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OBJECTIVES: This study assessed the quality of diabetes care in community health centers. METHODS: In 55 midwestern community health centers, we reviewed the charts of 2865 diabetic adults for American Diabetes Association measures of quality. RESULTS: On average, 70% of the patients in each community health center had measurements of glycosylated hemoglobin, 26% had dilated eye examinations, 66% had diet intervention, and 51% received foot care. The average glycosylated hemoglobin value per community health center was 8.6%. Practice guidelines were independently associated with higher quality of care. CONCLUSIONS: Rates of adherence to process measures of quality were relatively low among community health centers, compared with the targets established by the American Diabetes Association. [ABSTRACT FROM AUTHOR]

In animals, metabolic rates during ontogeny often scale differently from the way they do in cross-species or population comparisons, with near-isometric scaling patterns more often observed during juvenile growth. In multiple social insect taxa, colony metabolic rate scales hypometrically across species or populations at the same developmental stage, but metabolic patterns during ontogeny have not been examined for any social insect species. We performed the first ontogenetic study of social metabolic scaling in harvester ant colonies ( Pogonomyrmex californicus ) over 3.5 years as they grew from 52 [Formula: see text] 12 to 767 [Formula: see text] 380 workers. Our data reveal iso- and hypermetric metabolic scaling during the first 20 months of growth, transitioning to hypometric scaling (scaling exponent = 0.93) thereafter. We discovered that the fraction of colony as brood has dual effects on colonial metabolic rate. A higher fraction of the colony that is brood decreases colonial metabolic rate because brood has approximately half the mass-specific metabolic rates of adult ants. Conversely, metabolic rate and activity of adult ants increase as this fraction increases. We further developed a nonlinear composition model, which shows that the maximal colony metabolic rate occurs when 29% of the colony mass is brood, suggesting that demographic changes and colony size may interact to drive the metabolic scaling.

Solar radiation is an important environmental variable for terrestrial animals, but its impact on the heat balance of large flying insects has been poorly studied. Desert bees are critical to ecosystem function through their pollination services, and are exposed to high radiant loads. We assessed the role of solar radiation in the heat balance of flying desert Centris pallida bees by calculating heat budgets for individuals in a respirometer in shaded versus sunny conditions from 16 to 37°C air temperatures, comparing the large and small male morphs and females. Solar radiation was responsible for 43 to 54% of mean total heat gain. Bees flying in the sun had thorax temperatures 1.7°C warmer than bees flying in the shade, storing a very small fraction of incident radiation in body tissues. In most cases, flight metabolic rate was not suppressed for bees flying in the sun, but evaporative water loss rates more than doubled. The most dramatic response to solar radiation was an increase in convection, mediated by a more than doubling of convective conductance, allowing thermoregulation while conserving body water. In large morph males and females, the increased convective conductance in the sun was mediated by increased heat transfer from the thorax to abdomen. Because convection is limited as body temperatures approach air temperatures, solar radiation combined with warming air temperatures may cause endothermic flying bees to reach a tipping point at which increases in non-sustainable evaporation are necessary for survival., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2025. Published by The Company of Biologists.)

Organisms require dietary macronutrients in specific ratios to maximize performance, and variation in macronutrient requirements plays a central role in niche determination. Although it is well recognized that development and body size can have strong and predictable effects on many aspects of organismal function, we lack a predictive understanding of ontogenetic or scaling effects on macronutrient intake. We determined protein and carbohydrate intake throughout development on lab populations of locusts and compared to late instars of field populations. Self-selected protein:carbohydrate targets declined dramatically through ontogeny, due primarily to declines in mass-specific protein consumption rates which were highly correlated with declines in specific growth rates. Lab results for protein consumption rates partly matched results from field-collected locusts. However, field locusts consumed nearly double the carbohydrate, likely due to higher activity and metabolic rates. Combining our results with the available data for animals, both across species and during ontogeny, protein consumption scaled predictably and hypometrically, demonstrating a new scaling rule key for understanding nutritional ecology., Competing Interests: ST, JH, RF, JY, HM, RO, AC No competing interests declared, (© 2024, Talal et al.)

The developmental regulation of body size is a fundamental life-history characteristic that in most animals is tied to the transition from juvenile to adult form. In holometabolous insects, this transition is ostensibly initiated at the attainment of a critical weight in the final larval instar. It has been hypothesized that the size-sensing mechanism used to determine attainment of critical weight exploits oxygen limitation as a larvae grows beyond the oxygen-delivery capacity of its fixed tracheal system; that is, developmentally induced cellular hypoxia initiates the synthesis of the molting hormone ecdysone by the prothoracic gland. We tested this hypothesis in Drosophila by assaying cellular hypoxia throughout the third larval instar at 21 and 10 kPa O2, using the activity of the HIF (hypoxia inducible factor)-signaling pathway as a measure of hypoxia. While HIF signaling was elevated at low levels of environmental O2, it did not markedly increase during development at either oxygen level, and was only suppressed by hyperoxia after feeding had ceased. Further, changes in HIF signaling in the prothoracic gland alone did not alter body size or developmental time in a way that would be expected if cellular hypoxia in the prothoracic gland was part of the critical weight mechanism. Our data do show, however, that reduced HIF signaling in the prothoracic gland decreases survival and retards development at 10 kPa O2, suggesting that prothoracic HIF signaling is a necessary part of the beneficial plasticity mechanism that controls growth and development in response to low oxygen level., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Wolbachia is a widespread maternally transmitted endosymbiotic bacteria with diverse phenotypic effects on its insect hosts, ranging from parasitic to mutualistic. Wolbachia commonly infects social insects, where it faces unique challenges associated with its host's caste-based reproductive division of labor and colony living. Here, we dissect the benefits and costs of Wolbachia infection on life-history traits of the invasive pharaoh ant, Monomorium pharaonis, which are relatively short lived and show natural variation in Wolbachia infection status between colonies. We quantified the effects of Wolbachia infection on the lifespan of queen and worker castes, the egg-laying rate of queens across queen lifespan, and the metabolic rates of whole colonies and colony members. Infected queens laid more eggs than uninfected queens but had similar metabolic rates and lifespans. Interestingly, infected workers outlived uninfected workers. At the colony level, infected colonies were more productive as a consequence of increased queen egg-laying rates and worker longevity, and infected colonies had higher metabolic rates during peak colony productivity. While some effects of infection, such as elevated colony-level metabolic rates, may be detrimental in more stressful natural conditions, we did not find any costs of infection under relatively benign laboratory conditions. Overall, our study emphasizes that Wolbachia infection can have beneficial effects on ant colony growth and worker survival in at least some environments., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

The evolution of flight in an early winged insect ancestral lineage is recognized as a key adaptation explaining the unparalleled success and diversification of insects. Subsequent transitions and modifications to flight machinery, including secondary reductions and losses, also play a central role in shaping the impacts of insects on broadscale geographic and ecological processes and patterns in the present and future. Given the importance of insect flight, there has been a centuries-long history of research and debate on the evolutionary origins and biological mechanisms of flight. Here, we revisit this history from an interdisciplinary perspective, discussing recent discoveries regarding the developmental origins, physiology, biomechanics, and neurobiology and sensory control of flight in a diverse set of insect models. We also identify major outstanding questions yet to be addressed and provide recommendations for overcoming current methodological challenges faced when studying insect flight, which will allow the field to continue to move forward in new and exciting directions. By integrating mechanistic work into ecological and evolutionary contexts, we hope that this synthesis promotes and stimulates new interdisciplinary research efforts necessary to close the many existing gaps about the causes and consequences of insect flight evolution., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

AbstractGlobal warming trends, human-assisted transport, and urbanization have allowed poleward expansion of many tropical vector species, but the specific mechanisms responsible for thermal mediation of range changes and ecological success of invaders remain poorly understood. Aedes aegypti (Diptera: Culicidae) is a tropical mosquito currently expanding into many higher-latitude regions, including the urban desert region of Maricopa County, Arizona. Here, adult populations virtually disappear in winter and spring and then increase exponentially through summer and fall, indicating that winter conditions remain a barrier to the development of some life stages of A. aegypti . To determine whether cold limits the winter development of A. aegypti larvae in Maricopa County, we surveyed for larval abundance and tested their capacity to develop in ambient and warmed conditions. Aedes aegypti larvae were not observed in artificial aquatic habitats in winter and spring but were abundant in summer and fall, suggesting winter suppression of adults, larvae, or both. Water temperatures in winter months fluctuated strongly; larvae were usually cold paralyzed at night but active during the day. Despite daytime temperatures that allowed activity and achieving similar degree-days as warmed mesocosms, larvae reared under ambient winter conditions were unable to develop to adulthood, perhaps due to repetitive cold damage. However, warming average temperature by 1.7°C allowed many larvae to successfully develop to adults. Because daytime highs in winter will often allow adult flight, it is likely that relatively minor additional winter warming may allow A. aegypti populations to develop and reproduce year-round in Maricopa County.

Many theoretical treatments of foraging use energy as currency, with carbohydrates and lipids considered interchangeable as energy sources. However, herbivores must often synthesize lipids from carbohydrates since they are in short supply in plants, theoretically increasing the cost of growth. We tested whether a generalist insect herbivore ( Locusta migratoria ) can improve its growth efficiency by consuming lipids, and whether these locusts have a preferred caloric intake ratio of carbohydrate to lipid (C : L). Locusts fed pairs of isocaloric, isoprotein diets differing in C and L consistently selected a 2C : 1L target. Locusts reared on isocaloric, isoprotein 3C : 0L diets attained similar final body masses and lipid contents to locusts fed the 2C : 1L diet, but they ate more and had a ~12% higher metabolic rate, indicating an energetic cost for lipogenesis. These results demonstrate that some animals can selectively regulate carbohydrate-to-lipid intake and that consumption of dietary lipids can improve growth efficiency.

While multiple studies have shown that honey bees and some other flying insects lower their flight metabolic rates when flying at high air temperatures, critics have suggested such patterns result from poor experimental methods as, theoretically, air temperature should not appreciably affect aerodynamic force requirements. Here, we show that apparently contradictory studies can be reconciled by considering the thermal performance curve of flight muscle. We show that prior studies that found no effects of air temperature on flight metabolism of honey bees achieved flight muscle temperatures that were near or on equal, opposite sides of the thermal performance curve. Honey bees vary their wing kinematics and metabolic heat production to thermoregulate, and how air temperature affects the flight metabolic rate of honey bees is predictable using a non-linear thermal performance perspective of honey bee flight muscle., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Interactions between environmental stressors may contribute to ongoing pollinator declines, but have not been extensively studied. Here, we examined the interaction between the agricultural fungicide Pristine (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) and high temperatures on critical honeybee behaviours. We have previously shown that consumption of field-realistic levels of this fungicide shortens worker lifespan in the field and impairs associative learning performance in a laboratory-based assay. We hypothesized that Pristine would also impair homing and foraging behaviours in the field, and that an interaction with hot weather would exacerbate this effect. Both field-relevant Pristine exposure and higher air temperatures reduced the probability of successful return on their own. Together, the two factors synergistically reduced the probability of return and increased the time required for bees to return to the hive. Pristine did not affect the masses of pollen or volumes of nectar or water brought back to the hive by foragers, and it did not affect the ratio of forager types in a colony. However, Pristine-fed bees brought more concentrated nectar back to the hive. As both agrochemical usage and heat waves increase, additive and synergistic negative effects may pose major threats to pollinators and sustainable agriculture.

Heat waves are becoming increasingly common due to climate change, making it crucial to identify and understand the capacities for insect pollinators, such as honey bees, to avoid overheating. We examined the effects of hot, dry air temperatures on the physiological and behavioral mechanisms that honey bees use to fly when carrying nectar loads, to assess how foraging is limited by overheating or desiccation. We found that flight muscle temperatures increased linearly with load mass at air temperatures of 20 or 30 °C, but, remarkably, there was no change with increasing nectar loads at an air temperature of 40 °C. Flying, nectar-loaded bees were able to avoid overheating at 40 °C by reducing their flight metabolic rates and increasing evaporative cooling. At high body temperatures, bees apparently increase flight efficiency by lowering their wingbeat frequency and increasing stroke amplitude to compensate, reducing the need for evaporative cooling. However, even with reductions in metabolic heat production, desiccation likely limits foraging at temperatures well below bees' critical thermal maxima in hot, dry conditions., Competing Interests: Competing interests statement:The authors declare no competing interest.

Managed honey bees have experienced high rates of colony loss recently, with pesticide exposure as a major cause. While pesticides can be lethal at high doses, lower doses can produce sublethal effects, which may substantially weaken colonies. Impaired learning performance is a behavioral sublethal effect, and is often present in bees exposed to insecticides. However, the effects of other pesticides (such as fungicides) on honey bee learning are understudied, as are the effects of pesticide formulations versus active ingredients. Here, we investigated the effects of acute exposure to the fungicide formulation Pristine (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) on honey bee olfactory learning performance in the proboscis extension reflex (PER) assay. We also exposed a subset of bees to only the active ingredients to test which formulation component(s) were driving the learning effects. We found that the formulation produced negative effects on memory, but this effect was not present in bees fed only boscalid and pyraclostrobin. This suggests that the trade secret "other ingredients" in the formulation mediated the learning effects, either through exerting their own toxic effects or by increasing the toxicities of the active ingredients. These results show that pesticide co-formulants should not be assumed inert and should instead be included when assessing pesticide risks., (© 2023. The Author(s).)

Over decades, pesticide regulations have cycled between approval and implementation, followed by the discovery of negative effects on nontarget organisms that result in new regulations, pesticides, and harmful effects. This relentless pattern undermines the capacity to protect the environment from pesticide hazards and frustrates end users that need pest management tools. Wild pollinating insects are in decline, and managed pollinators such as honey bees are experiencing excessive losses, which threatens sustainable food security and ecosystem function. An increasing number of studies demonstrate the negative effects of field-realistic exposure to pesticides on pollinator health and fitness, which contribute to pollinator declines. Current pesticide approval processes, although they are superior to past practices, clearly continue to fail to protect pollinator health. In the present article, we provide a conceptual framework to reform cyclical pesticide approval processes and better protect pollinators., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Institute of Biological Sciences.)

The Proboscis Extension Reflex (PER) paradigm trains honey bees to associate an odor with a sugar reward and is commonly used to assess impacts on associative learning after exposure to pesticides. While the effects of some types of pesticides have been well-investigated, relatively little attention has been focused on fungicides that are applied to flowering crops. We have previously shown that consumption of field-relevant concentrations of the fungicide Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) impairs honey bee performance in an associative learning assay, but the mechanism of its action has not been investigated. We hypothesized that Pristine® interferes with carbohydrate absorption and/or regulation, thereby disrupting the post-ingestive feedback mechanisms necessary for robust learning. To test this hypothesis, we measured hemolymph glucose and trehalose levels at five time points during the ten minutes after bees consumed a sucrose solution. Pristine®-exposed bees had elevated baseline glucose concentrations in the hemolymph relative to control bees. Hemolymph glucose levels rose significantly within five minutes of feeding in control bees, but not in Pristine®-fed bees. These data suggest that the post-ingestive feedback mechanisms necessary for robust learning are disrupted in bees that have consumed this fungicide, providing a plausible mechanistic explanation for its effects on learning performance in the PER assay. Pristine®-exposed bees may have elevated hemolymph glucose levels because the fungicide elicits an inflammatory response. These results provide additional mechanistic understanding of the negative physiological effects of mitotoxic fungicides on this important pollinator., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Air sacs are a well-known aspect of insect tracheal systems, but have received little research attention. In this Commentary, we suggest that the study of the distribution and function of air sacs in tracheate arthropods can provide insights of broad significance. We provide preliminary phylogenetic evidence that the developmental pathways for creation of air sacs are broadly conserved throughout the arthropods, and that possession of air sacs is strongly associated with a few traits, including the capacity for powerful flight, large body or appendage size and buoyancy control. We also discuss how tracheal compression can serve as an additional mechanism for achieving advection in tracheal systems. Together, these patterns suggest that the possession of air sacs has both benefits and costs that remain poorly understood. New technologies for visualization and functional analysis of tracheal systems provide exciting approaches for investigations that will be of broad significance for understanding invertebrate evolution., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Migration allows animals to track favorable environments and avoid harmful conditions. However, migration is energetically costly, so migrating animals must prepare themselves by increasing their energy stores. Despite the importance of locust migratory swarms, we still understand little about the physiology of locust migration. During long-distance flight, locusts rely on lipid oxidation, despite the fact that lipids are relatively rare in their leaf-based diets. Therefore, locusts and other insect herbivores synthesize and store lipid from ingested carbohydrates, which are also important for initial flight. These data suggest that diets high in carbohydrate should increase lipid stores and the capacity for migratory flight in locusts. As predicted, locust lipid stores and flight performance increased with an increase in the relative carbohydrate content in their food. However, locust flight termination was not associated with complete lipid depletion. We propose potential testable mechanisms that might explain how macronutrient consumption can affect flight endurance., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Honey bee pollination services are of tremendous agricultural and economic importance. Despite this, honey bees and other pollinators face ongoing perils, including population declines due to a variety of environmental stressors. Fungicides may be particularly insidious stressors for pollinators due to their environmental ubiquity and widespread approval for application during crop bloom. The mechanisms by which fungicides affect honey bees are poorly understood and any seasonal variations in their impact are unknown. Here we assess the effects on honey bee colonies of four-week exposure (the approximate duration of the almond pollination season) of a fungicide, Pristine® (25.2% boscalid, 12.8% pyraclostrobin), that has been commonly used for almonds. We exposed colonies to Pristine® in pollen patties placed into the hive, in either summer or fall, and assessed colony brood and worker populations, colony pollen collection and consumption, and worker age of first foraging and longevity. During the summer, Pristine® exposure induced precocious foraging, and reduced worker longevity resulting in smaller colonies. During the fall, Pristine® exposure induced precocious foraging but otherwise had no significant measured effects. During the fall, adult and brood population levels, and pollen consumption and collection, were all much lower, likely due to preparations for winter. Fungicides and other pesticides may often have reduced effects on honey bees during seasons of suppressed colony growth due to bees consuming less pollen and pesticide., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Flying endothermic insects thermoregulate, likely to improve flight performance. Males of the Sonoran Desert bee, Centris caesalpiniae, seek females at aggregations beginning at sunrise and cease flight near midday when the air temperature peaks. To identify the thermoregulatory mechanisms for C. caesalpiniae males, we measured tagma temperature, wingbeat frequency, water loss rate, metabolic rate and tagma mass of flying bees across shaded air temperatures of 19-38°C. Surface area, wet mass and dry mass declined with air temperature, suggesting that individual bees do not persist for the entire morning. The largest bees may be associated with cool, early mornings because they are best able to warm themselves and/or because they run the risk of overheating in the hot afternoons. Thorax temperature was high (38-45°C) and moderately well regulated, while head and abdomen temperatures were cooler and less controlled. The abdominal temperature excess ratio increased as air temperature rose, indicating active heat transfer from the pubescent thorax to the relatively bare abdomen with warming. Mass-specific metabolic rate increased with time, and air and thorax temperatures, but wingbeat frequency did not vary. Mass-specific water loss rate increased with air temperature, but this was a minor mechanism of thermoregulation. Using a heat budget model, we showed that whole-body convective conductance more than doubled through the morning, providing strong evidence that the primary mechanism of regulating thorax temperature during flight for these bees is increased use of the abdomen as a convector at higher air temperatures., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

The scaling of respiratory structures has been hypothesized to be a major driving factor in the evolution of many aspects of animal physiology. Here, we provide the first assessment of the scaling of the spiracles in insects using 10 scarab beetle species differing 180× in mass, including some of the most massive extant insect species. Using X-ray microtomography, we measured the cross-sectional area and depth of all eight spiracles, enabling the calculation of their diffusive and advective capacities. Each of these metrics scaled with geometric isometry. Because diffusive capacities scale with lower slopes than metabolic rates, the largest beetles measured require 10-fold higher P O2 gradients across the spiracles to sustain metabolism by diffusion compared to the smallest species. Large beetles can exchange sufficient oxygen for resting metabolism by diffusion across the spiracles, but not during flight. In contrast, spiracular advective capacities scale similarly or more steeply than metabolic rates, so spiracular advective capacities should match or exceed respiratory demands in the largest beetles. These data illustrate a general principle of gas exchange: scaling of respiratory transport structures with geometric isometry diminishes the potential for diffusive gas exchange but enhances advective capacities; combining such structural scaling with muscle-driven ventilation allows larger animals to achieve high metabolic rates when active., Competing Interests: JW, CK, MD, JS, GC, HG, JH No competing interests declared, (© 2022, Wagner et al.)

Understanding the effect of body size on flight costs is critical for development of models of aerodynamics and animal energetics. Prior scaling studies that have shown that flight costs scale hypometrically have focused primarily on larger (> 100 mg) insects and birds, but most flying species are smaller. We studied the flight physiology of thirteen stingless bee species over a large range of body sizes (1-115 mg). Metabolic rate during hovering scaled hypermetrically (scaling slope = 2.11). Larger bees had warm thoraxes while small bees were nearly ecothermic; however, even controlling for body temperature variation, flight metabolic rate scaled hypermetrically across this clade. Despite having a lower mass-specific metabolic rate during flight, smaller bees could carry the same proportional load. Wingbeat frequency did not vary with body size, in contrast to most studies that find wingbeat frequency increases as body size decreases. Smaller stingless bees have greater relative forewing surface area which may help them reduce the energy requirements needed to fly. Further, we hypothesize that the relatively larger heads of smaller species may change their body pitch in flight. Synthesizing across all flying insects, we demonstrate that the scaling of flight metabolic rate changes from hypermetric to hypometric at approximately 58 mg body mass with hypermetic scaling below (slope = 1.2) and hypometric scaling (slope = 0.67) above 58 mg in body mass. The reduced cost of flight likely provides selective advantages for the evolution of small body size in insects. The biphasic scaling of flight metabolic rates and wingbeat frequencies in insects supports the hypothesis that the scaling of metabolic rate is closely related to the power requirements of locomotion and cycle frequencies., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Performance benefits of stable, warm muscles are believed to be important for the evolution of endothermy in mammals, birds and flying insects. However, thermal performance curves have never been measured for a free-flying endotherm, as it is challenging to vary body temperatures of these animals, and maximal flight performance is difficult to elicit. We varied air temperatures and gas densities to manipulate thoracic temperatures of flying honeybees from 29°C to 44°C, with low air densities used to increase flight metabolic rates to maximal values. Honeybees showed a clear thermal performance curve with an optimal temperature of 39°C. Maximal flight metabolic rates increased by approximately 2% per 1°C increase in thoracic temperature at suboptimal thoracic temperatures, but decreased approximately 5% per 1°C increase as the bees continued to heat up. This study provides the first quantification of the maximal metabolic performance benefit of thermoregulation in an endotherm. These data directly support aerobic capacity models for benefits of thermoregulation in honeybees, and suggest that improved aerobic capacity probably contributes to the multiple origins of endothermic heterothermy in bees and other insects.

Larger animals studied during ontogeny, across populations, or across species, usually have lower mass-specific metabolic rates than smaller animals (hypometric scaling). This pattern is usually observed regardless of physiological state (e.g. basal, resting, field, maximally-active). The scaling of metabolism is usually highly correlated with the scaling of many life history traits, behaviors, physiological variables, and cellular/molecular properties, making determination of the causation of this pattern challenging. For across-species comparisons of resting and locomoting animals (but less so for across populations or during ontogeny), the mechanisms at the physiological and cellular level are becoming clear. Lower mass-specific metabolic rates of larger species at rest are due to a) lower contents of expensive tissues (brains, liver, kidneys), and b) slower ion leak across membranes at least partially due to membrane composition, with lower ion pump ATPase activities. Lower mass-specific costs of larger species during locomotion are due to lower costs for lower-frequency muscle activity, with slower myosin and Ca++ ATPase activities, and likely more elastic energy storage. The evolutionary explanation(s) for hypometric scaling remain(s) highly controversial. One subset of evolutionary hypotheses relies on constraints on larger animals due to changes in geometry with size; for example, lower surface-to-volume ratios of exchange surfaces may constrain nutrient or heat exchange, or lower cross-sectional areas of muscles and tendons relative to body mass ratios would make larger animals more fragile without compensation. Another subset of hypotheses suggests that hypometric scaling arises from biotic interactions and correlated selection, with larger animals experiencing less selection for mass-specific growth or neurolocomotor performance. A additional third type of explanation comes from population genetics. Larger animals with their lower effective population sizes and subsequent less effective selection relative to drift may have more deleterious mutations, reducing maximal performance and metabolic rates. Resolving the evolutionary explanation for the hypometric scaling of metabolism and associated variables is a major challenge for organismal and evolutionary biology. To aid progress, we identify some variation in terminology use that has impeded cross-field conversations on scaling. We also suggest that promising directions for the field to move forward include: 1) studies examining the linkages between ontogenetic, population-level, and cross-species allometries, 2) studies linking scaling to ecological or phylogenetic context, 3) studies that consider multiple, possibly interacting hypotheses, and 4) obtaining better field data for metabolic rates and the life history correlates of metabolic rate such as lifespan, growth rate and reproduction., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

In almost all animals, physiologically low oxygen (hypoxia) during development slows growth and reduces adult body size. The developmental mechanisms that determine growth under hypoxic conditions are, however, poorly understood. Here we show that the growth and body size response to moderate hypoxia (10% O 2 ) in Drosophila melanogaster is systemically regulated via the steroid hormone ecdysone. Hypoxia increases level of circulating ecdysone and inhibition of ecdysone synthesis ameliorates the negative effect of low oxygen on growth. We also show that the effect of ecdysone on growth under hypoxia is through suppression of the insulin/IGF-signaling pathway, via increased expression of the insulin-binding protein Imp-L2. These data indicate that growth suppression in hypoxic Drosophila larvae is accomplished by a systemic endocrine mechanism that overlaps with the mechanism that slows growth at low nutrition. This suggests the existence of growth-regulatory mechanisms that respond to general environmental perturbation rather than individual environmental factors., (© 2022. The Author(s).)

Competing Interests: The author declares no competing interest.

Recent observations of many sublethal effects of pesticides on pollinators have raised questions about whether standard short-term laboratory tests of pesticide effects on survival are sufficient for pollinator protection. The fungicide Pristine® and its active ingredients (25.2% boscalid, 12.8% pyraclostrobin) have been reported to have low acute toxicity to caged honey bee workers, but many sublethal effects at field-relevant doses have been reported and Pristine® was recently found to increase worker pollen consumption, reduce worker longevity and colony populations at field relevant concentrations (Fisher et al. 2021). To directly compare these whole-colony field results to more standard laboratory toxicology tests, the effects of Pristine®, at a range of field-relevant concentrations, were assessed on the survival and pollen consumption of honey bee workers 0-14 days of age. Also, to separate the effects of the inert and two active ingredients, bees were fed pollen containing boscalid, pyraclostrobin, or pyraclostrobin plus boscalid, at concentrations matching those in the Pristine® treatments. Pyraclostrobin significantly reduced pollen consumption across the duration of the experiment, and dose-dependently reduced pollen consumption on days 12-14. Pristine® and boscalid significantly reduced pollen feeding rate on days 12-14. Boscalid reduced survival in a dose-dependent manner. Consumption of Pristine® or pyraclostrobin plus boscalid did not affect survival, providing evidence against strong negative effects of the inert ingredients in Pristine® and against negative synergistic effects of boscalid and pyraclostrobin. The stronger toxic effects of Pristine® observed in field colonies compared to this laboratory test, and the opposite responses of pollen consumption in the laboratory and field to Pristine®, show that standard laboratory toxicology tests can fail to predict responses of pollinators to pesticides and to provide protection., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Although fungicides were previously considered to be safe for important agricultural pollinators such as honey bees, recent evidence has shown that they can cause a number of behavioral and physiological sublethal effects. Here, we focus on the fungicide Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin), which is sprayed during the blooming period on a variety of crops and is known to affect honey bee mitochondria at field-relevant levels. To date, no study has tested the effects of a field-relevant concentration of a fungicide on associative learning ability in honey bees. We tested whether chronic, colony-level exposure at field-relevant and higher concentrations of Pristine® impairs performance on the proboscis extension reflex (PER) paradigm, an associative learning task. Learning performance was reduced at higher field-relevant concentrations of Pristine®. The reductions in learning performance could not be explained by effects on hunger or motivation, as sucrose responsiveness was not affected by Pristine® exposure. To determine whether Pristine®'s negative effects on learning performance were mediated at a specific life stage, we conducted a cross-fostering experiment that exposed bees to the fungicide either only as larvae, only as adults, or during both stages. We found that exposure across the entire life was necessary to significantly reduce learning performance, although non-significant reductions occurred when bees were exposed during just one stage. Our study provides strong evidence that Pristine® has significant sublethal effects on learning performance. As associative learning is a necessary ability for foraging, our results raise concerns that Pristine® could impair foraging abilities and substantially weaken colony health., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Pollinators and other insects are experiencing an ongoing worldwide decline. While various environmental stressors have been implicated, including pesticide exposure, the causes of these declines are complex and highly debated. Fungicides may constitute a particularly prevalent threat to pollinator health due to their application on many crops during bloom, and because pollinators such as bees may consume fungicide-tainted pollen or nectar. In a previous study, consumption of pollen containing the fungicide Pristine® at field-relevant concentrations by honey bee colonies increased pollen foraging, caused earlier foraging, lowered worker survival, and reduced colony population size. Because most pollen is consumed by young adults, we hypothesized that Pristine® (25.2% boscalid, 12.8% pyraclostrobin) in pollen exerts its negative effects on honey bee colonies primarily on the adult stage. To rigorously test this hypothesis, we used a cross-fostering experimental design, with bees reared in colonies provided Pristine® incorporated into pollen patties at a supra-field concentration (230 mg/kg), only in the larvae, only in the adult, or both stages. In contrast to our predictions, exposure to Pristine® in either the larval or adult stage reduced survival relative to control bees not exposed to Pristine®, and exposure to the fungicide at both larval and adult stages further reduced survival. Adult exposure caused precocious foraging, while larval exposure increased the tendency to forage for pollen. These results demonstrate that pollen containing Pristine® can induce significant negative effects on both larvae and adults in a hive, though the magnitude of such effects may be smaller at field-realistic doses. To further test the potential negative effects of direct consumption of Pristine® on larvae, we reared them in vitro on food containing Pristine® at a range of concentrations. Consumption of Pristine® reduced survival rates of larvae at all concentrations tested. Larval and adult weights were only reduced at a supra-field concentration. We conclude that consumption of pollen containing Pristine® by field honey bee colonies likely exerts impacts on colony population size and foraging behavior by affecting both larvae and adults., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Social groups form when the costs of breeding independently exceed fitness costs imposed by group living. The costs of independent breeding can often be energetic, especially for animals performing expensive behaviours, such as nest construction. To test the hypothesis that nesting costs can drive sociality by disincentivizing independent nest founding, we measured the energetics of nest construction and inheritance in a facultatively social carpenter bee ( Xylocopa sonorina Smith), which bores tunnel nests in wood. We measured metabolic rates of bees excavating wood and used computerized tomography images of nesting logs to measure excavation volumes. From these data, we demonstrate costly energetic investments in nest excavation of a minimum 4.3 kJ per offspring provisioned, an expense equivalent to nearly 7 h of flight. This high, potentially prohibitive cost of nest founding may explain why females compete for existing nests rather than constructing new ones, often leading to the formation of social groups. Further, we found that nest inheritors varied considerably in their investment in nest renovation, with costs ranging more than 12-fold (from 7.08 to 89.1 kJ energy), probably reflecting differences in inherited nest quality. On average, renovation costs were lower than estimated new nest construction costs, with some nests providing major savings. These results suggest that females may join social groups to avoid steep energetic costs, but that the benefits of this strategy are not experienced equally.

The fitness consequences of cooperation can vary across an organism's lifespan. For non-kin groups, especially, social advantages must balance intrinsic costs of cooperating with non-relatives. In this study, we asked how challenging life history stages can promote stable, long-term alliances among unrelated ant queens. We reared single- and multi-queen colonies of the primary polygynous harvester ant, Pogonomyrmex californicus, from founding through the first ten months of colony growth, when groups face high mortality risks. We found that colonies founded by multiple, unrelated queens experienced significant survival and growth advantages that outlasted the colony founding period. Multi-queen colonies experienced lower mortality than single-queen colonies, and queens in groups experienced lower mortality than solitary queens. Further, multi-queen colonies produced workers at a faster rate than did single-queen colonies, even while experiencing lower per-queen worker production costs. Additionally, we characterized ontogenetic changes in the organization of labor, and observed increasing and decreasing task performance diversity by workers and queens, respectively, as colonies grew. This dynamic task allocation likely reflects a response to the changing role of queens as they are increasingly able to delegate risky and costly tasks to an expanding workforce. Faster worker production in multi-queen colonies may beneficially accelerate this behavioral transition from a vulnerable parent-offspring group to a stable, growing colony. These combined benefits of cooperation may facilitate the retention of multiple unrelated queens in mature colonies despite direct fitness costs, providing insight into the evolutionary drivers of stable associations between unrelated individuals.

Commercial beekeepers in many locations are experiencing increased annual colony losses of honey bees (Apis mellifera), but the causes, including the role of agrochemicals in colony losses, remain unclear. In this study, we investigated the effects of chronic consumption of pollen containing a widely-used fungicide (Pristine®), known to inhibit bee mitochondria in vitro, which has recently been shown to reduce honey bee worker lifespan when field-colonies are provided with pollen containing field-realistic levels of Pristine®. We fed field colonies pollen with a field-realistic concentration of Pristine® (2.3 ppm) and a concentration two orders of magnitude higher (230 ppm). To challenge flight behavior and elicit near-maximal metabolic rate, we measured flight quality and metabolic rates of bees in two lower-than-normal air densities. Chronic consumption of 230 but not 2.3 ppm Pristine® reduced maximal flight performance and metabolic rates, suggesting that the observed decrease in lifespans of workers reared on field-realistic doses of Pristine®-laced pollen is not due to inhibition of flight muscle mitochondria. However, consumption of either the 230 or 2.3 ppm dose reduced thorax mass (but not body mass), providing the first evidence of morphological effects of Pristine®, and supporting the hypothesis that Pristine® reduces forager longevity by negatively impacting digestive or nutritional processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Honey bees (Apis mellifera) and other pollinator populations are declining worldwide, and the reasons remain controversial. Based on laboratory testing, fungicides have traditionally been considered bee-safe. However, there have been no experimental tests of the effects of fungicides on colony health under field conditions, and limited correlational data suggests there may be negative impacts on bees at levels experienced in the field. We tested the effects of one of the most commonly used fungicides on colony health by feeding honey bee colonies pollen containing Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) at four levels that bracketed concentrations we measured for pollen collected by bees in almond orchards. We also developed a method for calculating per-bee and per-larva dose. Pristine® consumption significantly and dose-dependently reduced worker lifespan and colony population size, with negative health effects observed even at the lowest doses. The lowest concentration we tested caused a 15% reduction in the worker population at an estimated dosage that was three orders of magnitude below the estimated LD 15 values for previous acute laboratory studies. The enhanced toxicity under field conditions is at least partially due to activation of colonial nutritional responses missed by lab tests. Pristine® causes colonies to respond to perceived protein malnutrition by increasing colony pollen collection. Additionally, Pristine induces much earlier transitioning to foraging in individual workers, which could be the cause of shortened lifespans. These findings demonstrate that Pristine® can negatively impact honey bee individual and colony health at concentrations relevant to what they experience from pollination behavior under current agricultural conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Locusts are major intermittent threats to food security and the ecological factors determining where and when these occur remain poorly understood. For many herbivores, obtaining adequate protein from plants is a key challenge. We tested how the dietary protein : non-structural carbohydrate ratio (p : c) affects the developmental and physiological performance of 4th-5th instar nymphs of the South American locust, Schistocerca cancellata, which has recently resurged in Argentina, Bolivia and Paraguay. Field marching locusts preferred to feed on high carbohydrate foods. Field-collected juveniles transferred to the laboratory selected artificial diets or local plants with low p : c. On single artificial diets, survival rate increased as foods became more carbohydrate-biased. On single local plants, growth only occurred on the plant with the lowest p : c. Most local plants had p : c ratios substantially higher than optimal, demonstrating that field marching locusts must search for adequate carbohydrate or their survival and growth will be carbohydrate-limited. Total body lipids increased as dietary p : c decreased on both artificial and plant diets, and the low lipid contents of field-collected nymphs suggest that obtaining adequate carbohydrate may pose a strong limitation on migration for S. cancellata . Anthropogenic influences such as conversions of forests to pastures, may increase carbohydrate availability and promote outbreaks and migration of some locusts.

Mammals regulate their brain tissue P O 2 tightly, and only small changes in brain P O 2 are required to elicit compensatory ventilation. However, unlike the flow-through cardiovascular system of vertebrates, insect tissues exchange gases through blind-ended tracheoles, which may involve a more prominent role for diffusive gas exchange. We tested the effect of progressive hypoxia on ventilation and the P O 2 of the metathoracic ganglion (neural site of control of ventilation) using microelectrodes in the American locust, Schistocerca americana . In normal air (21 kPa), P O 2 of the metathoracic ganglion was 12 kPa. The P O 2 of the ganglion dropped as air P O 2 dropped, with ventilatory responses occurring when ganglion P O 2 reached 3 kPa. Unlike vertebrates, insects tolerate relatively high resting tissue P O 2 levels and allow tissue P O 2 to drop during hypoxia, activity and discontinuous gas exchange before activating convective or spiracular gas exchange. Tracheated animals, and possibly pancrustaceans in general, seem likely to generally experience wide spatial and temporal variation in tissue P O 2 compared with vertebrates, with important implications for physiological function and the evolution of oxygen-using proteins.

Growth rate, development time, and response to environmental stressors vary tremendously across organisms, suggesting trade-offs that are affected by evolutionary or ecological factors, but such trade-offs are poorly understood. Prior studies using artificially selected lines of Manduca sexta suggest that insects with high growth rates, long development time, and large body size are more sensitive to hypoxic or hyperoxic stresses, such as reactive oxygen species (ROS) production, but the mechanisms and specific life-history associations remain unclear. Here, we manipulated the social environment to differentiate the effects of size, growth rate, and development time on oxygen sensitivity of the giant mealworm, Zophobas morio . Crowding reduced growth rates but yielded larger adults as a result of supernumerary molts and longer development times. The juvenile performance (growth rate, development time, adult mass) of crowd-reared mealworms was less sensitive to variation in atmospheric oxygen than it was for individually reared animals, consistent with the hypothesis that high growth rates are associated with increased sensitivity to ROS. Life span in normoxia was extended by crowd rearing, perhaps due to the larger size and/or increased resources of the larger adults. Life spans of crowd-reared animals were more negatively affected by hypoxia or hyperoxia than life spans of individually reared animals, possibly due to the longer total stress exposure of crowd-reared animals. These data suggest that animals with high growth rates experience a negative trade-off of performance with greater sensitivity to stress during the juvenile phase, while animals with long development times or life spans experience a negative trade-off of greater susceptibility of life span to environmental stress.

Gravity is one of the most ubiquitous environmental effects on living systems: Cellular and organismal responses to gravity are of central importance to understanding the physiological function of organisms, especially eukaryotes. Gravity has been demonstrated to have strong effects on the closed cardiovascular systems of terrestrial vertebrates, with rapidly responding neural reflexes ensuring proper blood flow despite changes in posture. Invertebrates possess open circulatory systems, which could provide fewer mechanisms to restrict gravity effects on blood flow, suggesting that these species also experience effects of gravity on blood pressure and distribution. However, whether gravity affects the open circulatory systems of invertebrates is unknown, partly due to technical measurement issues associated with small body size. Here we used X-ray imaging, radio-tracing of hemolymph, and micropressure measurements in the American grasshopper, Schistocerca americana , to assess responses to body orientation. Our results show that during changes in body orientation, gravity causes large changes in blood and air distribution, and that body position affects ventilation rate. Remarkably, we also found that insects show similar heart rate responses to body position as vertebrates, and contrasting with the classic understanding of open circulatory systems, have flexible valving systems between thorax and abdomen that can separate pressures. Gravitational effects on invertebrate cardiovascular and respiratory systems are likely to be widely distributed among invertebrates and to have broad influence on morphological and physiological evolution., Competing Interests: The authors declare no competing interest.

As the genetic bases to variation in anoxia tolerance are poorly understood, we used the Drosophila Genetics Reference Panel (DGRP) to conduct a genome-wide association study (GWAS) of anoxia tolerance in adult and larval Drosophila melanogaster Survival ranged from 0-100% in adults exposed to 6 h of anoxia and from 20-98% for larvae exposed to 1 h of anoxia. Anoxia tolerance had a broad-sense heritability of 0.552 in adults and 0.433 in larvae. Larval and adult phenotypes were weakly correlated but the anoxia tolerance of adult males and females were strongly correlated. The GWA identified 180 SNPs in adults and 32 SNPs in larvae associated with anoxia tolerance. Gene ontology enrichment analysis indicated that many of the 119 polymorphic genes associated with adult anoxia-tolerance were associated with ionic transport or immune function. In contrast, the 22 polymorphic genes associated with larval anoxia-tolerance were mostly associated with regulation of transcription and DNA replication. RNAi of mapped genes generally supported the hypothesis that disruption of these genes reduces anoxia tolerance. For two ion transport genes, we tested predicted directional and sex-specific effects of SNP alleles on adult anoxia tolerance and found strong support in one case but not the other. Correlating our phenotype to prior DGRP studies suggests that genes affecting anoxia tolerance also influence stress-resistance, immune function and ionic balance. Overall, our results provide evidence for multiple new potential genetic influences on anoxia tolerance and provide additional support for important roles of ion balance and immune processes in determining variation in anoxia tolerance., (Copyright © 2019 Campbell et al.)

Animals vary tremendously in their capacities to survive anoxia, and the mechanisms responsible are poorly understood. Adult Drosophila melanogaster are rapidly paralyzed and survive up to 12 h of anoxia, whereas larvae vigorously attempt escape but then die if anoxia exceeds 2 h. Here we use nuclear magnetic resonance methods to compare the metabolome of larvae and adult D. melanogaster under normoxic conditions and after various anoxic durations up to 1 h. Glucose increased during anoxia in both larvae and adults, so anoxic death by carbohydrate limitation is unlikely for either stage. Lactate and alanine were the primary anaerobic end products in both adults and larvae. During the first 30 min of anoxia, larvae accumulated anaerobic end products (predominately lactate) at a higher rate, suggesting that larvae may experience greater initial acid-base disruption during anoxic exposures. Adult Drosophila did not possess higher levels of putative protective metabolites; however, these increased during anoxia in adults and decreased in larvae. Metabolites that decreased during anoxia in larvae included mannitol, xylitol, glycerol, betaine, serine, and tyrosine, perhaps due to use as fuels, antioxidants, or binding to denatured proteins. Adults showed significant increases in glycine, taurine, and the polyols glycerol, mannitol, and xylitol, suggesting that adults upregulate protective metabolites to prevent damage. Our results suggest that lower initial metabolic demand due to paralytic hypometabolism and capacities to upregulate protective metabolites may assist the better anoxia tolerance of adult Drosophila .

When exposed to anoxia, insects rapidly go into a hypometabolic coma from which they can recover when exposed to normoxia again. However, prolonged anoxic bouts eventually lead to death in most insects, although some species are surprisingly tolerant. Anoxia challenges ATP, ion, pH and water homeostasis, but it is not clear how fast and to what degree each of these parameters is disrupted during anoxia, nor how quickly they recover. Further, it has not been investigated which disruptions are the primary source of the tissue damage that ultimately causes death. Here, we show, in the migratory locust ( Locusta migratoria ), that prolonged anoxic exposures are associated with increased recovery time, decreased survival, rapidly disrupted ATP and pH homeostasis and a slower disruption of ion ([K + ] and [Na + ]) and water balance. Locusts could not fully recover after 4 h of anoxia at 30°C, and at this point hemolymph [K + ] was elevated 5-fold and [Na + ] was decreased 2-fold, muscle [ATP] was decreased to ≤3% of normoxic values, hemolymph pH had dropped 0.8 units from 7.3 to 6.5, and hemolymph water content was halved. These physiological changes are associated with marked tissue damage in vivo and we show that the isolated and combined effects of hyperkalemia, acidosis and anoxia can all cause muscle tissue damage in vitro to equally large degrees. When locusts were returned to normoxia after a moderate (2 h) exposure of anoxia, ATP recovered rapidly (15 min) and this was quickly followed by recovery of ion balance (30 min), while pH recovery took 2-24 h. Recovery of [K + ] and [Na + ] coincided with the animals exiting the comatose state, but recovery to an upright position took ∼90 min and was not related to any of the physiological parameters examined., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Body size influences many traits including those that affect host competence, the propensity to cause new infections. Here, we employ a new framework to reveal that, for at least two infections, West Nile virus and Lyme disease, large hosts should be more competent than small ones, but their lower abundance could mitigate their impacts on local risk. By contrast, for rabies, small hosts will be disproportionately more competent than large ones, an effect amplified by the higher densities of small species. These outcomes differ quite a bit from previous approaches that incorporate allometries into epidemiological models. Subsequently, we advocate for future integrative work to resolve how interspecific variation in body size influences the emergence and spread of infections., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Hypometric scaling of aerobic metabolism [larger organisms have lower mass-specific metabolic rates (MR/g)] is nearly universal for interspecific comparisons among animals, yet we lack an agreed upon explanation for this pattern. If physiological constraints on the function of larger animals occur and limit MR/g, these should be observable as direct constraints on animals of extant species and/or as evolved responses to compensate for the proposed constraint. There is evidence for direct constraints and compensatory responses to O 2 supply constraint in skin-breathing animals, but not in vertebrates with gas-exchange organs. The duration of food retention in the gut is longer for larger birds and mammals, consistent with a direct constraint on nutrient uptake across the gut wall, but there is little evidence for evolving compensatory responses to gut transport constraints in larger animals. Larger placental mammals (but not marsupials or birds) show evidence of greater challenges with heat dissipation, but there is little evidence for compensatory adaptations to enhance heat loss in larger endotherms, suggesting that metabolic rate (MR) more generally balances heat loss for thermoregulation in endotherms. Size-dependent patterns in many molecular, physiological, and morphological properties are consistent with size-dependent natural selection, such as stronger selection for neurolocomotor performance and growth rate in smaller animals and stronger selection for safety and longevity in larger animals. Hypometric scaling of MR very likely arises from different mechanisms in different taxa and conditions, consistent with the diversity of scaling slopes for MR.

Oxygen limitation plays a key role in many pathologies; yet, we still lack a fundamental understanding of the mechanisms responsible for variation in anoxia tolerance. Most vertebrate studies suggest that anoxia tolerance involves the ability to maintain cellular ATP despite the loss of aerobic metabolism. However, insects such as adult Drosophila melanogaster are able to survive long periods of anoxia (LT 50 : ∼8 h) in a hypo-energetic state characterized by low [ATP]. In this study, we tested for possible mechanisms that allow D. melanogaster adults to survive long periods of anoxia. Adults are paralyzed within 30 s, and after 2 h of anoxia, ATP was 3% of normal, extracellular potassium concentration ([K + ] o ) increased threefold, pH dropped 1 unit, yet survival was 100%. With 0.5-6 h of anoxia, adults maintained low but constant ATP levels while [K + ] o and pH o continued to change. When returned to normoxia, adults restored [K + ] o and activity. With longer durations of anoxia, ATP levels decreased and [K + ] o rose further, and both correlated tightly with decreased survival. This response contrasts with the anoxia-sensitive larval stage (LT 50 : ∼1 h). During anoxia, larvae attempted escape for up to 30 min and after 2 h of anoxia, ATP was <1% of resting, [K + ] o increased by 50%, hemolymph pH fell by 1 unit, and survival was zero. The superior anoxia tolerance of adult D. melanogaster appears to be due to the capacity to maintain a paralytic hypometabolic state with low but non-zero ATP levels, and to be able to tolerate extreme extracellular ionic variability., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)