A Step Forward: Functional Diversity and Emerging Themes of Slow-Speed Locomotion in Vertebrates.

Walking can be defined broadly as a slow-speed movement produced when appendages interact with the ground to generate forward propulsion. Until recently, most studies of walking have focused on humans and a handful of domesticated vertebrates moving at a steady rate over highly simplified, static surfaces, which may bias our understanding of the unifying principles that underlie vertebrate locomotion. In the last few decades, studies have expanded to include a range of environmental contexts (e.g., uneven terrain, perturbations, deformable substrates) and greater phylogenetic breadth (e.g., non-domesticated species, small and/or ectothermic tetrapods and fishes); these studies have revealed that even a gait as superficially simple as walking is far more complex than previously thought. In addition, technological advances and accessibility of imaging systems and computational power have recently expanded our capabilities to test hypotheses about the locomotor movements of extant and extinct organisms in silico. In this symposium, scientists showcased diverse taxa (from extant fishes to extinct dinosaurs) moving through a range of variable conditions (speed perturbations, inclines, and deformable substrates) to address the causes and consequences of functional diversity in locomotor systems and discuss nascent research areas and techniques. From the symposium contributions, several themes emerged: (1) slow-speed, appendage-based movements in fishes are best described as walking-like movements rather than true walking gaits, (2) environmental variation (e.g., deformable substrates) and dynamic stimuli (e.g., perturbations) trigger kinematic and neuromuscular changes in animals that make defining a single gait or the transition between gaits more complicated than originally thought, and (3) computational advances have increased the ability to process large data sets, emulate the 3D motions of extant and extinct taxa, and even model species interactions in ancient ecosystems. Although this symposium allowed us to make great strides forward in our understanding of vertebrate walking, much ground remains to be covered. First, there is a much greater range of vertebrate appendage-based locomotor behaviors than has been previously recognized and existing terminology fails to accurately capture and describe this diversity. Second, despite recent efforts, the mechanisms that vertebrates use modify locomotor behaviors in response to predictable and unpredictable locomotor challenges are still poorly understood. Third, while computer-based models and simulations facilitate a greater understanding of the kinetics and kinematics of movement in both extant and extinct animals, a universal, one-size-fits-all, predictive model of appendage-based movement in vertebrates remains elusive.

The fast and the tendinous? Locomotor modifications of the caudal peduncle in Gila spp. from the American Southwest.

In the American Southwest, the fishes within the genus Gila evolved in an environment with seasonal rainstorms that caused stochastic flooding. Some species within this genus, such as bonytail (Gila elegans), possess locomotor morphologies that are similar to those seen in high-performance swimmers such as tuna and lamnid sharks. These shared features include a shallow caudal peduncle, lunate tail, and mechanisms to transmit force from the anterior musculature to the tail fin. We compared the skeletal anatomy of the caudal region of bonytail to roundtail chub (Gila robusta) and humpback chub (Gila cypha) to determine which vertebral elements have been modified to create a shallow peduncle. We also tested the tensile strength of the red (slow oxidative) axial muscle by performing a standard stress test. If the muscle can withstand a large load, this suggests it may play a tendon-like role in transmitting force from the anterior muscle to the hypural plate of the tail. Lastly, we measured the collagen content of the red axial muscle (visualized using serial sections and Masson's trichrome stain) to determine if increased tensile strength is associated with increased collagen content. We found bonytail caudal peduncles are characterized by acute vertebral spines and have red axial muscle that can resist tearing under tension. Roundtail chub peduncles are characterized by relatively more obtuse angles and the red muscle tears easily under tension. Humpback chub possess an intermediate morphology, with relatively obtuse vertebral spine angles and the red muscle can resist tearing under tension. Bonytail have increased collagen content in posterior red axial muscle compared to the anterior musculature also suggesting a tendon-like role of the posterior red muscle. In combination with previous studies of swimming performance, our findings suggest that the axial musculature of bonytail may play a role in transmitting force directly to the shallow peduncle in a manner similar to that of the great lateral tendon of scombrids.

Integrating Studies of Anatomy, Physiology, and Behavior into Conservation Strategies for the Imperiled Cyprinid Fishes of the Southwestern United States.

Over the last 100 years, fishes native to the Southwestern United States have faced a myriad of biotic and abiotic pressures which has resulted in most being federally listed as endangered or threatened. Most notably, water diversions and the introduction of non-native fishes have been the primary culprits in causing the downfall of native fish populations. We describe how recent studies of morphology, physiology, and behavior yield insights into the failed (occasionally successful) management of this vanishing biota. We describe how understanding locomotor morphologies, physiologies, and behaviors unique to Southwestern native fishes can be used to create habitats that favor native fishes. Additionally, through realizing differences in morphologies and behaviors between native and non-native fishes, we describe how understanding predator-prey interactions might render greater survivorship of native fishes when stocked into the wild from repatriation programs. Understanding fundamental form-function relationships is imperative for managers to make educated decisions on how to best recover species of concern in the Southwestern United States and worldwide.

Are Extreme Anatomical Modifications Required for Fish to Move Effectively on Land? Comparative Anatomy of the Posterior Axial Skeleton in the Cyprinodontiformes.

Many teleost fishes with no apparent modifications for life on land are able to produce effective terrestrial locomotor behaviors, including a ballistic behavior called the "tail-flip" jump. Cyprinodontiformes (killifishes, Teleostei: Atherinomorpha) that live at the water's edge vary in morphology and inclination to emerge onto land. Do fish with an amphibious predisposition have extensive modification of the propulsive region of the body when compared to fully aquatic relatives? We quantified body shape and anatomy of the caudal peduncle and tail (the propulsive organ on land and in water) in 11 cyprinodontiform species and two outgroup taxa (Atherinomorpha). We hypothesized that amphibious species would have longer, "shallower" bodies (larger body fineness ratios), deeper (proportionally larger) caudal peduncles, and more robust bones in the tail fin (larger ossified area of the hypural/epural bones) to facilitate locomotor movements on land. We found no evidence of convergence in body shape or skeletal anatomy among species known to make voluntary sojourns onto land. In fact, deep-bodied species, shallow-bodied species, and species with intermediate morphologies all are able to emerge from the water and move on land. It is possible that there are as-yet-undocumented subtle soft-tissue (muscle, tendon, and ligament) modifications that enhance terrestrial locomotor performance in species known to spend large periods of time on land. However, it is also possible that extreme anatomical changes are not required for aquatic cyprinodontiform species to produce effective locomotor movements when they emerge out of the water and move across the land. Anat Rec, 2019. © 2019 American Association for Anatomy.

A walking behavior generates functional overland movements in the tidepool sculpin, Oligocottus maculosus.

Tidepool sculpins (Oligocottus maculosus) have been observed moving overland in the rocky intertidal, and we documented the terrestrial walking behavior that they use to accomplish this. We quantified the terrestrial movements of O. maculosus and compared them to (1) their aquatic locomotion, (2) terrestrial locomotion of closely-related subtidal species (Leptocottus armatus and Icelinus borealis), and (3) terrestrial movements of walking catfishes (Clarias spp.). We recorded sculpin movements (210 fps) on a terrestrial platform and in a water tank and tracked body landmarks for kinematic analysis. The axial-appendage-based terrestrial locomotion of O. maculosus is driven by cyclic lateral oscillations of the tail, synchronized with alternating rotations about the base of the pectoral fins, a behavior that appears similar to a military "army crawl." The pectoral fins do not provide propulsion, but act as stable points for the body to rotate around. In contrast, individuals of O. maculosus use primarily axial undulation during slow-speed swimming. The army crawl is a more effective terrestrial behavior (greater distance ratio) than the movements produced by L. armatus and I. borealis, which use rapid, cyclic oscillations of the tail, without coordinated pectoral fin movements. Relative to Clarias spp., O. maculosus rotated the body about the base of the pectoral fin, rather than the tip of the fin, which may cause O. maculosus to have a lower distance ratio. Since O. maculosus lack major morphological adaptations for terrestrial locomotion, instead relying on behavioral adaptations, we propose behavioral adaptations may evolutionarily predate morphological adaptations for terrestrial locomotion in vertebrates.

Effects of organism and substrate size on burial mechanics of English sole, Parophrys vetulus.

Flatfishes use cyclic body undulations to force water into the sediment and fluidize substrate particles, displacing them into the water column. When water velocity decreases, suspended particles settle back onto the fish, hiding it from view. Burial may become more challenging as flatfishes grow because the area to be covered increases exponentially with the second power of length. In addition, particle size is not uniform in naturally occurring substrates, and larger particles require higher water velocities for fluidization. We quantified the effects of organism and particle-size scaling on burial behavior of English sole, Parophrys vetulus We recorded burial events from a size range of individuals (5-32 cm total length, TL), while maintaining constant substrate grain size. Larger fish used lower cycle frequencies and took longer to bury, but overall burial performance was maintained (∼100% coverage). To test the effect of particle size on burial performance, individuals of similar lengths (5.7-8.1 cm TL) were presented with different substrate sizes (0.125-0.710 mm). Particle size did not affect cycle frequency or time to burial, but fish did not achieve 100% coverage with the largest particles because they could not fluidize this substrate. Taken together, these results suggest that both body size and substrate grain size can potentially limit the ability of flatfishes to bury: a very large fish (>150 cm) may move too slowly to fluidize all but the smallest substrate particles and some particles are simply too large for smaller individuals to fluidize.

Behavioral and physiological adaptations to high-flow velocities in chubs (Gila spp.) native to Southwestern USA.

Morphological streamlining is often associated with physiological advantages for steady swimming in fishes. Though most commonly studied in pelagic fishes, streamlining also occurs in fishes that occupy high-flow environments. Before the installation of dams and water diversions, bonytail (Cyprinidae, Gila elegans), a fish endemic to the Colorado River (USA), regularly experienced massive, seasonal flooding events. Individuals of G. elegans display morphological characteristics that may facilitate swimming in high-flow conditions, including a narrow caudal peduncle and a high aspect ratio caudal fin. We tested the hypothesis that these features improve sustained swimming performance in bonytail by comparing locomotor performance in G. elegans with that of the closely related roundtail chub (Gila robusta) and two non-native species, rainbow trout (Oncorhynchus mykiss) and smallmouth bass (Micropterus dolomieu), using a Brett-style respirometer and locomotor step-tests. Gila elegans had the lowest estimated drag coefficient and the highest sustained swimming speeds relative to the other three species. There were no detectible differences in locomotor energetics during steady swimming among the four species. When challenged by high-velocity water flows, the second native species examined in this study, G. robusta, exploited the boundary effects in the flow tank by pitching forward and bracing the pelvic and pectoral fins against the acrylic tank bottom to 'hold station'. Because G. robusta can station hold to prevent being swept downstream during high flows and G. elegans can maintain swimming speeds greater than those of smallmouth bass and rainbow trout with comparable metabolic costs, we suggest that management agencies could use artificial flooding events to wash non-native competitors downstream and out of the Colorado River habitat.

Why does Gila elegans have a bony tail? A study of swimming morphology convergence.

Caudal-fin-based swimming is the primary form of locomotion in most fishes. As a result, many species have developed specializations to enhance performance during steady swimming. Specializations that enable high swimming speeds to be maintained for long periods of time include: a streamlined body, high-aspect-ratio (winglike) caudal fin, a shallow caudal peduncle, and high proportions of slow-twitch ("red") axial muscle. We described the locomotor specializations of a fish species native to the Colorado River and compared those specializations to other fish species from this habitat, as well as to a high-performance marine swimmer. The focal species for this study was the bonytail (Gila elegans), which has a distinct morphology when compared with closely related species from the Southwestern United States. Comparative species used in this study were the roundtail chub (Gila robusta), a closely related species from low-flow habitats; the common carp (Cyprinus carpio), an invasive cyprinid also found in low-flow habitats; and the chub mackerel (Scomber japonicus), a model high-performance swimmer from the marine environment. The bonytail had a shallow caudal peduncle and a high-aspect-ratio tail that were similar to those of the chub mackerel. The bonytail also had a more streamlined body than the roundtail chub and the common carp, although not as streamlined as the chub mackerel. The chub mackerel had a significantly higher proportion of red muscle than the other three species, which did not differ from one another. Taken together, the streamlined body, narrow caudal peduncle, and high-aspect-ratio tail of the bonytail suggest that this species has responded to the selection pressures of the historically fast-flowing Colorado River, where flooding events and base flows may have required native species to produce and sustain very high swimming speeds to prevent being washed downstream.

Look before you leap: Visual navigation and terrestrial locomotion of the intertidal killifish Fundulus heteroclitus.

Mummichogs (Fundulus heteroclitus; Cyprinodontiformes) are intertidal killifish that can breathe air and locomote on land. Our goals were to characterize the terrestrial locomotion of mummichogs and determine their method of navigation towards water in a terrestrial environment. We used high-speed video to record behavior during stranding experiments and found that mummichogs use a tail-flip jump to move overland, similarly to other Cyprinodontiformes. However, mummichogs also prop themselves upright into a prone position between each jump, a previously undescribed behavior. After becoming prone, mummichogs rotate about their vertical axis, directing the caudal fin towards the water. Then, they roll back onto their lateral aspect and use a tail-flip behavior to leap into a caudally-directed, ballistic flight path. We conducted experiments to determine the sensory stimulus used to locate a body of water by placing mummichogs on a square platform with one side adjacent to a sea table. Under artificial light, mummichogs moved towards the sea table with a higher frequency than towards the other sides. Under dark conditions, mummichogs did not show a preference for moving towards the sea table. When the surface of the water was covered with reflective foil, mummichogs moved towards it as if it were a body of water. These results suggest that mummichogs primarily use visual cues, specifically reflected light, to orient towards the water. The uprighting behavior that mummichogs perform between terrestrial jumps may provide an opportunity for these fish to receive visual information that allows them to safely return to the water. J. Exp. Zool. 325A:57-64, 2016. © 2015 Wiley Periodicals, Inc.

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.

Suction, Ram, and Biting: Deviations and Limitations to the Capture of Aquatic Prey.

When feeding, most aquatic organisms generate suction that draws prey into the mouth. The papers in this volume are a demonstration of this fact. However, under what circumstances is suction ineffective as a feeding mechanism? Here we consider the interplay between suction, ram, and biting, and analyze the contribution of each to the capture of prey by a wide variety of species of fish. We find, not surprisingly, that ram is the dominant contributor to feeding because suction, and biting, are only effective when very close to the prey. As species utilize more strongly ram-dominated modes of feeding, they may be released from the morphological and behavioral constraints associated with the need to direct a current of water into the head. Morphological and behavioral changes that facilitate larger gapes and stronger jaws are explored here, including predators that lack a protrusile upper jaw, predators with elongate jaws, predators that rely on suspension feeding, and predators that bite. Interestingly, while the mobility of the jaws and the shape of the opening of the mouth are modified in species that have departed from a primary reliance on suction feeding, the anterior-to-posterior wave of expansion persists. This wave may be greatly slowed in ram and biting species, but its retention suggests a fundamental importance to aquatic feeding.

The Teleost Intramandibular Joint: A mechanism That Allows Fish to Obtain Prey Unavailable to Suction Feeders.

Although the majority of teleost fishes possess a fused lower jaw (or mandible), some lineages have acquired a secondary joint in the lower jaw, termed the intramandibular joint (IMJ). The IMJ is a new module that formed within the already exceptionally complex teleost head, and disarticulation of two bony elements of the mandible potentially creates a "double-jointed" jaw. The apparent independent acquisition of this new functional module in divergent lineages raises a suite of questions. (1) How many teleostean lineages contain IMJ-bearing species? (2) Does the IMJ serve the same purpose in all teleosts? (3) Is the IMJ associated with altered feeding kinematics? (4) Do IMJ-bearing fishes experience trade-offs in other aspects of feeding performance? (5) Is the IMJ used to procure prey that are otherwise unavailable? The IMJ is probably under-reported, but has been documented in at least 10 lineages within the Teleostei. Across diverse IMJ-bearing lineages, this secondary joint in the lower jaw serves a variety of functions, including: generating dynamic out-levers that allow fish to apply additional force to a food item during jaw closing; allowing fish to "pick" individual prey items with pincer-like jaws; and facilitating contact with the substrate by altering the size and orientation of the gape. There are no consistent changes in feeding kinematics in IMJ-bearing species relative to their sister taxa; however, some IMJ-bearing taxa produce very slow movements during the capture of food, which may compromise their ability to move prey into the mouth via suction. Despite diversity in behavior, all IMJ-bearing lineages have the ability to remove foods that are physically attached to the substrate or to bite off pieces from sessile organisms. Because such prey cannot be drawn into the mouth by suction, the IMJ provides a new mechanism that enables fish to obtain food that otherwise would be unavailable.

Heads or tails: do stranded fish (mosquitofish, Gambusia affinis) know where they are on a slope and how to return to the water?

Aquatic vertebrates that emerge onto land to spawn, feed, or evade aquatic predators must return to the water to avoid dehydration or asphyxiation. How do such aquatic organisms determine their location on land? Do particular behaviors facilitate a safe return to the aquatic realm? In this study, we asked: will fully-aquatic mosquitofish (Gambusia affinis) stranded on a slope modulate locomotor behavior according to body position to facilitate movement back into the water? To address this question, mosquitofish (n = 53) were placed in four positions relative to an artificial slope (30° inclination) and their responses to stranding were recorded, categorized, and quantified. We found that mosquitofish may remain immobile for up to three minutes after being stranded and then initiate either a "roll" or a "leap". During a roll, mass is destabilized to trigger a downslope tumble; during a leap, the fish jumps up, above the substrate. When mosquitofish are oriented with the long axis of the body at 90° to the slope, they almost always (97%) initiate a roll. A roll is an energetically inexpensive way to move back into the water from a cross-slope body orientation because potential energy is converted back into kinetic energy. When placed with their heads toward the apex of the slope, most mosquitofish (>50%) produce a tail-flip jump to leap into ballistic flight. Because a tail-flip generates a caudually-oriented flight trajectory, this locomotor movement will effectively propel a fish downhill when the head is oriented up-slope. However, because the mass of the body is elevated against gravity, leaps require more mechanical work than rolls. We suggest that mosquitofish use the otolith-vestibular system to sense body position and generate a behavior that is "matched" to their orientation on a slope, thereby increasing the probability of a safe return to the water, relative to the energy expended.

Does feeding behavior facilitate trophic niche partitioning in two sympatric sucker species from the American Southwest?

We examined two sympatric desert fishes, Sonora suckers (Catostomus insignis) and desert suckers (Pantosteus clarkii), and asked, does feeding behavior facilitate trophic niche partitioning? To answer this question, we conducted laboratory-based feeding trials to determine whether morphology alone facilitates the diet separation between the relatively unspecialized, omnivorous Sonora sucker and the more morphologically specialized, algivorous desert sucker or whether behavioral differences accompany morphological specialization. We predicted that (1) algivorous desert suckers would maximize contact between jaws and substrate and produce a large mouth-gape to facilitate scraping attached food-material; (2) omnivorous Sonora suckers would be more effective suction feeders when consuming unattached food items from the benthos; and (3) because they are anatomically specialized for scraping, desert suckers could not alter their feeding behavior when presented with different prey types, whereas relatively unspecialized Sonora suckers could vary behavior with prey type. We found that both species maximized jaw contact when feeding on benthic-attached food, although desert suckers produced a greater gape area. We also found that Sonora suckers were more effective suction feeders when feeding on benthic-unattached prey. Counter to our initial predictions, both species altered key aspects of feeding behavior in response to different prey types/locations. It appears that both sucker species can function as generalist feeders to exploit a variety of prey types within their natural habitat; indeed, this behavioral versatility may allow desert and Sonora suckers to respond to the cyclic environmental changes that are characteristic of the aquatic habitats of the American Southwest.

Jumping sans legs: does elastic energy storage by the vertebral column power terrestrial jumps in bony fishes?

Despite having no obvious anatomical modifications to facilitate movement over land, numerous small fishes from divergent teleost lineages make brief, voluntary terrestrial forays to escape poor aquatic conditions or to pursue terrestrial prey. Once stranded, these fishes produce a coordinated and effective "tail-flip" jumping behavior, wherein lateral flexion of the axial body into a C-shape, followed by contralateral flexion of the body axis, propels the fish into a ballistic flight-path that covers a distance of multiple body lengths. We ask: how do anatomical structures that evolved in one habitat generate effective movement in a novel habitat? Within this context, we hypothesized that the mechanical properties of the axial skeleton play a critical role in producing effective overland movement, and that tail-flip jumping species demonstrate enhanced elastic energy storage through increased body flexural stiffness or increased body curvature, relative to non-jumping species. To test this hypothesis, we derived a model to predict elastic recoil work from the morphology of the vertebral (neural and hemal) spines. From ground reaction force (GRF) measurements and high-speed video, we calculated elastic recoil work, flexural stiffness, and apparent material stiffness of the body for Micropterus salmoides (a non-jumper) and Kryptolebias marmoratus (adept tail-flip jumper). The model predicted no difference between the two species in work stored by the vertebral spines, and GRF data showed that they produce the same magnitude of mass-specific elastic recoil work. Surprisingly, non-jumper M. salmoides has a stiffer body than tail-flip jumper K. marmoratus. Many tail-flip jumping species possess enlarged, fused hypural bones that support the caudal peduncle, which suggests that the localized structures, rather than the entire axial skeleton, may explain differences in terrestrial performance.

A forceful upper jaw facilitates picking-based prey capture: biomechanics of feeding in a butterflyfish, Chaetodon trichrous.

Biomechanical models of feeding mechanisms elucidate how animals capture food in the wild, which, in turn, expands our understanding of their fundamental trophic niche. However, little attention has been given to modeling the protrusible upper jaw apparatus that characterizes many teleost species. We expanded existing biomechanical models to include upper jaw forces using a generalist butterflyfish, Chaetodon trichrous (Chaetodontidae) that produces substantial upper jaw protrusion when feeding on midwater and benthic prey. Laboratory feeding trials for C. trichrous were recorded using high-speed digital imaging; from these sequences we quantified feeding performance parameters to use as inputs for the biomechanical model. According to the model outputs, the upper jaw makes a substantial contribution to the overall forces produced during mouth closing in C. trichrous. Thus, biomechanical models that only consider lower jaw closing forces will underestimate total bite force for this and likely other teleost species. We also quantified and subsequently modeled feeding events for C. trichrous consuming prey from the water column versus picking attached prey from the substrate to investigate whether there is a functional trade-off between prey capture modes. We found that individuals of C. trichrous alter their feeding behavior when consuming different prey types by changing the timing and magnitude of upper and lower jaw movements and that this behavioral modification will affect the forces produced by the jaws during prey capture by dynamically altering the lever mechanics of the jaws. In fact, the slower, lower magnitude movements produced during picking-based prey capture should produce a more forceful bite, which will facilitate feeding on benthic attached prey items, such as corals. Similarities between butterflyfishes and other teleost lineages that also employ picking-based prey capture suggest that a suite of key behavioral and morphological innovations enhances feeding success for benthic attached prey items.

Thrash, flip, or jump: the behavioral and functional continuum of terrestrial locomotion in teleost fishes.

Moving on land versus in water imposes dramatically different requirements on the musculoskeletal system. Although many limbed vertebrates, such as salamanders and prehistoric tetrapodomorphs, have an axial system specialized for aquatic locomotion and an appendicular system adapted for terrestrial locomotion, diverse extant teleosts use the axial musculoskeletal system (body plus caudal fin) to move in these two physically disparate environments. In fact, teleost fishes living at the water's edge demonstrate diversity in natural history that is reflected in a variety of terrestrial behaviors: (1) species that have only incidental contact with land (such as largemouth bass, Micropterus) will repeatedly thrash, which can roll an individual downhill, but cannot produce effective overland movements, (2) species that have occasional contact with land (like Gambusia, the mosquitofish, which evade predators by stranding themselves) will produce directed terrestrial movement via a tail-flip jump, and (3) species that spend more than half of their lives on land (like the mudskipper, Periopthalmus) will produce a prone-jump, a behavior that allows the fish to anticipate where it will land at the end of the flight phase. Both tail-flip and prone jumps are characterized by a two-phase movement consisting of body flexion followed by extension-a movement pattern that is markedly similar to the aquatic fast-start. Convergence in kinematic pattern between effective terrestrial behaviors and aquatic fast starts suggests that jumps are an exaptation of a neuromuscular system that powers unsteady escape behaviors in the water. Despite such evidence that terrestrial behaviors evolved from an ancestral behavior that is ubiquitous among teleosts, some teleosts are unable to move effectively on land-possibly due to morphological trade-offs, wherein specialization for one environment comes at a cost to performance in the other. Indeed, upon emergence onto land, gravity places an increased mechanical load on the body, which may limit the maximum size of fish that can produce terrestrial locomotion via jumping. In addition, effective terrestrial locomotor performance may require a restructuring of the musculoskeletal system that directly conflicts with the low-drag, fusiform body shape that enhances steady swimming performance. Such biomechanical trade-offs may constrain which teleost species are able to make the evolutionary transition to life on land. Here, we synthesize the current knowledge of intermittent terrestrial locomotion in teleosts and demonstrate that extant fishes represent an important model system for elucidating fundamental evolutionary mechanisms and defining the physiological constraints that must be overcome to permit life in both the aquatic and terrestrial realms.

Vertebrate land invasions-past, present, and future: an introduction to the symposium.

The transition from aquatic to terrestrial habitats was a seminal event in vertebrate evolution because it precipitated a sudden radiation of species as new land animals diversified in response to novel physical and biological conditions. However, the first stages of this environmental transition presented numerous challenges to ancestrally aquatic organisms, and necessitated changes in the morphological and physiological mechanisms that underlie most life processes, among them movement, feeding, respiration, and reproduction. How did solutions to these functional challenges evolve? One approach to this question is to examine modern vertebrate species that face analogous demands; just as the first tetrapods lived at the margins of bodies of water and likely moved between water and land regularly, many extant fishes and amphibians use their body systems in both aquatic and terrestrial habitats on a daily basis. Thus, studies of amphibious vertebrates elucidate the functional demands of two very different habitats and clarify our understanding of the initial evolutionary challenges of moving onto land. A complementary approach is to use studies of the fossil record and comparative development to gain new perspectives on form and function of modern amphibious and non-amphibious vertebrate taxa. Based on the synthetic approaches presented in the symposium, it is clear that our understanding of aquatic-to-terrestrial transitions is greatly improved by the reciprocal integration of paleontological and neontological perspectives. In addition, common themes and new insights that emerged from this symposium point to the value of innovative approaches, new model species, and cutting-edge research techniques to elucidate the functional challenges and evolutionary changes associated with vertebrates' invasion of the land.

Are kissing gourami specialized for substrate-feeding? Prey capture kinematics of Helostoma temminckii and other anabantoid fishes.

Helostoma temminckii are known for a "kissing" behavior, which is often used in intraspecific interactions, and an unusual cranial morphology that is characterized by an intramandibular joint (IMJ). The IMJ is located within the lower jaw and aids in generating the eponymous kissing movement. In other teleost linages the IMJ is associated with the adoption of a substrate-grazing foraging habit. However, because of anatomical modifications of the gill-rakers, Helostoma has been considered a midwater filter-feeding species. We offered midwater, benthic, and attached food to Helostoma, Betta, and two "true" osphronemid gouramis, to ask: (1) how do food capture kinematics differ in different foraging contexts; and (2) are Helostoma feeding kinematics distinct when compared with closely related anabantoids that lack an IMJ? For all anabantoid species except Helostoma, benthic prey were captured using a greater contribution of effective suction relative to midwater prey, though Helostoma was rarely willing to feed in the midwater. Helostoma individuals produced relatively less suction than other species regardless of the food type. Helostoma produced a much larger gape and more premaxillary protrusion than other species, but also took longer to do so. We suggest that the jaw morphology of Helostoma facilitates an extremely large mouth-gape to enhance substrate-scraping. The large amplitude mouth-opening that characterizes substrate-feeding may represent a functional trade-off, whereby the enhanced ability to procure food from the substrate is accompanied by a concomitant reduction in the ability to produce suction.