Comparative morphology of oral glands in snakes of the family Homalopsidae reveals substantial variation and additional independent origins of salt glands within Serpentes.

Using diffusible iodine-based contrast-enhanced computed tomography (diceCT), we examined the morphology of the oral glands of 12 species of the family Homalopsidae. Snakes of this family exhibit substantial interspecific morphological variation in their oral glands. Particular variables are the venom glands, ranging from large (e.g., Subsessor bocourti) to small (e.g., Erpeton tentaculatum). The supra- and infralabial glands are more uniform in morphology, being the second most developed in almost all the sampled species. Premaxillary glands distinct from the supralabial glands were observed in five species (Myron richardsonii, Bitia hydroides, Cantoria violacea, Fordonia leucobalia, and Gerarda prevostiana), in addition to Cerberus rynchops, the only species in which this condition was previously documented associated with the excretion of salt. In the three species of the saltwater group of homalopsids (C. violacea, F. leucobalia, and G. prevostiana), the premaxillary glands also extend posteriorly, occupying a large area above the supralabial gland, a condition not observed in any other species of snake studied thus far. Character evolution analyses indicate that premaxillary glands differentiated from the supralabial gland and evolved independently three or four times in the family, always in lineages that invaded marine habitats. Our results suggest that the differentiated premaxillary glands are likely salt glands, as is the case in C. rynchops. If corroborated, this increases to six or seven the number of independent evolutionary origins of salt glands in snakes that have undergone an evolutionary transition to marine life.

Show Me Your Teeth And I Will Tell You What You Eat: Differences in Tooth Enamel in Snakes with Different Diets.

Teeth are composed of the hardest tissues in the vertebrate body and have been studied extensively to infer diet in vertebrates. The morphology and structure of enamel is thought to reflect feeding ecology. Snakes have a diversified diet, some species feed on armored lizards, others on soft invertebrates. Yet, little is known about how tooth enamel, and specifically its thickness, is impacted by diet. In this study, we first describe the different patterns of enamel distribution and thickness in snakes. Then, we investigate the link between prey hardness and enamel thickness and morphology by comparing the dentary teeth of 63 species of snakes. We observed that the enamel is deposited asymmetrically at the antero-labial side of the tooth. Both enamel coverage and thickness vary a lot in snakes, from species with thin enamel, only at the tip of the tooth to a full facet covered with enamel. There variations are related with prey hardness: snakes feeding on hard prey have a thicker enamel and a lager enamel coverage while species. Snakes feeding on softer prey have a thin enamel layer confined to the tip of the tooth.

Armed to the teeth: The underestimated diversity in tooth shape in snakes and its relation to feeding behavior and diet.

The structure, composition, and shape of teeth have been related to dietary specialization in many vertebrate species, but comparative studies on snakes' teeth are lacking. Yet, snakes have diverse dietary habits that may impact the shape of their teeth. We hypothesize that prey properties, such as hardness and shape, as well as feeding behavior, such as aquatic or arboreal predation, or holding vigorous prey, impose constraints on the evolution of tooth shape in snakes. We compared the morphology of the dentary teeth of 63 species that cover the phylogenetic and dietary diversity of snakes, using 3D geometric morphometrics and linear measurements. Our results show that prey hardness, foraging substrate, and the main feeding mechanical challenge are important drivers of tooth shape, size, and curvature. Overall, long, slender, curved teeth with a thin layer of hard tissue are observed in species that need to maintain a grip on their prey. Short, stout, less curved teeth are associated with species that undergo high or repeated loads. Our study demonstrates the diversity of tooth morphology in snakes and the need to investigate its underlying functional implications to better understand the evolution of teeth in vertebrates.

Inside the head of snakes: influence of size, phylogeny, and sensory ecology on endocranium morphology.

Environmental properties, and the behavioral habits of species impact sensory cues available for foraging, predator avoidance and inter/intraspecific communication. Consequently, relationships have been discovered between the sensory ecology and brain morphology in many groups of vertebrates. However, these types of studies have remained scare on snake. Here, we investigate the link between endocranial shape and the sensory-related ecology of snakes by comparing 36 species of snakes for which we gathered six sensory-ecology characteristics. We use µCT scanning and 3D geometric morphometrics to compare their endocranium in a phylogenetically informed context. Our results demonstrate that size is a major driver of endocranial shape, with smaller species tending to maximize endocranial volume using a more bulbous shape, while larger species share an elongate endocranial morphology. Phylogeny plays a secondary role with more derived snakes diverging the most in endocranial shape, compared to other species. The activity period influences the shape of the olfactory and optic tract, while the foraging habitat impacts the shape of the cerebellum and cranial nerve regions: structures involved in orientation, equilibrium, and sensory information. However, we found that endocranial morphology alone is not sufficient to predict the activity period of a species without prior knowledge of its phylogenetic relationship. Our results thus demonstrate the value of utilizing endocranial shape as complementary information to size and volume in neurobiological studies.

Local Superimpositions Facilitate Morphometric Analysis of Complex Articulating Structures.

Articulating structures, such as the vertebrate skeleton or the segmented arthropod exoskeleton, comprise a majority of the morphological diversity across the eukaryotic tree of life. Quantifying the form of articulating structures is therefore imperative for a fuller understanding of the factors influencing biological form. A wealth of freely available 3D data capturing this morphological diversity is stored in online repositories such as Morphosource, but the geometric morphometric analysis of an articulating structure is impeded by arbitrary differences in the resting positions of its individual articulating elements. In complex articulating structures, where the angles between articulating elements cannot be standardized, landmarks on articulating elements must be Procrustes superimposed independently (locally) and then recombined to quantify variation in the entire articulating structure simultaneously. Here, we discuss recent advances in local superimposition techniques, namely the "matched local superimpositions" approach, which incorporates anatomically accurate relative sizes, positions, and orientations of locally-superimposed landmarks, enabling clearer biological interpretation. We also use simulations to evaluate the consequences of choice of superimposition approach. Our results show that local superimpositions will isolate shape variation within locally-superimposed landmark subsets by sacrificing size and positional variation. They may also create morphometric "modules" when there are none by increasing integration within the locally-superimposed subsets; however, this effect is no greater than the spurious between-module integration created when superimposing landmark subsets (i.e., articulating elements) together. Taken together, our results show that local superimposition techniques differ from conventional Procrustes superimpositions in predictable ways. Finally, we use empirical datasets of the skulls of wrasses and colubriform snakes to highlight the promise of local superimpositions and their utility. Complex articulating structures must be studied, and the only current solution to do so is local superimpositions.

Morphological integration and modularity in the hyperkinetic feeding system of aquatic-foraging snakes.

The kinetic skull is a key innovation that allowed snakes to capture, manipulate, and swallow prey exclusively using their heads using the coordinated movement of eight bones. Despite these unique feeding behaviors, patterns of evolutionary integration and modularity within the feeding bones of snakes in a phylogenetic framework have yet to be addressed. Here, we use a dataset of 60 µCT-scanned skulls and high-density geometric morphometric methods to address the origin and patterns of variation and integration in the feeding bones of aquatic-foraging snakes. By comparing alternate superimposition protocols allowing us to analyze the entire kinetic feeding system simultaneously, we find that the feeding bones are highly integrated, driven predominantly by functional selective pressures. The most supported pattern of modularity contains four modules, each associated with distinct functional roles: the mandible, the palatopterygoid arch, the maxilla, and the suspensorium. Further, the morphological disparity of each bone is not linked to its magnitude of integration, indicating that integration within the feeding system does not strongly constrain morphological evolution, and that adequate biomechanical solutions to a wide range of feeding ecologies and behaviors are readily evolvable within the constraint due to integration in the snake feeding system.

Exploring the functional meaning of head shape disparity in aquatic snakes.

Phenotypic diversity, or disparity, can be explained by simple genetic drift or, if functional constraints are strong, by selection for ecologically relevant phenotypes. We here studied phenotypic disparity in head shape in aquatic snakes. We investigated whether conflicting selective pressures related to different functions have driven shape diversity and explore whether similar phenotypes may give rise to the same functional output (i.e., many-to-one mapping of form to function). We focused on the head shape of aquatically foraging snakes as they fulfill several fitness-relevant functions and show a large amount of morphological variability. We used 3D surface scanning and 3D geometric morphometrics to compare the head shape of 62 species in a phylogenetic context. We first tested whether diet specialization and size are drivers of head shape diversification. Next, we tested for many-to-one mapping by comparing the hydrodynamic efficiency of head shape characteristic of the main axes of variation in the dataset. We 3D printed these shapes and measured the forces at play during a frontal strike. Our results show that diet and size explain only a small amount of shape variation. Shapes did not fully functionally converge as more specialized aquatic species evolved a more efficient head shape than others. The shape disparity observed could thus reflect a process of niche specialization.

Hydrodynamics of frontal striking in aquatic snakes: drag, added mass, and the possible consequences for prey capture success.

Transient locomotion under water is highly constrained by drag and added mass, yet some aquatic snakes catch their prey using a fast forward acceleration, with the mouth opened. These aquatic snakes show a convergence of their head shape in comparison with closely related species that do not forage under water. As both drag and added mass are related to some extent to the shape of the moving object, we explored how shape impacts the hydrodynamic forces applied to the head of a snake during a prey capture event. We compared two 3D-printed heads representing typical shapes of aquatically-foraging and non-aquatically-foraging snakes, and frontal strike kinematics based on in vivo observations. By using direct force measurements, we calculated the drag and added mass coefficient of the two models. Our results show that both drag and added mass are reduced in aquatic snakes. The drag coefficient of the aquatic model is 0.24, which is almost two times smaller than the non-aquatic model. The added mass coefficient of the aquatic model is 0.15 versus 0.24 for the non-aquatic model, showing that the convergence of head shape in aquatically foraging snakes is associated with a hydrodynamic advantage during frontal striking. The vorticity field measurements with particle image velocimetry show that a less intense recirculation bubble behind the jaw of the aquatic model, compared to the non-aquatic model, might be the basis of this advantage.

Does aquatic foraging impact head shape evolution in snakes?

Evolutionary trajectories are often biased by developmental and historical factors. However, environmental factors can also impose constraints on the evolutionary trajectories of organisms leading to convergence of morphology in similar ecological contexts. The physical properties of water impose strong constraints on aquatic feeding animals by generating pressure waves that can alert prey and potentially push them away from the mouth. These hydrodynamic constraints have resulted in the independent evolution of suction feeding in most groups of secondarily aquatic tetrapods. Despite the fact that snakes cannot use suction, they have invaded the aquatic milieu many times independently. Here, we test whether the aquatic environment has constrained head shape evolution in snakes and whether shape converges on that predicted by biomechanical models. To do so, we used three-dimensional geometric morphometrics and comparative, phylogenetically informed analyses on a large sample of aquatic snake species. Our results show that aquatic snakes partially conform to our predictions and have a narrower anterior part of the head and dorsally positioned eyes and nostrils. This morphology is observed, irrespective of the phylogenetic relationships among species, suggesting that the aquatic environment does indeed drive the evolution of head shape in snakes, thus biasing the evolutionary trajectory of this group of animals.

Impact of temperature on performance in two species of South African dwarf chameleon, Bradypodion pumilum and B. occidentale.

Temperature is an extrinsic factor that influences reptile behavior because of its impact on reptile physiology. Understanding the impact of temperature on performance traits is important as it may affect the ecology and fitness of ectothermic animals such as reptiles. Here, we examined the temperature dependence of performance in two species of South African dwarf chameleon (Bradypodion): one adapted to a semi-arid environment and one to a mesic environment. Ecologically relevant performance traits were tested at different temperatures to evaluate their thermal dependence, and temperature-performance breadths for 80% and 90% of each performance trait were calculated. Our results show distinct differences in the thermal dependence of speed- versus force-related performance traits. Moreover, our results show that the semi-arid species is better adapted to higher temperatures and as such has a better chance of coping with the predicted increases in environmental temperature. The mesic area-adapted species seems to be more sensitive to an increase in temperature and could therefore potentially be threatened by the predicted future climate change. However, further studies investigating the potential for acclimation in chameleons are needed to better understand how animals may respond to future climate change.