Stephen P. Roberts

Gene transcription during exposure to, and recovery from, cold and desiccation stress in Drosophila melanogaster

We exposed adult male Drosophila melanogaster to cold, desiccation or starvation, and examined expression of several genes during exposure and recovery. Frost was expressed during recovery from cold, and was up‐regulated during desiccation. Desiccation and starvation (but not cold) elicited increased expression of the senescence‐related gene smp‐30. Desat2 decreased during recovery from desiccation, but not in response to starvation or cold. Hsp70 expression increased after 1 h of recovery from cold exposure, but was unchanged in response to desiccation or starvation stress, and Hsp23 levels did not respond to any of the stressors. We conclude that D. melanogasters responses to cold and desiccation are quite different and that care must be taken to separate exposure and recovery when studying responses to environmental stress.

Achievement of Thermal Stability by Varying Metabolic Heat Production in Flying Honeybees

Thermoregulation of the thorax allows endothermic insects to achieve power outputs during flight that are among the highest in the animal kingdom. Flying endothermic insects, including the honeybee Apis mellifera, are believed to thermoregulate almost exclusively by varying heat loss. Here it is shown that a rise in air temperature from 20° to 40°C causes large decreases in metabolic heat production and wing-beat frequency in honeybees during hovering, agitated, or loaded flight. Thus, variation in heat production may be the primary mechanism for achieving thermal stability in flying honeybees, and this mechanism may occur commonly in endothermic insects.

Age and natural metabolically-intensive behavior affect oxidative stress and antioxidant mechanisms

Flying honey bees have among the highest mass-specific metabolic rates ever measured, suggesting that their flight muscles may experience high levels of oxidative stress during normal daily activities. We measured parameters of oxidative stress and antioxidant capacity in highly metabolic flight muscle and less active head tissue in cohorts of age-matched nurse bees, which rarely fly, and foragers, which fly several hours per a day. Naturally occurring foraging flight elicited an increase in flight muscle Hsp70 content in both young and old foragers; however catalase and total antioxidant capacity increased only in young flight muscle. Surprisingly, young nurse bees also showed a modest daily increase in Hsp70, catalase levels and antioxidant capacity, and these effects were likely due to collecting the young nurses soon after orientation flights. There were no differences in flight muscle carbonyl content over the course of daily activity and few differences in Hsp70, catalase, total antioxidant capacity and protein carbonyl levels in head tissue regardless of age or activity. In summary, honey bee flight likely produces high levels of reactive oxygen species in flight muscle that, when coupled with age-related decreases in antioxidant activity may be responsible for behavioral senescence and reduced longevity.

Ecological and Environmental Physiology of Insects

1. Introduction 2. Basic insect functional anatomy and physiological principles 3. Temperature 4. Water 5. Nutrition, growth, and size 6. Oxygen 7. Techniques and applications 8. Conclusions and future directions References Index

Cold rearing improves cold-flight performance in Drosophila via changes in wing morphology

SUMMARY We use a factorial experimental design to test whether rearing at colder temperatures shifts the lower thermal envelope for flight of Drosophila melanogaster Meigen to colder temperatures. D. melanogaster that developed in colder temperatures (15°C) had a significant flight advantage in cold air compared to flies that developed in warmer temperatures (28°C). At 14°C, cold-reared flies failed to perform a take-off flight∼ 47% of the time whereas warm-reared flies failed ∼94% of the time. At 18°C, cold- and warm-reared flies performed equally well. We also compared several traits in cold- and warm-developing flies to determine if cold-developing flies had better flight performance at cold temperatures due to changes in body mass, wing length, wing loading, relative flight muscle mass or wing-beat frequency. The improved ability to fly at low temperatures was associated with a dramatic increase in wing area and an increase in wing length (after controlling for wing area). Flies that developed at 15°C had∼ 25% more wing area than similarly sized flies that developed at 28°C. Cold-reared flies had slower wing-beat frequencies than similarly sized flies from warmer developmental environments, whereas other traits did not vary with developmental temperature. These results demonstrate that developmental plasticity in wing dimensions contributes to the improved flight performance of D. melanogaster at cold temperatures, and ultimately, may help D. melanogaster live in a wide range of thermal environments.

Effects of load type (pollen or nectar) and load mass on hovering metabolic rate and mechanical power output in the honey bee Apis mellifera.

SUMMARY In this study we tested the effect of pollen and nectar loading on metabolic rate (in mW) and wingbeat frequency during hovering, and also examined the effect of pollen loading on wing kinematics and mechanical power output. Pollen foragers had hovering metabolic rates approximately 10% higher than nectar foragers, regardless of the amount of load carried. Pollen foragers also had a more horizontal body position and higher inclination of stroke plane than measured previously for honey bees (probably nectar foragers). Thorax temperatures ranked pollen > nectar > water foragers, and higher flight metabolic rate could explain the higher thorax temperature of pollen foragers. Load mass did not affect hovering metabolic rate or wingbeat frequency in a regression-model experiment. However, using an analysis of variance (ANOVA) design, loaded pollen and nectar foragers (mean loads 27% and 40% of body mass, respectively) significantly increased metabolic rate by 6%. Mean pollen loads of 18% of body mass had no effect on wingbeat frequency, stroke amplitude, body angle or inclination of stroke plane, but increased the calculated mechanical power output by 16–18% (depending on the method of estimating drag). A rise in lift coefficient as bees carry loads without increasing wingbeat frequency or stroke amplitude (and only minimal increases in metabolic rate) suggests an increased use of unsteady power-generating mechanisms.

The effects of age and behavioral development on honey bee (Apis mellifera) flight performance

SUMMARY A critical but seldom-studied component of life history theory is how behavior and age affect whole-organism performance. To address this issue we compared the flight performance of honey bees (whose behavioral development and age can be assessed independently via simple manipulations of colony demographics) between distinct behavioral castes (in-hive nurse bees vs out-of-hive foragers) and across lifespan. Variable-density gases and high-speed video were used to determine the maximum hovering flight capacity and wing kinematics of age-matched nurse bees and foragers sampled from a single-cohort colony over a period of 34 days. The transition from hive work to foraging was accompanied by a 42% decrease in body mass and a proportional increase in flight capacity (defined as the minimum gas density allowing hovering flight). The lower flight capacity of hive bees was primarily due to the fact that in air they were functioning at a near-maximal wing angular velocity due to their high body masses. Foragers were lighter and when hovering in air required a much lower wing angular velocity, which they were able to increase by 32% during maximal flight performance. Flight performance of hive bees was independent of age, but in foragers the maximal wingbeat frequency and maximal average angular velocity were lowest in precocious (7–14 day old) foragers, highest in normal-aged (15–28 day old) foragers and intermediate in foragers older than 29 days. This pattern coincides with previously described age-dependent biochemical and metabolic properties of honey bee flight muscle.

Allometry of kinematics and energetics in carpenter bees (Xylocopa varipuncta) hovering in variable-density gases

SUMMARY We assessed the energetic and aerodynamic limits of hovering flight in the carpenter bee Xylocopa varipuncta. Using normoxic, variable-density mixtures of O2, N2 and He, we were able to elicit maximal hovering performance and aerodynamic failure in the majority of bees sampled. Bees were not isometric regarding thorax mass and wing area, both of which were disproportionately lower in heavier individuals. The minimal gas density necessary for hovering (MGD) increased with body mass and decreased with relative thoracic muscle mass. Only the four bees in our sample with the highest body mass-specific thorax masses were able to hover in pure heliox. Wingbeat frequency and stroke amplitude during maximal hovering were significantly greater than in normodense hovering, increased significantly with body mass during normodense hovering but were mass independent during maximal hovering. Reserve capacity for wingbeat frequency and stroke amplitude decreased significantly with increasing body mass, although reserve capacity in stroke amplitude (10–30%) exceeded that of wingbeat frequency (0–8%). Stroke plane angle during normodense hovering was significantly greater than during maximal hovering, whereas body angle was significantly greater during maximal hovering than during normodense hovering. Power production during normodense hovering was significantly less than during maximal hovering. Metabolic rates were significantly greater during maximal hovering than during normodense hovering and were inversely related to body mass during maximal and normodense hovering. Metabolic reserve capacity averaged 34% and was independent of body mass. Muscle efficiencies were slightly higher during normodense hovering. The allometry of power production, power reserve capacity and muscle efficiency were dependent on the assumed coefficient of drag (CD), with significant allometries most often at lower values of CD. Larger bees operate near the envelope of maximal performance even in normodense hovering due to smaller body mass-specific flight muscles and limited reserve capacities for kinematics and power production.

Muscle biochemistry and the ontogeny of flight capacity during behavioral development in the honey bee, Apis mellifera

SUMMARY A fundamental issue in physiology and behavior is understanding the functional and genetic mechanisms that underlie major behavioral shifts in organisms as they adopt new environments or life history tactics. Such transitions are common in nature and include the age-related switch from nest/hive work to foraging in social insects such as honey bees (Apis mellifera). Because of their experimental tractability, recently sequenced genome and well understood biology, honey bees are an ideal model system for integrating molecular, genetic, physiological and sociobiological perspectives to advance understanding of behavioral and life history transitions. When honey bees (Apis mellifera) transition from hive work to foraging, their flight muscles undergo changes that allow these insects to attain the highest rates of flight muscle metabolism and power output ever recorded in the animal kingdom. Here, we review research to date showing that honey bee flight muscles undergo significant changes in biochemistry and gene expression and that these changes accompany a significant increase in the capacity to generate metabolic and aerodynamic power during flight. It is likely that changes in muscle gene expression, biochemistry, metabolism and functional capacity may be driven primarily by behavior as opposed to age, as is the case for changes in honey bee brains.

The effect of selection for desiccation resistance on cold tolerance of Drosophila melanogaster

Abstract Low temperature and desiccation stress are thought to be mechanistically similar in insects, and several studies indicate that there is a degree of cross‐tolerance between them, such that increased cold tolerance results in greater desiccation tolerance and vice versa. This assertion is tested at an evolutionary scale by examining basal cold tolerance, rapid cold‐hardening (RCH) and chill coma recovery in replicate populations of Drosophila melanogaster selected for desiccation resistance (with controls for both selection and concomitant starvation) for over 50 generations. All of the populations display a RCH response, and there is no effect of selection regime on RCH or basal cold tolerance, although there are differences in basal cold tolerance between sampling dates, apparently related to inter‐individual variation in development time. Flies selected for desiccation tolerance recover from chill coma slightly, but significantly, faster than control and starvation‐control flies. These findings provide little support for cross‐tolerance between survival of near‐lethal cold and desiccation stress in D. melanogaster.