Knifefish turning control and hydrodynamics during forward swimming.

Rapid turning and swimming contribute to ecologically important behaviors in fishes such as predator avoidance, prey capture, mating and the navigation of complex environments. For riverine species, such as knifefishes, turning behaviors may also be important for navigating locomotive perturbations caused by turbulent flows. Most research on fish maneuvering focuses on fish with traditional fin and body morphologies, which primarily use body bending and the pectoral fins during turning. However, it is uncertain how fishes with uncommon morphologies are able to achieve sudden and controllable turns. Here, we studied the turning performance and the turning hydrodynamics of the black ghost knifefish (Apteronotus albifrons, N=6) which has an atypical elongated ribbon fin. Fish were filmed while swimming forward at ∼2 body lengths s-1 and feeding from a fixed feeder (control) and an oscillating feeder (75 Hz) at two different amplitudes. 3D kinematic analysis of the body revealed the highest pitch angles and lowest body bending coefficients during steady swimming. Low pitch angle, high maximum yaw angles and large body bending coefficients were characteristic of small and large turns. Asynchrony in pectoral fin use was low during turning; however, ribbon fin wavelength, frequency and wave speed were greatest during large turns. Digital particle image velocimetry (DPIV) showed larger counter-rotating vortex pairs produced during turning by the ribbon fin in comparison to vortices rotating in the same direction during steady swimming. Our results highlight the ribbon fin's role in controlled rapid turning through modulation of wavelength, frequency and wave speed.

Beyond the Kármán gait: knifefish swimming in periodic and irregular vortex streets.

Neotropical freshwater fishes such as knifefishes are commonly faced with navigating intense and highly unsteady streams. However, our knowledge on locomotion in apteronotids comes from laminar flows, where the ribbon fin dominates over the pectoral fins or body bending. Here, we studied the 3D kinematics and swimming control of seven black ghost knifefish (Apteronotus albifrons) moving in laminar flows (flow speed U∞≈1-5 BL s-1) and in periodic vortex streets (U∞≈2-4 BL s-1). Two different cylinders (∼2 and ∼3 cm diameter) were used to generate the latter. Additionally, fish were exposed to an irregular wake produced by a free oscillating cylinder (∼2 cm diameter; U∞≈2 BL s-1). In laminar flows, knifefish mainly used their ribbon fin, with wave frequency, speed and acceleration increasing with U∞. In contrast, knifefish swimming behind a fixed cylinder increased the use of pectoral fins, which resulted in changes in body orientation that mimicked steady backward swimming. Meanwhile, individuals behind the oscillating cylinder presented a combination of body bending and ribbon and pectoral fin movements that counteract the out-of-phase yaw oscillations induced by the irregular shedding of vortices. We corroborated passive out-of-phase oscillations by placing a printed knifefish model just downstream of the moving cylinder, but when placed one cylinder diameter downstream, the model oscillated in phase. Thus, the wake left behind an oscillating body is more challenging than a periodic vortex shedding for an animal located downstream, which may have consequences on inter- and intra-specific interactions.

Knifefish's suction makes water boil.

We discovered that knifefish (Apteronotus albifrons) during suction feeding can produce millimeter-sized cavitation bubbles and flow accelerations up to ~ 450 times the acceleration of gravity. Knifefish may use this powerful suction-induced cavitation to cause physical damage on prey hiding in narrow refuges, therefore facilitating capture.

Deploying Big Data to Crack the Genotype to Phenotype Code.

Mechanistically connecting genotypes to phenotypes is a longstanding and central mission of biology. Deciphering these connections will unite questions and datasets across all scales from molecules to ecosystems. Although high-throughput sequencing has provided a rich platform on which to launch this effort, tools for deciphering mechanisms further along the genome to phenome pipeline remain limited. Machine learning approaches and other emerging computational tools hold the promise of augmenting human efforts to overcome these obstacles. This vision paper is the result of a Reintegrating Biology Workshop, bringing together the perspectives of integrative and comparative biologists to survey challenges and opportunities in cracking the genotype to phenotype code and thereby generating predictive frameworks across biological scales. Key recommendations include promoting the development of minimum "best practices" for the experimental design and collection of data; fostering sustained and long-term data repositories; promoting programs that recruit, train, and retain a diversity of talent; and providing funding to effectively support these highly cross-disciplinary efforts. We follow this discussion by highlighting a few specific transformative research opportunities that will be advanced by these efforts.

Effects of hypoxia on swimming and sensing in a weakly electric fish.

Low dissolved oxygen (hypoxia) can severely limit fish performance, especially aerobically expensive behaviours including swimming and acquisition of sensory information. Fishes can reduce oxygen requirements by altering these behaviours under hypoxia, but the underlying mechanisms can be difficult to quantify. We used a weakly electric fish as a model system to explore potential effects of hypoxia on swim performance and sensory information acquisition, which enabled us to non-invasively record electric signalling activity used for active acquisition of sensory information during swimming. To quantify potential effects of hypoxia, we measured critical swim speed (Ucrit) and concurrent electric signalling activity under high- and low-dissolved oxygen concentrations in a hypoxia-tolerant African mormyrid fish, Marcusenius victoriae Fish were maintained under normoxia for 6 months prior to experimental treatments, and then acclimated for 8 weeks to normoxia or hypoxia and tested under both conditions (acute: 4 h exposure). Acute hypoxia exposure resulted in a significant reduction in both Ucrit and electric signalling activity in fish not acclimated to hypoxia. However, individuals acclimated to chronic hypoxia were characterized by a higher Ucrit under both hypoxia and normoxia than fish acclimated to normoxia. Following a 6 month re-introduction to normoxia, hypoxia-acclimated individuals still showed increased performance under acute hypoxic test conditions, but not under normoxia. Our results highlight the detrimental effects of hypoxia on aerobic swim performance and sensory information acquisition, and the ability of fish to heighten aerobic performance through acclimation processes that can still influence performance even months after initial exposure.

Kinematics of ribbon-fin locomotion in the bowfin, Amia calva.

An elongated dorsal and/or anal ribbon-fin to produce forward and backward propulsion has independently evolved in several groups of fishes. In these fishes, fin ray movements along the fin generate a series of waves that drive propulsion. There are no published data on the use of the dorsal ribbon-fin in the basal freshwater bowfin, Amia calva. In this study, frequency, amplitude, wavelength, and wave speed along the fin were measured in Amia swimming at different speeds (up to 1.0 body length/sec) to understand how the ribbon-fin generates propulsion. These wave properties were analyzed to (1) determine whether regional specialization occurs along the ribbon-fin, and (2) to reveal how the undulatory waves are used to control swimming speed. Wave properties were also compared between swimming with sole use of the ribbon-fin, and swimming with simultaneous use of the ribbon and pectoral fins. Statistical analysis of ribbon-fin kinematics revealed no differences in kinematic patterns along the ribbon-fin, and that forward propulsive speed in Amia is controlled by the frequency of the wave in the ribbon-fin, irrespective of the contribution of the pectoral fin. This study is the first kinematic analysis of the ribbon-fin in a basal fish and the model species for Amiiform locomotion, providing a basis for understanding ribbon-fin locomotion among a broad range of teleosts.

Prey processing in the Siamese fighting fish (Betta splendens).

We studied prey processing in the Siamese fighting fish (Betta splendens), involving slow, easily observed head-bobbing movements, which were compared with prey processing in other aquatic feeding vertebrates. We hypothesized that head-bobbing is a unique prey-processing behaviour, which alternatively could be structurally and functionally analogous with raking in basal teleosts, or with pharyngognathy in neoteleosts. Modulation of head-bobbing was elicited by prey with different motility and toughness. Head-bobbing involved sustained mouth occlusion and pronounced cranial elevation, similar to raking. However, the hyoid and pectoral girdle were protracted, and not retracted as in both raking and pharyngognathy. High-speed videofluoroscopy of hyoid movements confirmed that head-bobbing differs from other known aquatic prey-processing behaviours. Nevertheless, head-bobbing and other prey-processing behaviours converge on a recurrent functional theme in the trophic ecology of aquatic feeding vertebrates; the use of intraoral and oropharyngeal dentition surfaces to immobilize, reduce and process relatively large, tough or motile prey. Prey processing outside the pharyngeal region has not been described for neoteleosts previously, but morphological evidence suggests that relatives of Betta might use similar processing behaviours. Thus, our results suggest that pharyngognathy did not out-compete ancestral prey-processing mechanisms completely during the evolution of neoteleosts.

Evolution of muscle activity patterns driving motions of the jaw and hyoid during chewing in Gnathostomes.

Although chewing has been suggested to be a basal gnathostome trait retained in most major vertebrate lineages, it has not been studied broadly and comparatively across vertebrates. To redress this imbalance, we recorded EMG from muscles powering anteroposterior movement of the hyoid, and dorsoventral movement of the mandibular jaw during chewing. We compared muscle activity patterns (MAP) during chewing in jawed vertebrate taxa belonging to unrelated groups of basal bony fishes and artiodactyl mammals. Our aim was to outline the evolution of coordination in MAP. Comparisons of activity in muscles of the jaw and hyoid that power chewing in closely related artiodactyls using cross-correlation analyses identified reorganizations of jaw and hyoid MAP between herbivores and omnivores. EMG data from basal bony fishes revealed a tighter coordination of jaw and hyoid MAP during chewing than seen in artiodactyls. Across this broad phylogenetic range, there have been major structural reorganizations, including a reduction of the bony hyoid suspension, which is robust in fishes, to the acquisition in a mammalian ancestor of a muscle sling suspending the hyoid. These changes appear to be reflected in a shift in chewing MAP that occurred in an unidentified anamniote stem-lineage. This shift matches observations that, when compared with fishes, the pattern of hyoid motion in tetrapods is reversed and also time-shifted relative to the pattern of jaw movement.

Rhythmic chewing with oral jaws in teleost fishes: a comparison with amniotes.

Intra-oral prey processing (chewing) using the mandibular jaws occurs more extensively among teleost fishes than previously documented. The lack of muscle spindles, gamma-motoneurons and periodontal afferents in fishes makes them useful for testing hypotheses regarding the relationship between these sensorimotor components and rhythmic chewing in vertebrates. Electromyography (EMG) data from the adductor mandibulae (AM) were used to quantify variation in chew cycle duration in the bowfin Amia, three osteoglossomorphs (bony-tongues), four salmonids and one esocid (pike). All species chewed prey using their oral jaw in repetitive trains of between 3 and 30 consecutive chews, a pattern that resembles cyclic chewing in amniote vertebrates. Variance in rhythmicity was compared within and between lineages using coefficients of variation and Levene's test for homogeneity of variance. These comparisons revealed that some teleosts exhibit degrees of rhythmicity that are comparable to mammalian mastication and higher than in lepidosaurs. Moreover, chew cycle durations in fishes, as in mammals, scale positively with mandible length. Chewing among basal teleosts may be rhythmic because it is stereotyped and inflexible, the result of patterned interactions between sensory feedback and a central pattern generator, because the lack of a fleshy tongue renders jaw-tongue coordination unnecessary and/or because stereotyped opening and closing movements are important for controlling fluid flow in the oral cavity.

Functional morphology and biomechanics of the tongue-bite apparatus in salmonid and osteoglossomorph fishes.

The tongue-bite apparatus and its associated musculoskeletal elements of the pectoral girdle and neurocranium form the structural basis of raking, a unique prey-processing behaviour in salmonid and osteoglossomorph fishes. Using a quantitative approach, the functional osteology and myology of this system were compared between representatives of each lineage, i.e. the salmonid Salvelinus fontinalis (N = 10) and the osteoglossomorph Chitala ornata (N = 8). Divergence was found in the morphology of the novel cleithrobranchial ligament, which potentially relates to kinematic differences between the raking lineage representatives. Salvelinus had greater anatomical cross-sectional areas of the epaxial, hypaxial and protractor hyoideus muscles, whereas Chitala had greater sternohyoideus and adductor mandibulae mass. Two osteology-based biomechanical models (a third-order lever for neurocranial elevation and a modified four-bar linkage for hyoid retraction) showed divergent force/velocity priorities in the study taxa. Salvelinus maximizes both force (via powerful cranial muscles) and velocity (through mechanical amplification) during raking. In contrast, Chitala has relatively low muscle force but more efficient force transmission through both mechanisms compared with Salvelinus. It remains unclear if and how behavioural modulation and specializations in the post-cranial anatomy may affect the force/velocity trade-offs in Chitala. Further studies of tongue-bite apparatus morphology and biomechanics in a broader species range may help to clarify the role that osteology and myology play in the evolution of behavioural diversity.

Biomechanics of a convergently derived prey-processing mechanism in fishes: evidence from comparative tongue bite apparatus morphology and raking kinematics.

A tongue-bite apparatus (TBA) governs raking behaviors in two major and unrelated teleost lineages, the osteoglossomorph and salmoniform fishes. We present data on comparative morphology and kinematics from two representative species, the rainbow trout (Oncorhynchus mykiss) and the Australian arowana (Scleropages jardinii), which suggest that both the TBA and raking are convergently derived in these lineages. Similar TBA morphologies were present, except for differences in TBA dentition and shape of the novel cleithrobranchial ligament (CBL), which is arc-shaped in O. mykiss and straight in S. jardinii. Eight kinematic variables were used to quantify motion magnitude and maximum-timing in the kinematic input mechanisms of the TBA. Five variables differed inter-specifically (pectoral girdle retraction magnitude and timing, cranial and hyoid elevation and gape-distance timing), yet an incomplete taxon separation across multivariate kinematic space demonstrated an overall similarity in raking behavior. An outgroup analysis using bowfin (Amia calva) and pickerel (Esox americanus) to compare kinematics of raking with chewing and prey-capture provided robust quantitative evidence of raking being a convergently derived behavior. Support was also found for the notion that raking more likely evolved from the strike, a functionally distinct behavior, than from chewing, an alternative prey-processing behavior. Based on raking kinematic and muscle-activity data, we propose biomechanical models of the three input mechanisms that govern kinematics of the basihyal output mechanism during the raking power stroke: (1) cranial elevation protracts the upper TBA jaw from the lower (basihyal) TBA jaw; (2) basihyal retraction is caused directly by contraction of the sternohyoideus (SH); (3) hypaxial shortening, relayed via the pectoral girdle and SH-CBL complex, is an indirect basihyal retraction mechanism modeled as a four-bar linkage. These models will aid future analyses mapping structural and functional traits to the evolution of behaviors.

Fluid dynamics of feeding behaviour in white-spotted bamboo sharks.

Although the motor control of feeding is presumed to be generally conserved, some fishes are capable of modulating the feeding behaviour in response to prey type and or prey size. This led to the 'feeding modulation hypothesis', which states that rapid suction strikes are pre-programmed stereotyped events that proceed to completion once initiated regardless of sensory input. If this hypothesis holds true, successful strikes should be indistinguishable from unsuccessful strikes owing to a lack of feedback control in specialized suction feeding fishes. The hydrodynamics of suction feeding in white-spotted bamboo sharks (Chiloscyllium plagiosum) was studied in three behaviours: successful strikes, intraoral transports of prey and unsuccessful strikes. The area of the fluid velocity region around the head of feeding sharks was quantified using time-resolved digital particle image velocimetry (DPIV). The maximal size of the fluid velocity region is 56% larger in successful strikes than unsuccessful strikes (10.79 cm2 vs 6.90 cm2), but they do not differ in duration, indicating that strikes are modulated based on some aspect of the prey or simply as a result of decreased effort on the part of the predator. The hydrodynamic profiles of successful and unsuccessful strikes differ after 21 ms, a period probably too short to provide time to react through feedback control. The predator-to-prey distance is larger in missed strikes compared with successful strikes, indicating that insufficient suction is generated to compensate for the increased distance. An accuracy index distinguishes unsuccessful strikes (-0.26) from successful strikes (0.45 to 0.61). Successful strikes occur primarily between the horizontal axis of the mouth and the dorsal boundary of the ingested parcel of water, and missed prey are closer to the boundary or beyond. Suction transports are shorter in duration than suction strikes but have similar maximal fluid velocity areas to move the prey through the oropharyngeal cavity into the oesophagus (54 ms vs 67 ms).

Suction generation in white-spotted bamboo sharks Chiloscyllium plagiosum.

After the divergence of chondrichthyans and teleostomes, the structure of the feeding apparatus also diverged leading to alterations in the suction mechanism. In this study we investigated the mechanism for suction generation during feeding in white-spotted bamboo sharks, Chiloscyllium plagiosum and compared it with that in teleosts. The internal movement of cranial elements and pressure in the buccal, hyoid and pharyngeal cavities that are directly responsible for suction generation was quantified using sonomicrometry and pressure transducers. Backward stepwise multiple linear regressions were used to explore the relationship between expansion and pressure, accounting for 60-96% of the variation in pressure among capture events. The progression of anterior to posterior expansion in the buccal, hyoid and pharyngeal cavities is accompanied by the sequential onset of subambient pressure in these cavities as prey is drawn into the mouth. Gape opening triggers the onset of subambient pressure in the oropharyngeal cavities. Peak gape area coincides with peak subambient buccal pressure. Increased velocity of hyoid area expansion is primarily responsible for generating peak subambient pressure in the buccal and hyoid regions. Pharyngeal expansion appears to function as a sink to receive water influx from the mouth, much like that of compensatory suction in bidirectional aquatic feeders. Interestingly, C. plagiosum generates large suction pressures while paradoxically compressing the buccal cavity laterally, delaying the time to peak pressure. This represents a fundamental difference from the mechanism used to generate suction in teleost fishes. Interestingly, pressure in the three cavities peaks in the posterior to anterior direction. The complex shape changes that the buccal cavity undergoes indicate that, as in teleosts, unsteady flow predominates during suction feeding. Several kinematic variables function together, with great variation over long gape cycles to generate the low subambient pressures used by C. plagiosum to capture prey.

Is a convergently derived muscle-activity pattern driving novel raking behaviours in teleost fishes?

Behavioural differences across prey-capture and processing mechanisms may be governed by coupled or uncoupled feeding systems. Osteoglossomorph and salmonid fishes process prey in a convergently evolved tongue-bite apparatus (TBA), which is musculoskeletally coupled with the primary oral jaws. Altered muscle-activity patterns (MAPs) in these coupled jaw systems could be associated with the independent origin of a novel raking behaviour in these unrelated lineages. Substantial MAP changes in the evolution of novel behaviours have rarely been quantified so we examined MAP differences across strikes, chewing and rakes in a derived raking salmonid, the rainbow trout, Oncorhynchus mykiss. Electromyography, including activity onset timing, duration, mean amplitude and integrated area from five feeding muscles revealed significant differences between behaviour-specific MAPs. Specifically, early activity onset in the protractor hyoideus and adductor mandibularis muscles characterised raking, congruent with a recent biomechanical model of the component-mechanisms driving the raking preparatory and power-stroke phases. Oncorhynchus raking MAPs were then compared with a phylogenetically derived osteoglossomorph representative, the Australian arowana, Scleropages jardinii. In both taxa, early onset of protractor hyoideus and adductor mandibularis activity characterised the raking preparatory phase, indicating a convergently derived MAP, while more subtle inter-lineage divergence in raking MAPs resulted from onset-timing and duration differences in sternohyoideus and hypaxialis activity. Convergent TBA morphologies are thus powered by convergently derived MAPs, a phenomenon not previously demonstrated in feeding mechanisms. Between lineages, differences in TBA morphology and associated differences in the functional coupling of jaw systems appear to be important factors in shaping the diversification of raking behaviours.

Congruence between muscle activity and kinematics in a convergently derived prey-processing behavior.

Quantification of anatomical and physiological characteristics of the function of a musculoskeletal system may yield a detailed understanding of how the organizational levels of morphology, biomechanics, kinematics, and muscle activity patterns (MAPs) influence behavioral diversity. Using separate analyses of these organizational levels in representative study taxa, we sought patterns of congruence in how organizational levels drive behavioral modulation in a novel raking prey-processing behavior found in teleosts belonging to two evolutionarily distinct lineages. Biomechanically divergent prey (elusive, robust goldfish and sedentary, malleable earthworms) were fed to knifefish, Chitala ornata (Osteoglossomorpha) and brook trout, Salvelinus fontinalis (Salmoniformes). Electromyography recorded MAPs from the hyoid protractor, jaw adductor, sternohyoideus, epaxialis, and hypaxialis musculature, while sonomicrometry sampled deep basihyal kinesis and contractile length dynamics in the basihyal protractor and retractor muscles. Syntheses of our results with recent analyses of cranial morphology and raking kinematics showed that raking in Salvelinus relies on an elongated cranial out lever, extensive cranial elevation and a curved cleithrobranchial ligament (CBL), and that both raking MAPs and kinematics remain entirely unmodulated-a highly unusual trait, particularly among feeding generalists. Chitala had a shorter CBL and a raking power stroke involving increased retraction of the elongated pectoral girdle during raking on goldfish. The raking MAP was also modulated in Chitala, involving an extensive overlap between muscle activity of the preparatory and power stroke phases, driven by shifts in hypaxial timing and recruitment of the hyoid protractor muscle. Sonomicrometry revealed that the protractor hyoideus muscle stored energy from retraction of the pectoral girdle for ca. 5-20 ms after onset of the power stroke and then hyper-extended. This mechanism of elastic recoil in Chitala, which amplifies retraction of the basihyal during raking on goldfish without a significant increase in recruitment of the hypaxialis, suggests a unique mechanism of modulation based on performance-enhancing changes in the design and function of the musculoskeletal system.

Evolution and ecology of feeding in elasmobranchs.

Paleozoic chondrichthyans had a large gape, numerous spike-like teeth, limited cranial kinesis, and a non-suspensory hyoid, suggesting a feeding mechanism dominated by bite and ram. Modern sharks are characterized by a mobile upper jaw braced by a suspensory hyoid arch that is highly kinetic. In batoids, the upper jaw is dissociated from the cranium permitting extensive protrusion of the jaws. Similar to actinopterygians, the evolution of highly mobile mandibular and hyoid elements has been correlated with extensive radiation of feeding modes in elasmobranchs, particularly that of suction. Modern elasmobranchs possess a remarkable variety of feeding modes for a group containing so few species. Biting, suction or filter-feeding may be used in conjunction with ram to capture prey, with most species able to use a combination of behaviors during a strike. Suction-feeding has repeatedly arisen within all recent major elasmobranch clades and is associated with a suite of morphological and behavioral specializations. Prey capture in a diverse assemblage of purported suction-feeding elasmobranchs is investigated in this study. Drop in water pressure measured in the mouth and at the location of the prey shows that suction inflow drops off rapidly with distance from the predator's mouth. Elasmobranchs specializing in suction-feeding may be limited to bottom associated prey and because of their small gape may have a diet restricted to relatively small prey. Behavior can affect performance and overcome constraints imposed by the fluid medium. Suction performance can be enhanced by proximity to a substrate or by decreasing distance from predator to prey using various morphological and/or behavioral characteristics. Benthic suction-feeders benefit by the increased strike radius due to deflection of water flow when feeding close to a substrate, and perhaps require less accuracy when capturing prey. Suction and ram-suction-feeding elasmobranchs can also use suction inflow to draw prey to them from a short distance, while ram-feeding sharks must accelerate and overtake the prey. The relationship between feeding strategy and ecology may depend in part on ecological, mechanistic or evolutionary specialization. Mechanistic suction-feeding specialist elasmobranchs are primarily benthic, while most epibenthic and pelagic elasmobranchs are generalists and use ram, suction, and biting to catch a diversity of prey in various habitats. Some shark species are considered to be ecological specialists in choosing certain kinds of prey over others. Batoids are evolutionary specialists in having a flattened morphology and most are generalist feeders. Filter-feeding elasmobranchs are ecological, mechanistic, and evolutionary specialists.

Use of sonomicrometry demonstrates the link between prey capture kinematics and suction pressure in largemouth bass.

Suction feeding in fishes is the result of a highly coordinated explosive expansion of the buccal cavity that results in a rapid drop in pressure. Prey are drawn into the mouth by a flow of water that is generated by this expansion. At a gross level it is clear that the expansion of the buccal cavity is responsible for the drop in pressure. However, attempts using high-speed video recordings to demonstrate a tight link between prey capture kinematics and suction pressure have met with limited success. In a study with largemouth bass Micropterus salmoides, we adopted a new technique for studying kinematics, sonomicrometry, to transduce the movement of skeletal elements of the head during feeding, and synchronized pressure recordings at a sampling rate of 500 Hz. From the positional relationships of six piezoelectric crystals we monitored the internal movements of the buccal cavity and mouth in both mid-sagittal and transverse planes. We found that peak subambient pressure was reached very early in the kinematic expansion of the buccal cavity, occurring at the time when the rate of percentage change in buccal volume was at its peak. Using multiple regression analyses we were consistently able to account for over 90%, and in the best model 99%, of the variation in buccal pressure among strikes using kinematic variables. Sonomicrometry shows great promise as a method for documenting movements of biological structures that are not clearly visible in the external view provided by film and video recordings.

Functional morphology of the "tongue-bite" in the osteoglossomorph fish Notopterus.

Osteoglossomorph fishes are characterized by having three sets of jaws: a mandibular jaw apparatus (MJA) anteriorly, a pharyngeal jaw apparatus (PJA) posteriorly, and a tongue-bite apparatus (TBA) associated with basihyal and parasphenoid teeth. The TBA is a novel complex feature of the head that characterizes osteoglossomorph fishes and provides a case study in the origin of novel functions and roles in the vertebrate musculoskeletal system. The function of the tongue-bite in the osteoglossomorph fish Notopterus was characterized by using high-speed cinematography and electromyography. The tongue-bite is used during intraoral prey processing to shred and disable prey. Two distinct uses of the TBA were defined on the basis of kinematic and electromyographic profiles: raking and open-mouth chewing. During raking behavior, the prey is held fixed in the MJA, the neurocranium is elevated, and the pectoral girdle is retracted. The adductor mandibulae, hypaxialis, epaxialis, and posterior intermandibularis muscles are all highly active, but only very low activity is observed in the sternohyoideus muscle. During open-mouth chewing behavior, the prey is located within the oral cavity, posterior pectoral girdle rotation is less than during raking, and the levator operculi muscle shows relatively high activity. We propose that a shearing action of the basihyal (moved anteroposteriorly by the posterior intermandibularis and hypaxial muscles) with respect to the neurocranium (elevated by epaxial muscles) is the critical aspect of the tongue-bite in Notopterus. The body muscles (epaxials and hypaxials) provide the main power for the tongue-bite. We hypothesize that lack of sternohyoideus activity during intraoral prey processing, posterior pectoral girdle rotation, and a long-fibered posterior intermandibularis muscle are novel structures associated with the tongue-bite apparatus within osteoglossomorph fishes.