Nanoscale mesh acts as anti-adhesive surface against particulate contamination in eyes of whiteflies.

In many insects the surface of the eye is nanostructured by arrays of protuberances termed ommatidial gratings which provide the cuticle with anti-reflective, anti-wetting and self-cleaning properties. The hypothesised anti-contamination role of the gratings against dust and pollen results from theoretical predictions on grating geometry and experiments on synthetic replicas of ommatidia surfaces but has not yet been proven in an animal. Whiteflies are biological test beds for anti-contamination surfaces because they deliberately distribute wax particles extruded from abdominal plates over their entire bodies. The numerous particles protect the animal against water evaporation and radiation, but may severely impair vision. Using scanning electron microscopy (SEM) and CryoSEM, we here show that the cornea of whiteflies exhibits ~ 220 nm wide mesh-like structures forming hexagonal gratings with thin ~ 40 nm connecting walls. Quantitative measurements of wax particles on the eye show that the nanostructures reduce particle contamination by more than ~ 96% compared to other areas of the cuticle. Altogether, our study is the first description of a predicted optimized grating geometry for anti-contamination in an arthropod. The findings serve as evidence of the high effectiveness of nanostructured surfaces for reducing contact area and thus adhesion forces between biological surfaces and contaminating particles.

Tracheal hyperallometry and spatial constraints in a large beetle.

Insects exchange respiratory gases with their environment through their gas-filled tracheal system, a branched tracheal tree extending from segmental openings and terminating at fine tissue penetrating tracheoles. It was shown that the tracheal volume increases hyperallometrically with insect body size (Mb), both interspecifically and across developmental stages. In this study, we used the sixfold Mb variation in adult Batocera rufomaculata(Cerambicidae; Coleoptera) examining the allometry of adult tracheal volume (Vtr). We further explored the effect of sex and sexual maturity on tracheal gas conductance, testing the hypotheses that (i) larger body size and (ii) egg volume in gravid females would result in lower safety margins for tracheal oxygen transport due to structural restriction. We report a hyperallometric tracheal growth in both sexes of adult B. rufomaculata(mean mass exponent of 1.42 ± 0.09), similar in magnitude to previously reported values. Tracheal gas conductance was independent of Mb and reproductive state, but was significantly higher in females compared with males. We suggest that females may have pre-adapted a higher tracheal conductance required for the higher flight power output while gravid. Lack of compliant air sacs and rigid trachea may explain how gravid females retain their Vtr. However, we show that Vtr outgrows thoracic dimensions with increased B. rufomaculatasize. Hyperallometric growth of the giant cerambycid thoracic trachea could explain the previously reported hypometric scaling of flight muscles in B. rufomaculata, and the compromised long-distance flight performance of larger compared with smaller conspecifics.

Intraspecific scaling and early life history determine the cost of free-flight in a large beetle (Batocera rufomaculata).

The scaling of the energetic cost of locomotion with body mass is well documented at the interspecific level. However, methodological restrictions limit our understanding of the scaling of flight metabolic rate (MR) in free-flying insects. This is particularly true at the intraspecific level, where variation in body mass and flight energetics may have direct consequences for the fitness of an individual. We applied a 13C stable isotope method to investigate the scaling of MR with body mass during free-flight in the beetle Batocera rufomaculata. This species exhibits large intraspecific variation in adult body mass as a consequence of the environmental conditions during larval growth. We show that the flight-MR scales with body mass to the power of 0.57, with smaller conspecifics possessing up to 2.3 fold higher mass-specific flight MR than larger ones. Whereas the scaling exponent of free-flight MR was found to be like that determined for tethered-flight, the energy expenditure during free-flight was more than 2.7 fold higher than for tethered-flight. The metabolic cost of flight should therefore be studied under free-flight conditions, a requirement now enabled by the 13C technique described herein for insect flight.

Metabolic cost of flight and aerobic efficiency in the rose chafer, Protaetia cuprea (Cetoniinae).

Rose chafer beetles (Protetia cuprea) are pollinators as well as agricultural pests, flying between flowers and trees while foraging for pollen and fruits. Calculating the energy they expend on flying during foraging activity faces the challenge of measuring the metabolic rate (MR) of free-flying insects in an open space. We overcame this challenge by using the bolus injection of 13 C Na-bicarbonate technique to measure their metabolic energy expenditure while flying in a large flight arena. Concurrently, we tracked the insects with high-speed cameras to extract their flight trajectory, from which we calculated the mechanical power invested in flying for each flight bout. We found that the chemical (metabolic) energy input converted to mechanical flight energy output at a mean efficiency of 10.4% ± 5.2%, with a trend of increased efficiency in larger conspecifics (efficiency scaled with body mass to the power of 1.4). The transition in the summer from a diet of pollen to that of fruits may affect the energy budget available for foraging. Starved P. cuprea, feeding on apples ad libitum, increased their body mass by an average of 6% in 2 h. According to our calculations, such a meal can power a 630-m flight (assuming a carbohydrate assimilation efficiency of 90%). Pollen, with a low water and carbohydrate content but rich in proteins and lipids, has a higher caloric content and should assimilate differently when converting food to flight fuel. The high cost of aerial locomotion is inherent to the foraging behavior of rose chafers, explaining their short flight bouts followed by prolonged feeding activity.

The relationship between body size and flight power output in the mango stem borer (Batocera rufomaculata).

Adult body size in insects can be influenced by environmental conditions during larval growth. The effect of such intraspecific variation in body mass on flight performance is poorly understood. In Batocera rufomaculata, a large tree boring beetle, adults emerging from larvae that developed in a dying host tree, and therefore, under nutrient-deprived diet conditions, are smaller but have an elevated long-distance flight capability compared to larger conspecifics that developed in viable host trees. The improved endurance for long-distance flight in the smaller individuals appears to contradict the interspecific trend in flying animals of a decrease in Cost of Transport (CoT) with increased body mass. To explore the relationship between intraspecific variation in body size and power expended during steady forward flight, we flew these beetles tethered in a wind tunnel and compared the flapping kinematics and power output of individuals varying in body mass (1-7 gr). Concurrently, we measured the forces the insects applied on the tether allowing us to evaluate the tethering effects and correct for them. From the flapping kinematics we estimated the mechanical power expended using a quasi-steady blade-element model. We found that muscle mass-specific power did not differ between small and large individuals flying at the same wind (flight) speed in the tunnel. Consequently, the CoT of B. rufomaculata does not vary with body mass. Such invariance of mass-specific power with body mass may aid the dispersal of smaller individuals from deteriorating host trees to new ones.

Insect flight metabolic rate revealed by bolus injection of the stable isotope 13C.

Measuring metabolic rate (MR) poses a formidable challenge in free-flying insects who cannot breathe into masks or be trained to fly in controlled settings. Consequently, flight MR has been predominantly measured on hovering or tethered insects flying in closed systems. Stable isotopes such as labelled water allow measurement of MR in free-flying animals but integrates the measurement over long periods exceeding the average flight duration of insects. Here, we applied the 'bolus injection of isotopic 13C Na-bicarbonate' method to insects to measure their flight MR and report a 90% accuracy compared to respirometry. We applied the method on two beetle species, measuring MR during free flight and tethered flight in a wind tunnel. We also demonstrate the ability to repeatedly use the technique on the same individual. Therefore, the method provides a simple, reliable and accurate tool that solves a long-lasting limitation on insect flight research by enabling the measurement of MR during free flight.

The Aerodynamics and Power Requirements of Forward Flapping Flight in the Mango Stem Borer Beetle (Batocera rufomaculata).

The need for long dispersal flights can drive selection for behavioral, physiological, and biomechanical mechanisms to reduce the energy spent flying. However, some energy loss during the transfer of momentum from the wing to the fluid is inevitable, and inherent to the fluid-wing interaction. Here, we analyzed these losses during the forward flight of the mango stem borer (Batocera rufomaculata). This relatively large beetle can disperse substantial distances in search of new host trees, and laboratory experiments have demonstrated continuous tethered flights that can last for up to an hour. We flew the beetles tethered in a wind tunnel and used high-speed videography to estimate the aerodynamic power from their flapping kinematics and particle image velocimetry (PIV) to evaluate drag and kinetic energy from their unsteady wakes. To account for tethering effects, we measured the forces applied by the beetles on the tether arm holding them in place. The drag of the flying beetle over the flapping cycle, estimated from the flow fields in the unsteady wake, showed good agreement with direct measurement of mean horizontal force. Both measurements showed that total drag during flight is ∼5-fold higher than the parasite drag on the body. The aerodynamic power estimated from both the motion of the wings, using a quasi-steady blade-element model, and the kinetic energy in the wake, gave mean values of flight-muscle mass-specific power of 87 and 65 W kg muscle-1, respectively. A comparison of the two values suggests that ∼25% of the energy is lost within the fluid due to turbulence and heat. The muscle mass-specific power found here is low relative to the maximal power output reported for insect flight muscles. This can be attributed to reduce weight support during tethered flight or to operation at submaximal output that may ensure a supply of metabolic substrates to the flight muscles, thus delaying their fatigue during long-distance flights.

The effect of air resistance on the jump performance of a small parasitoid wasp, Anagyrus pseudococci (Encyrtidae).

The distance a small insect moves through the air during a jump is limited by the launch velocity at take-off and by air resistance. The launch velocity is limited by the length of the jumping legs and the maximum power that the jump apparatus can provide for pushing against the ground. The effect of air resistance is determined by the insect mass-to-area ratio. Both limitations are highly dependent on body size, making high jumps a challenge for smaller insects. We studied both effects in the tiny Encyrtid wasp Anagyrus pseudococci. Males are smaller than females (mean body length 1.2 and 1.8 mm, respectively), but both sexes take off in a powerful jump. Using high-speed cameras, we analyzed the relationship between take-off kinematics and distance traveled through the air. We show that the velocity, acceleration and mass-specific power when leaving the ground places A. pseudococci among the most prominent jumpers of the insect world. However, the absolute distance moved through the air is modest compared with other jumping insects, as a result of air resistance acting on the small body. A biomechanical model suggests that air resistance reduces the jump distance of these insects by 49% compared with jumping in the absence of air resistance. The effect of air resistance is more pronounced in the smaller males, resulting in a segregation of the jumping performance between sexes. The limiting effect of air resistance is inversely proportional to body mass, seriously constraining jumping as a form of moving through the air in these and other small insects.