ArborSim: Articulated, branching, OpenSim routing for constructing models of multi-jointed appendages with complex muscle-tendon architecture.

Computational models of musculoskeletal systems are essential tools for understanding how muscles, tendons, bones, and actuation signals generate motion. In particular, the OpenSim family of models has facilitated a wide range of studies on diverse human motions, clinical studies of gait, and even non-human locomotion. However, biological structures with many joints, such as fingers, necks, tails, and spines, have been a longstanding challenge to the OpenSim modeling community, especially because these structures comprise numerous bones and are frequently actuated by extrinsic muscles that span multiple joints-often more than three-and act through a complex network of branching tendons. Existing model building software, typically optimized for limb structures, makes it difficult to build OpenSim models that accurately reflect these intricacies. Here, we introduce ArborSim, customized software that efficiently creates musculoskeletal models of highly jointed structures and can build branched muscle-tendon architectures. We used ArborSim to construct toy models of articulated structures to determine which morphological features make a structure most sensitive to branching. By comparing the joint kinematics of models constructed with branched and parallel muscle-tendon units, we found that among various parameters-the number of tendon branches, the number of joints between branches, and the ratio of muscle fiber length to muscle tendon unit length-the number of tendon branches and the number of joints between branches are most sensitive to branching modeling method. Notably, the differences between these models showed no predictable pattern with increased complexity. As the proportion of muscle increased, the kinematic differences between branched and parallel models units also increased. Our findings suggest that stress and strain interactions between distal tendon branches and proximal tendon and muscle greatly affect the overall kinematics of a musculoskeletal system. By incorporating complex muscle-tendon branching into OpenSim models using ArborSim, we can gain deeper insight into the interactions between the axial and appendicular skeleton, model the evolution and function of diverse animal tails, and understand the mechanics of more complex motions and tasks.

Jumping over fences: why field- and laboratory-based biomechanical studies can and should learn from each other.

Locomotor biomechanics faces a core trade-off between laboratory-based and field-based studies. Laboratory conditions offer control over confounding factors, repeatability, and reduced technological challenges, but limit the diversity of animals and environmental conditions that may influence behavior and locomotion. This article considers how study setting influences the selection of animals, behaviors and methodologies for studying animal motion. We highlight the benefits of both field- and laboratory-based studies and discuss how recent work leverages technological advances to blend these approaches. These studies have prompted other subfields of biology, namely evolutionary biology and ecology, to incorporate biomechanical metrics more relevant to survival in natural habitats. The concepts discussed in this Review provide guidance for blending methodological approaches and inform study design for both laboratory and field biomechanics. In this way, we hope to facilitate integrative studies that relate biomechanical performance to animal fitness, determine the effect of environmental factors on motion, and increase the relevance of biomechanics to other subfields of biology and robotics.

Metatarsal fusion resisted bending as jerboas (Dipodidae) transitioned from quadrupedal to bipedal.

Hind limbs undergo dramatic changes in loading conditions during the transition from quadrupedal to bipedal locomotion. For example, the most early diverging bipedal jerboas (Rodentia: Dipodidae) are some of the smallest mammals in the world, with body masses that range between 2-4 g. The larger jerboa species exhibit developmental and evolutionary fusion of the central three metatarsals into a single cannon bone. We hypothesize that small body size and metatarsal fusion are mechanisms to maintain the safety factor of the hind limb bones despite the higher ground reaction forces associated with bipedal locomotion. Using finite-element analysis to model collisions between the substrate and the metatarsals, we found that body size reduction was insufficient to reduce bone stress on unfused metatarsals, based on the scaled dynamics of larger jerboas, and that fused bones developed lower stresses than unfused bones when all metatarsals are scaled to the same size and loading conditions. Based on these results, we conclude that fusion reinforces larger jerboa metatarsals against high ground reaction forces. Because smaller jerboas with unfused metatarsals develop higher peak stresses in response to loading conditions scaled from larger jerboas, we hypothesize that smaller jerboas use alternative dynamics of bipedal locomotion to reduce the impact of collisions between the foot and substrate.

Convergent metatarsal fusion in jerboas and chickens is mediated by similarities and differences in the patterns of osteoblast and osteoclast activities.

In many vertebrate animals that run or leap, the metatarsals and/or metacarpals of the distal limb are fused into a single larger element, likely to resist fracture due to high ground-reaction forces during locomotion. Although metapodial fusion evolved independently in modern birds, ungulates, and jerboas, the developmental basis has only been explored in chickens, which diverged from the mammalian lineage approximately 300 million years ago. Here, we use a bipedal rodent, the lesser Egyptian jerboa (Jaculus jaculus), to understand the cellular processes of metatarsal fusion in a mammal, and we revisit the developing chicken to assess similarities and differences in the localization of osteoblast and osteoclast activities. In both species, adjacent metatarsals align along flat surfaces, osteoblasts cross the periosteal membrane to unite the three elements in a single circumference, and osteoclasts resorb bone at the interfaces leaving a single marrow cavity. However, the pattern of osteoclast activity differs in each species; osteoclasts are highly localized to resorb bone at the interfaces of neighboring jerboa metatarsals and are distributed throughout the endosteum of chicken metatarsals. Each species, therefore, provides an opportunity to understand mechanisms that pattern osteoblast and osteoclast activities to alter bone shape during development and evolution.

Tail-assisted pitch control in lizards, robots and dinosaurs.

In 1969, a palaeontologist proposed that theropod dinosaurs used their tails as dynamic stabilizers during rapid or irregular movements, contributing to their depiction as active and agile predators. Since then the inertia of swinging appendages has been implicated in stabilizing human walking, aiding acrobatic manoeuvres by primates and rodents, and enabling cats to balance on branches. Recent studies on geckos suggest that active tail stabilization occurs during climbing, righting and gliding. By contrast, studies on the effect of lizard tail loss show evidence of a decrease, an increase or no change in performance. Application of a control-theoretic framework could advance our general understanding of inertial appendage use in locomotion. Here we report that lizards control the swing of their tails in a measured manner to redirect angular momentum from their bodies to their tails, stabilizing body attitude in the sagittal plane. We video-recorded Red-Headed Agama lizards (Agama agama) leaping towards a vertical surface by first vaulting onto an obstacle with variable traction to induce a range of perturbations in body angular momentum. To examine a known controlled tail response, we built a lizard-sized robot with an active tail that used sensory feedback to stabilize pitch as it drove off a ramp. Our dynamics model revealed that a body swinging its tail experienced less rotation than a body with a rigid tail, a passively compliant tail or no tail. To compare a range of tails, we calculated tail effectiveness as the amount of tailless body rotation a tail could stabilize. A model Velociraptor mongoliensis supported the initial tail stabilization hypothesis, showing as it did a greater tail effectiveness than the Agama lizards. Leaping lizards show that inertial control of body attitude can advance our understanding of appendage evolution and provide biological inspiration for the next generation of manoeuvrable search-and-rescue robots.

Vertical leaping mechanics of the Lesser Egyptian Jerboa reveal specialization for maneuverability rather than elastic energy storage.

BACKGROUND: Numerous historical descriptions of the Lesser Egyptian jerboa, Jaculus jaculus, a small bipedal mammal with elongate hindlimbs, make special note of their extraordinary leaping ability. We observed jerboa locomotion in a laboratory setting and performed inverse dynamics analysis to understand how this small rodent generates such impressive leaps. We combined kinematic data from video, kinetic data from a force platform, and morphometric data from dissections to calculate the relative contributions of each hindlimb muscle and tendon to the total movement. RESULTS: Jerboas leapt in excess of 10 times their hip height. At the maximum recorded leap height (not the maximum observed leap height), peak moments for metatarso-phalangeal, ankle, knee, and hip joints were 13.1, 58.4, 65.1, and 66.9 Nmm, respectively. Muscles acting at the ankle joint contributed the most work (mean 231.6 mJ / kg Body Mass) to produce the energy of vertical leaping, while muscles acting at the metatarso-phalangeal joint produced the most stress (peak 317.1 kPa). The plantaris, digital flexors, and gastrocnemius tendons encountered peak stresses of 25.6, 19.1, and 6.0 MPa, respectively, transmitting the forces of their corresponding muscles (peak force 3.3, 2.0, and 3.8 N, respectively). Notably, we found that the mean elastic energy recovered in the primary tendons of both hindlimbs comprised on average only 4.4% of the energy of the associated leap. CONCLUSIONS: The limited use of tendon elastic energy storage in the jerboa parallels the morphologically similar heteromyid kangaroo rat, Dipodomys spectabilis. When compared to larger saltatory kangaroos and wallabies that sustain hopping over longer periods of time, these small saltatory rodents store and recover less elastic strain energy in their tendons. The large contribution of muscle work, rather than elastic strain energy, to the vertical leap suggests that the fitness benefit of rapid acceleration for predator avoidance dominated over the need to enhance locomotor economy in the evolutionary history of jerboas.

Jointed tails enhance control of three-dimensional body rotation.

Tails used as inertial appendages induce body rotations of animals and robots-a phenomenon that is governed largely by the ratio of the body and tail moments of inertia. However, vertebrate tails have more degrees of freedom (e.g., number of joints, rotational axes) than most current theoretical models and robotic tails. To understand how morphology affects inertial appendage function, we developed an optimization-based approach that finds the maximally effective tail trajectory and measures error from a target trajectory. For tails of equal total length and mass, increasing the number of equal-length joints increased the complexity of maximally effective tail motions. When we optimized the relative lengths of tail bones while keeping the total tail length, mass, and number of joints the same, this optimization-based approach found that the lengths match the pattern found in the tail bones of mammals specialized for inertial maneuvering. In both experiments, adding joints enhanced the performance of the inertial appendage, but with diminishing returns, largely due to the total control effort constraint. This optimization-based simulation can compare the maximum performance of diverse inertial appendages that dynamically vary in moment of inertia in 3D space, predict inertial capabilities from skeletal data, and inform the design of robotic inertial appendages.

Polymorphism in the aggressive mimicry lure of the parasitic freshwater mussel Lampsilis fasciola.

Unionoid freshwater mussels (Bivalvia: Unionidae) are free-living apart from a brief, obligately parasitic, larval stage that infects fish hosts, and gravid female mussels have evolved a spectrum of strategies to infect fish hosts with their larvae. In many North American species, this involves displaying a mantle lure: a pigmented fleshy extension that acts as an aggressive mimic of a host fish prey, thereby eliciting a feeding response that results in host infection. The mantle lure of Lampsilis fasciola is of particular interest because it is apparently polymorphic, with two distinct primary lure phenotypes. One, described as "darter-like", has "eyespots", a mottled body coloration, prominent marginal extensions, and a distinct "tail". The other, described as "worm-like", lacks those features and has an orange and black coloration. We investigated this phenomenon using genomics, captive rearing, biogeographic, and behavioral analyses. Within-brood lure variation and within-population phylogenomic (ddRAD-seq) analyses of individuals bearing different lures confirmed that this phenomenon is a true polymorphism. The relative abundance of the two morphs appears stable over ecological timeframes: the ratio of the two lure phenotypes in a River Raisin (MI) population in 2017 was consistent with that of museum samples collected at the same site six decades earlier. Within the River Raisin, four main "darter-like" lure motifs visually approximated four co-occurring darter species (Etheostoma blennioides, E. exile, E. microperca, and Percina maculata), and the "worm-like" lure resembled a widespread common leech, Macrobdella decora. Darters and leeches are typical prey of Micropterus dolomieui (smallmouth bass), the primary fish host of L. fasciola. In situ field recordings of the L. fasciola "darter" and "leech" lure display behaviors, and the lure display of co-occurring congener L. cardium, were captured. Despite having putative models in distinct phyla, both L. fasciola lure morphs have largely similar display behaviors that differ significantly from that of sympatric L. cardium individuals. Some minor differences in the behavior between the two L. fasciola morphs were observed, but we found no clear evidence for a behavioral component of the polymorphism given the criteria measured. Discovery of discrete within-brood inheritance of the lure polymorphism implies potential control by a single genetic locus and identifies L. fasciola as a promising study system to identify regulatory genes controlling a key adaptive trait of freshwater mussels.

Instantaneous kinematic phase reflects neuromechanical response to lateral perturbations of running cockroaches.

Instantaneous kinematic phase calculation allows the development of reduced-order oscillator models useful in generating hypotheses of neuromechanical control. When perturbed, changes in instantaneous kinematic phase and frequency of rhythmic movements can provide details of movement and evidence for neural feedback to a system-level neural oscillator with a time resolution not possible with traditional approaches. We elicited an escape response in cockroaches (Blaberus discoidalis) that ran onto a movable cart accelerated laterally with respect to the animals' motion causing a perturbation. The specific impulse imposed on animals (0.50 [Formula: see text] 0.04 m s[Formula: see text]; mean, SD) was nearly twice their forward speed (0.25 [Formula: see text] 0.06 m s[Formula: see text]. Instantaneous residual phase computed from kinematic phase remained constant for 110 ms after the onset of perturbation, but then decreased representing a decrease in stride frequency. Results from direct muscle action potential recordings supported kinematic phase results in showing that recovery begins with self-stabilizing mechanical feedback followed by neural feedback to an abstracted neural oscillator or central pattern generator. Trials fell into two classes of forward velocity changes, while exhibiting statistically indistinguishable frequency changes. Animals pulled away from the side with front and hind legs of the tripod in stance recovered heading within 300 ms, whereas animals that only had a middle leg of the tripod resisting the pull did not recover within this period. Animals with eight or more legs might be more robust to lateral perturbations than hexapods.

A Template Model Explains Jerboa Gait Transitions Across a Broad Range of Speeds.

For cursorial animals that maintain high speeds for extended durations of locomotion, transitions between footfall patterns (gaits) predictably occur at distinct speed ranges. How do transitions among gaits occur for non-cursorial animals? Jerboas (Jaculus) are bipedal hopping rodents that frequently transition between gaits throughout their entire speed range. It has been hypothesized that these non-cursorial bipedal gait transitions are likely to enhance their maneuverability and predator evasion ability. However, it is difficult to use the underlying dynamics of these locomotion patterns to predict gait transitions due to the large number of degrees of freedom expressed by the animals. To this end, we used empirical jerboa kinematics and dynamics to develop a unified spring Loaded Inverted Pendulum model with defined passive swing leg motions. To find periodic solutions of this model, we formulated the gait search as a boundary value problem and described an asymmetrical running gait exhibited by the jerboas that emerged from the numerical search. To understand how jerboas change from one gait to another, we employed an optimization approach and used the proposed model to reproduce observed patterns of jerboa gait transitions. We then ran a detailed numerical study of the structure of gait patterns using a continuation approach in which transitions are represented by bifurcations. We found two primary mechanisms to increase the range of speeds at which gait transitions can occur. Coupled changes in the neutral leg swing angle alter leg dynamics. This mechanism generates changes in gait features (e.g., touchdown leg angle and timings of gait events) that have previously been shown to induce gait transitions. This mechanism slightly alters the speeds at which existing gait transitions occur. The model can also uncouple the left and right neutral leg swing angle, which generates asymmetries between left and right leg dynamics. New gait transitions emerge from uncoupled models across a broad range of speeds. In both the experimental observations and in the model, the majority of the gait transitions involve the skipping and asymmetrical running gaits generated by the uncoupled neutral leg swing angle mechanism. This simulated jerboa model is capable of systematically reproducing all biologically relevant gait transitions at a broad range of speeds.

Virtual Expeditions Facilitated by Open Source Solutions Broaden Student Participation in Natural History Research.

From its genesis in the Victorian era as an activity for the elite to today's emphasis on "Big Data" and continuous monitoring, natural history has a prominent role in scientific discoveries for many fields. However, participation in field expeditions is limited by funding, space, accessibility, and safety constraints. Others have detailed the active exclusion of minoritized groups from field expeditions and harm/discrimination faced by the few who do participate, but we provide one solution to broaden opportunities for participation in natural history: Virtual Expeditions. Virtual Expeditions are broadly defined as open source, web-facilitated research activities designed to analyze bulk-collected digital data from field expeditions that require visual human interpretation. We show two examples here of their use: an independent research-based analysis of snake behavior and a course-based identification of invertebrate species. We present a guide to their appropriate design, facilitation, and evaluation to result in research grade data. We highlight the importance of open source technology to allow for longevity in methodology and appropriate quality control measures necessary for projects that include dozens of researchers over multiple years. In this perspective, we specifically emphasize the prominent role that open source technology plays in making these experiences feasible and scalable. Even without explicit design as broadening participation endeavors, Virtual Expeditions allow for more inclusive participation of early career researchers with specific participatory limitations. Not only are Virtual Expeditions integral to the large-scale analysis necessary for field expeditions that generate impossibly enormous datasets, but they can also be effective facilitators of inclusivity in natural history research.

Batch-Mask: Automated Image Segmentation for Organisms with Limbless or Non-Standard Body Forms.

Efficient comparisons of biological color patterns are critical for understanding the mechanisms by which organisms evolve in nature, including sexual selection, predator-prey interactions, and thermoregulation. However, limbless, elongate, or spiral-shaped organisms do not conform to the standard orientation and photographic techniques required for many automated analyses. Currently, large-scale color analysis of elongate animals requires time-consuming manual landmarking, which reduces their representation in coloration research despite their ecological importance. We present Batch-Mask: an automated, customizable workflow to automatically analyze large photographic datasets to isolate non-standard biological organisms from the background. Batch-Mask is completely open-source and does not depend on any proprietary software. We also present a user guide for fine-tuning weights to a custom dataset and incorporating existing manual visual analysis tools (e.g., micaToolbox) into a single automated workflow for comparing color patterns across images. Batch-Mask was 60x faster than manual landmarking and produced masks that correctly identified 96% of all snake pixels. To validate our approach, we used micaToolbox to compare pattern energy in a sample set of snake photographs segmented by Batch-Mask and humans and found no significant difference in the output results. The fine-tuned weights, user guide, and automated workflow substantially decrease the amount of time and attention required to quantitatively analyze non-standard biological subjects. With these tools, biologists can compare color, pattern, and shape differences in large datasets that include significant morphological variation in elongate body forms. This advance is especially valuable for comparative analyses of natural history collections across a broad range of morphologies. Through landmark-free automation, Batch-Mask can greatly expand the scale of space, time, or taxonomic breadth across which color variation can be quantitatively examined.

Examining Cultural Structures and Functions in Biology.

Scientific culture and structure organize biological sciences in many ways. We make choices concerning the systems and questions we study. Our research then amplifies these choices into factors that influence the directions of future research by shaping our hypotheses, data analyses, interpretation, publication venues, and dissemination via other methods. But our choices are shaped by more than objective curiosity-we are influenced by cultural paradigms reinforced by societal upbringing and scientific indoctrination during training. This extends to the systems and data that we consider to be ethically obtainable or available for study, and who is considered qualified to do research, ask questions, and communicate about research. It is also influenced by the profitability of concepts like open-access-a system designed to improve equity, but which enacts gatekeeping in unintended but foreseeable ways. Creating truly integrative biology programs will require more than intentionally developing departments or institutes that allow overlapping expertise in two or more subfields of biology. Interdisciplinary work requires the expertise of large and diverse teams of scientists working together-this is impossible without an authentic commitment to addressing, not denying, racism when practiced by individuals, institutions, and cultural aspects of academic science. We have identified starting points for remedying how our field has discouraged and caused harm, but we acknowledge there is a long path forward. This path must be paved with field-wide solutions and institutional buy-in: our solutions must match the scale of the problem. Together, we can integrate-not reintegrate-the nuances of biology into our field.

Characterizing the limits of human stability during motion: perturbative experiment validates a model-based approach for the Sit-to-Stand task.

Falls affect a growing number of the population each year. Clinical methods to assess fall risk usually evaluate the performance of specific motions such as balancing or Sit-to-Stand. Unfortunately, these techniques have been shown to have poor predictive power, and are unable to identify the portions of motion that are most unstable. To this end, it may be useful to identify the set of body configurations that can accomplish a task under a specified control strategy. The resulting strategy-specific boundary between stable and unstable motion could be used to identify individuals at risk of falling. The recently proposed Stability Basin is defined as the set of configurations through time that do not lead to failure for an individual under their chosen control strategy. This paper presents a novel method to compute the Stability Basin and the first experimental validation of the Stability Basin with a perturbative Sit-to-Stand experiment involving forwards or backwards pulls from a motor-driven cable with 11 subjects. The individually-constructed Stability Basins are used to identify when a trial fails, i.e. when an individual must switch from their chosen control strategy (indicated by a step or sit) to recover from a perturbation. The constructed Stability Basins correctly predict the outcome of trials where failure was observed with over 90 % accuracy, and correctly predict the outcome of successful trials with over 95 % accuracy. The Stability Basin was compared to three other methods and was found to estimate the stable region with over 45 % more accuracy in all cases. This study demonstrates that Stability Basins offer a novel model-based approach for quantifying stability during motion, which could be used in physical therapy for individuals at risk of falling.

Unpredictability of escape trajectory explains predator evasion ability and microhabitat preference of desert rodents.

Mechanistically linking movement behaviors and ecology is key to understanding the adaptive evolution of locomotion. Predator evasion, a behavior that enhances fitness, may depend upon short bursts or complex patterns of locomotion. However, such movements are poorly characterized by existing biomechanical metrics. We present methods based on the entropy measure of randomness from Information Theory to quantitatively characterize the unpredictability of non-steady-state locomotion. We then apply the method by examining sympatric rodent species whose escape trajectories differ in dimensionality. Unlike the speed-regulated gait use of cursorial animals to enhance locomotor economy, bipedal jerboa (family Dipodidae) gait transitions likely enhance maneuverability. In field-based observations, jerboa trajectories are significantly less predictable than those of quadrupedal rodents, likely increasing predator evasion ability. Consistent with this hypothesis, jerboas exhibit lower anxiety in open fields than quadrupedal rodents, a behavior that varies inversely with predator evasion ability. Our unpredictability metric expands the scope of quantitative biomechanical studies to include non-steady-state locomotion in a variety of evolutionary and ecologically significant contexts.Biomechanical understanding of animal gait and maneuverability has primarily been limited to species with more predictable, steady-state movement patterns. Here, the authors develop a method to quantify movement predictability, and apply the method to study escape-related movement in several species of desert rodents.

Outrun or Outmaneuver: Predator-Prey Interactions as a Model System for Integrating Biomechanical Studies in a Broader Ecological and Evolutionary Context.

Behavioral studies performed in natural habitats provide a context for the development of hypotheses and the design of experiments relevant both to biomechanics and to evolution. In particular, predator-prey interactions are a model system for integrative study because success or failure of predation has a direct effect on fitness and drives the evolution of specialized performance in both predator and prey. Although all predators share the goal of capturing prey, and all prey share the goal of survival, the behavior of predators and prey are diverse in nature. This article presents studies of some predator-prey interactions sharing common predation strategies that reveal general principles governing the behaviors of predator and prey, even in distantly related taxa. Studies of predator-prey interactions also reveal that maximal performance observed in a laboratory setting is not necessarily the performance that determines fitness. Thus, considering locomotion in the context of predation ecology can aid in evolutionarily relevant experimental design. Classification by strategy reveals that displaying unpredictable trajectories is a relevant anti-predator behavior in response to multiple predation strategies. A predator's perception and pursuit of prey can be affected indirectly by divergent locomotion of similar animals that share an ecosystem. Variation in speed and direction of locomotion that directly increases the unpredictability of a prey's trajectory can be increased through genetic mutation that affects locomotor patterns, musculoskeletal changes that affect maneuverability, and physical interactions between an animal and the environment. By considering the interconnectedness of ecology, physical constraints, and the evolutionary history of behavior, studies in biomechanics can be designed to inform each of these fields.

Multiple phylogenetically distinct events shaped the evolution of limb skeletal morphologies associated with bipedalism in the jerboas.

Recent rapid advances in experimental biology have expanded the opportunity for interdisciplinary investigations of the evolution of form and function in non-traditional model species. However, historical divisions of philosophy and methodology between evolutionary/organismal biologists and developmental geneticists often preclude an effective merging of disciplines. In an effort to overcome these divisions, we take advantage of the extraordinary morphological diversity of the rodent superfamily Dipodoidea, including the bipedal jerboas, to experimentally study the developmental mechanisms and biomechanical performance of a remarkably divergent limb structure. Here, we place multiple limb character states in a locomotor and phylogenetic context. Whereas obligate bipedalism arose just once in the ancestor of extant jerboas, we find that digit loss, metatarsal fusion, between-limb proportions, and within-hindlimb proportions all evolved independently of one another. Digit loss occurred three times through at least two distinct developmental mechanisms, and elongation of the hindlimb relative to the forelimb is not simply due to growth mechanisms that change proportions within the hindlimb. Furthermore, we find strong evidence for punctuated evolution of allometric scaling of hindlimb elements during the radiation of Dipodoidea. Our work demonstrates the value of leveraging the evolutionary history of a clade to establish criteria for identifying the developmental genetic mechanisms of morphological diversification.