Regional differences in vertebral shape along the axial skeleton in caecilians (Amphibia: Gymnophiona).

Caecilians are elongate, limbless and annulated amphibians that, as far as is known, all have an at least partly fossorial lifestyle. It has been suggested that elongate limbless vertebrates show little morphological differentiation throughout the postcranial skeleton. However, relatively few studies have explored the axial skeleton in limbless tetrapods. In this study, we used µCT data and three-dimensional geometric morphometrics to explore regional differences in vertebral shape across a broad range of caecilian species. Our results highlight substantial differences in vertebral shape along the axial skeleton, with anterior vertebrae being short and bulky, whereas posterior vertebrae are more elongated. This study shows that despite being limbless, elongate tetrapods such as caecilians still show regional heterogeneity in the shape of individual vertebrae along the vertebral column. Further studies are needed, however, to understand the possible causes and functional consequences of the observed variation in vertebral shape in caecilians.

Is vertebral shape variability in caecilians (Amphibia: Gymnophiona) constrained by forces experienced during burrowing?

Caecilians are predominantly burrowing, elongate, limbless amphibians that have been relatively poorly studied. Although it has been suggested that the sturdy and compact skulls of caecilians are an adaptation to their head-first burrowing habits, no clear relationship between skull shape and burrowing performance appears to exist. However, the external forces encountered during burrowing are transmitted by the skull to the vertebral column, and, as such, may impact vertebral shape. Additionally, the muscles that generate the burrowing forces attach onto the vertebral column and consequently may impact vertebral shape that way as well. Here, we explored the relationships between vertebral shape and maximal in vivo push forces in 13 species of caecilian amphibians. Our results show that the shape of the two most anterior vertebrae, as well as the shape of the vertebrae at 90% of the total body length, is not correlated with peak push forces. Conversely, the shape of the third vertebrae, and the vertebrae at 20% and 60% of the total body length, does show a relationship to push forces measured in vivo. Whether these relationships are indirect (external forces constraining shape variation) or direct (muscle forces constraining shape variation) remains unclear and will require quantitative studies of the axial musculature. Importantly, our data suggest that mid-body vertebrae may potentially be used as proxies to infer burrowing capacity in fossil representatives.

The relationship between head shape, head musculature and bite force in caecilians (Amphibia: Gymnophiona).

Caecilians are enigmatic limbless amphibians that, with a few exceptions, all have an at least partly burrowing lifestyle. Although it has been suggested that caecilian evolution resulted in sturdy and compact skulls as an adaptation to their head-first burrowing habits, no relationship between skull shape and burrowing performance has been demonstrated to date. However, the unique dual jaw-closing mechanism and the osteological variability of their temporal region suggest a potential relationship between skull shape and feeding mechanics. Here, we explored the relationships between skull shape, head musculature and in vivo bite forces. Although there is a correlation between bite force and external head shape, no relationship between bite force and skull shape could be detected. Whereas our data suggest that muscles are the principal drivers of variation in bite force, the shape of the skull is constrained by factors other than demands for bite force generation. However, a strong covariation between the cranium and mandible exists. Moreover, both cranium and mandible shape covary with jaw muscle architecture. Caecilians show a gradient between species with a long retroarticular process associated with a large and pennate-fibered m. interhyoideus posterior and species with a short process but long and parallel-fibered jaw adductors. Our results demonstrate the complexity of the relationship between form and function of this jaw system. Further studies that focus on factors such as gape distance or jaw velocity will be needed in order to fully understand the evolution of feeding mechanics in caecilians.

Under pressure: the relationship between cranial shape and burrowing force in caecilians (Gymnophiona).

Caecilians are elongate, limbless and annulated amphibians that, with the exception of one aquatic family, all have an at least partly fossorial lifestyle. It has been suggested that caecilian evolution resulted in sturdy and compact skulls with fused bones and tight sutures, as an adaptation to their head-first burrowing habits. However, although their cranial osteology is well described, relationships between form and function remain poorly understood. In the present study, we explored the relationship between cranial shape and in vivo burrowing forces. Using micro-computed tomography (µCT) data, we performed 3D geometric morphometrics to explore whether cranial and mandibular shapes reflected patterns that might be associated with maximal push forces. The results highlight important differences in maximal push forces, with the aquatic Typhlonectes producing a lower force for a given size compared with other species. Despite substantial differences in head morphology across species, no relationship between overall skull shape and push force could be detected. Although a strong phylogenetic signal may partly obscure the results, our conclusions confirm previous studies using biomechanical models and suggest that differences in the degree of fossoriality do not appear to be driving the evolution of head shape.

Measurement error in µCT-based three-dimensional geometric morphometrics introduced by surface generation and landmark data acquisition.

Computed-tomography-derived (CT-derived) polymesh surfaces are widely used in geometric morphometric studies. This approach is inevitably associated with decisions on scanning parameters, resolution, and segmentation strategies. Although the underlying processing steps have been shown to potentially contribute artefactual variance to three-dimensional landmark coordinates, their effects on measurement error have rarely been assessed systematically in CT-based geometric morphometric studies. The present study systematically assessed artefactual variance in landmark data introduced by the use of different voxel sizes, segmentation strategies, surface simplification degrees, and by inter- and intra-observer differences, and compared their magnitude to true biological variation. Multiple CT-derived surface variants of the anuran (Amphibia: Anura) pectoral girdle were generated by systematic changes in the factors that potentially influence the surface geometries. Twenty-four landmarks were repeatedly acquired by different observers. The contribution of all factors to the total variance in the landmark data was assessed using random-factor nested permanovas. Selected sets of Euclidean distances between landmark sets served further to compare the variance among factor levels. Landmark precision was assessed by landmark standard deviation and compared among observers and days. Results showed that all factors, except for voxel size, significantly contributed to measurement error in at least some of the analyses performed. In total, 6.75% of the variance in landmark data that mimicked a realistic biological study was caused by measurement error. In this landmark dataset, intra-observer error was the major source of artefactual variance followed by inter-observer error; the factor segmentation contributed < 1% and slight surface simplification had no significant effect. Inter-observer error clearly exceeded intra-observer error in a different landmark dataset acquired by six partly inexperienced observers. The results suggest that intra-observer error can potentially be reduced by including a training period prior to the actual landmark acquisition task and by acquiring landmarks in as few sessions as possible. Additionally, the application of moderate and careful surface simplification and, potentially, also the use of case-specific optimal combinations of automatic local thresholding algorithms and parameters for segmentation can help reduce intra-observer error. If landmark data are to be acquired by several observers, it is important to ensure that all observers are consistent in landmark identification. Despite the significant amount of artefactual variance, we have shown that landmark data acquired from microCT-derived surfaces are precise enough to study the shape of anuran pectoral girdles. Yet, a systematic assessment of measurement error is advisable for all geometric morphometric studies.

A biomechanical view on stinger diversity in scorpions.

Scorpions have elongated metasomas that bear a telson, which is used as a stinger for venom injection. There is a remarkable diversity in the use of the stinger among scorpions, comprising defensive behavior, prey subjugation and mating. This diversity could be reflected by the shape of the telson, as different stinging behaviors will result in very different functional demands. Here we explored the diversity of telson shapes in scorpions by providing morphological measurements, such as curvature and tip angle, as well as by testing stingers under load using finite element analysis (FEA). FEA models were loaded with forces scaled to the surface area of the models, to allow comparison of the relative strain energy based on shape alone. Load force angle was rotated to identify the optimal stinging angle based on the lowest strain energy. Aculeus length and mean aculeus height correlated with minimal strain energy. Optimal stinging angle correlated with tip angle, and differed from the tip angle by about 28.4 ±â€…6.22 °. We found that species that are more venomous have long aculei (stinger barbs) with a larger radius of curvature. FEA models of these longer aculei showed basal stress concentrations, indicating a potential greater risk of basal breakage due to shape alone. Telsons with shorter and thicker aculeus shapes showed stress concentrations at the tip only. Despite these marked differences in shape, we found no difference in the scaled strain energy between groups of species that are more venomous and less venomous groups of species. These results show that scorpion stingers may be biomechanically optimized, and this may indicate different usage of the stinger in different species.

Oil Secretory System in Vegetative Organs of Three Arnica Taxa: Essential Oil Synthesis, Distribution and Accumulation.

Arnica, a genus including the medicinal species A. montana, in its Arbo variety, and A. chamissonis, is among the plants richest in essential oils used as pharmaceutical materials. Despite its extensive use, the role of anatomy and histochemistry in the internal secretory system producing the essential oil is poorly understood. Anatomical sections allowed differentiation between two forms of secretory structures which differ according to their distribution in plants. The first axial type is connected to the vascular system of all vegetative organs and forms canals lined with epithelial cells. The second cortical type is represented by elongated intercellular spaces filled with oil formed only between the cortex cells of roots and rhizomes at maturity, with canals lacking an epithelial layer.Only in A. montana rhizomes do secretory structures form huge characteristic reservoirs. Computed tomography illustrates their spatial distribution and fusiform shape. The axial type of root secretory canals is formed at the interface between the endodermis and cortex parenchyma, while, in the stem, they are located in direct contact with veinal parenchyma. The peripheral phloem parenchyma cells are arranged in strands around sieve tube elements which possess a unique ability to accumulate large amounts of oil bodies. The cells of phloem parenchyma give rise to the aforementioned secretory structures while the lipid components (triacylglycerols) stored there support the biosynthesis of essential oils by later becoming a medium in which these oils are dissolved. The results indicate the integrity of axial secretory structures forming a continuous system in vegetative plant organs.

Landing on branches in the frog Trachycephalus resinifictrix (Anura: Hylidae).

Frogs (Lissamphibia: Anura) are famous for their saltatory or hopping locomotion, which is related to numerous anatomical specialisations that are characteristic for the group. However, while the biomechanics of take-off in frogs have been studied in detail, much less is known on how frogs land after a jump. Besides terrestrial and aquatic species, several lineages of frogs adopted an arboreal lifestyle and especially the biomechanics of landing on challenging, small, and unpredictable substrates, such as leaves or branches, are virtually unknown. Here we studied the landing kinematics of the arboreal frog Trachycephalus resinifictrix (Hylidae) on a wooden stick that was used to mimic a small tree branch. We observed two different landing behaviours: (1) landing on the abdomen and (2) attachment with the toes of either the forelimb or the hindlimb. In the latter case, the frogs performed a cartwheel around the stick, while they were only attached by their adhesive toe pads. We estimated the forces that act on the toes during this behaviour to be up to fourteen times the body weight of the animals. This behaviour demonstrates the remarkable adhesive capabilities of the toe pads and the body control of the frogs.

The unique sound production of the Death's-head hawkmoth (Acherontia atropos (Linnaeus, 1758)) revisited.

When disturbed, adults of the Death's-head hawkmoth (Lepidoptera, Sphingidae: Acherontia atropos) produce short squeaks by drawing in and deflating air into and out of the pharynx as a defence mechanism. We took a new look at Prell's hypothesis of a two-phase mechanism by providing new insights into the functional morphology behind the pharyngeal sound production of this species. First, we compared the head anatomy of A. atropos with another sphingid species, Manduca sexta, by using micro-computed tomography (CT) and 3D reconstruction methods. Despite differences in feeding behaviour and capability of sound production in the two species, the musculature in the head is surprisingly similar. However, A. atropos has a much shorter proboscis and a modified epipharynx with a distinct sclerotised lobe projecting into the opening of the pharynx. Second, we observed the sound production in vivo with X-ray videography, mammography CT and high-speed videography. Third, we analysed acoustic pressure over time and spectral frequency composition of six A. atropos specimens, both intact and with a removed proboscis. Single squeaks of A. atropos last for ca. 200 ms and consist of an inflation phase, a short pause and a deflation phase. The inflation phase is characterised by a burst of ca. 50 pulses with decreasing pulse frequency and a major frequency peak at ca. 8 kHz, followed by harmonics ranging up to more than 60 kHz. The deflation phase is characterised by a less clear acoustic pattern, a lower amplitude and more pronounced peaks in the same frequency range. The removal of the proboscis resulted in a significantly shortened squeak, a lower acoustic pressure level and a slightly more limited frequency spectrum. We hypothesise that the uptake of viscous honey facilitated the evolution of an efficient valve at the opening of the pharynx (i.e. a modified epipharynx), and that sound production could relatively easily have evolved based on this morphological pre-adaptation.

Stick tight: suction adhesion on irregular surfaces in the northern clingfish.

The northern clingfish, Gobiesox maeandricus, is able to adhere to slippery, fouled and irregular surfaces in the marine intertidal environment. We have found that the fish can adhere equally well to surfaces with a broad range of surface roughness, from the finest sandpaper (R(a) = 15 µm) to textures suitable for removing finish from flooring (R(a) = 269 µm). The fishes outperform man-made suction cups, which only adhere to the smoothest surfaces. The adhesive forces of clingfish correspond to pressures 0.2-0.5 atm below ambient and are 80-230 times the body weight of the fish. The tenacity appears related to hierarchically structured microvilli around the edges of the adhesive disc that are similar in size and aspect ratio to the setae found on the feet of geckoes, spiders and insects. This points to a possible biomimetic solution to the problem of reversibly adhering to irregular, submerged surfaces.

Packing a pinch: functional implications of chela shapes in scorpions using finite element analysis.

Scorpions depend on their pedipalps for prey capture, defense, mating and sensing their environment. Some species additionally use their pedipalps for burrowing or climbing. Because the pincers or chelae at the end of the pedipalps vary widely in shape, they have been used as part of a suite of characters to delimit ecomorphotypes. We here evaluate the influence of the different chela cuticular shapes on their performance under natural loading conditions. Chelae of 20 species, representing seven families and spanning most of the range of chela morphologies, were assigned to clusters based on chela shape parameters using hierarchical cluster analysis. Several clusters were identified corresponding approximately to described scorpion ecomorphotypes. Finite element models of the chela cuticulae were constructed from CT scans and loaded with estimated pinch forces based on in vivo force measurements. Chela shape clusters differed significantly in mean Von Mises stress and strain energy. Normalized FEA showed that chela shape significantly influenced Von Mises stress and strain energy in the chela cuticula, with Von Mises stress varying up to an order of magnitude and strain energy up to two orders of magnitude. More elongate, high-aspect ratio chela forms showed significantly higher mean stress compared with more robust low-aspect ratio forms. This suggests that elongate chelae are at a higher risk of failure when operating near the maximum pinch force. Phylogenetic independent contrasts (PIC) were calculated based on a partly resolved phylogram with branch lengths based on an alignment of the 12S, 16S and CO1 mitochondrial genes. PIC showed that cuticular stress and strain in the chela were correlated with several shape parameters, such as aspect ratio, movable finger length, and chela height, independently of phylogenetic history. Our results indicate that slender chela morphologies may be less suitable for high-force functions such as burrowing and defense. Further implications of these findings for the ecology and evolution of the different chela morphologies are discussed.

Is solid always best? Cranial performance in solid and fenestrated caecilian skulls.

Caecilians (Lissamphibia: Gymnophiona) are characterized by a fossorial lifestyle that appears to play a role in the many anatomical specializations in the group. The skull, in particular, has been the focus of previous studies because it is driven into the substrate for burrowing. There are two different types of skulls in caecilians: (1) stegokrotaphic, where the squamosal completely covers the temporal region and the jaw closing muscles, and (2) zygokrotaphic, with incomplete coverage of the temporal region by the squamosal. We used 3-D imaging and modeling techniques to explore the functional consequences of these skull types in an evolutionary context. We digitally converted stegokrotaphic skulls into zygokrotaphic skulls and vice versa. We also generated a third, akinetic skull type that was presumably present in extinct caecilian ancestors. We explored the benefits and costs of the different skull types under frontal loading at different head angles with finite element analysis (FEA). Surprisingly, the differences in stress distributions and bending between the three tested skull types were minimal and not significant. This suggests that the open temporal region in zygokrotaphic skulls does not lead to poorer performance during burrowing. However, the results of the FEA suggest a strong relationship between the head angle and skull performance, implying there is an optimal head angle during burrowing.

Ecomorphology of the pectoral girdle in anurans (Amphibia, Anura): Shape diversity and biomechanical considerations.

Frogs and toads (Lissamphibia: Anura) show a diversity of locomotor modes that allow them to inhabit a wide range of habitats. The different locomotor modes are likely to be linked to anatomical specializations of the skeleton within the typical frog Bauplan. While such anatomical adaptations of the hind limbs and the pelvic girdle are comparably well understood, the pectoral girdle received much less attention in the past. We tested for locomotor-mode-related shape differences in the pectoral girdle bones of 64 anuran species by means of micro-computed-tomography-based geometric morphometrics. The pectoral girdles of selected species were analyzed with regard to the effects of shape differences on muscle moment arms across the shoulder joint and stress dissipation within the coracoid. Phylogenetic relationships, size, and locomotor behavior have an effect on the shape of the pectoral girdle in anurans, but there are differences in the relative impact of these factors between the bones of this skeletal unit. Remarkable shape diversity has been observed within locomotor groups indicating many-to-one mapping of form onto function. Significant shape differences have mainly been related to the overall pectoral girdle geometry and the shape of the coracoid. Most prominent shape differences have been found between burrowing and nonburrowing species with headfirst and backward burrowing species significantly differing from one another and from the other locomotor groups. The pectoral girdle shapes of burrowing species have generally larger moment arms for (simulated) humerus retractor muscles across the shoulder joint, which might be an adaptation to the burrowing behavior. The mechanisms of how the moment arms were enlarged differed between species and were associated with differences in the reaction of the coracoid to simulated loading by physiologically relevant forces.

The insect unguitractor plate in action: Force transmission and the micro CT visualizations of inner structures.

The unguitractor plate (UT) within insect tarsus was previously assumed to hold claws in a bent position with reduced muscular efforts due to the specific interlocking mechanism. In this study, the functional morphology of the unguitractor plate in the beetle Pachnoda marginata (Coleoptera, Scarabaeidae) was examined using force measurements and the micro CT visualization of the UT position at different straining states of the retractor unguis muscle tendon. Pulling forces were applied in a controlled manner to the tendon and forces elicited by the claws to the stiff substrate were simultaneously recorded, in order to understand the force transmission mechanism between the tendon and claws through the UT. After claw bending and entanglement with the substrate, the claws were not released, until the tendon was relaxed to an average of 22% of the original applied force. The time delay in the returning of the claws to their original position was observed due to the frictional mechanism between the UT and corresponding microstructures of the pretarsus. This mechanism provides energy saving, when claws are engaged with the substrate. However, physical contact between the UT and the inner pretarsal wall was not observed in preparations of prestrained tendons in the micro CT, presumably due to the deformations caused by fixation and drying procedures.

Zooplankters' nightmare: The fast and efficient catching basket of larval phantom midges (Diptera: Chaoborus).

Filter feeding zooplankton are a crucial component of limnic food webs. Copepods and cladocerans are important prey organisms for first-level predators like the common and abundant larvae of phantom midges (Chaoborus sp.). The latter possess a complex catching basket built of head appendages specialized to capture small crustaceans. The predator-prey-relationship of Chaoborus (Diptera, Nematocera) and Daphnia (Crustacea, Cladocera) has been studied in particular detail owing to the daphniids' ability to react upon the threat of predation with inducible defenses. Daphnia pulex expresses so-called 'neckteeth' in the presence of Chaoborus larvae that are discussed as a defensive trait that interferes with the larval head appendages and their effectiveness has been shown in several studies. Nonetheless, mode of function of these neckteeth is not understood and the hypothesis that they interfere with the predator's head appendages still has to be confirmed. To clarify the role of neckteeth in Daphnia, an understanding of the Chaoborus capture apparatus is essential. Here, we present a detailed three-dimensional analysis of Chaoborus obscuripes' larval head morphology as well as a kinematic analysis of the attack motion, which revealed an impressive strike velocity (14 ms to prey contact). The movement of the larvae's head appendages is reconstructed in the three-dimensional space using a combination of high-speed videography, micro-computed tomography and computer animation. Furthermore, we provide predation trial data to distinguish between pre- and post-attack defensive effects in D. pulex. Our findings suggest a combination of pre- and post-attack defenses with an average effectiveness of 50% each. With this study, we quantitatively describe prey capture kinematics of C. obscuripes and take a further step to reveal the neckteeth' mode of function in D. pulex.

Caecilian jaw-closing mechanics: integrating two muscle systems.

Caecilians (Lissamphibia: Gymnophiona) are unique among vertebrates in having two sets of jaw-closing muscles, one on either side of the jaw joint. Using data from high-resolution X-ray radiation computed tomography scans, we modelled the effect of these two muscle groups (mm. levatores mandibulae and m. interhyoideus posterior) on bite force over a range of gape angles, employing a simplified lever arm mechanism that takes into account muscle cross-sectional area and fibre angle. Measurements of lever arm lengths, muscle fibre orientations and physiological cross-sectional area of cranial muscles were available from three caecilian species: Ichthyophis cf. kohtaoensis; Siphonops annulatus; and Typhlonectes natans. The maximal gape of caecilians is restricted by a critical gape angle above which the mm. levatores mandibulae will open the jaw and destabilize the mandibular joint. The presence of destabilizing forces in the caecilian jaw mechanism may be compensated for by a mandibular joint in that the fossa is wrapped around the condyle to resist dislocation. The caecilian skull is streptostylic; the quadrate-squamosal complex moves with respect to the rest of the skull. This increases the leverage of the jaw-closing muscles. We also demonstrate that the unusual jaw joint requires streptostyly because there is a dorsolateral movement of the quadrate-squamosal complex when the jaw closes. The combination of the two jaw-closing systems results in high bite forces over a wide range of gape angles, an important advantage for generalist feeders such as caecilians. The relative sizes and leverage mechanics of the two closing systems allow one to exert more force when the other has a poor mechanical advantage. This effect is seen in all three species we examined. In the aquatic T. natans, with its less well-roofed skull, there is a larger contribution of the mm. levatores mandibulae to total bite force than in the terrestrial I. cf. kohtaoensis and S. annulatus.

Competing with barnacle cement: wetting resistance of a re-entrant surface reduces underwater adhesion of barnacles.

Surfaces with re-entrant topographies can repel liquids even of extremely low surface tension, almost independently of the material's inherent wettability. We show that this topography-based wetting resistance can also be applied to underwater applications, reducing the permanent adhesion of marine hardfouling organisms. Having combined a biofouling assay in the marine environment with microscopic analyses, we demonstrate how a synergistic effect of a soft silicone-based material with a re-entrant mushroom-shaped surface topography strongly increases the fouling release ability of such coatings compared with a smooth control made from the same material. Our coating inhibited the complete wetting of the solidifying glues produced by marine organisms, which resulted in a decreased contact area and, consequently, low adhesion. Our work suggests that topography-induced wetting resistance of surfaces may become a viable approach in preventing permanent adhesion of marine hardfouling organisms.

Surface chemistry of the frog sticky-tongue mechanism.

Frogs capture their prey with a highly specialized tongue. Recent studies indicate this tongue is covered with fibril-forming mucus that acts as a pressure sensitive adhesive. However, no analysis of the interfacial chemistry of frog tongue mucus has been performed. The goal of this study is to examine the chemical structure of the surface of mucus after a tongue strike. Previous studies of mucus from other animals suggest that mucus from a frog's tongue consists of mucins-serine-, threonine-, and proline-rich glycoproteins. Therefore, the authors expect to observe chemical bonds associated with glycoproteins, as well as fibrils formed at the mucus-tongue interface. To test this hypothesis, they collected both near-edge x-ray absorption fine structure (NEXAFS) microscopy images and sum frequency generation (SFG) vibrational spectra from layers of mucus left after frog tongue strikes on cleaned glass slides. NEXAFS imaging demonstrates a uniform distribution of amide, hydroxyl, and carbon-carbon bonds across the mucus surface. Difference spectra of individual N1s and C1s K-edge spectra pulled from these images indicate a structure consistent with fibril formation as well as disorder of oligosaccharide groups near the mucus surface. C-H region SFG spectra reveal surface active modes which likely stem from serine and threonine within the mucin protein. Combined, this work suggests that glycoproteins are well-ordered at the mucus-tongue interface.

Cranial musculature in the larva of the caecilian, Ichthyophis kohtaoensis (Lissamphibia: Gymnophiona).

Within the Gymnophiona (caecilians) oviparous species with biphasic life-cycles possess a free living semi-aquatic larval stage that feeds in aquatic habitats. The larvae pass through a metamorphosis to a purely terrestrial adult stage. It is likely that the cranial morphology of caecilian larvae has specializations for aquatic feeding. However, little is known about the cranial morphology, and the cranial musculature is especially neglected in the literature. This study provides a detailed description of the jaw and hyobranchial musculature in larval stages of a caecilian. We studied late embryonic and early larval specimens of Ichthyophis kohtaoensis. Furthermore, we compared and homologized the cranial muscles found in larval I. kohtaoensis with the muscles described for adult caecilians. Most cranial muscles of larval I. kohtaoensis are also present in the adult, except for the m. levator mandibulae externus and the m. subarcualis obliquus II. Our results were compared with the data available for larval frogs and salamanders in order to hypothesize the cranial musculature in the larva of the most recent common ancestor of the Lissamphibia. Larval caecilians, frog tadpoles, and salamander larvae share many characters in their cranial musculature, which, consequently, can be assigned to the lissamphibian ground pattern. However, the m. pterygoideus and the m. levator quadrati are unique to the Gymnophiona.

Frog tongue surface microstructures: functional and evolutionary patterns.

Frogs (Lissamphibia: Anura) use adhesive tongues to capture fast moving, elusive prey. For this, the tongues are moved quickly and adhere instantaneously to various prey surfaces. Recently, the functional morphology of frog tongues was discussed in context of their adhesive performance. It was suggested that the interaction between the tongue surface and the mucus coating is important for generating strong pull-off forces. However, despite the general notions about its importance for a successful contact with the prey, little is known about the surface structure of frog tongues. Previous studies focused almost exclusively on species within the Ranidae and Bufonidae, neglecting the wide diversity of frogs. Here we examined the tongue surface in nine different frog species, comprising eight different taxa, i.e., the Alytidae, Bombinatoridae, Megophryidae, Hylidae, Ceratophryidae, Ranidae, Bufonidae, and Dendrobatidae. In all species examined herein, we found fungiform and filiform papillae on the tongue surface. Further, we observed a high degree of variation among tongues in different frogs. These differences can be seen in the size and shape of the papillae, in the fine-structures on the papillae, as well as in the three-dimensional organization of subsurface tissues. Notably, the fine-structures on the filiform papillae in frogs comprise hair-like protrusions (Megophryidae and Ranidae), microridges (Bufonidae and Dendrobatidae), or can be irregularly shaped or absent as observed in the remaining taxa examined herein. Some of this variation might be related to different degrees of adhesive performance and may point to differences in the spectra of prey items between frog taxa.