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Background: Wheat grain containers or silos can be perfect habitats for insects, which generate large economic losses to grain production. Natural alternatives to synthetic insecticides have grown in popularity because of health, economic and ecological issues. Diatomaceous earth is a natural compound that has an insecticide effect by enhancing an insect's dehydration with no toxicity on mammals including humans. The aim of this study is to confirm the effect of diatomaceous earth as an insecticide for the wheat grain pest, the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae) and demonstrate its underlying mechanisms as an insecticide by open-flow respirometry and scanning electron microscopy., Results: Survival bioassays of T. castaneum revealed a dose-dependent insecticide effect of diatomaceous earth. Gravimetric measurements showed that 2 days exposure to diatomaceous earth produces a significant increase of mass loss. Open-flow respirometry measurements showed an increase of total water emission rate on insects due to an increase of both, respiratory and cuticular water loss. Our study revealed that diatomaceous earth produces an increase of insect's cuticle permeability, which is responsible for elevated cuticular water loss. Scanning electron microscopy images provided visual evidence of the lipid absorbent properties of diatomaceous earth particles, and showed a tendency for higher, although not significant, damaged area of the cuticle's surface from diatomaceous earth treated insects compared to control ones., Conclusion: With state-of-the art techniques like open-flow respirometry and scanning electron microscopy, we demonstrated the underlying mechanism of diatomaceous earth as an insecticide and provided new cues for understanding the properties of the cuticle and its ecological importance. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Worldwide, with the decline of natural habitats, species with reduced niche breadth (specialists) are at greater risk of extinction as they cannot colonise or persist in disturbed habitat types. However, the role of thermal tolerance as a critical trait in understanding changes in species diversity in disturbed habitats, e.g., due to forest replacement by tree plantations, is still understudied. To examine the role of thermal tolerance on the responses of specialist and generalist species to habitat disturbances, we measured and compared local temperature throughout the year and thermotolerance traits [upper (CTmax) and lower (CTmin) thermal limits] of the most abundant species of spiders from different guilds inhabiting pine tree plantations and native Atlantic Forests in South America. Following the thermal adaptation hypothesis, we predicted that generalist species would show a wider thermal tolerance range (i.e., lower CTmin and higher CTmax) than forest specialist species. As expected, generalist species showed significantly higher CTmax and lower CTmin values than specialist species with wider thermal tolerance ranges than forest specialist species. These differences are more marked in orb weavers than in aerial hunter spiders. Our study supports the specialisation disturbance and thermal hypotheses. It highlights that habitat-specialist species are more vulnerable to environmental changes associated with vegetation structure and microclimatic conditions. Moreover, thermal tolerance is a key response trait to explain the Atlantic Forest spider's ability (or inability) to colonise and persist in human-productive land uses., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Immature mosquitoes are thought to breathe only atmospheric air through their siphons despite reports of prolonged submerged survival. We studied the survival of last-instar larvae of Aedes aegypti fully submerged at different temperatures and measured the oxygen consumption from air and water-dissolved larvae and pupae of this species under different conditions. Larvae survived much longer than expected, reaching 50% mortality only after 58, 10, and 5 days at 15°, 25°, and 35 °C, respectively. Larval to pupa molt was only observed in larvae with access to air, whereas individuals kept submerged never molted. Although most of the oxygen was obtained from the air, larvae obtained 12.72% of their oxygen from the water, while pupae took only 5.32%. In both media, temperature affected the respiration rate of the larvae, with relatively close Q 10 values (1.56 and 1.83 for water and air, respectively). A similar pattern of O 2 consumption was observed in Ae. albopictus , whose larvae obtained 12.14% of their oxygen from the water. The detailed quantification of oxygen consumption by mosquito larvae showed that water-dissolved oxygen is not negligible and physiologically relevant, challenging the idea that mosquito larvae only breathe atmospheric oxygen.

The spotted-wing fly, Drosophila suzukii, is a world-wide pest insect for which there is increasing interest in its physiological traits including metabolism and thermotolerance. Most studies focus only on survival to different time exposures to extreme temperatures, mainly in female flies. In addition, it has not been tested yet how anesthesia affects these measurements. We analyzed the effects of anesthesia by brief exposures to cold, anoxia by CO 2 or N 2 on three standard thermotolerance assays, as well as the aerobic metabolic rate in both sexes. For heat tolerance we measured CTmax by thermolimit respirometry, and CTmin and chill-coma recovery time for cold tolerance. Aerobic metabolism was calculated by CO 2 production of individual flies in real time by open flow respirometry. Results showed that females have a significantly higher V̇CO 2 for inactive (at 25 °C) and maximum metabolic rate than males. This difference is mainly explained by body mass and disappears after mass correction. Males had a more sensitive MR to temperature than females showed by a significantly higher Q 10 (2.19 vs. 1.98, for males and females, respectively). We observed a significantly lower CTmin (X 2  = 4.27, P = 0.03) in females (3.68 ± 0.38 °C) than males (4.56 ± 0.39 °C), although we did not find significant effects of anesthesia. In contrast, anesthesia significantly modifies CTmax for both sexes (F 3,62  = 7.86, P < 0.001) with a decrease of the CTmax in cold-anesthetized flies. Finally, we found a significantly higher CTmax in females (37.87 ± 0.07 °C) than males (37.36 ± 0.09 °C). We conclude that cold anesthesia seems to have detrimental effects on heat tolerance, and females have broader thermotolerance range than males, which could help them to establish in invaded temperate regions with more variable environmental temperatures., 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. Published by Elsevier Ltd.)

Temperate species, contrary to their tropical counterparts, are exposed not only to thermally variable environments with low temperatures but also to long winters. Different selective pressures may have driven divergent physiological adaptations in closely related species with different biogeographic origins. To survive unfavourable winter conditions, Drosophila species in temperate areas generally undergo a period of reproductive dormancy, associated with a cold-induced cessation of oogenesis and metabolic reorganization. This work aims to compare cold tolerance and metabolic signatures of cold-exposed females exhibiting different reproductive maturity status (mature and immature females) of four Drosophila species from tropical vs. temperate origins. We expected that the capacity for delayed reproduction of immature females could result in the redirection of the energy-related metabolites to be utilized for surviving the cold season. To do so, we studied an array of 45 metabolites using quantitative target GC-MS profiling. Reproductively immature females of temperate species showed the lower CT min and the faster chill coma recovery time (i.e. the most cold-tolerant group). Principal component analysis captured differences across species, but also between reproductive maturity states. Notably, temperate species exhibited significantly higher levels of glucose, alanine, and gluconolactone than tropical ones. As proline and glycerol showed higher abundances in immature females of temperate species compared to the levels exhibited by the rest of the groups, we reasoned that glucose and alanine could serve as intermediates in the synthesis of these compatible solutes. All in all, our findings suggest that cold-exposed females of temperate species accumulate energy-related and protective metabolites (e.g. glycerol and proline) while delaying reproduction, and that these metabolites are relevant to cold tolerance even at modest concentrations., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Species Distribution Modelling (SDM) determines habitat suitability of a species across geographic areas using macro-climatic variables; however, micro-habitats can buffer or exacerbate the influence of macro-climatic variables, requiring links between physiology and species persistence. Experimental approaches linking species physiology to micro-climate are complex, time consuming and expensive. E.g., what combination of exposure time and temperature is important for a species thermal tolerance is difficult to judge a priori. We tackled this problem using an active learning approach that utilized machine learning methods to guide thermal tolerance experimental design for three kissing-bug species: Triatoma infestans, Rhodnius prolixus, and Panstrongylus megistus (Hemiptera: Reduviidae: Triatominae), vectors of the parasite causing Chagas disease. As with other pathogen vectors, triatomines are well known to utilize micro-habitats and the associated shift in microclimate to enhance survival. Using a limited literature-collected dataset, our approach showed that temperature followed by exposure time were the strongest predictors of mortality; species played a minor role, and life stage was the least important. Further, we identified complex but biologically plausible nonlinear interactions between temperature and exposure time in shaping mortality, together setting the potential thermal limits of triatomines. The results from this data led to the design of new experiments with laboratory results that produced novel insights of the effects of temperature and exposure for the triatomines. These results, in turn, can be used to better model micro-climatic envelope for the species. Here we demonstrate the power of an active learning approach to explore experimental space to design laboratory studies testing species thermal limits. Our analytical pipeline can be easily adapted to other systems and we provide code to allow practitioners to perform similar analyses. Not only does our approach have the potential to save time and money: it can also increase our understanding of the links between species physiology and climate, a topic of increasing ecological importance., Competing Interests: The authors have declared that no competing interests exist.

It is well known that viscosity reduces the intake rates in nectar-feeding insects, such as nectivorous ants, though it remains unclear whether viscosity imposes a higher energy investment in these insects, and how this affects their feeding motivation. To address this issue, we studied feeding behavior, metabolism, and pharyngeal pump activity in the carpenter ant Camponotus mus during ingestion of ad libitum sucrose solutions. In some solutions tylose was added to modify viscosity without changing its sucrose concentration, in a way that allowed comparing: (1) two solutions with the same viscosity and different sucrose concentration (10 T and 50), and (2) two solutions with different viscosity and the same sucrose concentration (50 and 50 T). The viscosity increase was detrimental to the metabolic rate and energy balance. Ants feeding on a solution with high sucrose concentration and increased viscosity (50 T) spent extra-time until reaching a crop load similar to that reached by ingesting the solution without tylose (50). For all solutions offered, ants started feeding with the same pharyngeal pump frequencies, reflecting a similar motivation. Interesting, when ants fed on a low sucrose concentration and increased viscosity solution (10 T), their pump frequencies dropped rapidly respect to the pure-sucrose solution (50). On the contrary, pump frequencies for 50 and 50 T remained similar until the end of the intake. Since the pump frequency is strongly modulated by the ant motivation, an increase in viscosity with low sucrose content, demotivates the ants rapidly, suggesting a rapid integration of different kinds of information about the food value. Our results helped to understand how nectivorous ants could modulate their foraging decision-making., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Leaf-cutting ants (LCAs) are amongst the most important forestry pests in South America. Currently, their control is carried out almost exclusively through the application of toxic baits of restricted use. Here we evaluate a push-pull strategy (i.e., the simultaneous use of attractant and repellent stimuli in order to divert pests) to manage LCAs Acromyrmex spp. in young willow plantations in the area of Delta of the Parana River, Argentina, a wetland ecosystem. First, we surveyed ants' selection of farmland vegetation during one year. Then, we estimated ants' preferences between the willow Salix babylonica and a subsample of plant species from farmland vegetation under laboratory conditions. Finally, we designed and performed a fully crossed experimental field assay to evaluate a push-pull strategy by using farmland vegetation as pull stimulus., Results: We surveyed 39 plant species in the area, 19 of which had been foraged by LCAs along the year. Plants were selected by species, not by abundance. In the lab, ants showed similar preference for the cultivated willow and the subsample of plant species. Push-pull was the only treatment that maintained willow remaining vegetation above 60-80% at the end of the growing season., Conclusions: For the first time the push-pull strategy was evaluated in social insects. We demonstrated that it can be successfully used to manage LCAs in young willow plantations. Our strategy generates biodiversity, which can improve the ecosystem functioning, and it can be easily implemented by producers since its design is based on regular willow plantations., (© 2020 Society of Chemical Industry.)

Triatoma infestans (Klug, 1834), the main vector of Chagas disease in Latin America, is regularly controlled by spraying the pyrethroid deltamethrin, to which some populations have developed resistance. The three main mechanisms of resistance are 1) metabolic resistance by overexpression or increased activity of detoxifying enzymes, 2) target site mutations, and 3) cuticle thickening/modification. We use open-flow respirometry to measure real-time H2O loss rate (V˙H2O) and CO2 production rate (V˙CO2), on nymphs from susceptible and resistant populations before and after exposure to the insecticide to understand the underlying mechanisms of resistance in live insects. Lack of differences in V˙H2O between populations suggested that cuticular thickness/composition is not acting as a relevant resistance mechanism. Similarly, there was no difference in resting V˙CO2, suggesting a trade-off between resistance mechanisms and other physiological processes. The increment in V˙CO2 after application of deltamethrin was similar in both populations, which suggested that while enhanced enzymatic detoxification may play a role in resistance expression in this population, the main mechanism involved should be a passive one such as target site mutations. Open-flow respirometry provided useful evidence for evaluating the mechanisms involved in deltamethrin resistance. Using this technique could improve efficiency of scientific research in the area of insecticide resistance management, leading to a faster decision making and hence improved control results., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The sense of taste provides information about the "good" or "bad" quality of a food source, which may be potentially nutritious or toxic. Most alkaloids taste bitter to humans, and because bitter taste is synonymous of noxious food, they are generally rejected. This response may be due to an innate low palatability or due to a malaise that occurs after food ingestion, which could even lead to death. We investigated in the kissing bug Rhodnius prolixus, whether alkaloids such as quinine, caffeine and theophylline, are merely distasteful, or if anti-appetitive responses are caused by a post-ingestion physiological effect, or both of these options. Although anti-appetitive responses were observed for the three alkaloids, only caffeine and theophylline affect metabolic and respiratory parameters that reflected an underlying physiological stress following their ingestion. Furthermore, caffeine caused the highest mortality. In contrast, quinine appears to be a merely unpalatable compound. The sense of taste helps insects to avoid making wrong feeding decisions, such as the intake of bitter/toxic foods, and thus avoid potentially harmful effects on health, a mechanism preserved in obligate hematophagous insects.

Noonan syndrome and related disorders are caused by mutations in genes encoding for proteins of the RAS-ERK1/2 signaling pathway, which affect development by enhanced ERK1/2 activity. However, the mutations' effects throughout adult life are unclear. In this study, we identify that the protein most commonly affected in Noonan syndrome, the phosphatase SHP2, known in Drosophila as corkscrew (CSW), controls life span, triglyceride levels, and metabolism without affecting ERK signaling pathway. We found that CSW loss-of-function mutations extended life span by interacting with components of the insulin signaling pathway and impairing AKT activity in adult flies. By expressing csw-RNAi in different organs, we determined that CSW extended life span by acting in organs that regulate energy availability, including gut, fat body and neurons. In contrast to that in control animals, loss of CSW leads to reduced homeostasis in metabolic rate during activity. Clinically relevant gain-of-function csw allele reduced life span, when expressed in fat body, but not in other tissues. However, overexpression of a wild-type allele did not affect life span, showing a specific effect of the gain-of-function allele independently of a gene dosage effect. We concluded that CSW normally regulates life span and that mutations in SHP2 are expected to have critical effects throughout life by insulin-dependent mechanisms in addition to the well-known RAS-ERK1/2-dependent developmental alterations., (Copyright © 2020 Ruzzi, Schilman, San Martin, Lew, Gelb and Pagani.)

Temperature is recognized as the most influential abiotic factor on the distribution and dispersion of most insect species including Rhodnius prolixus (Stål, 1859) and Triatoma infestans (Klug, 1834), the two most important Chagas disease vectors. Although, these species thermotolerance range is well known their plasticity has never been addressed in these or any other triatomines. Herein, we investigate the effects of acclimation on thermotolerance range and resistance to stressful low temperatures by assessing thermal critical limits and 'chill-coma recovery time' (CCRT), respectively. We found positive effects of acclimation on thermotolerance range, especially on the thermal critical minimum of both species. In contrast, CCRT did not respond to acclimation in either. Our results reveal the plasticity of these Triatomines thermal tolerance in response to a wide range of acclimation temperatures. This presumably represents a physiological adaptation to daily or seasonal temperature variation with concomitant improvement in dispersion potential., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The range of thermal tolerance is one of the main factors influencing the geographic distribution of species. Climate change projections predict increases in average and extreme temperatures over the coming decades; hence, the ability of living beings to resist these changes will depend on physiological and adaptive responses. On an evolutionary scale, changes will occur as the result of selective pressures on individual heritable differences. In this work, we studied the genetic basis of tolerance to high temperatures in the fly Drosophila melanogaster and whether this species presents sufficient genetic variability to allow expansion of its upper thermo-tolerance limit. To do so, we used adult flies derived from a natural population belonging to the Drosophila Genetic Reference Panel, for which genomic sequencing data are available. We characterized the phenotypic variation of the upper thermal limit in 34 lines by measuring knockdown temperature (i.e., critical thermal maximum [CTmax]) by exposing flies to a ramp of increasing temperature (0.25°C/min). Fourteen percent of the variation in CTmax is explained by the genetic variation across lines, without a significant sexual dimorphism. Through a genomewide association study, 12 single nucleotide polymorphisms associated with the CTmax were identified. In most of these SNPs, the less frequent allele increased the upper thermal limit suggesting that this population harbors raw genetic variation capable of expanding its heat tolerance. This potential upper thermal tolerance increase has implications under the global warming scenario. Past climatic records show a very low incidence of days above CTmax (10 days over 25 years); however, future climate scenarios predict 243 days with extreme high temperature above CTmax from 2045 to 2070. Thus, in the context of the future climate warming, rising temperatures might drive the evolution of heat tolerance in this population by increasing the frequency of the alleles associated with higher CTmax.

Environmental temperature is an abiotic factor with great influence on biological processes of living beings. Jensen's inequality states that for non-lineal processes, such as most biological phenomena, the effects of thermal fluctuations cannot be predicted from mean constant temperatures. We studied the effect of daily temperature fluctuation (DTF) on Rhodnius prolixus, a model organism in insect physiology, and an important vector of Chagas disease. We measured development time from egg to adult, fecundity, fertility, body mass reduction rate (indirect measurement of nutrient consumption rates) and survival after a single blood meal. Insects were reared at constant temperature (24 °C), or with a DTF (17-32 °C; mean = 24 °C). Taking into account Jensen's inequality as well as the species tropical distribution, we predict that living in a variable thermal environment will have higher costs than inhabiting a stable one. Development time and fertility were not affected by DTF. However, fecundity was lower in females reared at DTF than at constant temperature, and males had higher body mass reduction rate and lower survival in the DTF regime, suggesting higher costs associated to fluctuating thermal environments. At a population and epidemiological level, higher energetic costs would imply an increase in nutrient consumption rate, biting frequency, and, consequently increasing disease transmission from infected insects. On the contrary, lower fecundity could be associated with a decrease in population growth. This knowledge will not only provide basic information to the field of insect ecophysiology, but also could be a useful background to develop population and disease transmission models., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In order to assess how triatomines (Hemiptera, Reduviidae), Chagas disease vectors, are distributed through Latin America, we analysed the relationship between the ecological niche and the limits of the physiological thermal niche in seven species of triatomines. We combined two methodological approaches: species distribution models, and physiological tolerances. First, we modelled the ecological niche and identified the most important abiotic factor for their distribution. Then, thermal tolerance limits were analysed by measuring maximum and minimum critical temperatures, upper lethal temperature, and 'chill-coma recovery time'. Finally, we used phylogenetic independent contrasts to analyse the link between limiting factors and the thermal tolerance range for the assessment of ecological hypotheses that provide a different outlook for the geo-epidemiology of Chagas disease. In triatomines, thermo-tolerance range increases with increasing latitude mainly due to better cold tolerances, suggesting an effect of thermal selection. In turn, physiological analyses show that species reaching southernmost areas have a higher thermo-tolerance than those with tropical distributions, denoting that thermo-tolerance is limiting the southern distribution. Understanding the latitudinal range along its physiological limits of disease vectors may prove useful to test ecological hypotheses and improve strategies and efficiency of vector control at the local and regional levels., (© 2017 The Royal Entomological Society.)

Small ectotherms, such as insects, with high surface area-to-volume ratios are usually at risk of dehydration in arid environments. We hypothesize that desiccation tolerance in insects could be reflected in their distribution, which is limited by areas with high relative values of water vapor pressure deficit (VPD) (e.g., hot and dry). The main goal of this study was to explore whether incorporation of eco-physiological traits such as desiccation tolerance in arid environments can improve our understanding of species distribution models (SDM). We use a novel eco-physiological approach to understand the distribution and the potential overlap with their fundamental niche in triatomine bugs, Chagas disease vectors. The desiccation dimension for T. infestans, T. delpontei, T. dimidiata, and T. sordida niches seems to extend to very dry areas. For T. vitticeps, xeric areas seem to limit the geographical range of their realized niche. The maximum VPD limits the western and southern distributions of T. vitticeps, T. delpontei, and T. patagonica. All species showed high tolerance to desiccation with survival times (35 °C-RH ~ 15%) ranging from 24 to 38 days, except for T. dimidiata (9 days), which can be explained by a higher water-loss rate, due to a higher cuticular permeability along with a higher critical water content. This approach indicates that most of these triatomine bugs could be exploiting the dryness dimension of their fundamental niche. Incorporating such species-specific traits in studies of distribution, range, and limits under scenarios of changing climate could enhance predictions of movement of disease-causing vectors into novel regions.

Females of the haematophagous bug Rhodnius prolixus attach their eggs in clusters on substrates related to their hosts, such as nests or avian feathers. Because the hosts are an enormous food resource as well as potential predators, the choice of the site and pattern of oviposition could have an important adaptive value. Here we investigated proximate and a potential ultimate cause of this aggregated pattern of laid eggs. First, we studied proximal causes by analyzing the use of chemical or physical cues associated with aggregated oviposition in R. prolixus. For all terrestrial organisms there is a trade-off between exchange of respiratory gases and water loss. Particularly, insect eggs are highly susceptible to this trade-off because they do not obtain water from the environment, hence our second objective is to study the possible mechanisms involved in dehydration resistance in this species. Therefore we examined the dynamics of change in CO 2 release rate (ṀCO 2 ), and water loss rate (ṀH 2 O) in relation to embryo development as energetic demands increase, and tested the energetic or hygric efficiency hypothesis as a potential ultimate cause of aggregated oviposition. This hypothesis states that grouped eggs consume less energy or lose less water than equal numbers of isolated eggs, the latter being more susceptible to dehydration. Results indicated the use of physical external cues such as dummy eggs or edges of the oviposition substrates, but we did not find any chemical cues associated with the aggregated pattern of oviposition. There are no energetic or hygric benefits associated with egg's aggregated pattern. However, when we analyzed the ṀCO 2 and ṀH 2 O change in relation to embryo development, we found a fairly constant and low ṀH 2 O albeit a clear increase in ṀCO 2 , suggesting a tightly control of egg's desiccation tolerance. This high resistance to desiccation coupled with a temporal strategy of hatching allows R. prolixus embryos to successfully develop and hatch under harsh environmental conditions., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Long-term exposure to low temperatures during adult maturation might decrease fertility after cold recovery as a consequence of carry-over effects on reproductive tissues. This pattern should be more pronounced in tropical than in temperate species as protective mechanisms against chilling injuries are expected to be more effective in the latter. We initially determined the lower thermal thresholds to induce ovarian maturation in four closely related Drosophila species, two inhabiting temperate regions and the other two tropical areas of South America. As expected, only temperate species regularly experience cold-inducing conditions for reproductive arrest during winter in their natural environment. Subsequently, we exposed reproductively arrested and mature females to cold-inducing conditions for reproductive arrest over a long period. Following cold exposure, tropical species exhibited a dramatic fertility decline, irrespective of reproductive status. In contrast, not only were temperate females fecund and fertile but also fertility was superior in females that underwent cold-induced reproductive arrest, suggesting that it might act as a protecting mechanism ensuring fertility after cold recovery. Based on these findings, we decided to evaluate the extent to which reproductive status affects cold tolerance and energy metabolism at low temperature. We found a lower metabolic rate and a higher cold tolerance in reproductively arrested females, although only temperate species attained high levels of chill tolerance. These findings highlight the role of cold-induced reproductive arrest as part of an integrated mechanism of cold adaptation that could potentially contribute to the spread of temperate species into higher latitudes or altitudes., (© 2017. Published by The Company of Biologists Ltd.)

Insect's metabolic rate and patterns of gas-exchange varies according to different factors such as: species, activity, mass, and temperature among others. One particular striking pattern of gas-exchange in insects is discontinuous gas-exchange cycles, for which many different hypotheses regarding their evolution have been stated. This article does not pretend to be an extensive review on the subject, rather to focus on the work performed on the haematophagous bug Rhodnius prolixus, a model organism used from the mid XX century until present days, with the great influence of Wigglesworth and his students/collaborator's work. I have no doubt that the renovated field of insect gas-exchange has a bright future and will advance at large gaits thank to the help of this model organism, R. prolixus, whose entire genome has recently being unraveled., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Haematophagous insects suffer big changes in water needs under different levels of starvation. Rhodnius prolixus is the most important haematophagous vector of Chagas disease in the north of South America and a model organism in insect physiology. Although there have been some studies on patterns of gas exchange and metabolic rates, there is little information regarding water loss in R. prolixus. We investigated whether there is any modulation of water loss and metabolic rate under different requirements for saving water. We measured simultaneously CO2 production, water emission and activity in individual insects in real time by open-flow respirometry at different temperatures (15, 25 and 35°C) and post-feeding days (0, 5, 13 and 29). We found: (1) a clear drop in metabolic rate between 5 and 13 days after feeding that cannot be explained by activity and (2) a decrease in water loss rate with increasing starvation level, by a decrease in cuticular water loss during the first 5 days after feeding and a drop in the respiratory component thereafter. We calculated the surface area of the insects and estimated cuticular permeability. In addition, we analysed the pattern of gas exchange; the change from a cyclic to a continuous pattern was affected by temperature and activity, but it was not affected by the level of starvation. Modulation of metabolic and water loss rates with temperature and starvation could help R. prolixus to be more flexible in tolerating different periods of starvation, which is adaptive in a changing environment with the uncertainty of finding a suitable host., (© 2014. Published by The Company of Biologists Ltd.)

Insects in general, and Drosophila in particular, are much more capable of surviving anoxia than vertebrates, and the mechanisms involved are of considerable biomedical and ecological interest. Temperature is likely to strongly affect both the rates of damage occurring in anoxia and the recovery processes in normoxia, but as yet there is no information on the effect of this crucial variable on recovery rates from anoxia in any animal. We studied the effects of temperature, and thus indirectly of metabolic flux rates, on survival and recovery times of individual male Drosophila melanogaster following anoxia and O(2) reperfusion. Individual flies were reared at 25°C and exposed to an anoxic period of 7.5, 25, 42.5 or 60 min at 20, 25 or 30°C. Before, during and after anoxic exposure the flies' metabolic rates (MRs), rates of water loss and activity indices were recorded. Temperature strongly affected the MR of the flies, with a Q(10) of 2.21. Temperature did not affect the slope of the relationship between time to recovery and duration of anoxic exposure, suggesting that thermal effects on damage and repair rates were similar. However, the intercept of that relationship was significantly lower (i.e. recovery was most rapid) at 25°C, which was the rearing temperature. When temperatures during exposure to anoxia and during recovery were switched, recovery times matched those predicted from a model in which the accumulation and clearance of metabolic end-products share a similar dependence on temperature.

In fasted mammals, glucose homeostasis is maintained through induction of the cAMP response element-binding protein (CREB) coactivator transducer of regulated CREB activity 2 (TORC2), which stimulates the gluconeogenic program in concert with the forkhead factor FOXO1. Here we show that starvation also triggers TORC activation in Drosophila, where it maintains energy balance through induction of CREB target genes in the brain. TORC mutant flies have reduced glycogen and lipid stores and are sensitive to starvation and oxidative stress. Neuronal TORC expression rescued stress sensitivity as well as CREB target gene expression in TORC mutants. During refeeding, increases in insulin signaling inhibited TORC activity through the salt-inducible kinase 2 (SIK2)-mediated phosphorylation and subsequent degradation of TORC. Depletion of neuronal SIK2 increased TORC activity and enhanced stress resistance. As disruption of insulin signaling also augmented TORC activity in adult flies, our results illustrate the importance of an insulin-regulated pathway that functions in the brain to maintain energy balance.

In nectar-feeding insects, sugars are an important source of fuel and energy storage. Here, we analyzed the haemolymph sugar levels in foragers of the ant Camponotus rufipes trained to collect nectar from an artificial feeder, and their dependence on the metabolic rate during feeding. The main sugar found was trehalose, followed by glucose and traces of fructose and sucrose. In foragers, trehalose level was independent of their activity and metabolic rate while feeding. Carbohydrate deprivation of the colony had a strong effect on the haemolymph sugar levels of workers, with a significant decrease in trehalose and glucose with increasing starvation. We also found a correlation between haemolymph sugar levels and behavioral states, with immobile workers having higher trehalose and fructose levels than active ones. It is suggested that under food deprivation, inside-nest workers initially stay completely immobile as a strategy to save energy, and only become active and start to search for food when the trehalose levels decrease even more. Based on a conservative estimation, well-fed ants could travel up to 500 m, or spend more than 20 h inactive at 25 degrees C, using only the energy provided by the haemolymph trehalose, before reaching the levels found in starved nest-mates.

Flightless, diurnal tenebrionid beetles are commonly found in deserts. They possess a curious morphological adaptation, the subelytral cavity (an air space beneath the fused elytra) the function of which is not completely understood. In the tenebrionid beetle Eleodes obscura, we measured abdominal movements within the subelytral cavity, and the activity of the pygidial cleft (which seals or unseals the subelytral cavity), simultaneously with total CO2 release rate and water loss rate. First, we found that E. obscura has the lowest cuticular permeability measured in flow-through respirometry in an insect (0.90 microg H2O cm(-2) Torr(-1) h(-1)). Second, it does not exhibit a discontinuous gas exchange cycle. Third, we describe the temporal coupling between gas exchange, water loss, subelytral space volume, and the capacity of the subelytral space to exchange gases with its surroundings as indicated by pygidial cleft state. Fourth, we suggest possible mechanisms that may reduce respiratory water loss rates in E. obscura. Finally, we suggest that E. obscura cannot exchange respiratory gases discontinuously because of a morphological constraint (small tracheal or spiracular conductance). This "conductance constraint hypothesis" may help to explain the otherwise puzzling phylogenetic patterns of continuous vs. discontinuous gas exchange observed in tracheate arthropods.

The deleterious effects of anoxia followed by reperfusion with oxygen in higher animals including mammals are well known. A convenient and genetically well characterized small-animal model that exhibits reproducible, quantifiable oxygen reperfusion damage is currently lacking. Here we describe the dynamics of whole-organism metabolic recovery from anoxia in an insect, Drosophila melanogaster, and report that damage caused by oxygen reperfusion can be quantified in a novel but straightforward way. We monitored CO(2) emission (an index of mitochondrial activity) and water vapor output (an index of neuromuscular control of the spiracles, which are valves between the outside air and the insect's tracheal system) during entry into, and recovery from, rapid-onset anoxia exposure with durations ranging from 7.5 to 120 minutes. Anoxia caused a brief peak of CO(2) output followed by knock-out. Mitochondrial respiration ceased and the spiracle constrictor muscles relaxed, but then re-contracted, presumably powered by anaerobic processes. Reperfusion to sustained normoxia caused a bimodal re-activation of mitochondrial respiration, and in the case of the spiracle constrictor muscles, slow inactivation followed by re-activation. After long anoxia durations, both the bimodality of mitochondrial reactivation and the recovery of spiracular control were impaired. Repeated reperfusion followed by episodes of anoxia depressed mitochondrial respiratory flux rates and damaged the integrity of the spiracular control system in a dose-dependent fashion. This is the first time that physiological evidence of oxygen reperfusion damage has been described in an insect or any invertebrate. We suggest that some of the traditional approaches of insect respiratory biology, such as quantifying respiratory water loss, may facilitate using D. melanogaster as a convenient, well-characterized experimental model for studying the underlying biology and mechanisms of ischemia and reperfusion damage and its possible mitigation.

We examined the quantitative relationship between the energetic costs and benefits of nectar collection by nectar-feeding ants, Camponotus rufipes. In the laboratory, individual workers were trained to visit an artificial feeder that provided a sucrose solution of 1%, 5%, 10%, 30% or 50% at controlled flows, in a similar span range to those observed in natural nectar sources. We measured foraging times, nectar loads collected, and CO(2) production during actual feeding, as an indication of the energy expenditure for a single forager. Results show an increase in individual metabolic rates with increasing flow rate of sugar solution, but no dependence on sucrose concentration. This increase in metabolic expenditure does not depend on the crop load attained while feeding, as intuitively expected, and is therefore a result of an increased activity brought about by the food-source profitability experienced by the forager. The energy gained during collection of sugar solution is always higher than the energy spent by the ant. Even with a food source of lower quality than a natural source, the ants gain ca. tenfold of what they spend. Based on a simplified model, we calculated that foragers of C. rufipes could travel from 0.5 to 9 km with the energy gained in a single foraging trip only. These results suggest that decreasing foraging time is more important than increasing individual energetic efficiency when workers of the nectar-feeding ant C. rufipes decide to stop drinking and return to the nest with partial crop loads.

Respiratory water loss (RWL) in insects showing continuous emission of CO(2) is poorly studied because few methodologies can measure it. Comparisons of RWL between insects showing continuous and discontinuous gas exchange cycles (DGC) are therefore difficult. We used two recently developed methodologies (the hyperoxic switch and correlation between water-loss and CO(2) emission rates) to compare cuticular permeabilities and rates of RWL in five species of ants, the Argentine ant (Linepithema humile) and four common native ant competitors. Our results showed that RWL in groups of ants with moderate levels of activity and continuous gas exchange were similar across the two measurement methods, and were similar to published values on insects showing the DGC. Furthermore, ants exposed to anoxia increased their total water loss rates by 50-150%. These results suggest that spiracular control under continuous gas exchange can be as effective as the DGC in reducing RWL. Finally, the mesic-adapted Argentine ant showed significantly higher rates of water loss and cuticular permeability compared to four ant species native to dry environments. Physiological limitations may therefore be responsible for restricting the distribution of this invasive species in seasonally dry environments.

Partitioning the relative contributions of cuticular and respiratory water loss in a tracheate arthropod is relatively easy if it undergoes discontinuous gas exchange cycles or DGCs, leaving its rate of cuticular water loss in primary evidence while its spiracles are closed. Many arthropods are not so obliging and emit CO(2) continuously, making cuticular and respiratory water losses difficult or impossible to partition. We report here that by switching ambient air from 21 to 100% O(2), marked spiracular constriction takes place, causing a transient but substantial - up to 90% - reduction in CO(2) output. A reduction in water loss rate occurs at the same time. Using this approach, we investigated respiratory water loss in Drosophila melanogaster and in two ant species, Forelius mccooki and Pogonomyrmex californicus. Our results - respiratory water loss estimates of 23%, 7.6% and 5.6% of total water loss rates, respectively - are reasonable in light of literature estimates, and suggest that the 'hyperoxic switch' may allow straightforward estimation of respiratory water loss rates in arthropods lacking discontinuous gas exchange. In P. californicus, which we were able to measure with and without a DGC, presence or absence of a DGC did not affect respiratory vs total water loss rates.

The present work examines the thermal preference of adult Rhodnius prolixus along a temperature gradient. The mean preferred temperature differed slightly between sexes: 25.0 degrees C for males versus 25.4 degrees C for females. This preference was not constant, but varied daily by about 0.2 degrees C for both sexes, and reached its highest value at the onset of the dark phase and was lowest during the light phase. A change in the preferred temperature with the level of starvation was also observed (about 1 degrees C lower after 20 days of starvation). Changes in environmental temperature strongly affected the rate of weight loss for both sexes. When insects were maintained for 20 days in a chamber at 32 degrees C, they lost significantly more weight than when kept at 24 degrees C; both water loss and nutrient conversion processes are involved. This increase in weight loss rate with increasing temperature would cause a higher biting rate and consequently higher probability of Chagas' disease transmission. Females oviposit across a range of temperatures from 22 to 33 degrees C with a peak at 25-26 degrees C. These results are compared with patterns of thermopreference in other species of triatomine, as related to differences in their distribution and tolerance to starvation.

The thermopreference of resting P. megistus (Burmeister, 1834) adults was studied in a temperature gradient. We also determined the thermopreference for oviposition and ecdysis. Thermopreference of resting individuals depended on the feeding state of the insects. Results demonstrated an initial resting preference of approximately 290 degrees C, which gradually changed to 26-27 degrees C with increasing starvation. A daily variation in thermopreference was also observed with this species. The insects showed a daily maximum preference at the beginning of the dark phase, and a minimum preference during the first half of the light phase. Ecdysis took place all along the gradient, but exhibited a maximum frequency at 30 degrees C. Most eggs were found between 25 and 29 degrees C. Results are discussed in relation to the ecology of P. megistus as compared with other related species, and the existence of mechanisms of behavioral thermoregulation in the group.

It has been observed that Triatoma infestans and Rhodnius prolixus females stridulate to reject copulatory attempts performed by males. In addition, triatomines stridulate when disturbed or handled. In the present study, the temporal structure and frequency spectra of vibrational signals produced by mechanically disturbed T. infestans, T. sordida, T. guasayana, R. prolixus and Dipetalogaster maxima were analysed and compared. The inter-ridge distances of the prosternal stridulatory organ of the same species were also measured. The frequency spectra and repetition rates were similar, despite individuals of these five species have different sizes, their stridulatory grooves have different inter-ridge distances, and also their vibratory signals exhibited different temporal patterns. The hypothesis that disturbance stridulations are non-specific signals and could function to deter predators was discussed.

In this study the substrate-borne stridulatory vibrations produced by Rhodnius prolixus females were recorded and analysed in two different behavioural contexts. In the context of sexual communication females spontaneously stridulated to reject copulatory attempts performed by males. These male-deterring stridulations were fully effective: out of 61 attempts, no copulation occurred. These stridulations consisted of short series of repetitive syllables, each one composed by a single chirp. In the context of defensive behaviour, bugs stridulated if they were clasped or restrained. These disturbance stridulations consisted of long series of repetitive syllables, each one composed by a series of short chirps and a long one. Male-deterring and disturbance stridulations differed in their temporal pattern and frequency spectra, having a main carrier frequency of about 1500 and 2200 Hz, respectively. As no differences in the inter-ridge distances along the whole stridulatory organ were found, the differences in the frequency between both signals could be explained on the basis of a different velocity of rubbing of the proboscis against the prosternal stridulatory organ. It was found that R. prolixus and the related species Triatoma infestans rubbed only the central region of the stridulatory groove (around 1/3 of the total length) to produce disturbance stridulations. The results are discussed in relation to previous work on vibrational sensitivity in R. prolixus and are also compared with results reported for T. infestans.