Elucidating the complex organization of neural micro-domains in the locust Schistocerca gregaria using dMRI.

To understand brain function it is necessary to characterize both the underlying structural connectivity between neurons and the physiological integrity of these connections. Previous research exploring insect brain connectivity has typically used electron microscopy techniques, but this methodology cannot be applied to living animals and so cannot be used to understand dynamic physiological processes. The relatively large brain of the desert locust, Schistercera gregaria (Forksȧl) is ideal for exploring a novel methodology; micro diffusion magnetic resonance imaging (micro-dMRI) for the characterization of neuronal connectivity in an insect brain. The diffusion-weighted imaging (DWI) data were acquired on a preclinical system using a customised multi-shell diffusion MRI scheme optimized to image the locust brain. Endogenous imaging contrasts from the averaged DWIs and Diffusion Kurtosis Imaging (DKI) scheme were applied to classify various anatomical features and diffusion patterns in neuropils, respectively. The application of micro-dMRI modelling to the locust brain provides a novel means of identifying anatomical regions and inferring connectivity of large tracts in an insect brain. Furthermore, quantitative imaging indices derived from the kurtosis model that include fractional anisotropy (FA), mean diffusivity (MD) and kurtosis anisotropy (KA) can be extracted. These metrics could, in future, be used to quantify longitudinal structural changes in the nervous system of the locust brain that occur due to environmental stressors or ageing.

Do the enlarged hind legs of male thick-legged flower beetles contribute to take-off or mating?

The volume of the hind femora in the adult male flower beetle Oedemera nobilis is 38 times greater than in adult females. To determine what advantage limbs with swollen femora might provide, the behaviour of these insects was analysed with high-speed videography. First, because large hind legs are often associated with jumping and take-off, the performance of this behaviour by the two sexes was determined. Take-off was generated by a series of small-amplitude wing beats followed by larger ones, with the hind legs contributing little or no propulsion. The mean acceleration time to take-off was not significantly different in males (46.2 ms) and females (45.5 ms), but the mean take-off velocity of males was 10% higher than in females. Second, to determine if enlarged hind legs were critical in specifically male behaviour, interactions between males and females, and between males were videoed. The male mounted a female and then encircled her abdomen between the enlarged femora and tibiae of both his hind legs. The joint between these leg parts acted like a mole wrench (vice grip) so that when the tibia was fully flexed, a triangular space of 0.3 mm2 remained, in which a female abdomen (cross-sectional area 0.9 mm2) could be compressed and restrained firmly without inflicting damage. The flexor tibiae muscle in a male hind femur was 5.9 times larger than the extensor. In interactions between males, attempts to achieve a similar entrapment were frequently thwarted by the pursued male extending his hind legs vertically.

Effectiveness and efficiency of two distinct mechanisms for take-off in a derbid planthopper insect.

Analysis of the kinematics of take-off in the planthopper Proutista moesta (Hemiptera, Fulgoroidea, family Derbidae) from high-speed videos showed that these insects used two distinct mechanisms involving different appendages. The first was a fast take-off (55.7% of 106 take-offs by 11 insects) propelled by a synchronised movement of the two hind legs and without participation of the wings. The body was accelerated in 1 ms or less to a mean take-off velocity of 1.7 m s-1 while experiencing average forces of more than 150 times gravity. The power required from the leg muscles implicated a power-amplification mechanism. Such take-offs propelled the insect along its trajectory a mean distance of 7.9 mm in the first 5 ms after take-off. The second and slower take-off mechanism (44.3% of take-offs) was powered by beating movements of the wings alone, with no discernible contribution from the hind legs. The resulting mean acceleration time was 16 times slower at 17.3 ms, the mean final velocity was six times lower at 0.27 m s-1, the g forces experienced were 80 times lower and the distance moved in 5 ms after take-off was 7 times shorter. The power requirements could be readily met by direct muscle contraction. The results suggest a testable hypothesis that the two mechanisms serve distinct behavioural actions: the fast take-offs could enable escape from predators and the slow take-offs that exert much lower ground reaction forces could enable take-off from more flexible substrates while also displacing the insect in a slower and more controllable trajectory.

Take-off mechanisms in parasitoid wasps.

High-speed video analyses of the natural behaviour of parasitoid wasps revealed three strategies used to launch the insects into the air. Which strategy is the most energy efficient? In Pteromalus puparum, 92% of take-offs by were propelled entirely by movements of the middle and hind legs, which were depressed at their coxo-trochanteral and extended at their femoro-tibial joints. The front legs left the ground first, followed by the hind legs, so that the middle legs provided the final propulsion. Second, in other species of a similar mass, Cotesia glomerata and Leptopilina boulardi, all take-offs were propelled by a mean of 2.8 and 3.8 wingbeats, respectively, with little or no contribution from the legs. The first strategy resulted in take-off times that were four times shorter (5 versus 22.8 ms) and take-off velocities that were four times faster (0.8 versus 0.2 m s-1). Calculations from the kinematics indicate that propulsion by the legs was the most energy-efficient strategy, because more energy is put into propulsion of the body, whereas in take-off propelled by repetitive wing movements energy is lost to generating these movements and moving the air. In heavier species such as Netelia testacea and Amblyteles armatorius, take-off was propelled by the combined movements of the middle and hind legs and wingbeats. In A. armatorius, this resulted in the longest mean take-off time of 33.8 ms but an intermediate take-off velocity of 0.4 m s-1 In all three strategies the performance could be explained without invoking energy storage and power amplification mechanisms.

Three dimensional reconstruction of energy stores for jumping in planthoppers and froghoppers from confocal laser scanning microscopy.

Jumping in planthopper and froghopper insects is propelled by a catapult-like mechanism requiring mechanical storage of energy and its quick release to accelerate the hind legs rapidly. To understand the functional biomechanics involved in these challenging movements, the internal skeleton, tendons and muscles involved were reconstructed in 3-D from confocal scans in unprecedented detail. Energy to power jumping was generated by slow contractions of hind leg depressor muscles and then stored by bending specialised elements of the thoracic skeleton that are composites of the rubbery protein resilin sandwiched between layers of harder cuticle with air-filled tunnels reducing mass. The images showed that the lever arm of the power-producing muscle changed in magnitude during jumping, but at all joint angles would cause depression, suggesting a mechanism by which the stored energy is released. This methodological approach illuminates how miniaturized components interact and function in complex and rapid movements of small animals.

Jumping without slipping: leafhoppers (Hemiptera: Cicadellidae) possess special tarsal structures for jumping from smooth surfaces.

Many hemipteran bugs can jump explosively from plant substrates, which can be very smooth. We therefore analysed the jumping performance of froghoppers (Philaenus spumarius, Aphrophoridae) and leafhoppers (Aphrodes bicinctus/makarovi, Cicadellidae) taking off from smooth (glass) and rough (sandpaper, 30 µm asperity size) surfaces. On glass, the propulsive hind legs of Philaenus froghoppers slipped, resulting in uncontrolled jumps with a fast forward spin, a steeper angle and only a quarter of the velocity compared with jumps from rough surfaces. By contrast, Aphrodes leafhoppers took off without their propulsive hind legs slipping, and reached low take-off angles and high velocities on both substrates. This difference in jumping ability from smooth surfaces can be explained not only by the lower acceleration of the long-legged leafhoppers, but also by the presence of 2-9 soft pad-like structures (platellae) on their hind tarsi, which are absent in froghoppers. High-speed videos of jumping showed that platellae contact the surface briefly (approx. 3 ms) during the acceleration phase. Friction force measurements on individual hind tarsi on glass revealed that at low sliding speeds, both pushing and pulling forces were small, and insufficient to explain the recorded jumps. Only when the tarsi were pushed with higher velocities did the contact area of the platellae increase markedly, and high friction forces were produced, consistent with the observed jumps. Our findings show that leafhoppers have special adhesive footpads for jumping from smooth surfaces, which achieve firm grip and rapid control of attachment/detachment by combining anisotropic friction with velocity dependence.

Development and deposition of resilin in energy stores for locust jumping.

Locusts jump by using a catapult mechanism in which energy produced by slow contractions of the extensor tibiae muscles of the hind legs is stored in distortions of the exoskeleton, most notably (1) the two semi-lunar processes at each knee joint and (2) the tendons of the extensor muscles themselves. The energy is then suddenly released from these stores to power the rapid, propulsive movements of the hind legs. The reliance on the mechanical storage of energy is likely to impact on jumping because growth occurs by a series of five moults, at each of which the exoskeleton is replaced by a new one. All developmental stages (instars) nevertheless jump as a means of forward locomotion, or as an escape movement. Here, I show that in each instar, resilin is added to the semi-lunar processes and to the core of the extensor tendons so that their thickness increases. As the next moult approaches, a new exoskeleton forms within the old one, with resilin already present in the new semi-lunar processes. The old exoskeleton, the tendons and their resilin are discarded at moulting. The resilin of the semi-lunar processes and tendons of the new instar is initially thin, but a similar pattern of deposition results in an increase of their thickness. In adults, resilin continues to be deposited so that at 4 weeks old the thickness in the semi-lunar processes has increased fourfold. These changes in the energy stores accompany changes in jumping ability and performance during each moulting cycle.

Increased muscular volume and cuticular specialisations enhance jump velocity in solitarious compared with gregarious desert locusts, Schistocerca gregaria.

The desert locust, Schistocerca gregaria, shows a strong phenotypic plasticity. It can develop, depending upon population density, into either a solitarious or gregarious phase that differs in many aspects of behaviour, physiology and morphology. Prominent amongst these differences is that solitarious locusts have proportionately longer hind femora than gregarious locusts. The hind femora contain the muscles and energy-storing cuticular structures that propel powerful jumps using a catapult-like mechanism. We show that solitarious locusts jump on average 23% faster and 27% further than gregarious locusts, and attribute this improved performance to three sources: first, a 17.5% increase in the relative volume of their hind femur, and hence muscle volume; second, a 24.3% decrease in the stiffness of the energy-storing semi-lunar processes of the distal femur; and third, a 4.5% decrease in the stiffness of the tendon of the extensor tibiae muscle. These differences mean that solitarious locusts can generate more power and store more energy in preparation for a jump than can gregarious locusts. This improved performance comes at a cost: solitarious locusts expend nearly twice the energy of gregarious locusts during a single jump and the muscular co-contraction that energises the cuticular springs takes twice as long. There is thus a trade-off between achieving maximum jump velocity in the solitarious phase against the ability to engage jumping rapidly and repeatedly in the gregarious phase.

Jumping mechanisms in adult caddis flies (Insecta, Trichoptera).

To understand the jumping mechanisms and strategies of adult caddis flies, leg morphology and movements were analysed in three species with mean masses of 3.9 to 38 mg. Two distinct jumping strategies were found. First (67% of 90 jumps), take-off was propelled solely by the middle and hind legs while the wings remained closed. Second (33% of jumps), the same leg movements were combined with wing movements before take-off. The hind legs were 70% and the middle legs were 50% longer than the front legs and represented 105% and 88%, respectively, of body length. Both hind and middle trochantera were depressed together, approximately 15 ms before take-off. The front legs apparently did not contribute to thrust in either strategy and were the first to be lifted from the ground. The hind legs were the next to lose contact, so that the middle legs alone provided the final thrust before take-off. Jumping performance did not differ significantly in the two jumping strategies or between species, in acceleration times (range of means for the three species 14.5-15.4 ms), take-off velocities (range 0.7-1 m s(-1)) and trajectory angles. A significant difference in jumps propelled only by the legs was the lower angle (9.3 ± 1.9 deg) of the body relative to the horizontal at take-off compared with jumps involving wing movements (35.3 ± 2.5 deg). Calculations from the kinematics indicated that jumps were produced by direct muscle contractions and did not require power amplification or energy storage.

Jumping mechanisms and strategies in moths (Lepidoptera).

To test whether jumping launches moths into the air, take-off by 58 species, ranging in mass from 0.1 to 220 mg, was captured in videos at 1000 frames s(-1). Three strategies for jumping were identified. First, rapid movements of both middle and hind legs provided propulsion while the wings remained closed. Second, middle and hind legs again provided propulsion but the wings now opened and flapped after take-off. Third, wing and leg movements both began before take-off and led to an earlier transition to powered flight. The middle and hind legs were of similar lengths and were between 10 and 130% longer than the front legs. The rapid depression of the trochantera and extension of the middle tibiae began some 3 ms before similar movements of the hind legs, but their tarsi lost contact with the ground before take-off. Acceleration times ranged from 10 ms in the lightest moths to 25 ms in the heaviest ones. Peak take-off velocities varied from 0.6 to 0.9 m s(-1) in all moths, with the fastest jump achieving a velocity of 1.2 m s(-1). The energy required to generate the fastest jumps was 1.1 µJ in lighter moths but rose to 62.1 µJ in heavier ones. Mean accelerations ranged from 26 to 90 m s(-2) and a maximum force of 9 G: was experienced. The highest power output was within the capability of normal muscle so that jumps were powered by direct contractions of muscles without catapult mechanisms or energy storage.

Mantises exchange angular momentum between three rotating body parts to jump precisely to targets.

Flightless animals have evolved diverse mechanisms to control their movements in air, whether falling with gravity or propelling against it. Many insects jump as a primary mode of locomotion and must therefore precisely control the large torques generated during takeoff. For example, to minimize spin (angular momentum of the body) at takeoff, plant-sucking bugs apply large equal and opposite torques from two propulsive legs [1]. Interacting gear wheels have evolved in some to give precise synchronization of these legs [2, 3]. Once airborne, as a result of either jumping or falling, further adjustments may be needed to control trajectory and orient the body for landing. Tails are used by geckos to control pitch [4, 5] and by Anolis lizards to alter direction [6, 7]. When falling, cats rotate their body [8], while aphids [9] and ants [10, 11] manipulate wind resistance against their legs and thorax. Falling is always downward, but targeted jumping must achieve many possible desired trajectories. We show that when making targeted jumps, juvenile wingless mantises first rotated their abdomen about the thorax to adjust the center of mass and thus regulate spin at takeoff. Once airborne, they then smoothly and sequentially transferred angular momentum in four stages between the jointed abdomen, the two raptorial front legs, and the two propulsive hind legs to produce a controlled jump with a precise landing. Experimentally impairing abdominal movements reduced the overall rotation so that the mantis either failed to grasp the target or crashed into it head first.

Jumping mechanisms in lacewings (Neuroptera, Chrysopidae and Hemerobiidae).

Lacewings launch themselves into the air by simultaneous propulsive movements of the middle and hind legs as revealed in video images captured at a rate of 1000 s(-1). These movements were powered largely by thoracic trochanteral depressor muscles but did not start from a particular preset position of these legs. Ridges on the lateral sides of the meso- and metathorax fluoresced bright blue when illuminated with ultraviolet light, suggesting the presence of the elastic protein resilin. The middle and hind legs were longer than the front legs but their femora and tibiae were narrow tubes of similar diameter. Jumps were of two types. First, those in which the body was oriented almost parallel to the ground (-7±8 deg in green lacewings, 13.7±7 deg in brown lacewings) at take-off and remained stable once animals were airborne. The wings did not move until 5 ms after take-off when flapping flight ensued. Second, were jumps in which the head pointed downwards at take-off (green lacewings, -37±3 deg; brown lacewings, -35±4 deg) and the body rotated in the pitch plane once airborne without the wings opening. The larger green lacewings (mass 9 mg, body length 10.3 mm) took 15 ms and the smaller brown lacewings (3.6 mg and 5.3 mm) 9 ms to accelerate the body to mean take-off velocities of 0.6 and 0.5 m s(-1). During their fastest jumps green and brown lacewings experienced accelerations of 5.5 or 6.3 G: , respectively. They required an energy expenditure of 5.6 or 0.7 µJ, a power output of 0.3 or 0.1 mW and exerted a force of 0.6 or 0.2 mN. The required power was well within the maximum active contractile limit of normal muscle, so that jumping could be produced by direct muscle contractions without a power amplification mechanism or an energy store.

Jumping mechanisms in flatid planthoppers (Hemiptera, Flatidae).

The jumping performance of three species of hemipterans from Australia and Europe belonging to the family Flatidae was analysed from images captured at a rate of 5000 s(-1). The shape of a flatid was dominated by large triangular or wedge-shaped front wings, which, when folded, covered and extended above and behind the body to give a laterally compressed and possibly streamlined appearance. The body lengths of the three species of adults ranged from 7 to 9 mm and their mass from 8 to 19 mg. The propulsive hind legs were 30% longer than the front legs but only 36-54% of the body length. Jumps with the fastest take-off velocities of 2.8-3.2 m s(-1) had acceleration times of 1.4-1.8 ms. During such jumps, adults experienced an acceleration of 174-200 G: . These jumps required an energy expenditure of 76-225 µJ, a power output of 13-60 mW and exerted a force of 9-37 mN. The required power output per mass of jumping muscle in adults ranged from 24,000 to 27,000 W kg(-1) muscle, 100 times greater than the maximum active contractile limit of normal muscle. The free-living nymphs were also proficient jumpers, reaching take-off velocities of 2.2 m s(-1). To achieve such a jumping performance requires a power amplification mechanism. The energy store for such a mechanism was identified as the internal skeleton linking a hind coxa to the articulation of a hind wing. These pleural arches fluoresced bright blue when illuminated with UV light, indicating the presence of the elastic protein resilin. The energy generated by the prolonged contractions of the trochanteral depressor muscles was stored in distortions of these structures, and the rapid elastic recoil of these muscles powered the synchronous propulsive movements of the hind legs.

Rapid behavioural gregarization in the desert locust, Schistocerca gregaria entails synchronous changes in both activity and attraction to conspecifics.

Desert Locusts can change reversibly between solitarious and gregarious phases, which differ considerably in behaviour, morphology and physiology. The two phases show many behavioural differences including both overall levels of activity and the degree to which they are attracted or repulsed by conspecifics. Solitarious locusts perform infrequent bouts of locomotion characterised by a slow walking pace, groom infrequently and actively avoid other locusts. Gregarious locusts are highly active with a rapid walking pace, groom frequently and are attracted to conspecifics forming cohesive migratory bands as nymphs and/or flying swarms as adults. The sole factor driving the onset of gregarization is the presence of conspecifics. In several previous studies concerned with the mechanism underlying this transformation we have used an aggregate measure of behavioural phase state, Pgreg, derived from logistic regression analysis, which combines and weights several behavioural variables to characterise solitarious and gregarious behaviour. Using this approach we have analysed the time course of behavioural change, the stimuli that induce gregarization and the key role of serotonin in mediating the transformation. Following a recent critique that suggested that using Pgreg may confound changes in general activity with genuine gregarization we have performed a meta-analysis examining the time course of change in the individual behaviours that we use to generate Pgreg. We show that the forced crowding of solitarious locusts, tactile stimulation of the hind femora, and the short-term application of serotonin each induce concerted changes in not only locomotion-related variables but also grooming frequency and attraction to other locusts towards those characteristic of long-term gregarious locusts. This extensive meta-analysis supports and extends our previous conclusions that solitarious locusts undergo a rapid behavioural gregarization upon receiving appropriate stimulation for a few hours that is mediated by serotonin, at the end of which their behaviour is largely indistinguishable from locusts that have been in the gregarious phase their entire lives.

Jumping mechanisms in dictyopharid planthoppers (Hemiptera, Dicytyopharidae).

The jumping performance of four species of hemipterans belonging to the family Dictyopharidae, from Europe, South Africa and Australia, were analysed from high-speed images. The body shape in all was characterised by an elongated and tapering head that gave a streamlined appearance. The body size ranged from 6 to 9 mm in length and from 6 to 23 mg in mass. The hind legs were 80-90% of body length and 30-50% longer than the front legs, except in one species in which the front legs were particularly large so that all legs were of similar length. Jumping was propelled by rapid and simultaneous depression of the trochantera of both hind legs, powered by large muscles in the thorax, and was accompanied by extension of the tibiae. In the best jumps, defined as those with the fastest take-off velocity, Engela minuta accelerated in 1.2 ms to a take-off velocity of 5.8 m s(-1), which is the fastest achieved by any insect described to date. During such a jump, E. minuta experienced an acceleration of 4830 m s(-2) or 490 g, while other species in the same family experienced 225-375 g. The best jumps in all species required an energy expenditure of 76-225 µJ, a power output of 12-80 mW and exerted a force of 12-29 mN. The required power output per mass of jumping muscle ranged from 28,000 to 140,200 W kg(-1) muscle and thus greatly exceeded the maximum active contractile limit of normal muscle. To achieve such a jumping performance, these insects must be using a power amplification mechanism in a catapult-like action. It is suggested that their streamlined body shape improves jumping performance by reducing drag, which, for a small insect, can substantially affect forward momentum.

Interacting gears synchronize propulsive leg movements in a jumping insect.

Gears are found rarely in animals and have never been reported to intermesh and rotate functionally like mechanical gears. We now demonstrate functional gears in the ballistic jumping movements of the flightless planthopper insect Issus. The nymphs, but not adults, have a row of cuticular gear (cog) teeth around the curved medial surfaces of their two hindleg trochantera. The gear teeth on one trochanter engaged with and sequentially moved past those on the other trochanter during the preparatory cocking and the propulsive phases of jumping. Close registration between the gears ensured that both hindlegs moved at the same angular velocities to propel the body without yaw rotation. At the final molt to adulthood, this synchronization mechanism is jettisoned.

Jumping mechanisms in gum treehopper insects (Hemiptera, Eurymelinae).

Jumping in a species of Australian gum treehopper was analysed from high-speed images. Pauroeurymela amplicincta adults and nymphs lived together in groups that were tended by ants, but only adults jumped. The winged adults with a body mass of 23 mg and a body length of 7 mm had some morphological characteristics intermediate between those of their close relatives the leafhoppers (Cicadellidae) and the treehoppers (Membracidae). They, like leafhoppers, lacked the prominent prothoracic helmets of membracid treehoppers, and their large hind coxae were linked by press studs (poppers), that are present in leafhoppers but not treehoppers. The hindlegs were only 30-40% longer than the other legs and 67% of body length. They are thus of similar proportion to the hindlegs of treehoppers but much shorter than those of most leafhoppers. Jumping was propelled by the hindlegs, which moved in the same plane as each other beneath and almost parallel to the longitudinal axis of the body. A jump was preceded by full levation of the coxo-trochanteral joints of the hindlegs. In its best jumps, the rapid depression of these joints then accelerated the insect in 1.4 ms to a take-off velocity of 3.8 m s(-1) so that it experienced a force of almost 280 g. In 22% of jumps, the wings opened before take-off but did not flap until the gum treehopper was airborne, when the body rotated little in any plane. The energy expended was 170 µJ, the power output was 122 mW and the force exerted was 64 mN. Such jumps are predicted to propel the insect forwards 1450 mm (200 times body length) and to a height of 430 mm if there is no effect of wind resistance. The power output per mass of jumping muscle far exceeded the maximum active contractile limit of muscle and indicates that a catapult-like action must be used. This eurymelid therefore out-performs both leafhoppers and treehoppers in i ts faster acceleration and in its higher take-off velocity.

Jumping from the surface of water by the long-legged fly Hydrophorus (Diptera, Dolichopodidae).

The fly Hydrophorus alboflorens (4 mm long, 4.7 mg mass) moves around upon and jumps from water without its tarsi penetrating the surface. All six tarsi have a surface area of 1.3 mm(-2) in contact with the water, but they did not dimple its surface when standing. Jumping was propelled by depression of the trochantera of both hind and middle legs, which are 40% longer than the front legs and 170% longer than the body. As these four legs progressively propelled the insect to take-off, they each created dimples on the water surface that expanded in depth and area. No dimples were associated with the front legs, which were not moved in a consistent sequence. The wings opened while the legs were moving and then flapped at a frequency of 148 Hz. The body was accelerated in a mean time of 21 ms to a mean take-off velocity of 0.7 m s(-1). The best jumps reached velocities of 1.6 m s(-1), and required an energy output of 7 µJ and a power output of 0.6 mW, with the fly experiencing a force of 140 g. The required power output indicates that direct muscle contractions could propel the jump without the need for elaborate mechanisms for energy storage. Take-off trajectories were steep, with a mean of 87 deg to the horizontal. Take-off velocity fell if a propulsive tarsus penetrated the surface of the water. If more tarsi became submerged, take-off was not successful. A second strategy for take-off was powered only by the wings and was associated with slower (1 deg ms(-1) compared with 10 deg ms(-1) when jumping) and less extensive movements of the propulsive joints of the middle and hind legs. No dimples were then created on the surface of the water. When jumping was combined with wing flapping, the acceleration time to take-off was reduced by 84% and the take-off velocity was increased by 168%. Jumping can potentially therefore enhance survival when threatened by a potential predator.