Vertebral trabecular bone mechanical properties vary among functional groups of cetaceans

Since their appearance in the fossil record 34 Mya, modern cetaceans (dolphins, whales, and porpoises) have radiated into diverse habitats circumglobally, developing vast phenotypic variations among species. Traits such as skeletal morphology and ecologically-linked behaviors denote swimming activity; trade-offs in flexibility and rigidity along the vertebral column determine patterns of caudal oscillation. Here, we categorized 10 species of cetaceans (Families Delphinidae and Kogiidae; N = 21 animals) into functional groups based on vertebral centra morphology, swimming speeds, diving behavior, and inferred swimming patterns. We quantified trabecular bone mechanical properties (yield strength, apparent stiffness, and resilience) among functional groups and regions of the vertebral column (thoracic, lumbar, and caudal). We extracted 6 mm3 samples from vertebral bodies and tested them in compression in three orientations (rostrocaudal, dorsoventral, and mediolateral) at 2 mm min−1. Overall, bone from the pre-fluke/fluke boundary had the greatest yield strength and resilience, indicating that the greatest forces are translated to the tail during caudal oscillatory swimming. Group 1, composed of five shallow-diving delphinid species, had the greatest vertebral trabecular bone yield strength, apparent stiffness, and resilience of all functional groups. Conversely, Group 3, composed of two deep-diving kogiid species, had the least strong, stiff, and resilient bone, while Group 2 (three deep-diving delphinid species) exhibited intermediate values. These data suggest that species that incorporate prolonged glides during deep descents in the water column actively swim less, and place relatively smaller loads on their vertebral columns, compared with species that execute shallower dives. We found that cetacean vertebral trabecular bone properties differed from the properties of terrestrial mammals; for every given bone strength, cetacean bone was less stiff by comparison. This relative lack of material rigidity within vertebral bone may be attributed to the non-weight bearing locomotor modes of fully aquatic mammals.

Only the good die old? Ontogenetic determinants of locomotor performance in Eastern cottontail rabbits (Sylvilagus floridanus)

For many animals, the juvenile stage of life can be particularly perilous. Once independent, immature animals must often complete the same basic survival functions as adults despite smaller body size and other growth-related limits on performance. Because, by definition, juveniles have yet to reproduce, we should expect strong selection for mechanisms to offset these ontogenetic limitations, allowing individuals to reach reproductive adulthood and maintain Darwinian fitness. We use an integrated ontogenetic dataset on morphology, locomotor performance, and longevity in wild cottontail rabbits (Sylvilagus floridanus, Allen 1848) to test the hypothesis that prey animals are under selective pressure to maximize juvenile performance. We predicted that 1) juveniles would accelerate more quickly than adults, allowing them to reach adult-like escape speeds, and 2) juveniles with greater levels of performance should survive for longer durations in the wild, thus increasing their reproductive potential. Using high speed video and force platform measurements, we quantified burst acceleration, escape speed, and mechanical power production in 42 wild-caught S. floridanus (29 juveniles, 13 adults; all rabbits >1kg in body mass were designated to be adults, based on published growth curves and evidence of epiphyseal fusion). A subsample of 22 rabbits (16 juveniles, 6 adults) were fitted with radio-telemetry collars for documenting survivorship in the wild. We found that acceleration and escape speed peaked in the late juvenile period in S. floridanus, at an age range that coincides with a period of pronounced demographic attrition in wild populations. Differences in mass-specific mechanical power production explained ∼75% of the variation in acceleration across the dataset, indicating that juvenile rabbits outpace adults by producing more power per unit body mass. We found a positive, though non-significant, association between peak escape speed and survivorship duration in the wild, suggesting a complex relationship between locomotor performance and fitness in growing S. floridanus.

Flight and dietary antioxidants influence antioxidant expression and activity in a migratory bird

Ecologically-relevant factors such as exercise and diet quality can directly influence how physiological systems work including those involved in maintaining oxidative balance; however, to our knowledge, no studies to date have focused on how such factors directly affect expression of key components of the endogenous antioxidant system (i.e., transcription factors, select antioxidant genes, and corresponding antioxidant enzymes) in several metabolically active tissues of a migratory songbird. We conducted a 3-factor experiment that tested the following hypotheses: (H1) Daily flying over several weeks increases the expression of transcription factors NRF2 and PPARs as well as endogenous antioxidant genes (i.e., CAT, SOD1, SOD2, GPX1, GPX4), and upregulates endogenous antioxidant enzyme activities (i.e., CAT, SOD, GPx). (H2) Songbirds fed diets composed of more 18:2n-6 PUFA are more susceptible to oxidative damage and thus upregulate their endogenous antioxidant system compared to when fed diets with less PUFA. (H3) Songbirds fed dietary anthocyanins gain additional antioxidant protection and thus upregulate their endogenous antioxidant system less compared to songbirds not fed anthocyanins. Flight training increased the expression of 3 of the 6 antioxidant genes and transcription factors measured in the liver, consistent with H1, but for only one gene (SOD2) in the pectoralis. Dietary fat quality had no effect on antioxidant pathways (H2) whereas dietary anthocyanins increased the expression of select antioxidant enzymes in the pectoralis, but not in the liver (H3). These tissue-specific differences in response to flying and dietary antioxidants are likely explained by functional differences between tissues as well as fundamental differences in their turnover rates. The consumption of dietary antioxidants along with regular flying enables birds during migration to stimulate the expression of genes involved in antioxidant protection likely through increasing the transcriptional activity of NRF2 and PPARs, and thereby demonstrates for the first time that these relevant ecological factors affect the regulation of key antioxidant pathways in wild birds. What remains to be demonstrated is how the extent of these ecological factors (i.e., intensity or duration of flight, amounts of dietary antioxidants) influences the regulation of these antioxidant pathways and thus oxidative balance.

Dermal denticle diversity in sharks: novel patterns on the interbranchial skin

Shark skin is covered in dermal denticles – tooth-like structures consisting of enameloid, dentine, and a central pulp cavity. Previous studies have demonstrated differences in denticle morphology both among species and across different body regions within a species, including one report of extreme morphological variation within a 1 cm distance on the skin covering the branchial pouches, a region termed “interbranchial skin”. We used gel-based profilometry, histology, and scanning electron microscopy to quantify differences in denticle morphology and surface topography of interbranchial skin denticles among 13 species of sharks to better understand the surface structure of this region. We show that 1) interbranchial skin denticles differ across shark species, and 2) denticles on the leading edge of the skin covering each gill pouch have different morphology and surface topography compared to denticles on the trailing edge. Across all species studied, there were significant differences in denticle length (P = 0.01) and width (P = 0.002), with shorter and wider leading edge denticles compared to trailing edge denticles. Surface skew was also higher in leading edge denticles (P = 0.009), though most values were still negative, indicating more valleys than peaks. Overall, leading edge denticles were smoother-edged than trailing edge denticles in all of the species studied. These data suggest two hypotheses: 1) smoother-edged leading edge denticles protect the previous gill flap from abrasion during respiration, and 2) ridged denticle morphology at the trailing edge might alter water turbulence exiting branchial pouches after passing over the gills. Future studies will focus on determining the relationship between denticle morphology and water flow by visualizing fluid motion over interbranchial denticles during in vivo respiration.

Environmental Thermal Stress Induces Neuronal Cell Death and Developmental Malformations in Reptiles

Every stage of organismal life history is being challenged by global warming. Many species are already experiencing temperatures approaching their physiological limits; this is particularly true for ectothermic species, such as lizards. Embryos are markedly sensitive to thermal insult. Here, we demonstrate that temperatures currently experienced in natural nesting areas can modify gene expression levels and induce neural and craniofacial malformations in embryos of the lizard Anolis sagrei. Developmental abnormalities ranged from minor changes in facial structure to significant disruption of anterior face and forebrain. The first several days of post-oviposition development are particularly sensitive to this thermal insult. These results raise new concern over the viability of ectothermic species under contemporary climate change. Herein, we propose and test a novel developmental hypothesis that describes the cellular and developmental origins of those malformations: cell death in the developing forebrain and disruption of normal facial induction due to disrupted Hedgehog signaling. Based on similarities in the embryonic response to thermal stress among distantly related species, we propose that this developmental hypothesis represents a common embryonic response to thermal insult among amniote embryos. Our results emphasize the importance of adopting a broad, multi-disciplinary approach that includes both lab and field perspectives when trying to understand the future impacts of anthropogenic change on animal development.

Social Group Size and Shelter Availability Influence Individual Metabolic Traits in a Social Fish

Group living is widespread among animal species and yields both costs and benefits. Presence of conspecifics can restrict or enhance the expression of individual behaviour, and the recent social environment is thought to affect behavioural responses in later contexts, even when individuals are alone. However, little is known about how social group size influences the expression of individual physiological traits, including metabolic rates. There is some evidence that shoaling can reduce fish metabolic rates but this variable may be affected by habitat conditions such as shelter availability via density-dependent processes. We investigated how social group size and shelter availability influence Eurasian minnow Phoxinus phoxinus metabolic rates estimated by respirometry. Respirometry trials were conducted on fish in isolation before and after they were housed for three weeks in a social treatment consisting in a specific group size (n = 4 or 8) and shelter availability (presence or absence of plant shelter in the experimental tank). Plant shelter was placed over respirometers for half of the duration of the respirometry trials, allowing estimation of minimum day-time and night-time metabolic rates in both conditions (in the presence or absence of plant shelter). Standard metabolic rate (SMR), maximum metabolic rate (MMR), and aerobic scope (AS) were also estimated over the entire trial. Minimum day-time and night-time metabolic rates estimated while in presence of plant shelter were lower than when estimated in absence of plant shelter, both before and after individuals were housed in their social treatment. After the social treatment, SMR were higher for fish that were held in groups of four as compared to that of fish held in groups of eight while MMR showed no difference. Plant shelter availability during the social treatments did not influence SMR or MMR. Our results suggest that social group size may directly influence energy demands of individuals, highlighting the importance of understanding the role of group size on variations in physiological traits associated with energy expenditure.

Turtling the salamander: tail movements mitigate need for kinematic limb changes during walking in tiger salamanders (Ambystoma tigrinum) with restricted lateral movement

Lateral undulation and trunk flexibility offer performance benefits to maneuverability, stability, and stride length (via speed and distance traveled). These benefits make them key characteristics of the locomotion of tetrapods with sprawling posture, with the exception of turtles. Despite their bony carapace preventing lateral undulations, turtles are able to improve their locomotor performance by increasing stride length via greater limb protraction. The goal of this study was to quantify the effect of reduced lateral flexibility in a generalized sprawling tetrapod, the tiger salamander (Ambystoma tigrinum). We had two potential predictions: (1) either salamanders completely compensate by changing their limb kinematics, or (2) their performance (i.e., speed) will suffer due to the reduced lateral flexibility. This reduction was performed by artificially limiting trunk flexibility by attaching a 2-piece shell around the body between the pectoral and pelvic girdles. Adult tiger salamanders (n = 3, SVL = 9 cm-14.5 cm) walked on a 1 m trackway under three different conditions: unrestricted, flexible shell (Tygon tubing), and rigid shell (PVC tubing). Trials were filmed in a single, dorsal view, and kinematics of entire midline and specific body regions (head, trunk, tail), as well as the fore and hindlimbs, were calculated. Tygon individuals had significantly higher curvature than both PVC and unrestricted individuals for the body, but this trend was primarily driven by changes in tail movements. PVC individuals had significantly lower curvature in the trunk region compared to unrestricted individuals or Tygon; however, there was no difference between unrestricted and Tygon individuals suggesting the shells performed as expected. PVC and Tygon individuals had significantly higher curvature in the tails compared to unrestricted individuals. There were no significant differences for any limb kinematic variables among treatments including average, minimum, maximum angles. Thus, salamanders respond to decreased lateral movement in their trunk by increasing movements in their tail, without changes in limb kinematics. These results suggest that tail undulations may be a more critical component to sprawling-postured tetrapod locomotion than previously recognized.

Sex differences in how territory quality affects aggression in convict cichlids

In animal contests, the value an individual assigns to limited resources can directly impact the level of aggression they demonstrate. For territorial species, individuals often assess their territory quality and appropriately modify the time and energy invested in its defense. In this study, male and female convict cichlids, Amatitlania nigrofasciata, were acclimated to one of three territorial treatments representing either a Low, Medium, or High resource value. Territories with a “Low Value” included substrate alone, “Medium Value” territories included substrate and a nest site, and a “High Value” territory included substrate, a nest site, and constant food source. After three days of acclimation, a size-matched intruder was introduced to elicit territorial aggression and behaviors were observed. Territory quality affected one measure of low-intensity aggression (displays) in residents but had no effect on high-intensity aggression (bites and chases). Moreover, there was a significant effect of sex, with males and females differing in the types of aggressive behaviors demonstrated across all treatments. Females showed more low-intensity aggressive behaviors toward intruders than males did. Additionally, a significant interaction of sex and territory quality was observed on two measures of high-intensity aggressive behavior (bites and chases), with females more likely than males to increase aggressive behaviors along with increasing territory quality. This suggests that females may be more sensitive and/or responsive to changes in the quality of a territory, possibly due to the necessity of a suitable nest site for egg deposition within a territory.

Not so fast: strike kinematics of the araneoid trap-jaw spider Pararchaea alba (Malkaridae: Pararchaeinae)

To capture prey otherwise unattainable by muscle function alone, some animal lineages have evolved movements that are driven by stored elastic energy, producing movements of remarkable speed and force. One such example that has evolved multiple times is a trap-jaw mechanism, in which the mouthparts of an animal are loaded with energy as they open to a wide gape and then, when triggered to close, produce a terrific force. Within the spiders (Araneae), this type of attack has thus far solely been documented in the palpimanoid family Mecysmaucheniidae but a similar morphology has also been observed in the distantly related araneoid subfamily Pararchaeinae, leading to speculation of a trap-jaw attack in that lineage as well. Here, using high-speed videography, we test whether cheliceral strike power output suggests elastic-driven movements in the pararchaeine Pararchaea alba. The strike speed attained place P. alba as a moderately fast strikers exceeding the slowest mecysmaucheniids, but failing to the reach the most extreme high-speed strikers that have elastic-driven mechanisms. Using micro-Computed-Tomography, we compare the morphology of P. alba chelicerae in the resting and open positions, and their related musculature, and based on results propose a mechanism for cheliceral strike function that includes a torque reversal latching mechanism. Similar to the distantly related trap-jaw mecysmaucheniid spiders, the unusual prosoma morphology in P. alba seemingly allows for highly maneuverable chelicerae with a much wider gape than typical spiders, suggestive that increasingly maneuverable joints coupled with a latching mechanism may serve as a precursor to elastic-driven movements.

AutoBend: An automated approach for estimating intervertebral joint function from bone-only digital models

Deciphering the biological function of rare or extinct species is key to understanding evolutionary patterns across the tree of life. While soft tissues are vital determinants of joint function, they are rarely available for study. Therefore, extracting functional signals from skeletons, which are more widely available via museum collections, has become a priority for the field of comparative biomechanics. While most work has focused on the limb skeleton, the axial skeleton plays a critical role in body support, respiration, and locomotion, and is therefore of central importance for understanding broad-scale functional evolution. Here, we describe and experimentally validate AutoBend, an automated approach to estimating intervertebral joint function from bony vertebral columns. AutoBend calculates osteological range of motion (oROM) by automatically manipulating digitally articulated vertebrae while incorporating multiple constraints on motion, including both bony intersection and the role of soft tissues by restricting excessive strain in both centrum and zygapophyseal articulations. Using AutoBend and biomechanical data from cadaveric experiments on cats and tegus, we validate important modeling parameters required for oROM estimation, including the degree of zygapophyseal disarticulation, and the location of the center of rotation. Based on our validation, we apply a model with the center of rotation located within the vertebral disc, no joint translation, around 50% strain permitted in both zygapophyses and discs, and a small amount of vertebral intersection permitted. Our approach successfully reconstructs magnitudes and craniocaudal patterns of motion obtained from ex vivo experiments, supporting its potential utility. It also performs better than more typical methods that rely solely on bony intersection, emphasizing the importance of accounting for soft tissues. We estimated the sensitivity of the analyses to vertebral model reconstruction by varying joint spacing, degree of overlap, and the impact of landmark placement. The effect of these factors was small relative to biological variation craniocaudally and between bending directions. Within, we also present a new approach for estimating joint stiffness directly from oROM and morphometric measurements that can successfully reconstruct the craniocaudal patterns, but not magnitudes, derived from experimental data. Together, this work represents a significant step forward for understanding vertebral function in difficult-to-study (e.g., rare or extinct) species, paving the way for a broader understanding of patterns of functional evolution in the axial skeleton.