Showing papers in "The Journal of Experimental Biology in 2007"

Modulation of locomotor activity in larval zebrafish during light adaptation.

Abstract: The neural basis of behavioral choice in vertebrates remains largely unknown. Zebrafish larvae have a defined locomotor repertoire as well as a simple nervous system and are therefore an attractive vertebrate system in which to study this process. Here we describe a high-throughput system for quantifying the kinematics of motor events in zebrafish larvae in order to measure the initiation frequency of different maneuvers. We use this system to analyze responses to photic stimuli and find that larvae respond to changes in illumination with both acute responses and extended behavioral programs. Reductions in illumination elicit large angle turns, distinct from startle responses, which orient larvae toward the source of light. In continuing darkness, larvae are transiently hyperactive before adopting a quiescent state. Indeed, locomotor activity is controlled by the state of light or dark adaptation similar to masking phenomena in higher vertebrates where light directly regulates motor activity. We propose that regulation of motor activity by photic stimuli in zebrafish larvae serves a behavioral goal of maximizing exposure to well lit environments optimal for feeding.

Advanced sequencing technologies and their wider impact in microbiology.

Abstract: In the past 10 years, microbiology has undergone a revolution that has been driven by access to cheap high-throughput DNA sequencing. It was not long ago that the cloning and sequencing of a target gene could take months or years, whereas now this entire process has been replaced by a 10 min Internet search of a public genome database. There has been no single innovation that has initiated this rapid technological change; in fact, the core chemistry of DNA sequencing is the same as it was 30 years ago. Instead, progress has been driven by large sequencing centers that have incrementally industrialized the Sanger sequencing method. A side effect of this industrialization is that large-scale sequencing has moved out of small research labs, and the vast majority of sequence data is now generated by large genome centers. Recently, there have been advances in technology that will enable high-throughput genome sequencing to be established in research labs using bench-top instrumentation. These new technologies are already being used to explore the vast microbial diversity in the natural environment and the untapped genetic variation that can occur in bacterial species. It is expected that these powerful new methods will open up new questions to genomic investigation and will also allow high-throughput sequencing to be more than just a discovery exercise but also a routine assay for hypothesis testing. While this review will concentrate on microorganisms, many of the important arguments about the need to measure and understand variation at the species, population and ecosystem level will hold true for many other biological systems.

Fish biorobotics: kinematics and hydrodynamics of self-propulsion

Abstract: As a result of years of research on the comparative biomechanics and physiology of moving through water, biologists and engineers have made considerable progress in understanding how animals moving underwater use their muscles to power movement, in describing body and appendage motion during propulsion, and in conducting experimental and computational analyses of fluid movement and attendant forces. But it is clear that substantial future progress in understanding aquatic propulsion will require new lines of attack. Recent years have seen the advent of one such new avenue that promises to greatly broaden the scope of intellectual opportunity available to researchers: the use of biorobotic models. In this paper we discuss, using aquatic propulsion in fishes as our focal example, how using robotic models can lead to new insights in the study of aquatic propulsion. We use two examples: (1) pectoral fin function, and (2) hydrodynamic interactions between dorsal and caudal fins. Pectoral fin function is characterized by considerable deformation of individual fin rays, as well as spanwise (along the length) and chordwise (across the fin) deformation and area change. The pectoral fin can generate thrust on both the outstroke and instroke. A robotic model of the pectoral fin replicates this result, and demonstrates the effect of altering stroke kinematics on the pattern of force production. The soft dorsal fin of fishes sheds a distinct vortex wake that dramatically alters incoming flow to the tail: the dorsal fin and caudal fin act as dual flapping foils in series. This design can be replicated with a dual-foil flapping robotic device that demonstrates this phenomenon and allows examination of regions of the flapping performance space not available to fishes. We show how the robotic flapping foil device can also be used to better understand the significance of flexible propulsive surfaces for locomotor performance. Finally we emphasize the utility of self-propelled robotic devices as a means of understanding how locomotor forces are generated, and review different conceptual designs for robotic models of aquatic propulsion.

Adaptational responses of the human Achilles tendon by modulation of the applied cyclic strain magnitude.

Abstract: Tendons are able to remodel their mechanical and morphological properties in response to mechanical loading. However, there is little information about the effects of controlled modulation in cyclic strain magnitude applied to the tendon on the adaptation of tendon's properties in vivo. The present study investigated whether the magnitude of the mechanical load induced as cyclic strain applied to the Achilles tendon may have a threshold in order to trigger adaptation effects on tendon mechanical and morphological properties. Twenty-one adults (experimental group, N=11; control group, N=10) participated in the study. The participants of the experimental group exercised one leg at low-magnitude tendon strain (2.85+/-0.99%) and the other leg at high-magnitude tendon strain (4.55+/-1.38%) of similar frequency and volume. After 14 weeks of exercise intervention we found a decrease in strain at a given tendon force, an increase in tendon-aponeurosis stiffness and tendon elastic modulus and a region-specific hypertrophy of the Achilles tendon only in the leg exercised at high strain magnitude. These findings provide evidence of the existence of a threshold or set-point at the applied strain magnitude at which the transduction of the mechanical stimulus may influence the tensional homeostasis of the tendons. The results further show that the mechanical load exerted on the Achilles tendon during the low-strain-magnitude exercise is not a sufficient stimulus for triggering further adaptation effects on the Achilles tendon than the stimulus provided by the mechanical load applied during daily activities.

Going wild: what a global small-animal tracking system could do for experimental biologists

Abstract: Tracking animals over large temporal and spatial scales has revealed invaluable and spectacular biological information, particularly when the paths and fates of individuals can be monitored on a global scale. However, only large animals (greater than approximately 300 g) currently can be followed globally because of power and size constraints on the tracking devices. And yet the vast majority of animals is small. Tracking small animals is important because they are often part of evolutionary and ecological experiments, they provide important ecosystem services and they are of conservation concern or pose harm to human health. Here, we propose a small-animal satellite tracking system that would enable the global monitoring of animals down to the size of the smallest birds, mammals (bats), marine life and eventually large insects. To create the scientific framework necessary for such a global project, we formed the ICARUS initiative (www.IcarusInitiative.org), the International Cooperation for Animal Research Using Space. ICARUS also highlights how small-animal tracking could address some of the ;Grand Challenges in Environmental Sciences' identified by the US National Academy of Sciences, such as the spread of infectious diseases or the relationship between biological diversity and ecosystem functioning. Small-animal tracking would allow the quantitative assessment of dispersal and migration in natural populations and thus help solve enigmas regarding population dynamics, extinctions and invasions. Experimental biologists may find a global small-animal tracking system helpful in testing, validating and expanding laboratory-derived discoveries in wild, natural populations. We suggest that the relatively modest investment into a global small-animal tracking system will pay off by providing unprecedented insights into both basic and applied nature. Tracking small animals over large spatial and temporal scales could prove to be one of the most powerful techniques of the early 21st century, offering potential solutions to a wide range of biological and societal questions that date back two millennia to the Greek philosopher Aristotle's enigma about songbird migration. Several of the more recent Grand Challenges in Environmental Sciences, such as the regulation and functional consequences of biological diversity or the surveillance of the population ecology of zoonotic hosts, pathogens or vectors, could also be addressed by a global small-animal tracking system. Our discussion is intended to contribute to an emerging groundswell of scientific support to make such a new technological system happen.

Tribute to P. L. Lutz: putting life on `pause' – molecular regulation of hypometabolism

Abstract: Entry into a hypometabolic state is an important survival strategy for many organisms when challenged by environmental stress, including low oxygen, cold temperatures and lack of food or water. The molecular mechanisms that regulate transitions to and from hypometabolic states, and stabilize long-term viability during dormancy, are proving to be highly conserved across phylogenic lines. A number of these mechanisms were identified and explored using anoxia-tolerant turtles as the model system, particularly from the research contributions made by Dr Peter L. Lutz in his explorations of the mechanisms of neuronal suppression in anoxic brain. Here we review some recent advances in understanding the biochemical mechanisms of metabolic arrest with a focus on ideas such as the strategies used to reorganize metabolic priorities for ATP expenditure, molecular controls that suppress cell functions (e.g. ion pumping, transcription, translation, cell cycle arrest), changes in gene expression that support hypometabolism, and enhancement of defense mechanisms (e.g. antioxidants, chaperone proteins, protease inhibitors) that stabilize macromolecules and promote long-term viability in the hypometabolic state.

Mechanical power and efficiency of level walking with different stride rates.

Abstract: Walking humans prefer to use the stride rate that results in the lowest rate of metabolic energy expenditure. Mechanical power requirements have been suggested to underlie the metabolic response, but mechanical power is consistently reported to be minimal at stride rates 20-30% lower than preferred. This may be due to limitations in how total mechanical power has been computed, as well as a failure to account for the efficiency with which muscular work is done. We investigated how mechanical power and efficiency depend on stride rate in walking, with both quantities computed from the work done by the hip, knee and ankle joint moments. Our hypotheses were that mechanical power and efficiency are both optimized at the preferred stride rate, which would explain why metabolic energy expenditure is minimized at this rate. Contrary to our hypotheses, mechanical power curves exhibited plateaus that spanned stride rates lower than preferred (predicted optima: 11-12% below preferred), while net mechanical efficiency exhibited a plateau that spanned stride rates higher than preferred (predicted optimum: 8% above preferred). As expected, preferred stride rate (54.3 strides min(-1)) was not different from the stride rate that minimized net metabolic energy expenditure (predicted optimum: 0.2% above preferred). Given that mechanical power and mechanical efficiency exhibited plateaus on opposite sides of the preferred stride rate, the preferred rate in walking likely represents a compromise between these two factors. This may also explain the relative flatness of the curve for metabolic rate in the vicinity of the preferred stride rate.

Three-dimensional kinematics of hummingbird flight

Abstract: Hummingbirds are specialized for hovering flight, and substantial research has explored this behavior. Forward flight is also important to hummingbirds, but the manner in which they perform forward flight is not well documented. Previous research suggests that hummingbirds increase flight velocity by simultaneously tilting their body angle and stroke-plane angle of the wings, without varying wingbeat frequency and upstroke: downstroke span ratio. We hypothesized that other wing kinematics besides stroke-plane angle would vary in hummingbirds. To test this, we used synchronized high-speed (500 Hz) video cameras and measured the three-dimensional wing and body kinematics of rufous hummingbirds (Selasphorus rufus, 3 g, N=5) as they flew at velocities of 0-12 m s(-1) in a wind tunnel. Consistent with earlier research, the angles of the body and the stroke plane changed with velocity, and the effect of velocity on wingbeat frequency was not significant. However, hummingbirds significantly altered other wing kinematics including chord angle, angle of attack, anatomical stroke-plane angle relative to their body, percent of wingbeat in downstroke, wingbeat amplitude, angular velocity of the wing, wingspan at mid-downstroke, and span ratio of the wingtips and wrists. This variation in bird-centered kinematics led to significant effects of flight velocity on the angle of attack of the wing and the area and angles of the global stroke planes during downstroke and upstroke. We provide new evidence that the paths of the wingtips and wrists change gradually but consistently with velocity, as in other bird species that possess pointed wings. Although hummingbirds flex their wings slightly at the wrist during upstroke, their average wingtip-span ratio of 93% revealed that they have kinematically ;rigid' wings compared with other avian species.

Gill remodeling in fish – a new fashion or an ancient secret?

Abstract: SUMMARY While a large respiratory surface area is good for gas exchange, it also poses several problems, including energetically unfavorable fluxes of water and ions. As a result, fishes appear to have a respiratory surface area that is matched to their oxygen demands. When faced with changes in their need for oxygen uptake, e.g. through altered physical activity or altered ambient oxygen levels, fishes have long been known to make two different adjustments: (1) to change the water flow over the gills or (2) to change the blood flow inside the gills. It has recently become clear that at least some teleosts have a third option: to reversibly remodel the gill morphology. Studies have shown that the lamellae of crucian carp Carassius carassius gills are embedded in a cell mass during normoxic conditions or at low temperature, while much of this cell mass dies off in hypoxia and at higher temperatures, thereby exposing a much larger respiratory surface area. Gill remodeling has subsequently been seen in two more cyprinids and in the mangrove killifish Kryptolebias marmoratus . In the latter case it appears to be an adaptation to periods of air exposure. Gill remodeling in response to changing respiratory requirements could be an ancient mechanism, occurring in many more teleosts than presently known. It is tempting to suggest that gill remodeling has been overlooked in many fishes, either because it is relatively subtle in some species, or because fishes are often kept at the warmer end of their temperature range where they need fully protruding lamellae.

A new paradigm for developmental biology.

Abstract: It is usually thought that the development of complex organisms is controlled by protein regulatory factors and morphogenetic signals exchanged between cells and differentiating tissues during ontogeny. However, it is now evident that the majority of all animal genomes is transcribed, apparently in a developmentally regulated manner, suggesting that these genomes largely encode RNA machines and that there may be a vast hidden layer of RNA regulatory transactions in the background. I propose that the epigenetic trajectories of differentiation and development are primarily programmed by feed-forward RNA regulatory networks and that most of the information required for multicellular development is embedded in these networks, with cell-cell signalling required to provide important positional information and to correct stochastic errors in the endogenous RNA-directed program.

Magnetic maps in animals: nature's GPS.

Abstract: Diverse animals detect the Earth's magnetic field and use it as a cue in orientation and navigation. Most research on magnetoreception has focused on the directional or \`compass' information that can be extracted from the Earth's field. Because the field varies predictably across the surface of the globe, however, it also provides a potential source of positional or \`map' information, which some animals use to steer themselves along migratory pathways or to navigate toward specific target areas. The use of magnetic positional information has been demonstrated in several diverse animals including sea turtles, spiny lobsters, newts and birds, suggesting that such systems are phylogenetically widespread and can function over a wide range of spatial scales. These `magnetic maps' have not yet been fully characterized. They may be organized in several fundamentally different ways, some of which bear little resemblance to human maps, and they may also be used in conjunction with unconventional navigational strategies.

Differential expression of gill Na+,K+-ATPase alpha- and beta-subunits, Na+,K+,2Cl- cotransporter and CFTR anion channel in juvenile anadromous and landlocked Atlantic salmon Salmo salar.

Abstract: This study examines changes in gill Na(+),K(+)-ATPase (NKA) alpha- and beta-subunit isoforms, Na(+),K(+),2Cl(-) cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR I and II) in anadromous and landlocked strains of Atlantic salmon during parr-smolt transformation, and after seawater (SW) transfer in May/June. Gill NKA activity increased from February through April, May and June among both strains in freshwater (FW), with peak enzyme activity in the landlocked salmon being 50% below that of the anadromous fish in May and June. Gill NKA-alpha1b, -alpha3, -beta(1) and NKCC mRNA levels in anadromous salmon increased transiently, reaching peak levels in smolts in April/May, whereas no similar smolt-related upregulation of these transcripts occurred in juvenile landlocked salmon. Gill NKA-alpha1a mRNA decreased significantly in anadromous salmon from February through June, whereas alpha1a levels in landlocked salmon, after an initial decrease in April, remained significantly higher than those of the anadromous smolts in May and June. Following SW transfer, gill NKA-alpha1b and NKCC mRNA increased in both strains, whereas NKA-alpha1a decreased. Both strains exhibited a transient increase in gill NKA alpha-protein abundance, with peak levels in May. Gill alpha-protein abundance was lower in SW than corresponding FW values in June. Gill NKCC protein abundance increased transiently in anadromous fish, with peak levels in May, whereas a slight increase was observed in landlocked salmon in May, increasing to peak levels in June. Gill CFTR I mRNA levels increased significantly from February to April in both strains, followed by a slight, though not significant increase in May and June. CFTR I mRNA levels were significantly lower in landlocked than anadromous salmon in April/June. Gill CFTR II mRNA levels did not change significantly in either strain. Our findings demonstrates that differential expression of gill NKA-alpha1a, -alpha1b and -alpha3 isoforms may be important for potential functional differences in NKA, both during preparatory development and during salinity adjustments in salmon. Furthermore, landlocked salmon have lost some of the unique preparatory upregulation of gill NKA, NKCC and, to some extent, CFTR anion channel associated with the development of hypo-osmoregulatory ability in anadromous salmon.

Whole-animal metabolic rate is a repeatable trait: a meta-analysis

Abstract: Repeatability studies are gaining considerable interest among physiological ecologists, particularly in traits affected by high environmental/residual variance, such as whole-animal metabolic rate (MR). The original definition of repeatability, known as the intraclass correlation coefficient, is computed from the components of variance obtained in a one-way ANOVA on several individuals from which two or more measurements are performed. An alternative estimation of repeatability, popular among physiological ecologists, is the Pearson product-moment correlation between two consecutive measurements. However, despite the more than 30 studies reporting repeatability of MR, so far there is not a definite synthesis indicating: (1) whether repeatability changes in different types of animals; (2) whether some kinds of metabolism are more repeatable than others; and most important, (3) whether metabolic rate is significantly repeatable. We performed a meta-analysis to address these questions, as well as to explore the historical trend in repeatability studies. Our results show that metabolic rate is significantly repeatable and its effect size is not statistically affected by any of the mentioned factors (i.e. repeatability of MR does not change in different species, type of metabolism, time between measurements, and number of individuals). The cumulative meta-analysis revealed that repeatability studies in MR have already reached an asymptotical effect size with no further change either in its magnitude and/or variance (i.e. additional studies will not contribute significantly to the estimator). There was no evidence of strong publication bias.

Effects of ship noise on the detectability of communication signals in the Lusitanian toadfish

Abstract: Underwater noise pollution is an increasing environmental problem which might affect communication, behaviour, fitness and consequently species' survival. The most common anthropogenic noises in aquatic habitats derive from shipping. In the present study we investigated the implications of noise pollution from a ship on the sound detectability, namely of conspecific vocalizations in the Lusitanian toadfish, Halobatrachus didactylus. Ambient and ferry-boat noises were recorded in the Tagus River estuary (Portugal), as well as toadfish sounds, and their sound pressure levels determined. Hearing sensitivities were measured under quiet lab conditions and in the presence of these masking noises at levels encountered in the field, using the auditory evoked potentials (AEP) recording technique. The Lusitanian toadfish is a hearing generalist, with best hearing sensitivity at low frequencies between 50 and 200 Hz (below 100 dB re. 1 microPa). Under ambient noise conditions, hearing was only slightly masked at lower frequencies. In the presence of ship noise, auditory thresholds increased considerably, by up to 36 dB, at most frequencies tested. This is mainly because the main energies of ferry-boat noise were within the most sensitive hearing range of this species. Comparisons between masked audiograms and sound spectra of the toadfish's mating and agonistic vocalizations revealed that ship noise decreased the ability to detect conspecific acoustic signals. This study provides the first evidence that fishes' auditory sensitivity can be impaired by ship noise and that acoustic communication, which is essential during agonistic encounters and mate attraction, might be restricted in coastal environments altered by human activity.

Biomechanics of bird flight.

Abstract: SUMMARY Power output is a unifying theme for bird flight and considerable progress has been accomplished recently in measuring muscular, metabolic and aerodynamic power in birds. The primary flight muscles of birds, the pectoralis and supracoracoideus, are designed for work and power output, with large stress (force per unit cross-sectional area) and strain (relative length change) per contraction. U-shaped curves describe how mechanical power output varies with flight speed, but the specific shapes and characteristic speeds of these curves differ according to morphology and flight style. New measures of induced, profile and parasite power should help to update existing mathematical models of flight. In turn, these improved models may serve to test behavioral and ecological processes. Unlike terrestrial locomotion that is generally characterized by discrete gaits, changes in wing kinematics and aerodynamics across flight speeds are gradual. Take-off flight performance scales with body size, but fully revealing the mechanisms responsible for this pattern awaits new study. Intermittent flight appears to reduce the power cost for flight, as some species flap–glide at slow speeds and flap–bound at fast speeds. It is vital to test the metabolic costs of intermittent flight to understand why some birds use intermittent bounds during slow flight. Maneuvering and stability are critical for flying birds, and design for maneuvering may impinge upon other aspects of flight performance. The tail contributes to lift and drag; it is also integral to maneuvering and stability. Recent studies have revealed that maneuvers are typically initiated during downstroke and involve bilateral asymmetry of force production in the pectoralis. Future study of maneuvering and stability should measure inertial and aerodynamic forces. It is critical for continued progress into the biomechanics of bird flight that experimental designs are developed in an ecological and evolutionary context.

Echolocation signals of wild harbour porpoises, Phocoena phocoena.

Abstract: Field recordings of harbour porpoises (Phocoena phocoena) were made in the inner Danish waters with a vertical array of three or four hydrophones. The back-calculated source level ranged from 178 to 205 dB re 1 muPa pp @ 1 m with a mean source level of 191 dB re 1 muPa pp @ 1 m. The maximum source level was more than 30 dB above what has been measured from captive animals, while the spectral and temporal properties were comparable. Calculations based on the sonar equation indicate that harbour porpoises, using these high click intensities, should be capable of detecting fish and nets and should be detectable by porpoise detectors over significantly larger distances than had previously been assumed. Harbour porpoises in this study preferred a relatively constant inter-click interval of about 60 ms, but intervals up to 200 ms and down to 30 ms were also recorded.

Running stability is enhanced by a proximo-distal gradient in joint neuromechanical control

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The role of larval fat cells in adult Drosophila melanogaster.

Abstract: In the life history of holometabolous insects, distinct developmental stages are tightly linked to feeding and non-feeding periods. The larval stage is characterized by extensive feeding, which supports the rapid growth of the animal and allows accumulation of energy stores, primarily in the larval fat body. In Drosophila melanogaster access to these stores during pupal development is possible because the larval fat body is preserved in the pupa as individual fat cells. These larval fat cells are refractive to autophagic cell death that removes most of the larval cells during metamorphosis. The larval fat cells are thought to persist into the adult stage and thus might also have a nutritional role in the young adult. We used cell markers to demonstrate that the fat cells in the young adult are in fact dissociated larval fat body cells, and we present evidence that these cells are eventually removed in the adult by a caspase cascade that leads to cell death. By genetically manipulating the lifespan of the larval fat cells, we demonstrate that these cells are nutritionally important during the early, non-feeding stage of adulthood. We experimentally blocked cell death of larval fat cells using the GAL4/UAS system and found that in newly eclosed adults starvation resistance increased from 58 h to 72 h. Starvation survival was highly correlated with the number of remaining larval fat cells. We discuss the implications of these results in terms of the overall nutritional status of the larva as an important factor in adult survival in environmental stresses such as starvation.

The desert ant odometer: a stride integrator that accounts for stride length and walking speed.

Abstract: Desert ants, Cataglyphis, use path integration as a major means of navigation. Path integration requires measurement of two parameters, namely, direction and distance of travel. Directional information is provided by a celestial compass, whereas distance measurement is accomplished by a stride integrator, or pedometer. Here we examine the recently demonstrated pedometer function in more detail. By manipulating leg lengths in foraging desert ants we could also change their stride lengths. Ants with elongated legs ('stilts') or shortened legs ('stumps') take larger or shorter strides, respectively, and misgauge travel distance. Travel distance is overestimated by experimental animals walking on stilts, and underestimated by animals walking on stumps - strongly indicative of stride integrator function in distance measurement. High-speed video analysis was used to examine the actual changes in stride length, stride frequency and walking speed caused by the manipulations of leg length. Unexpectedly, quantitative characteristics of walking behaviour remained almost unaffected by imposed changes in leg length, demonstrating remarkable robustness of leg coordination and walking performance. These data further allowed normalisation of homing distances displayed by manipulated animals with regard to scaling and speed effects. The predicted changes in homing distance are in quantitative agreement with the experimental data, further supporting the pedometer hypothesis.

Muscles do more positive than negative work in human locomotion.

Abstract: Summary Muscle work during level walking and ascent and descent ramp and stairway walking was assessed in order to explore the proposition that muscles perform more positive than negative work during these locomotion tasks. Thirty four healthy human adults were tested while maintaining a constant average walking velocity in the five gait conditions. Ground reaction force and sagittal plane kinematic data were obtained during the stance phases of these gaits and used in inverse dynamic analyses to calculate joint torques and powers at the hip, knee and ankle. Muscle work was derived as the area under the joint power vs time curves and was partitioned into positive, negative and net components. Dependent t-tests were used to compare positive and negative work in level walking and net joint work between ascent and descent gaits on the ramp and stairs (P<0.010). Total negative and positive work in level walking was −34 J and 50 J, respectively, with the difference in magnitude being statistically significant (P<0.001). Level walking was therefore performed with 16 J of net positive muscle work per step. The magnitude of the net work in ramp ascent was 25% greater than the magnitude of net work in ramp descent (89 vs −71 J m −1 , P<0.010). Similarly, the magnitude of the net work in stair ascent was 43% greater than the magnitude of net work in stair descent (107 vs −75 J step −1 , P<0.000). We identified three potential causes for the reduced negative vs positive work in these locomotion tasks: (1) the larger magnitude of the accelerations induced by the larger ground reaction forces in descending compared to ascending gaits elicited greater energy dissipation in non-muscular tissues, (2) the ground reaction force vector was directed closer to the joint centers in ramp and stair descent compared to ascent, which reduced the load on the muscular tissues and their energy dissipating response, and (3) despite the need to produce negative muscle work in descending gaits, both ramp and stair descent also had positive muscle work to propel the lower extremity upward and forward into the swing phase movement trajectory. We used these data to formulate two novel hypotheses about human locomotion. First, level walking requires muscles to generate a net positive amount of work per gait cycle to overcome energy losses by other tissues. Second, skeletal muscles generate more mechanical energy in gait tasks that raise the center of mass compared to the mechanical energy they dissipate in gait tasks that lower the center of mass, despite equivalent changes in total mechanical energy.

Temperature adaptation in two bivalve species from different thermal habitats: energetics and remodelling of membrane lipids.

Abstract: We compared lipid dynamics and the physiological responses of blue mussels Mytilus edulis, a cold-adapted species, and oysters Crassostrea virginica, a warmer-water species, during simulated overwintering and passage to spring conditions. To simulate overwintering, animals were held at 0 degrees C, 4 degrees C and 9 degrees C for 3 months and then gradually brought to and maintained at 20 degrees C for 5 weeks to simulate spring-summer conditions. Changes in lipid class and fatty acid composition were related to clearance rate and oxygen consumption. We found major differences between species in triglyceride (TAG) metabolism during overwintering. Mussels used digestive gland TAG stores for energy metabolism or reproductive processes during the winter, whereas oysters did not accumulate large TAG stores prior to overwintering. Mussel TAG contained high levels of 20:5n-3 compared to levels in oysters and in the diet. This may help to counteract the effect of low temperature by reducing the melting point of TAG and thus increasing the availability of storage fats at low temperature. Mussels seemed better able to mobilise 20:5n-3 and 18:4n-3 than other fatty acids. We also found that both bivalves underwent a major remodelling of membrane phospholipids. The unsaturation index decreased in the gills and digestive glands of both species during the early stages of warming, principally due to decreases in 22:6n-3 and 20:5n-3. In digestive glands, the unsaturation index did not increase with decreasing temperature beyond a threshold attained at 9 degrees C whereas a perfect negative relationship was observed in gills, as predicted by homeoviscous adaptation. The presence of digestive enzymes and acids in the digestive gland microenvironment may lead to specific requirements for membrane stability. That oysters had lower metabolic rates than mussels coincides with a lower unsaturation index of their lipids, as predicted by Hulbert's theory of membranes as metabolic pacemakers. Both species showed increased 20:4n-6 levels in their tissues as temperature rose, suggesting an increasing availability of this fatty acid for eicosanoid biosynthesis during stress responses. The contrast between the species in TAG dynamics and the similarity of their phospholipid remodelling emphasises the essential functional role of membrane phospholipid structure and the contrasting use of TAG by oysters and mussels during overwintering.

Honeybees perform optimal scale-free searching flights when attempting to locate a food source.

Abstract: The foraging strategies used by animals are key to their success in spatially and temporally heterogeneous environments. We hypothesise that when a food source at a known location ceases to be available, flying insects will exhibit search patterns that optimise the rediscovery of such resources. In order to study these searching patterns, foraging honeybees were trained to an artificial feeder that was then removed, and the subsequent flight patterns of the bees were recorded using harmonic radar. We show that the flight patterns have a scale-free (Levy-flight) characteristic that constitutes an optimal searching strategy for the location of the feeder. It is shown that this searching strategy would remain optimal even if the implementation of the Levy-flights was imprecise due, for example, to errors in the bees' path integration system or difficulties in responding to variable wind conditions. The implications of these findings for animal foraging in general are discussed.

Regulation of feeding behaviour and locomotor activity by takeout in Drosophila.

Abstract: The hormonal regulation of feeding behaviour is well known in vertebrates, whereas it remains poorly understood in insects. Here, we report that the takeout gene is an essential component of nutritional homeostasis in Drosophila. takeout encodes a putative juvenile hormone (JH) binding protein and has been described as a link between circadian rhythm and feeding behaviour. However, the physiological role of takeout and its putative link to JH remain unknown. In this study, we show that takeout (to(1)) flies failed to adapt their food intake according to food availability and that most defects could be genetically rescued. When food is abundant, to(1) are hyperphagic, yielding to hypertrophy of the fat body. When food reappears after a starvation period, to(1) flies do not increase their food intake as much as wild-type flies. This defect in food intake regulation is partly based on the action of Takeout on taste neurons, because the sensitivity of to(1) gustatory neurons to sugars does not increase after starvation, as in wild-type neurons. This lack of regulation is also evident at the locomotor activity, which normally increases during starvation, a behaviour related to food foraging. In addition, to(1) flies lack sexual dimorphism of locomotor activity, which has previously been linked to the JH circulating level. Moreover, application of the JH analog methoprene rescues the phenotype. These results suggest that takeout plays a central role as a feeding regulator and may act by modulating the circulating JH level.

Holdfast heroics: comparing the molecular and mechanical properties of Mytilus californianus byssal threads.

Abstract: SUMMARY The marine mussel Mytilus californianus Conrad inhabits the most wave-exposed regions of the rocky intertidal by dint of its extraordinary tenacity. Tenacity is mediated in large part by the byssus, a fibrous holdfast structure. M. californianus byssal threads, which are mechanically superior to the byssal threads of other mytilids, are composed almost entirely of a consortium of three modular proteins known as the preCols. In this study, the complete primary sequence of preCols from M. californianus was deduced and compared to that of two related species with mechanically inferior byssal threads, M. edulis Linnaeus and M. galloprovincialis Lamarck in order to explore structure–function relationships. The preCols from M. californianus are more divergent from the other two species than they are from one another. However, the degree of divergence is not uniform among the various domains of the preCols, allowing us to speculate on their mechanical role. For instance, the extra spider silk-like runs of alanine-rich sequence in the flanking domains of M. californianus may increase crystalline order, enhancing strength and stiffness. Histidine-rich domains at the termini, in contrast, are highly conserved between species, suggesting a mechanical role common to all three. Mechanical testing of pH-treated and chemically derivatized distal threads strongly suggests that histidine side chains are ligands in reversible, metal-mediated cross-links in situ. By combining the mechanical and sequence data, yield and self-healing in the distal region of threads have been modeled to emphasize the intricate interplay of enthalpic and entropic effects during tensile load and recovery.

Unsteady locomotion: integrating muscle function with whole body dynamics and neuromuscular control.

Abstract: By integrating studies of muscle function with analysis of whole body and limb dynamics, broader appreciation of neuromuscular function can be achieved. Ultimately, such studies need to address non-steady locomotor behaviors relevant to animals in their natural environments. When animals move slowly they likely rely on voluntary coordination of movement involving higher brain centers. However, when moving fast, their movements depend more strongly on responses controlled at more local levels. Our focus here is on control of fast-running locomotion. A key observation emerging from studies of steady level locomotion is that simple spring-mass dynamics, which help to economize energy expenditure, also apply to stabilization of unsteady running. Spring-mass dynamics apply to conditions that involve lateral impulsive perturbations, sudden changes in terrain height, and sudden changes in substrate stiffness or damping. Experimental investigation of unsteady locomotion is challenging, however, due to the variability inherent in such behaviors. Another emerging principle is that initial conditions associated with postural changes following a perturbation define different context-dependent stabilization responses. Distinct stabilization modes following a perturbation likely result from proximo-distal differences in limb muscle architecture, function and control strategy. Proximal muscles may be less sensitive to sudden perturbations and appear to operate, in such circumstances, under feed-forward control. In contrast, multiarticular distal muscles operate, via their tendons, to distribute energy among limb joints in a manner that also depends on the initial conditions of limb contact with the ground. Intrinsic properties of these distal muscle–tendon elements, in combination with limb and body dynamics, appear to provide rapid initial stabilizing mechanisms that are often consistent with spring-mass dynamics. These intrinsic mechanisms likely help to simplify the neural control task, in addition to compensating for delays inherent to subsequent force- and length-dependent neural feedback. Future work will benefit from integrative biomechanical approaches that employ a combination of modeling and experimental techniques to understand how the elegant interplay of intrinsic muscle properties, body dynamics and neural control allows animals to achieve stability and agility over a variety of conditions.

Effective limb length and the scaling of locomotor cost in terrestrial animals

Abstract: Relative to body size, smaller animals use more energy to travel a given distance than larger animals, but the anatomical variable driving this negative allometry remains the subject of debate. Here, I report a simple inverse relationship between effective limb length (i.e. hip height) and the energy cost of transport (COT; J kg(-1) m(-1)) for terrestrial animals. Using published data for a diverse set of terrestrial species including birds, mammals, reptiles and arthropods, I show that between-species differences in locomotor cost are driven by differences in limb length. Notably, there is no independent effect of body mass on cost. Remarkably, effective limb length explains 98% of the observed variance in locomotor cost across a wide range of terrestrial species including mammals, birds, reptiles and arthropods. Variation about the limb-length/COT scaling relationship is attributable to taxonomic differences in limb design, with birds and arthropods exhibiting greater residuals than mammals. Differences in COT between semi-aquatic, generalist and cursorial species also corresponds to differences in leg length between these groups. These results are discussed in light of previous investigations of the limb length and locomotor cost.

Automatic aeroelastic devices in the wings of a steppe eagle Aquila nipalensis.

Abstract: SUMMARY Here we analyse aeroelastic devices in the wings of a steppe eagle Aquila nipalensis during manoeuvres. Chaotic deflections of the upperwing coverts observed using video cameras carried by the bird (50 frames s –1 ) indicate trailing-edge separation but attached flow near the leading edge during flapping and gust response, and completely stalled flows upon landing. The underwing coverts deflect automatically along the leading edge at high angle of attack. We use high-speed digital video (500 frames s –1 ) to analyse these deflections in greater detail during perching sequences indoors and outdoors. Outdoor perching sequences usually follow a stereotyped three-phase sequence comprising a glide, pitch-up manoeuvre and deep stall. During deep stall, the spread-eagled bird has aerodynamics reminiscent of a cross-parachute. Deployment of the underwing coverts is closely phased with wing sweeping during the pitch-up manoeuvre, and is accompanied by alula protraction. Surprisingly, active alula protraction is preceded by passive peeling from its tip. Indoor flights follow a stereotyped flapping perching sequence, with deployment of the underwing coverts closely phased with alula protraction and the end of the downstroke. We propose that the underwing coverts operate as an automatic high-lift device, analogous to a Kruger flap. We suggest that the alula operates as a strake, promoting formation of a leading-edge vortex on the swept hand-wing when the arm-wing is completely stalled, and hypothesise that its active protraction is stimulated by its initial passive deflection. These aeroelastic devices appear to be used for flow control to enhance unsteady manoeuvres, and may also provide sensory feedback.

Gill morphology of the mangrove killifish (Kryptolebias marmoratus) is plastic and changes in response to terrestrial air exposure

Abstract: Amphibious mangrove killifish, Kryptolebias marmoratus (formerly Rivulus marmoratus), are frequently exposed to aerial conditions in their natural environment. We tested the hypothesis that gill structure is plastic and that metabolic rate is maintained in response to air exposure. During air exposure, when gills are no longer functional, we predicted that gill surface area would decrease. In the first experiment, K. marmoratus were exposed to either water (control) or air for 1 h, 1 day, 1 week, or 1 week followed by a return to water for 1 week (recovery). Scanning electron micrographs (SEM) and light micrographs of gill sections were taken, and morphometric analyses of lamellar width, lamellar length and interlamellar cell mass (ILCM) height were performed. Following 1 week of air exposure, SEM indicated that there was a decrease in lamellar surface area. Morphometric analysis of light micrographs revealed that there were significant changes in the height of the ILCM, but there were no significant differences in lamellae width and length between any of the treatments. Following 1 week of recovery in water, the ILCM regressed and gill lamellae were similar to control fish, indicating that the morphological changes were reversible. In the second experiment, V(CO(2)) was measured in fish continuously over a 5-day period in air and compared with previous measurements of oxygen uptake (V(O(2))) in water. V(CO(2)) varied between 6 and 10 micromol g(-1) h(-1) and was significantly higher on days 3, 4 and 5 relative to days 1 and 2. In contrast to V(O(2)) in water, V(CO(2)) in air showed no diurnal rhythm over a 24 h period. These findings indicate that K. marmoratus remodel their gill structures in response to air exposure and that these changes are completely reversible. Furthermore, over a similar time frame, changes in V(CO(2)) indicate that metabolic rate is maintained at a rate comparable to that of fish in water, underlying the remarkable ability of K. marmoratus to thrive in both aquatic and terrestrial habitats.

Extreme anoxia tolerance in embryos of the annual killifish Austrofundulus limnaeus: insights from a metabolomics analysis

Abstract: SUMMARY The annual killifish Austrofundulus limnaeus survives in ephemeral pond habitats by producing drought-tolerant diapausing embryos. These embryos probably experience oxygen deprivation as part of their normal developmental environment. We assessed the anoxia tolerance of A. limnaeus embryos across the duration of embryonic development. Embryos develop a substantial tolerance to anoxia during early development, which peaks during diapause II. This extreme tolerance of anoxia is retained during the first 4 days of post-diapause II development and is then lost. Metabolism during anoxia appears to be supported mainly by production of lactate, with alanine and succinate production contributing to a lesser degree. Anoxic embryos also accumulate large quantities of γ-aminobutyrate (GABA), a potential protector of neural function. It appears that the suite of characters associated with normal development and entry into diapause II in this species prepares the embryos for long-term survival in anoxia even while the embryos are exposed to aerobic conditions. This is the first report of such extreme anoxia tolerance in a vertebrate embryo, and introduces a new model for the study of anoxia tolerance in vertebrates.

Changes in mitochondrial oxidative capacities during thermal acclimation of rainbow trout Oncorhynchus mykiss: roles of membrane proteins, phospholipids and their fatty acid compositions.

Abstract: Changes in the properties of mitochondria from oxidative muscle of rainbow trout Oncorhynchus mykiss were examined during warm (5 degrees C to 15 degrees C) acclimation. Trout were studied shortly after the initial thermal change and after 8 weeks acclimation to 15 degrees C. To identify potential mechanisms by which oxidative capacities change, the modifications of phospholipid composition, membrane proteins and functional capacities of red muscle mitochondria were examined. Marked functional changes of isolated muscle mitochondria during warm acclimation of rainbow trout were reflected by a host of modifications in phospholipid composition, but by few shifts in protein components. Shortly after transfer of trout from 5 degrees C to 15 degrees C, the maximal oxidative capacity of mitochondria measured at 15 degrees C increased slightly, but rates at both assay temperatures (5 degrees C and 15 degrees C) decreased markedly after warm acclimation. The increase in capacity in short-term warm exposed trout was most pronounced when rates at 15 degrees C were expressed relative to cytochrome a and c(1) levels. Non-phosphorylating (State 4) rates of oxygen uptake increased with short-term warm exposure before returning to initial levels after warm acclimation. Cytochrome c oxidase (CCO) activity in the mitochondrial preparations decreased with warm acclimation. The thermal sensitivity of the ADP affinity was markedly modified during short-term warm exposure, when the ADP/O ratio increased, but warm acclimation returned these values to those observed initially. ADP affinity increased after warm acclimation. Changes in the mitochondrial content of cytochromes and adenine nucleotide translocase (ANT) could not explain these patterns. On the other hand, changes in the proportions of the lipid classes and in the acyl chain composition of certain phospholipid classes mirror the modifications in functional properties. Short-term exposure to 15 degrees C decreased the ratio of diacylphosphatidylethanolamine/diacylphosphatidylcholine (diacylPE/diacylPC), whereas warm acclimation led to restructuring of fatty acids (FA) and to increases of plasmalogen forms of PE and PC. Modification of overall membrane unsaturation did not appear to be the primary aim of restructuring membrane FA during warm acclimation, as total mitochondrial phospholipids and the major phospholipid classes only showed slight shifts of their acyl composition with warm acclimation. On the other hand, natural lysophosphatidylcholine (LysoPC) showed dramatic changes in FA content, as 16:0 and 18:1n-9 doubled whereas 22:6n-3 decreased from around 50% to 32% in warm acclimated trout. Similarly, in cardiolipin (CL), the levels of 16:0 and 18:1n-7 halved while 18:2n-6 increased to over 20% of the FA with warm acclimation. Given the central role of CL in modulating the activity of CCO, F(0)F(1)-ATPase and ANT, these changes suggest that specific compositional changes in CL are important modulators of mitochondrial capacities. The many structural changes in membrane lipids contrast with the limited modifications of the membrane protein components examined and support the concept of lipid structure modulating mitochondrial capacities.