among the samples as well. CONCLUSION: Our data confirm that a large inter-individual variability exists in the properties of the TM-malleus connection in humans in terms of its dimensions, tissue composition and configuration. Average data and their variability margins will be useful input for testing the importance of the TMC in finite element models.