Gas Exchange Cycles Research Articles

Gas exchange patterns for a small, stored-grain insect pest, Tribolium castaneum.

Insects breathe using one or a combination of three gas exchange patterns; continuous, cyclic and discontinuous, which vary in their rates of exchange of oxygen, carbon dioxide and water. In general, there is a trade-off between lowering gas exchange using discontinuous exchange that limits water loss at the cost of lower metabolic rate. These patterns and hypotheses for the evolution of discontinuous exchange have been examined for relatively large insects (>20 mg) over relatively short periods (<4 h), but smaller insects and longer time periods have yet to be examined. We measured gas exchange patterns and metabolic rates for adults of a small insect pest of grain, the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), using flow-through respirometry in dry air for 48 h. All adults survived the desiccating measurement period; initially they used continuous gas exchange, then after 24 h switched to cyclic gas exchange with a 27% decrease in metabolic rate, and then after 48 h switched to discontinuous gas exchange with increased interburst duration and further decrease in metabolic rate. The successful use of the Qubit, a lower cost and so more common gas analyser, to measure respiration in the very small T. castaneum, may prompt more flow-through respirometry studies of small insects. Running such studies over long durations may help to better understand the evolution of respiration physiology and thus suggest new methods of pest management.

Body size, metabolic rate and diapause in the oriental hornet (Vespa orientalis), in two extreme climatic regions

Abstract Temperature and precipitation are major components of climate that have a pervasive effect on the phenotype and physiology of insects. The Oriental hornet has a wide geographical distribution and is the only vespid species found in a desert environment. Recently, it has extended its geographic distribution due to climate change and human activities. We compared the standard metabolic rate, diapause metabolic rate and body size of workers and gynes in two different climatic regions, 150 km apart along the northern part of the Great Rift Valley in Israel: In the north, high elevation Mediterranean habitat and in the south, low elevation, warm and dry desert. The brood in the desert region developed in higher ambient temperatures than in the Mediterranean, but only the workers of the desert had a smaller body size as adults. Worker hornets from the Mediterranean region had a higher standard metabolic rate (SMR) at the environmental temperature of main activity compared to desert hornets. Queens had a typical discontinuous gas exchange cycles (DGC) pattern, with Mediterranean queens having a 30% lower SMR. Such lower SMR might reflect an adaptation for prolonged and more profound diapause typical to the cold and long winter conditions in this region.

Why Do Insects Close Their Spiracles? A Meta-Analytic Evaluation of the Adaptive Hypothesis of Discontinuous Gas Exchange in Insects.

Simple SummaryInsects breathe with the aid of thin capillary tubes that open out to the exterior of their body as spiracles. These spiracles are often modulated in a rhythmic gas pattern known as the discontinuous gas exchange cycle. During this cycle, spiracles are either firmly shut to allow no gaseous exchange or slightly open/fully open to allow for gaseous exchange. Two explanations are put forward to rationalize this process, namely, the rhythmic pattern is to (1) reduce water loss or (2) facilitate gaseous exchange in environments with high carbon dioxide and low oxygen. Interestingly, certain insects (such as some desert insects) do not use this rhythmic pattern where it would have been most beneficial and logical. Such an observation has led to the questioning of the explanations of the discontinuous gas exchange cycle. Consequently, we attempt to resolve this controversy by conducting a meta-analysis by synthesizing apposite data from across all insects where a discontinuous gas exchange cycle has been reported. A meta-analysis allows for a shift from viewing data through the lens of a single species to an order view. Thus, our goal is to use this holistic view of data to examine the explanations of the discontinuous gas exchange cycle across multiple groups of insects.The earliest description of the discontinuous gas exchange cycle (DGC) in lepidopterous insects supported the hypothesis that the DGC serves to reduce water loss (hygric hypothesis) and facilitate gaseous exchange in hyperoxia/hypoxia (chthonic hypothesis). With technological advances, other insect orders were investigated, and both hypotheses were questioned. Thus, we conducted a meta-analysis to evaluate the merit of both hypotheses. This included 46 insect species in 24 families across nine orders. We also quantified the percent change in metabolic rates per °C change of temperature during the DGC. The DGC reduced water loss (−3.27 ± 0.88; estimate ± 95% confidence limits [95% CI]; p < 0.0001) in insects. However, the DGC does not favor gaseous exchange in hyperoxia (0.21 ± 0.25 [estimate ± 95% CI]; p = 0.12) nor hypoxia, but did favor gaseous exchange in normoxia (0.27 ± 0.26 [estimate ± 95% CI]; p = 0.04). After accounting for variation associated with order, family, and species, a phylogenetic model reflected that metabolic rate exhibited a significant, non-zero increase of 8.13% (± 3.48 95% CI; p < 0.0001) per °C increase in temperature. These data represent the first meta-analytic attempt to resolve the controversies surrounding the merit of adaptive hypotheses in insects.

Discontinuous gas exchange in Madagascar hissing cockroaches is not a consequence of hysteresis around a fixed PCO2threshold.

It has been hypothesised that insects display discontinuous gas-exchange cycles (DGCs) as a result of hysteresis in their ventilatory control, where CO2-sensitive respiratory chemoreceptors respond to changes in haemolymph PCO2only after some delay. If correct, DGCs would be a manifestation of an unstable feedback loop between chemoreceptors and ventilation, causing PCO2to oscillate around some fixed threshold value: PCO2above this ventilatory threshold would stimulate excessive hyperventilation, driving PCO2below the threshold and causing a subsequent apnoea. This hypothesis was tested by implanting micro-optodes into the haemocoel of Madagascar hissing cockroaches and measuring haemolymph PO2and PCO2simultaneously during continuous and discontinuous gas exchange. The mean haemolymph PCO2of 1.9 kPa measured during continuous gas exchange was assumed to represent the threshold level stimulating ventilation, and this was compared with PCO2levels recorded during DGCs elicited by decapitation. Cockroaches were also exposed to hypoxic (PO210 kPa) and hypercapnic (PCO22 kPa) gas mixtures to manipulate haemolymph PO2and PCO2. Decapitated cockroaches maintained DGCs even when their haemolymph PCO2was forced above or below the putative ∼2 kPa ventilation threshold, demonstrating that the characteristic oscillation between apnoea and gas exchange is not driven by a lag between changing haemolymph PCO2and a PCO2chemoreceptor with a fixed ventilatory threshold. However, it was observed that the gas exchange periods within the DGC were altered to enhance O2 uptake and CO2 release during hypoxia and hypercapnia exposure. This indicates that while respiratory chemoreceptors do modulate ventilatory activity in response to haemolymph gas levels, their role in initiating or terminating the gas exchange periods within the DGC remains unclear.

An aromatic ionomer in the anode catalyst layer improves the start-up durability of polymer electrolyte fuel cells

The sulfonated polyphenylene ionomer, SPP-QP, being used as the anode binder, improves the durability of polymer electrolyte fuel cells in the gas exchange cycle test simulating start-up conditions.

Respiratory Strategies in Relation to Ecology and Behaviour in Three Diurnal Namib Desert Tenebrionid Beetles.

Simple SummaryThe Namib Desert has a large diversity of darkling beetle species. All these beetles are flightless and feed on plant detritus. The aim of this study was to investigate whether the respiration strategies used by these beetles are linked to their ecology and behaviour. Three beetle species, which are all active during the day, in direct sunshine, running across the sand surface, but found in different parts of the desert, were chosen for this study. All three beetle species used intermittent breathing. They held their breath for several minutes and released CO2 in pulses. The large beetle species, which runs rapidly on the dune slip face when air temperatures are high, used evaporative cooling to prevent over-heating. The water used in the cooling comes from their respiratory surfaces and is replenished from metabolising food and drinking water droplets left on vegetation after a fog event. The two smaller beetle species, which inhabit the gravel plains, limit the area of the respiratory surface exposed to the atmosphere, which reduces body water loss. These two beetle species are not known to drink water, and thus have a greater need to conserve their body water.The respiratory physiology of three diurnal ultraxerophilous tenebrionid beetles inhabiting either the dune slipface or gravel plain in the Namib Desert was investigated. The role of the mesothoracic spiracles and subelytral cavity in gas exchange was determined by flow-through respirometry. All three species exhibited the discontinuous gas exchange cycles with a distinct convection based flutter period and similar mass specific metabolic rates. There was variation in their respiration mechanics that related to the ecology of the species. The largest beetle species, Onymacris plana, living on the dune slipface, has a leaky subelytral cavity and used all its spiracles for gas exchange. Thus, it could use evaporative cooling from its respiratory surface. This species is a fog harvester as well as able to replenish water through metabolising fats while running rapidly. The two smaller species inhabiting the gravel plains, Metriopus depressus and Zophosis amabilis, used the mesothoracic spiracles almost exclusively for gas exchange as well as increasing the proportional length of the flutter period to reduce respiratory water loss. Neither species have been reported to drink water droplets, and thus conserving respiratory water would allow them to be active longer.

Metabolic responses to starvation and feeding contribute to the invasiveness of an emerging pest insect

Metabolic rate, and the flexibility thereof, is a complex trait involving several inter-linked variables that can influence animal energetics, behavior, and ultimately, fitness. Metabolic traits respond readily to ambient temperature variation, in some cases increasing relative or absolute energetic costs, while in other cases, depending on the organism’s metabolic and behavioral responses to changing conditions, resulting in substantial energy savings. To gain insight into the rapid recent emergence of the indigenous South African longhorn beetle Cacosceles newmannii as a crop pest in sugarcane, a better understanding of its metabolic rate, feeding response, digestion times, and aerobic scope is required, in conjunction with any behavioral responses to food availability or limitation thereof. Here, we therefore experimentally determined metabolic rate, estimated indirectly as CO2 production using flow-through respirometry, in starved, fasted, and fed C. newmannii larvae, at 20 °C and 30 °C. We estimated multiple parameters of metabolic rate (starved, standard, active, and maximum metabolic rates) as well as aerobic scope (AS), specific dynamic action (SDA), and the percentage time active during respirometry trials. Additionally, in individuals that showed cyclic or discontinuous gas exchange patterns, we compared rate, volume, and duration of cycles, and how these were influenced by temperature. Standard and active metabolic rate, and AS and SDA were significantly higher in the larvae measured at 30 °C than those measured at 20 °C. By contrast, starved and maximum metabolic rates and percentage time active were unaffected by temperature. At rest and after digestion was complete, 35% of larvae showed cyclic gas exchange at both temperatures; 5% and 15% showed continuous gas exchange at 20 °C and 30 °C respectively, and 10% and 0% showed discontinuous gas exchange at 20 °C and 30 °C respectively. We propose that the ability of C. newmannii larvae to survive extended periods of resource limitation, combined with a rapid ability to process food upon securing resources, even at cooler conditions that would normally suppress digestion in tropical insects, may have contributed to their ability to feed on diverse low energy resources typical of their host plants, and become pests of, and thrive on, a high energy host plant like sugarcane.

The Effect of Oxygen Limitation on a Xylophagous Insect's Heat Tolerance Is Influenced by Life-Stage Through Variation in Aerobic Scope and Respiratory Anatomy.

Temperature has a profound impact on insect fitness and performance via metabolic, enzymatic or chemical reaction rate effects. However, oxygen availability can interact with these thermal responses in complex and often poorly understood ways, especially in hypoxia-adapted species. Here we test the hypothesis that thermal limits are reduced under low oxygen availability – such as might happen when key life-stages reside within plants – but also extend this test to attempt to explain that the magnitude of the effect of hypoxia depends on variation in key respiration-related parameters such as aerobic scope and respiratory morphology. Using two life-stages of a xylophagous cerambycid beetle, Cacosceles (Zelogenes) newmannii we assessed oxygen-limitation effects on metabolic performance and thermal limits. We complement these physiological assessments with high-resolution 3D (micro-computed tomography scan) morphometry in both life-stages. Results showed that although larvae and adults have similar critical thermal maxima (CTmax) under normoxia, hypoxia reduces metabolic rate in adults to a greater extent than it does in larvae, thus reducing aerobic scope in the former far more markedly. In separate experiments, we also show that adults defend a tracheal oxygen (critical) setpoint more consistently than do larvae, indicated by switching between discontinuous gas exchange cycles (DGC) and continuous respiratory patterns under experimentally manipulated oxygen levels. These effects can be explained by the fact that the volume of respiratory anatomy is positively correlated with body mass in adults but is apparently size-invariant in larvae. Thus, the two life-stages of C. newmannii display key differences in respiratory structure and function that can explain the magnitude of the effect of hypoxia on upper thermal limits.

In-situ 3D CT-XAFS Imaging of Pt/C Cathode Catalysts in Polymer Electrolyte Fuel Cell during Degradation Processes by Anode Gas Exchange Cycles

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Variability in metabolic rate and gas exchange patterns of the Colorado potato beetle of winter and prolonged diapauses

AbstractWe compare standard metabolic rate (VCO2) and gas exchange patterns in the Colorado potato beetle (Leptinotarsa decemlineata Say) in winter diapause (i.e. lasting only one overwintering period) with those of beetles in prolonged diapause (i.e. diapause lasting 2 or 3 years). The length of diapause is estimated by the behaviour of the beetles: burrowing into soil as a sign of the beginning of diapause and emergence from the soil as the ending of diapause. Measurement with a flow‐through carbon dioxide (CO2) infrared gas analyzer reveals that most beetles in winter or prolonged diapause display distinct discontinuous gas exchange cycles at 23 °C. Beetles with cyclic gas exchange and continuous breathing do not survive the winter. Beetles in prolonged diapause are characterized by a three‐ to five‐fold lower standard metabolic rates, longer discontinuous gas exchange periods and shorter CO2 releases (open phase) than those in winter diapause.

Why do models of insect respiratory patterns fail?

Insects exchange respiratory gases using an astonishing diversity of patterns. Of these, discontinuous gas exchange cycles (DGCs) have received the most study, but there are many other patterns exhibited intraspecifically and interspecifically. Moreover, some individual insects transition between patterns based on poorly understood combinations of internal and external factors. Why have biologists failed, so far, to develop a framework capable of explaining this diversity? Here, we propose two answers. The first is that the framework will have to be simultaneously general and highly detailed. It should describe, in a universal way, the physical and chemical processes that any insect uses to exchange gases through the respiratory system (i.e. tracheal tubes and spiracles) while simultaneously containing enough morphological, physiological and neural detail that it captures the specifics of patterns exhibited by any species or individual. The second difficulty is that the framework will have to provide ultimate, evolutionary explanations for why patterns vary within and among insects as well as proximate physiological explanations for how different parts of the respiratory system are modified to produce that diversity. Although biologists have made significant progress on all of these problems individually, there has been little integration among approaches. We propose that renewed efforts be undertaken to integrate across levels and approaches with the goal of developing a new class of general, flexible models capable of explaining a greater fraction of the observed diversity of respiratory patterns.

Respiratory patterns in field collected brown locust, Locustana pardalina, in the gregarious phase

In this paper we report on the metabolic rates and respiratory patterns measured from gregarious brown locusts, Locustana pardalina, collected from the Nama Karoo region in South Africa. All five instar hopper stages and adults were collected over a three year period when significant numbers of locust swarms were seen. Flow-through respirometry was used to measure the CO2 emission from individual locusts from all the developmental stages and adults within a week of collection. Carbon dioxide emission scaled hypometrically with mass, 0.863 ± 0.026. Except in the 1st and 5th instar stage there was no difference in the mass specific rate of CO2 emission (V̇<ce:small-caps>CO2</ce:small-caps>). These had significantly higher metabolic rates compared to the other stages which reflects their biology, with the 1st instar undergoing rapid growth and the 5th instar also undergoing rapid growth and development in preparation for becoming an adult. The 1st instars used a form of continuous gas exchange while all the other stages showed discontinuous gas exchange cycles. A clear burst phase and interburst periods could be seen. The 2nd and 3rd instars use mainly diffusion to expel CO2 and so exhibited an open form of the burst phase. There was an increase in CO2 volleys seen in the burst phase from the 4th instar stage onwards thus indicating an increased use of convection. There was no change in the duration or frequency of the discontinuous gas exchange cycles through the locust development or with body mass.

Intricate but tight coupling of spiracular activity and abdominal ventilation during locust discontinuous gas exchange cycles.

Discontinuous gas exchange (DGE) is the best studied among insect gas exchange patterns. DGE cycles comprise three phases, which are defined by their spiracular state: closed, flutter and open. However, spiracle status has rarely been monitored directly; rather, it is often assumed based on CO2 emission traces. In this study, we directly recorded electromyogram (EMG) signals from the closer muscle of the second thoracic spiracle and from abdominal ventilation muscles in a fully intact locust during DGE. Muscular activity was monitored simultaneously with CO2 emission, under normoxia and under various experimental oxic conditions. Our findings indicate that locust DGE does not correspond well with the commonly described three-phase cycle. We describe unique DGE-related ventilation motor patterns, coupled to spiracular activity. During the open phase, when CO2 emission rate is highest, the thoracic spiracles do not remain open; rather, they open and close rapidly. This fast spiracle activity coincides with in-phase abdominal ventilation, while alternating with the abdominal spiracle and thus facilitating a unidirectional air flow along the main trachea. A change in the frequency of rhythmic ventilation during the open phase suggests modulation by intra-tracheal CO2 levels. A second, slow ventilatory movement pattern probably serves to facilitate gas diffusion during spiracle closure. Two flutter-like patterns are described in association with the different types of ventilatory activity. We offer a modified mechanistic model for DGE in actively ventilating insects, incorporating ventilatory behavior and changes in spiracle state.

The mechanisms underlying the production of discontinuous gas exchange cycles in insects.

This review examines the control of gas exchange in insects, specifically examining what mechanisms could explain the emergence of discontinuous gas exchange cycles (DGCs). DGCs are gas exchange patterns consisting of alternating breath-hold periods and bouts of gas exchange. While all insects are capable of displaying a continuous pattern of gas exchange, this episodic pattern is known to occur within only some groups of insects and then only sporadically or during certain phases of their life cycle. Investigations into DGCs have tended to emphasise the role of chemosensory thresholds in triggering spiracle opening as critical for producing these gas exchange patterns. However, a chemosensory basis for episodic breathing also requires an as-of-yet unidentified hysteresis between internal respiratory stimuli, chemoreceptors, and the spiracles. What has been less appreciated is the role that the insect's central nervous system (CNS) might play in generating episodic patterns of ventilation. The active ventilation displayed by many insects during DGCs suggests that this pattern could be the product of directed control by the CNS rather than arising passively as a result of self-sustaining oscillations in internal oxygen and carbon dioxide levels. This paper attempts to summarise what is currently known about insect gas exchange regulation, examining the location and control of ventilatory pattern generators in the CNS, the influence of chemoreceptor feedback in the form of O2 and CO2/pH fluctuations in the haemolymph, and the role of state-dependent changes in CNS activity on ventilatory control. This information is placed in the context of what is currently known regarding the production of discontinuous gas exchange patterns.

Palladium composite membrane fabricated on rough porous alumina tube without intermediate layer for hydrogen separation

A thin palladium composite membrane without any modified layer was successfully obtained on a rough porous alumina substrate. Prior to the fabrication of palladium membrane, a poly(vinyl) alcohol (PVA) layer was first coated onto the porous substrate by dip-coating technique to improve its surface roughness and pore size. After deposition of palladium membrane on the PVA modified substrate, the polymer layer can be completely removed from the composite membrane by heat treatment. The microstructure of the palladium composite membrane was characterized in detail using SEM, EDXS and XRD analysis. Permeation measurements were carried out using H2 and N2 at temperatures of 623 K, 673 K, 723 K and 773 K. The results indicated that the hydrogen permeation flux of 0.238 mol m−2 s−1 with H2 separation factor α(H2/N2) of 956 for the as-prepared palladium membrane was obtained at 773 K and 100 kPa. Furthermore, the good membrane stability was proven during the total operation time of 160 h at the temperature range of 623 K–773 K and gas exchange cycles of 30 between hydrogen and nitrogen at 723 K.

Metabolism and gas exchange patterns in Rhodnius prolixus

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.

Gas exchange pattern in the two‐spot ladybird beetle, Adalia bipunctata, differs in dry vs. moist air

AbstractGas exchange patterns in the ladybird beetle, Adalia bipunctata (L.) (Coleoptera: Coccinellidae), were investigated using an infrared gaseous analyser (IRGA) and a coulometric O2 respirometer (manometric–volumetric system). Before testing, the beetles were kept either in dry (dehydrated) or moist (hydrated) conditions for 1 day. Their subsequent gas exchange patterns did not depend on their state of humidity but rather were controlled by the humidity of the insect chamber during gas exchange measurement. If this chamber contained dry air, the beetles exhibited CO2 release by burst, which we interpreted as cyclic gas exchange (CGE) with inter‐burst periods, but if the chamber was switched to contain moist air, then cyclic CO2 release was soon abandoned and a pattern of continuous gas exchange appeared. Measurements with the coulometric respirometer in moist air showed that continuous gas exchange was often associated with weak abdominal pulsations, which we interpreted as active ventilation. Their metabolic rate was lower during gas exchange cycles than during continuous gas exchange. We revealed that in the ladybird beetle metabolic rate increased in moist air when the gas exchange pattern transitioned from cyclic to continuous.

The opening-closing rhythms of the subelytral cavity associated with gas exchange patterns in diapausing Colorado potato beetle, Leptinotarsa decemlineata

The opening-closing rhythms of the subelytral cavity and associated gas exchange patterns were monitored in diapausing Leptinotarsa decemlineata beetles. Measurements were made by means of a flow-through CO2 analyser and a coulometric respirometer. Under the elytra of these beetles there is a more or less tightly enclosed space, the subelytral cavity (SEC). When the cavity was tightly closed, air pressure inside was sub-atmospheric, as a result of oxygen uptake into the tracheae by the beetle. In about half of the beetles, regular opening-closing rhythms of the SEC were observed visually and also recorded; these beetles displayed a discontinuous gas exchange pattern. The SEC opened at the start of the CO2 burst and was immediately closed. On opening, a rapid passive suction inflow of atmospheric air into the SEC occurred, recorded coulometrically as a sharp upward peak. As the CO2 burst lasted beyond the closure of the SEC, we suggest that most of the CO2 was expelled through the mesothoracic spiracles. In the remaining beetles, the SEC was continually semi-open, and cyclic gas exchange was exhibited. The locking mechanisms and structures between the elytra and between the elytra and the body were examined under a stereomicroscope and by means of microphotography. We conclude that at least some of the L. decemlineata diapausing beetles were able to close their subelytral cavity tightly, and that the cavity then served as a water-saving device.

Effects of Repeated Feed-Gas Exchange Processes on Degradation Processes of Pt-Based PEFC Cathode Catalysts Studied by In-situ XAFS, STEM-EDS and Electrochemical Methods

Repeated anode gas exchange (AGEX) cycles and cathode gas exchange (CGEX) cycles were applied to MEAs with Pt3Co/C cathode catalysts to investigate the degradation processes and to compare with the case of Pt/C. The structural and electronic changes of the catalysts during the feed gas-exchange cycles were examined at the top position area of the MEAs by in-situ XAFS and at the top and bottom positions by STEM-EDS. The ORR performance decreased steadily by the AGEX cycles though STEM revealed neither large cracks nor voids nor detached particles in the top position area of the MEA by the AGEX cycles where the electrochemical surface area (ECSA) remained unchanged. On the other hand, neither ORR performance nor ECSA decreased by the CGEX cycles though the particle size only in the bottom position area increased a little after the CGEX treatment. In-situ time-resolved XAFS spectra at Pt LIII-edge showed a decrease of the XANES white line peak height in case of the AGEX, which is not due to particle growth.

DGC not necessarily saving water

Equipped with a delicate network of tubules that deliver oxygen directly to every tissue in the body, insects open and close the valves (spiracles) at the ends of the tubules (tracheae) to admit oxygen to the body and release carbon dioxide in a series of well-established patterns matched to their activity. One such pattern, known as the discontinuous gas exchange cycle (DGC), occurs when some insects are inactive. The spiracles are initially closed for extended periods as the insect consumes oxygen from the air in the closed tracheae. However, once the oxygen level has fallen sufficiently, the spiracles begin to open and close rapidly (flutter phase), to draw in more air. The carbon dioxide levels in the body then continue rising until they are high enough to trigger the final phase of the cycle, when the spiracles open for an extended period as the insect releases the accumulated carbon dioxide and takes in more oxygen ready to begin the cycle again. Yet, how this particular gas exchange pattern evolved is a mystery. Several competing theories had been proposed, but Eran Gefen says, ‘None of the existing hypotheses… is backed by unequivocal support’. Explaining that one theory suggests that DGC evolved to protect insects from dehydration, Shu-Ping Huang, Stav Talal, Amir Ayali and Gefen decided to test whether grasshoppers from three dramatically different environments saved water when using DGC.‘We wanted to know whether xeric grasshoppers [that live in dry conditions] “do DGC better”’, says Gefen, so Huang and Talal headed out to three locations in Israel – ranging from the desiccated Negev desert to a hill overlooking the Sea of Galilee (where the rainfall is higher), to the wettest location, Mount Hermon on the Syrian border – to collect grasshoppers endemic to each environment. Back in the lab, Huang began the laborious process of measuring the insects’ gas exchange patterns and water losses over periods of several hours before scrutinising the carbon dioxide traces to identify examples of DGC and continuous breathing for analysis.Noticing that 60% of the desert dwelling Tmethis pulchripennis grasshoppers used DGC while only 19% of the grasshoppers from the most humid environment – Ocneropsis lividipes – used DGC, Huang then compared the insects’ respiratory water loss rates when they were breathing discontinuously and continuously. If the grasshoppers were breathing discontinuously to protect themselves from desiccation, they would all reduce their respiratory water losses when they used DGC. However, the team was impressed to see that even though the desert species (T. pulchripennis) reduced its respiratory water losses, neither of the species that were adapted to more humid conditions (Ocneropsis bethlemita and O. lividipes) did. ‘DGC in the two Ocneropsis species did not result in any measureable water savings,’ says Gefen, adding ‘Our results suggest that DGC has not necessarily evolved as a water-saving mechanism’.Instead, Gefen suggests that the ability to reduce water loss through DGC in some, but not all, species may be related to differences in tracheal dimensions. He explains that although T. pulchripennis may have developed a large tracheal system to conserve water during DGC, there is another possible explanation: that T. pulchripennis – which is the only insect in the study that can fly – evolved a larger tracheal system to deliver sufficient oxygen for the high metabolic demands of flight. ‘It would be extremely interesting to tease apart the two in the future’, says Gefen.