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Understanding how the global climate impacts the physiology of wildlife animals is of importance. Amphibians are particularly sensitive to climate change, and it is hypothesized that rising temperatures impair their neurodevelopment. Temperature influences the composition of the gut microbiota, which is critical to host neurodevelopment through the microbiota-gut-brain (MGB) axis. Most research investigating the link between the gut microbiota and neurodevelopment occurs in germ-free mammalian model systems, leaving the nature of the MGB axis in non-mammalian wildlife unclear. Here, we tested the hypothesis that the temperature and the microbial environment in which tadpoles were raised shapes neurodevelopment, possibly through the MGB axis. Newly hatched Green Frog tadpoles (Lithobates clamitans) were raised in natural pond water or autoclaved pond water, serving as an experimental manipulation of the microbiota by reducing colonizing microbes, at three different water temperatures: 14°C, 22°C, and 28°C. Neurodevelopment was analyzed through measures of relative brain mass and morphology of brain structures of interest. We found that tadpole development in warmer temperatures increased relative brain mass and optic tectum width and length. Further, tadpole development in autoclaved pond water increased relative optic tectum width and length. Additionally, the interaction of treatments altered relative diencephalon length. Lastly, we found that variation in brain morphology was associated with gut microbial diversity and the relative abundance of individual bacterial taxa. Our results indicate that both environmental temperature and microbial communities influence relative brain mass and shape. Furthermore, we provide some of the first evidence for the MGB axis in amphibians.

Like many other animals, insects are capable of returning to previously visited locations using path integration, a memory of travelled direction and distance. Recent studies suggest that also Drosophila can use path integration to return to a food reward. However, the existing experimental evidence for path integration in Drosophila has a potential confound: pheromones deposited at the site of reward might enable flies to find previously rewarding locations even without memory. Here we show that pheromones indeed can cause naïve flies to accumulate where previous flies had been rewarded in a navigation task. Therefore, we designed an experiment to determine if flies can use path integration memory despite potential pheromonal cues by displacing the flies shortly after an optogenetic reward. We found that rewarded flies returned to the location predicted by a memory-based model. Several analyses are consistent with path integration as the mechanism by which flies returned to the reward. We conclude that while pheromones may often be important in fly navigation and must be carefully controlled in future experiments, Drosophila may indeed be capable of performing path integration.

Axon regeneration helps maintain lifelong function of neurons in many animals. Depending on the site of injury, new axons can grow either from the axon stump (after distal injury) or from the tip of a dendrite (after proximal injury). However, some neuron types do not have dendrites to be converted to a regenerating axon after proximal injury. For example, many sensory neurons receive information from a specialized sensory cilium rather than a branched dendrite arbor. We hypothesized that the lack of traditional dendrites would limit the ability of ciliated sensory neurons to respond to proximal axon injury. We tested this hypothesis by performing laser microsurgery on ciliated lch1 neurons in Drosophila larvae and tracking cells over time. These cells survived proximal axon injury as well as distal axon injury, and, like many other neurons, initiated growth from the axon stump after distal injury. After proximal injury, neurites regrew in a surprisingly flexible manner. Most cells initiated outgrowth directly from the cell body, but neurite growth could also emerge from the short axon stump or base of the cilium. New neurites were often branched. Although outgrowth after proximal axotomy was variable, it depended on the core DLK axon injury signaling pathway. Moreover, each cell had at least one new neurite specified as an axon based on microtubule polarity and accumulation of the endoplasmic reticulum. We conclude that ciliated sensory neurons are not intrinsically limited in their ability to grow a new axon after proximal axon removal.

Evolutionary change in diapause timing can be an adaptive response to changing seasonality, and even result in ecological speciation. However, the molecular and cellular mechanisms regulating shifts in diapause timing remain poorly understood. One of the hallmarks of diapause is a massive slowdown in the cell cycle of target organs such as brains and primordial imaginal structures, and resumption of cell cycle proliferation is an indication of diapause termination and resumption of development. Characterizing cell cycle parameters between lineages differing in diapause life history timing may help identify molecular mechanisms associated with alterations of diapause timing. We tested the extent to which progression of the cell cycle differs across diapause between two genetically distinct European corn borer strains that differ in their seasonal diapause timing. We show the cell cycle slows down during larval diapause with a significant decrease in the proportion of cells in the S phase. Brain-suboesophageal complex cells slow primarily in the G0/G1 phase whereas most wing disc cells are in the G2 phase. Diapausing larvae of the earlier-emerging, bivoltine E-strain (BE) suppressed cell cycle progression less than the later-emerging, univoltine Z-strain (UZ) individuals, with greater proportions of cells in S phase across both tissues during diapause. Additionally, resumption of cell cycle proliferation occurred earlier in the BE strain compared to the UZ strain after exposure to diapause-terminating conditions. We propose regulation of cell cycle progression rates ultimately drives differences in larval diapause termination, and adult emergence timing, between early and late emerging European corn borer strains.

Turbidity challenges the visual performance of aquatic animals. During development, environments with limited visibility may affect the fine-tuning of visual systems and thus the perception of, and response to, risk. While turbidity has frequently been used to characterise permanent aquatic habitats, it has been an overlooked feature of ephemeral ones.Here, we use the natural diversity of ephemeral rearing sites (phytotelmata) in which the tadpoles of two poison frog species are deposited and confined until metamorphosis to explore the relationship between environments with limited visibility and response to perceived risk.We sampled wild tadpoles ofDendrobates tinctorius, a rearing-site generalist with facultatively cannibalistic tadpoles, andOophaga(formerlyDendrobates)pumilio, a small-phytotelm specialist dependent on maternal food-provisioning, to investigate how the visual environment in rearing sites influences tadpole behaviour. We hypothesised that turbid rearing conditions negatively impact both species’ ability to perceive risk, decreasing response strength to predatory visual stimuli. Using experimental arenas, we measured tadpole activity and space first on a black and white background, and then on either black or white backgrounds where tadpoles were exposed to visual stimuli of (potentially cannibalistic) conspecifics or potential predators.When placed in a novel arena, the effects of rearing environment onD. tinctoriustadpoles were clear: tadpoles from darker pools were less active than tadpoles from brighter pools, and did not respond to either visual stimuli, whereas tadpoles from brighter pools swam more when paired with conspecifics versus odonate larvae, suggesting that tadpoles can visually discriminate between predators. ForO. pumilio, tadpoles were more active on experimental backgrounds that more closely matched the luminosity of their rearing sites, but their responses to the two visual stimuli did not differ.Larval specialisation associated with species-specific microhabitat use may underlie the observed responses to visual stimuli, which has implications for the stability of species interactions and trophic dynamics in pool communities. Together, our findings demonstrate that light availability of wild larval rearing conditions influences the perception of risk in novel contexts, and provide insight into how visually guided animals may respond to sudden environmental disturbances.

Rapid movements of limbs and appendages, faster than those produced by simple muscle contraction alone, are generated through mechanical networks consisting of springs and latches. The latch plays a central role in these spring-loaded mechanisms, but the structural details of the latch are not always known. The mandibles of the trap-jaw ant Odontomachus kuroiwae closes the mandible extremely quickly to capture prey or to perform mandible-powered defensive jumps to avoid potential threats. The jump is mediated by a mechanical spring and latch system embodied in the mandible. An ant can strike the tip of the mandible onto the surface of an obstacle (prey, predator or ground) in order to bounce its body away from potential threats. The angular velocity of the closing mandible was 2.3×104 rad s−1 (1.3×106 deg s−1). Latching of the joint is a key mechanism to aid the storage of energy required to power the ballistic movements of the mandibles. We have identified the fine structure of two latch systems on the mandible forming a ‘ball joint’ using an X-ray micro-computational tomography system (X-ray micro-CT) and X-ray live imaging with a synchrotron. Here, we describe the surface of the inner section of the socket and a projection on the lip of the ball. The X-ray live imaging and movements of the 3D model show that the ball with a detent ridge slipped into a socket and over the socket ridge before snapping back at the groove edge. Our results give insight into the complex spring-latch systems that underpin ultra-fast movements in biological systems.

Climate change is leading to rapid change in aquatic environments, increasing the mean and variability of temperatures, and increasing the incidence of hypoxia. We investigated how acclimation to constant temperatures or to diel temperature fluctuations affects hypoxia tolerance in mummichog killifish (Fundulus heteroclitus). Killifish were acclimated to constant cool (15°C), constant warm (25°C) or a diel temperature cycle (15°C at night, 25°C during day) for 6 weeks. We then measured hypoxia tolerance (time to loss of equilibrium in severe hypoxia, tLOE; critical O2 tension, Pcrit), whole-animal metabolism, gill morphology, haematology and tissue metabolites at 15°C and 25°C in a full factorial design. Among constant temperature groups, tLOE was highest and Pcrit was lowest in fish tested at their acclimation temperature. Warm-acclimated fish had lower metabolic rate at 25°C and greater gill surface area (less coverage of lamellae by interlamellar cell mass, ILCM), but cool-acclimated fish had greater brain glycogen stores. Therefore, effects of constant temperature acclimation on hypoxia tolerance were temperature specific and not exhibited broadly across test temperatures, and they were associated with different underlying mechanisms. Hypoxia tolerance was less sensitive to test temperature in fish acclimated to fluctuating temperatures compared with fish acclimated to constant temperature. Acclimation to fluctuating temperatures also increased haemoglobin–O2 affinity of the blood (decreased P50) compared with constant temperature groups. Therefore, acclimation to fluctuating temperatures helps maintain hypoxia tolerance across a broader range of temperatures, and leads to some distinct physiological adjustments that are not exhibited by fish acclimated to constant temperatures.

Across vertebrate diversity, limb bone morphology is typically expected to reflect differences in the habitats and functional tasks that species utilize. Arboreal vertebrates are often recognized to have longer limbs than terrestrial relatives, a feature thought to help extend the reach of limbs across gaps between branches. Among terrestrial vertebrates, longer limbs can experience greater bending moments that might expose bones to a greater risk of failure. However, changes in habitat or behavior can impose changes in the forces that bones experience. If locomotion imposed lower loads in trees than on the ground, such a release from loading demands might have produced conditions under which potential constraints on the evolution of long limbs were removed, making it easier for them to evolve in arboreal species. We tested for such environmental differences in limb bone loading using the green iguana (Iguana iguana), a species that readily walks over ground and climbs trees. We implanted strain gauges on the humerus and femur, and then compared loads between treatments modeling substrate conditions of arboreal habitats. For hindlimbs, inclined substrate angles were most correlated with strain increases, whereas the forelimbs had a similar pattern but of lesser magnitude. Unlike some other habitat transitions, these results do not support biomechanical release as a mechanism likely to have facilitated limb elongation. Instead, limb bone adaptations in arboreal habitats were likely driven by selective pressures other than responses to skeletal loading.

Larval stomatopods have generally been described as having a typical larval crustacean compound eye, which lacks the visual pigment diversity and morphological specializations of the well-studied stomatopod adult eye. However, recent work has suggested that larval stomatopod eyes are more complex than previously described. In this study, we provide physiological and behavioral evidence of at least three distinct photoreceptor classes in three species of larval stomatopods: Gonodactylellus n. sp., Gonodactylaceus falcatus and Pullosquilla n. sp. First, electroretinogram recordings were used to measure the spectral sensitivity of each species. Evidence for at least three spectral classes were identified in each: an ultraviolet, peaking at 340–376 nm; a short-wavelength blue, peaking at 455–464 nm; and a long-wavelength orange, peaking at 576–602 nm. Next, the behavioral response to light was investigated. We found that each species demonstrated positive phototactic responses to monochromatic stimuli across the UV–visible spectrum. In wavelength preference trials, distinct preferences among species were identified when different colored light stimuli were presented simultaneously. All species displayed a strong response to the UV stimulus, as well as responses to blue and orange stimuli, although at different response strengths, but no response to green. The results of this study demonstrate that larval stomatopods not only have multiple physiologically active spectral classes but they also display clear and distinct responses to wavelengths across the spectrum. We propose that the spectral classes demonstrated in each are related to visually guided ecological tasks of the larvae, which may differ between species.

Given that all interactions between an animal and its environment are mediated by movement, questions of how animals inherit, refine and execute trajectories through space are fundamental to our understanding of biology. As with any behavioural trait, navigation can be thought of on many conceptual levels – from the mechanistic to the functional, and from the static to the dynamic – as laid out by Niko Tinbergen in his four questions of animal behaviour. Here, we use a navigation-centric interpretation of Tinbergen's questions to summarise and critique advances in the field of animal navigation. We discuss the ‘state of the art’; consider how a proximal/mechanistic understanding of navigation is not a prerequisite to understanding ultimate questions of evolutionary/adaptive importance; propose that certain aspects of animal navigation research – and certain taxa – are being neglected; and suggest that extreme experimental manipulations might lead to the mischaracterisation of non-adaptive ‘spandrels’ as functional navigational mechanisms. More generally, we highlight pressing questions within the field, the answers to which we believe are within reach, and highlight the important role that novel methods will have in helping us elucidate them.

Foot structures define the leverage in which the ankle muscles push off against the ground during locomotion. While prior studies have indicated that inter-individual variation in anthropometry (e.g. heel and hallux lengths) can directly affect force production of ankle plantar flexor muscles, its effect on the metabolic energy cost of locomotion has been inconclusive. Here, we tested the hypotheses that shorter heels and longer halluces are associated with slower plantar flexor (soleus) shortening velocity and greater ankle plantar flexion moment, indicating enhanced force potential as a result of the force–velocity relationship. We also hypothesized that such anthropometry profiles would reduce the metabolic energy cost of walking at faster walking speeds. Healthy young adults (N=15) walked at three speeds (1.25, 1.75 and 2.00 m s−1), and we collected in vivo muscle mechanics (via ultrasound), activation (via electromyography) and whole-body metabolic energy cost of transport (via indirect calorimetry). Contrary to our hypotheses, shorter heels and longer halluces were not associated with slower soleus shortening velocity or greater plantar flexion moment. Additionally, longer heels were associated with reduced metabolic cost of transport, but only at the fastest speed (2.00 m s−1, R2=0.305, P=0.033). We also found that individuals with longer heels required less increase in plantar flexor (soleus and gastrocnemius) muscle activation to walk at faster speeds, potentially explaining the reduced metabolic cost.

Air sacs are a well-known aspect of insect tracheal systems, but have received little research attention. In this Commentary, we suggest that the study of the distribution and function of air sacs in tracheate arthropods can provide insights of broad significance. We provide preliminary phylogenetic evidence that the developmental pathways for creation of air sacs are broadly conserved throughout the arthropods, and that possession of air sacs is strongly associated with a few traits, including the capacity for powerful flight, large body or appendage size and buoyancy control. We also discuss how tracheal compression can serve as an additional mechanism for achieving advection in tracheal systems. Together, these patterns suggest that the possession of air sacs has both benefits and costs that remain poorly understood. New technologies for visualization and functional analysis of tracheal systems provide exciting approaches for investigations that will be of broad significance for understanding invertebrate evolution.

The sensory systems of crustaceans (aquatic decapods and stomatopods) have adapted to a diverse range of aquatic ecosystems. Sound production in aquatic crustaceans is more widespread than previously thought, and has been shown to play a major role in many of their life-history strategies; however, there are still many gaps in our understanding of their sound reception abilities. Crustaceans have three main sensory receptors for sound – the statocyst, superficial hair cells and chordotonal organs – which are all sensitive to the particle motion component of the sound field, rather than the pressure component. Our current understanding of these receptors is that they are sensitive to low-frequency sounds (

Regional heterothermy is a pattern whereby different body regions are maintained at different temperatures, often to prioritize function of certain body parts over others, or to maximize function of organs and tissues that vary in thermal sensitivity. Regional heterothermy is relatively well understood in endotherms, where physiological mechanisms maintain heterogeneity. However, less is known about regional heterothermy in ectotherms, where behavioral mechanisms are more important for generating thermal variation. In particular, whether small and elongate ectotherms with high surface area to volume ratios such as diminutive snakes can maintain regional heterothermy, despite rapid thermal equilibration, is not yet known. We measured regional variation in body temperature and tested whether environmental heterogeneity is used to generate regional heterothermy in the ring-necked snake (Diadophis punctatus) using both field and laboratory studies. We found that ring-necked snakes have robust regional heterothermy in a variety of contexts, despite their small body size and elongate body shape. Temperature variation along the length of their bodies was not detectable when measured externally. However, snakes had higher mouth than cloacal temperatures both in the field and in laboratory thermal gradients. Further, this regional heterothermy was maintained even in ambient laboratory conditions, where the thermal environment was relatively homogeneous. Our results indicate that regional heterothermy in ring-necked snakes is not solely driven by environmental variation, but is instead linked to physiological or morphological mechanisms that maintain regional variation in body temperature irrespective of environmental context.

Eight juvenile European seabass were exposed to two thermal ramping protocols with different levels of aerobic activity and tolerance endpoint: the critical thermal maximum for swimming (CTSmax) while exercising aerobically until fatigue, and the critical thermal maximum (CTmax) under static conditions until loss of equilibrium (LOE). In the CTSmax, warming caused a profound increase in oxygen uptake rate (M˙O2) culminating in a gait transition, from steady aerobic towards unsteady anaerobic swimming, then fatigue at 30.3±0.4°C (mean±SE). Gait transition and fatigue presumably indicate an oxygen limitation, an inability to meet the combined demands of swimming plus warming. The CTmax also elicited an increase in M˙O2, culminating in LOE at 34.0±0.4°C, significantly warmer than fatigue at CTSmax. The maximum M˙O2 achieved in the CTmax was, however, less than 30% of that achieved in the CTSmax. Therefore, the static CTmax did not exploit full cardiorespiratory capacity for oxygen supply, indicating that LOE was not caused by systemic oxygen limitation. Consequently, systemic oxygen supply can be significant for tolerance of acute warming in seabass but this depends upon the physiological context and the endpoint used.

Scaling between specific organs and overall body size has long fascinated biologists, being a primary mechanism by which organ shapes evolve. Yet, the genetic mechanisms that underlie the evolution of scaling relationships remain elusive. Here, we compared wing and fore tibia lengths (the latter as a proxy of body size) in Drosophila melanogaster, Drosophila simulans, Drosophila ananassae and Drosophila virilis, and show that the first three of these species have roughly a similar wing-to-tibia scaling behavior. In contrast, D. virilis exhibits much smaller wings relative to their body size compared with the other species and this is reflected in the intercept of the wing-to-tibia allometry. We then asked whether the evolution of this relationship could be explained by changes in a specific cis-regulatory region or enhancer that drives expression of the wing selector gene, vestigial (vg), whose function is broadly conserved in insects and contributes to wing size. To test this hypothesis directly, we used CRISPR/Cas9 to replace the DNA sequence of the predicted Quadrant Enhancer (vgQE) from D. virilis for the corresponding vgQE sequence in the genome of D. melanogaster. Strikingly, we discovered that D. melanogaster flies carrying the D. virilis vgQE sequence have wings that are significantly smaller with respect to controls, partially shifting the intercept of the wing-to-tibia scaling relationship towards that observed in D. virilis. We conclude that a single cis-regulatory element in D. virilis contributes to constraining wing size in this species, supporting the hypothesis that scaling could evolve through genetic variations in cis-regulatory elements.

Chronic exposure to high temperatures may leave freshwater fishes vulnerable to opportunistic pathogens, particularly during early life stages. Lake sturgeon, Acipenser fulvescens, populations within the northern expanse of their range in Manitoba, Canada, may be susceptible to high temperature stress and pathogenic infection. We acclimated developing lake sturgeon for 22 days to two ecologically relevant, summer temperatures (16 and 20°C). Individuals from both acclimation treatments were then exposed to 0, 30 and 60 µg ml−1 bacterial lipopolysaccharides (endotoxins), as an immune stimulus, for 48 h and sampled 4 and 48 h during trial exposures and following a 7 day recovery period. We then measured whole-body transcriptional (mRNA) responses involved in the innate immune, stress and fatty acid responses following acute exposure to the bacterial endotoxins. Data revealed that overall levels of mRNA transcript abundance were higher in 20°C-reared sturgeon under control conditions. However, following exposure to a bacterial stimulus, lake sturgeon acclimated to 16°C produced a more robust and persistent transcriptional response with higher mRNA transcript abundance across innate immune, stress and fatty acid responses than their 20°C-acclimated counterparts. Additional whole-animal performance metrics (critical thermal maximum, metabolic rate, cortisol concentration and whole-body and mucosal lysozyme activity) demonstrated acclimation-specific responses, indicating compromised metabolic, stress and enzymatic capacity following the initiation of immune-related responses. Our study showed that acclimation to 20°C during early development impaired the immune capacity of developing lake sturgeon as well as the activation of molecular pathways involved in the immune, stress and fatty acid responses. The present study highlights the effects of ecologically relevant, chronic thermal stress on seasonal pathogen susceptibility in this endangered species.

The wings of the purple spotted swallowtail Graphium weiskei are marked by an unusual bright colour pattern. Spectrophotometry on G. weiskei wings demonstrated the presence of a pigment with an absorption spectrum (peak wavelength λmax=676 nm) similar to that of the bile pigment sarpedobilin in the wings of the congeneric Graphium sarpedon (λmax=672 nm). Sarpedobilin alone causes cyan–blue wing areas, but the green-coloured areas of G. sarpedon wings result from subtractive colour mixing with the carotenoid lutein. Reflectance spectra of the blue-coloured areas of G. weiskei wings indicate that sarpedobilin is mixed with the short-wavelength-absorbing papiliochrome II. An enigmatic pigment, tentatively called weiskeipigment (λmax=580 nm), enhances the saturation of the blue colour. Weiskeipigment causes a purple colour in areas where the sarpedobilin concentration is low. The wings of the related papilionid Papilio phorcas contain the bile pigment pharcobilin (λmax=604 nm), as well as another sarpedobilin (λmax=663 nm). The cyan to greenish wings of P. phorcas are due to phorcabilin and sarpedobilin mixed with papiliochrome II. A survey of known subspecies of G. weiskei as well as of congeneric Graphium species of the ‘weiskei’ group shows various degrees of subtractive colour mixing of bilins and short-wavelength absorbers (carotenoids and/or papiliochromes) in their wings. This study illuminates the underestimated role of bile pigments in butterfly wing colouration.

Skin provides functions such as protection and prevention of water loss. In some taxa, the outer surface of skin has been modified to form structures that enable attachment to various surfaces. Constant interaction with surfaces is likely to cause damage to these attachment systems and reduce function. It seems logical that when skin is shed via ecdysis, its effectiveness will increase, through repair of damage or other rejuvenating mechanisms. We address two questions using three diplodactylid geckos as model species. (1) Does repeated mechanical damage affect clinging ability in geckos to the point that they cannot support their own body weight? (2) Does use without induced damage reduce effectiveness of the attachment system, and if so, does ecdysis restore clinging ability? We found that repeated damage reduced clinging ability in all three species, although at different rates. Additionally, use reduced clinging ability over time when no apparent damage was incurred. Clinging ability increased after ecdysis in all three species, both when damage was specially induced, and when it was not. After normal use without induced damage, the increase in clinging ability after ecdysis was statistically significant in two of three species. Our findings show that use decreases clinging ability, and mechanical damage also effects geckos' capacity to exert shear forces consistently. Thus, ecdysis improves clinging ability both in scenarios where damage is induced and more generally. In addition to the physiological functions provided by skin, our study highlights an important function of ecdysis in a speciose vertebrate group.

In the past century, several authors investigated the allometry of hematological parameters in mammals. Since hematocrit and hemoglobin concentration are almost constant within the Mammalia (although with notable exceptions), differences in other hematological parameters are mainly reducible to red blood cell size (MCV). Past studies testing for correlation between MCV and body mass have given contradictory results. Using phylogenetically-informed regressions, here is demonstrated that correlation between MCV and body mass is indirect, and is in reality due to correlation between MCV and basal metabolic rates. This could be explained by the fact that small erythrocytes allow a fast release of oxygen in tissues with high metabolic demand. Nonetheless, hypoxia-adapted species show MCV greater than those predicted by their metabolic rates, while Ruminantia show the inverse. Interestingly, these species show the highest and lowest, respectively, hemoglobin affinity for oxygen. In the present paper, it is suggested that hemoglobin affinity, acting as a biological constrain for oxygen exchange, determines the size of red blood cells. Hemoglobin intrinsic affinity showed little variations during evolution and modifying the levels of allosteric factors can be viewed as an adaption to adjust hemoglobin affinity to metabolic demands (the same happens also during ontogeny). Nonetheless, in some lineages, mutations raising hemoglobin affinity allowed some species to colonize hypoxic environments; in Ruminantia, instead, there was a drastic decrease, which cannot be adaptively explained.

Ocean acidification and warming are key stressors for many marine organisms. Some organisms display physiological acclimatisation or plasticity, but this may vary across species ranges, especially if populations are adapted to local climatic conditions. Understanding how acclimatisation potential varies among populations is therefore important in predicting species responses to climate change. We carried out a common garden experiment to investigate how different populations of the economically important great scallop (Pecten maximus) from France and Norway responded to variation in temperature andpCO2concentration. After acclimation, post-larval scallops (spat) were reared for 31 days at one of two temperatures (13°C and 19°C) under either ambient or elevatedpCO2(pH 8.0 and pH 7.7). We combined measures of proteomic, metabolic, and phenotypic traits to produce an integrative picture of how physiological plasticity varies between the populations. The proteome of French spat showed significant sensitivity to environmental variation, with 12 metabolic, structural and stress-response proteins responding to temperature and/orpCO2. Principal component analysis revealed seven energy metabolism proteins in French spat that were consistent with countering ROS stress under elevated temperature. Oxygen uptake in French spat did not change under elevated temperature, but increased under elevatedpCO2. In contrast, Norwegian spat reduced oxygen uptake under both elevated temperature andpCO2. Metabolic plasticity seemingly allowed French scallops to maintain greater energy availability for growth than Norwegian spat. However, increased physiological plasticity and growth in French spat may come at a cost, as French (but not Norwegian) spat showed reduced survival under elevated temperature.Summary StatementJuvenile scallops from France and Norway differ in their response to warming and acidification. French scallops show more physiological plasticity, adjusting their proteome and metabolism in order to maintain growth.

Changing ocean temperatures are predicted to challenge marine organisms, especially when combined with other factors, such as ocean acidification. Acclimation, as a form of phenotypic plasticity, can however, moderate the consequences of changing environments for biota. Our understanding of how altered temperature and acidification together influence species acclimation responses is, however, limited compared to responses to single stressors. This study investigated how temperature and acidification affected the thermal tolerance and righting speed of the Girdled Dogwhelk, Trochia cingulata (Linnaeus, 1771). Whelks were acclimated for two weeks to combinations of three temperatures (11°C: cold, 13°C: moderate and 15°C: warm) and two pH regimes (8.0: moderate and 7.5: acidic). We measured the temperature sensitivity of righting response by generating thermal performance curves from individual data collected at seven test temperatures and determined critical thermal minima (CTmin) and maxima (CTmax). We found that T. cingulata has a broad basal thermal tolerance range (∼38°C) and after acclimation to the warm temperature regime, both the optimal temperature for maximum righting speed and CTmax increased. Contrary to predictions, acidification did not narrow this population's thermal tolerance but increased CTmax. These plastic responses are likely driven by the predictable exposure to temperature extremes measured in the field which originate from the local tidal cycle and the periodic acidification associated with ocean upwelling in the region. This acclimation ability suggests that T. cingulata has at least some capacity to buffer the thermal changes and increased acidification predicted to occur with climate change.

Many animals use body parts such as tails to stabilize posture while moving at high speed. In flying insects, leg or abdominal inertia can influence flight posture. In the hawkmoth Manduca sexta, the abdomen contributes ∼50% of the total body weight and it can therefore serve to inertially redirect flight forces. How do torques generated by the wings and abdomen interact for flight control? We studied the yaw optomotor response of M. sexta by using a torque sensor attached to their thorax. In response to yaw visual motion, the abdomen moved antiphase with the stimulus, head and total torque. By studying moths with ablated wings and a fixed abdomen, we resolved abdomen and wing torques and revealed their individual contribution to total yaw torque production. Frequency-domain analysis revealed that the abdomen torque is overall smaller than wing torque, although the abdomen torque is ∼80% of the wing torque at higher visual stimulus temporal frequency. Experimental data and modeling revealed that the wing and abdomen torque are transmitted linearly to the thorax. By modeling the thorax and abdomen as a two-link system, we show that abdomen flexion can inertially redirect the thorax to add constructively to wing steering efforts. Our work argues for considering the role of the abdomen in tethered insect flight experiments that use force/torque sensors. Taken together, the hawkmoth abdomen can regulate wing torques in free flight, which could modulate flight trajectories and increase maneuverability.

Journal of Experimental Biology has a long history of reporting research discoveries on animal echolocation, the subject of this Centenary Review. Echolocating animals emit intense sound pulses and process echoes to localize objects in dynamic soundscapes. More than 1100 species of bats and 70 species of toothed whales rely on echolocation to operate in aerial and aquatic environments, respectively. The need to mitigate acoustic clutter and ambient noise is common to both aerial and aquatic echolocating animals, resulting in convergence of many echolocation features, such as directional sound emission and hearing, and decreased pulse intervals and sound intensity during target approach. The physics of sound transmission in air and underwater constrains the production, detection and localization of sonar signals, resulting in differences in response times to initiate prey interception by aerial and aquatic echolocating animals. Anti-predator behavioral responses of prey pursued by echolocating animals affect behavioral foraging strategies in air and underwater. For example, many insect prey can detect and react to bat echolocation sounds, whereas most fish and squid are unresponsive to toothed whale signals, but can instead sense water movements generated by an approaching predator. These differences have implications for how bats and toothed whales hunt using echolocation. Here, we consider the behaviors used by echolocating mammals to (1) track and intercept moving prey equipped with predator detectors, (2) interrogate dynamic sonar scenes and (3) exploit visual and passive acoustic stimuli. Similarities and differences in animal sonar behaviors underwater and in air point to open research questions that are ripe for exploration.

The bodies of most swimming fishes are very flexible and deform as result of both external fluid dynamic forces and internal musculoskeletal forces. If fluid forces change, the body motion will also change unless the fish senses the change and alters its muscle activity to compensate. Lampreys and other fishes have mechanosensory cells in their spinal cords that allow them to sense how their body is bending. We hypothesized that lampreys (Petromyzon marinus) actively regulate body curvature to maintain a fairly constant swimming waveform even as swimming speed and fluid dynamic forces change. To test this hypothesis, we measured the steady swimming kinematics of lampreys swimming in normal water, and water in which the viscosity was increased by 10 or 20 times by adding methylcellulose. Increasing the viscosity over this range increases the drag coefficient, potentially increasing fluid forces up to 40%. Previous computational results suggested that if lampreys did not compensate for these forces, the swimming speed would drop by about 52%, the amplitude would drop by 39%, and posterior body curvature would increase by about 31%, while tail beat frequency would remain the same. Five juvenile sea lampreys were filmed swimming through still water, and midlines were digitized using standard techniques. Although swimming speed dropped by 44% from 1× to 10× viscosity, amplitude only decreased by 4%, and curvature increased by 7%, a much smaller change than the amount we estimated if there was no compensation. To examine the waveform overall, we performed a complex orthogonal decomposition and found that the first mode of the swimming waveform (the primary swimming pattern) did not change substantially, even at 20× viscosity. Thus, it appears that lampreys are compensating, at least partially, for the changes in viscosity, which in turn suggests that sensory feedback is involved in regulating the body waveform.

Oxidative metabolism is a key component of organismal physiology and it is primarily determined by aerobic capacity, which depends on the capacity of blood to carry oxygen. However, experimental manipulations of blood oxygen-carrying capacity are rarely implemented to test ecophysiological hypotheses in vertebrate populations. Here, we combined an experimental manipulation of blood oxygen-carrying capacity with GPS tracking to test whether suboptimal (reduced) haematological performance affects foraging behaviour in a colonial waterbird, the black-headed gull, Chroicocephalus ridibundus. First, a validation of phenylhydrazine (PHZ) treatment in gulls revealed a 9–18% reduction in haematocrit and blood haemoglobin concentration (via oxidative denaturation and haemolysis of erythrocytes). Then, GPS tracking of experimental (PHZ-treated) and control (saline-treated) gulls during the incubation period provided no support for reduced or suspended engagement in energetically costly activities (long-distance foraging trips) by experimental birds. Instead, we found evidence for fine-scale alterations in foraging behaviour of PHZ-treated individuals, which resulted in fewer foraging trips per unit time, but trips that were longer in duration and distance compared with those of control birds. This suggests reduced foraging performance of experimental birds (e.g. lower capacity to find and collect food during trips) or evasion of social competition, although no differences in the total investment in foraging may also suggest compensatory physiological responses to haemolytic anaemia. Our study contributes to a better understanding of the physio-ecological nexus in non-diving colonial avian species. Whether behavioural effects of reduced aerobic capacity have any implications for gull condition and reproductive performance should be the subject of further investigation.

Environmental challenges early in development can result in complex phenotypic trade-offs and long-term effects on individual physiology, performance and behavior, with implications for disease and predation risk. We examined the effects of simulated pond drying and elevated water temperatures on development, growth, thermal physiology and behavior in a North American amphibian, Rana sphenocephala. Tadpoles were raised in outdoor mesocosms under warming and drying regimes based on projected climatic conditions in 2070. We predicted that amphibians experiencing the rapid pond drying and elevated pond temperatures associated with climate change would accelerate development, be smaller at metamorphosis and demonstrate long-term differences in physiology and exploratory behavior post-metamorphosis. Although both drying and warming accelerated development and reduced survival to metamorphosis, only drying resulted in smaller animals at metamorphosis. Around 1 month post-metamorphosis, animals from the control treatment jumped relatively farther at high temperatures in jumping trials. In addition, across all treatments, frogs with shorter larval periods had lower critical thermal minima and maxima. We also found that developing under warming and drying resulted in a less exploratory behavioral phenotype, and that drying resulted in higher selected temperatures in a thermal gradient. Furthermore, behavior predicted thermal preference, with less exploratory animals selecting higher temperatures. Our results underscore the multi-faceted effects of early developmental environments on behavioral and physiological phenotypes later in life. Thermal preference can influence disease risk through behavioral thermoregulation, and exploratory behavior may increase risk of predation or pathogen encounter. Thus, climatic stressors during development may mediate amphibian exposure and susceptibility to predators and pathogens into later life stages.

Control of locomotion involves the interplay of sensory signals and motor commands. Sensory information is essential for adjusting locomotion in response to environmental changes. A previous study using mathematical modelling of lamprey swimming has shown that, in the absence of sensory feedback, increasing fluid viscosity constrains swimming kinematics, limiting tail amplitude and body wavelength, resulting in decreased swimming speed. In contrast, previous experiments with Polypterus senegalus reported increased magnitude swimming kinematics (increased body curvature, body wave speed and frequency, and pectoral fin frequency) in high viscosity water suggesting that sensory information is used to adjust swimming form. It is not known what sensory systems are providing the necessary information to respond to these environmental changes. We tested the hypothesis that lateral line and visual input are responsible for the sensory-driven increase in swimming kinematics in response to experimentally increased fluid viscosity. The kinematics of five P. senegalus were recorded in two different viscosities of water while removing lateral line and visual sensory feedback. Unlike the mathematical model devoid of sensory feedback, P. senegalus with lateral line and/or visual senses removed did not reduce the magnitude of swimming kinematic variables, suggesting that additional sensory feedback mechanisms are present in these fish to help overcome increased fluid viscosity. Increases in swimming speed when both lateral line and visual sensory feedback were removed suggest that lateral line and visual information may be used to regulate swimming speed in P. senegalus, possibly using an internal model of predictions to adjust swimming form.

Multiple variables determine the success of an escape response of an animal, and the rapidity of the escape manoeuvre is often the most important. Fan worms (Annelida: Sabellidae) can rapidly withdraw their tentacles, which are covered in heavily ciliated ramifications called pinnules, into their tubes to protect them from approaching threats. Here, we explore the dynamic and mechanistic features behind this escape manoeuvre. The escape responses of fan worms were recorded by high-speed videography and quantified by computerized motion analysis, showing an ultrahigh retraction speed of 272±135 mm s−1 (8±4 body lengths s−1). We found that fan worms possess powerful muscle-driven systems, which can generate contractive forces up to 36 times their body weight. In order to achieve these rapid, forceful movements through seawater without damaging their tentacles, fan worms have developed functional morphological adaptations to reduce fluidic drag, including the flattening of their radiolar pinnules and the deformation of bodily segmental ridges. Our hydrodynamic models indicate that these mechanical processes can decrease fluidic drag by 47%, trapped mass by 75% and friction coefficient by 89%. These strategies allow fan worms to execute rapid escape responses and could inspire the design of fast in-pipe robots.

Vertebrates capable of powered flight rely on wings, muscles that drive their flapping and sensory inputs to the brain allowing for control of the motor output. In birds, the wings are formed of arrangements of adjacent flight feathers (remiges), whereas the wings of bats consist of double-layered skin membrane stretched out between the forelimb skeleton, body and legs. Bird feathers become worn from use and brittle from UV exposure, which leads to loss of function; to compensate, they are renewed (moulted) at regular intervals. Bird feathers and the wings of bats can be damaged by accident. Wing damage and loss of wing surface due to moult almost invariably cause reduced flight performance in measures such as take-off angle and speed. During moult in birds, this is partially counteracted by concurrent mass loss and enlarged flight muscles. Bats have sensory hairs covering their wing surface that provide feedback information about flow; thus, wing damage affects flight speed and turning ability. Bats also have thin, thread-like muscles, distributed within the wing membrane and, if these are damaged, the control of wing camber is lost. Here, I review the effects of wing damage and moult on flight performance in birds, and the consequences of wing damage in bats. I also discuss studies of life-history trade-offs that make use of experimental trimming of flight feathers as a way to handicap parent birds feeding their young.

Nearly all fish have flexible bodies that bend as a result of internal muscular forces and external fluid forces that are dynamically coupled with the mechanical properties of the body. Swimming is therefore strongly influenced by the body's flexibility, yet we do not know how fish species vary in their flexibility and in their ability to modulate flexibility with muscle activity. A more fundamental problem is our lack of knowledge about how any of these differences in flexibility translate into swimming performance. Thus, flexibility represents a hidden axis of diversity among fishes that may have substantial impacts on swimming performance. Although engineers have made substantial progress in understanding these fluid–structure interactions using physical and computational models, the last biological review of these interactions and how they give rise to fish swimming was carried out more than 20 years ago. In this Review, we summarize work on passive and active body mechanics in fish, physical models of fish and bioinspired robots. We also revisit some of the first studies to explore flexural stiffness and discuss their relevance in the context of more recent work. Finally, we pose questions and suggest future directions that may help reveal important links between flexibility and swimming performance.

Muscular hydrostats are organs composed entirely of packed arrays of incompressible muscles and lacking any skeletal support. Found in both vertebrates and invertebrates, they are of great interest for comparative biomechanics from engineering and evolutionary perspectives. The arms of cephalopods (e.g. octopus and squid) are particularly interesting muscular hydrostats because of their flexibility and ability to generate complex behaviors exploiting elaborate nervous systems. Several lines of evidence from octopus studies point to the use of both brain and arm-embedded motor control strategies that have evolved to simplify the complexities associated with the control of flexible and hyper-redundant limbs and bodies. Here, we review earlier and more recent experimental studies on octopus arm biomechanics and neural motor control. We review several dynamic models used to predict the kinematic characteristics of several basic motion primitives, noting the shortcomings of the current models in accounting for behavioral observations. We also discuss the significance of impedance (stiffness and viscosity) in controlling the octopus's motor behavior. These factors are considered in light of several new models of muscle biomechanics that could be used in future research to gain a better understanding of motor control in the octopus. There is also a need for updated models that encompass stiffness and viscosity for designing and controlling soft robotic arms. The field of soft robotics has boomed over the past 15 years and would benefit significantly from further progress in biomechanical and motor control studies on octopus and other muscular hydrostats.

Outside laboratory conditions and human-made structures, animals rarely encounter flat surfaces. Instead, natural substrates are uneven surfaces with height variation that ranges from the microscopic scale to the macroscopic scale. For walking animals (which we define as encompassing any form of legged movement across the ground, such as walking, running, galloping, etc.), such substrate ‘roughness’ influences locomotion in a multitude of ways across scales, from roughness that influences how each toe or foot contacts the ground, to larger obstacles that animals must move over or navigate around. Historically, the unpredictability and variability of natural environments has limited the ability to collect data on animal walking biomechanics. However, recent technical advances, such as more sensitive and portable cameras, biologgers, laboratory tools to fabricate rough terrain, as well as the ability to efficiently store and analyze large variable datasets, have expanded the opportunity to study how animals move under naturalistic conditions. As more researchers endeavor to assess walking over rough terrain, we lack a consistent approach to quantifying roughness and contextualizing these findings. This Review summarizes existing literature that examines non-human animals walking on rough terrain and presents a metric for characterizing the relative substrate roughness compared with animal size. This framework can be applied across terrain and body scales, facilitating direct comparisons of walking over rough surfaces in animals ranging in size from ants to elephants.

Locomotor biomechanics faces a core trade-off between laboratory-based and field-based studies. Laboratory conditions offer control over confounding factors, repeatability, and reduced technological challenges, but limit the diversity of animals and environmental conditions that may influence behavior and locomotion. This article considers how study setting influences the selection of animals, behaviors and methodologies for studying animal motion. We highlight the benefits of both field- and laboratory-based studies and discuss how recent work leverages technological advances to blend these approaches. These studies have prompted other subfields of biology, namely evolutionary biology and ecology, to incorporate biomechanical metrics more relevant to survival in natural habitats. The concepts discussed in this Review provide guidance for blending methodological approaches and inform study design for both laboratory and field biomechanics. In this way, we hope to facilitate integrative studies that relate biomechanical performance to animal fitness, determine the effect of environmental factors on motion, and increase the relevance of biomechanics to other subfields of biology and robotics.

Studying the motion of cheetahs – especially in the wild – is a technically challenging endeavour that pushes the limits of field biomechanics methodology. Consequently, it provides an interesting example of the scientific symbiosis that exists between experimental biology and the technological disciplines that support it. This article uses cheetah motion research as a basis to review the past, present and likely future of field biomechanics. Although the focus is on a specific animal, the methods and challenges discussed are broadly relevant to the study of terrestrial locomotion. We also highlight the external factors contributing to the evolution of this technology, including recent advancements in machine learning, and the influx of interest in cheetah biomechanics from the legged robotics community.

Foraging insects fly over long distances through complex aerial environments, and many can maintain constant ground speeds in wind, allowing them to gauge flight distance. Although insects encounter winds from all directions in the wild, most lab-based studies have employed still air or headwinds (i.e. upwind flight); additionally, insects are typically compelled to fly in a single, fixed environment, so we know little about their preferences for different flight conditions. We used automated video collection and analysis methods and a two-choice flight tunnel paradigm to examine thousands of foraging flights performed by hundreds of bumblebees flying upwind and downwind. In contrast to the preference for flying with a tailwind (i.e. downwind) displayed by migrating insects, we found that bees prefer to fly upwind. Bees maintained constant ground speeds when flying upwind or downwind in flow velocities from 0 to 2 m s−1 by adjusting their body angle, pitching down to raise their air speed above flow velocity when flying upwind, and pitching up to slow down to negative air speeds (flying backwards relative to the flow) when flying downwind. Bees flying downwind displayed higher variability in body angle, air speed and ground speed. Taken together, bees' preference for upwind flight and their increased kinematic variability when flying downwind suggest that tailwinds may impose a significant, underexplored flight challenge to bees. Our study demonstrates the types of questions that can be addressed with newer approaches to biomechanics research; by allowing bees to choose the conditions they prefer to traverse and automating filming and analysis to examine massive amounts of data, we were able to identify significant patterns emerging from variable locomotory behaviors, and gain valuable insight into the biomechanics of flight in natural environments.

The study of breathing in fishes has featured prominently in Journal of Experimental Biology (JEB), particularly during the latter half of the past century. Indeed, many of the seminal discoveries in this important sub-field of comparative respiratory physiology were reported first in JEB. The period spanning 1960–1990 (the ‘golden age of comparative respiratory physiology’) witnessed intense innovation in the development of methods to study the control of breathing. Many of the guiding principles of piscine ventilatory control originated during this period, including our understanding of the dominance of O2 as the driver of ventilation in fish. However, a critical issue – the identity of the peripheral O2 chemoreceptors – remained unanswered until methods for cell isolation, culture and patch-clamp recording established that gill neuroepithelial cells (NECs) respond to hypoxia in vitro. Yet, the role of the NECs and other putative peripheral or central chemoreceptors in the control of ventilation in vivo remains poorly understood. Further progress will be driven by the implementation of genetic tools, most of which can be used in zebrafish (Danio rerio). These tools include CRISPR/Cas9 for selective gene knockout, and Tol2 systems for transgenesis, the latter of which enables optogenetic stimulation of cellular pathways, cellular ablation and in vivo cell-specific biosensing. Using these methods, the next period of discovery will see the identification of the peripheral sensory pathways that initiate ventilatory responses, and will elucidate the nature of their integration within the central nervous system and their link to the efferent motor neurons that control breathing.

Spatially invariant feature detection is a property of many visual systems that rely on visual information provided by two eyes. However, how information across both eyes is integrated for invariant feature detection is not fully understood. Here, we investigated spatial invariance of looming responses in descending neurons (DNs) of Drosophila melanogaster. We found that multiple looming responsive DNs integrate looming information across both eyes, even though their dendrites are restricted to a single visual hemisphere. One DN, the giant fiber (GF), responds invariantly to looming stimuli across tested azimuthal locations. We confirmed visual information propagates to the GF from the contralateral eye, through an unidentified pathway, and demonstrated that the absence of this pathway alters GF responses to looming stimuli presented to the ipsilateral eye. Our data highlight a role for bilateral visual integration in generating consistent, looming-evoked escape responses that are robust across different stimulus locations and parameters.