Respiratory strategies of tenebrionid beetles in arid Australia: does physiology beget nocturnality?

Abstract The respiratory patterns and role of mesothoracic and abdominal spiracles in gas exchange are examined using flow‐through respirometry in three species of Heleini beetles from the Simpson Desert, central Australia. Two species, Helea (Heleus) waitei and Helea sp., show a form of continuous respiration with 70% and 75% of the CO₂ being emitted from the mesothoracic spiracles, respectively. Their mass specific metabolic rates are similar and similar to other nocturnally active desert‐dwelling tenebrionid beetles. Brises blairi also shows a continuous form of respiration, with 66% of CO₂ being emitted from the mesothoracic spiracle but has a significantly higher mass specific metabolic rate. Unusually for arid‐dwelling tenebrionid beetles, all three of the study species are confirmed to be exclusively nocturnal. There is no evidence that this activity pattern is driven by current ecological factors, such as competition or predation, and it is proposed instead that nocturnality arises from physiological constraint. Heleini probably evolved under mesic conditions and lack some of the key physiological adaptations to reduce water loss that characterize day‐active tenebrionid beetles in arid environments elsewhere. In consequence, they are able to exploit arid conditions only by restricting their activity to the most benign phase of the 24‐h cycle.     

The role of the spiracles in gas exchange during development of Samia cynthia (Lepidoptera, Saturniidae).

Spiracles and the tracheal system of insects allow effective delivery of respiratory gases. During development, holometabolous insects encounter large changes in the functional morphology of gas exchange structures. To investigate changes in respiratory patterns during development, CO2-release was measured in larvae, pre-pupae and pupae of Samia cynthia (Lepidoptera, Saturniidae). Gas exchange patterns showed great variability. Caterpillars had high metabolic rates and released carbon dioxide continuously. Pre-pupae and pupae showed typical discontinuous gas exchange cycles (DGC) at reduced metabolic rates. Changes in gas exchange patterns can partly be explained with low metabolic rates during pupation. Sequential blocking of spiracles in pre-pupae and pupae reduced spiracle conductance with tracheal conductance remaining unaffected. Analysis of gas exchange patterns indicates that caterpillars and pre-pupae use more than 14 spiracles simultaneously while pupae only use 8 to 10 spiracles. Total conductance is not a simple multiple of single spiracles, but may be gradually adaptable to gas exchange demands. Surprisingly, moth pupae showed a DGC if all except one spiracle were blocked. The huge conductance of single spiracles is discussed as a pre-adaptation to high metabolic demands at the beginning and the end of the pupal as well as in the adult stage.     

Breathe softly, beetle: continuous gas exchange, water loss and the role of the subelytral space in the tenebrionid beetle, Eleodes obscura

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 role of the spiracles in gas exchange during development of Samia cynthia (Lepidoptera, Saturniidae)

Spiracles and the tracheal system of insects allow effective delivery of respiratory gases. During development, holometabolous insects encounter large changes in the functional morphology of gas exchange structures. To investigate changes in respiratory patterns during development, CO₂-release was measured in larvae, pre-pupae and pupae of Samia cynthia (Lepidoptera, Saturniidae). Gas exchange patterns showed great variability. Caterpillars had high metabolic rates and released carbon dioxide continuously. Pre-pupae and pupae showed typical discontinuous gas exchange cycles (DGC) at reduced metabolic rates. Changes in gas exchange patterns can partly be explained with low metabolic rates during pupation. Sequential blocking of spiracles in pre-pupae and pupae reduced spiracle conductance with tracheal conductance remaining unaffected. Analysis of gas exchange patterns indicates that caterpillars and pre-pupae use more than 14 spiracles simultaneously while pupae only use 8 to 10 spiracles. Total conductance is not a simple multiple of single spiracles, but may be gradually adaptable to gas exchange demands. Surprisingly, moth pupae showed a DGC if all except one spiracle were blocked. The huge conductance of single spiracles is discussed as a pre-adaptation to high metabolic demands at the beginning and the end of the pupal as well as in the adult stage.     

Novel case of a tenebrionid beetle using discontinuous gas exchange cycle when dehydrated

Abstract In this study we show a link between the respiratory method and state of hydration in an arid dwelling tenebrionid beetle ( Pimelia grandis ). Dehydrated beetles use discontinuous gas exchange cycles with a flutter period consisting of several discrete bursts of CO₂ release, whereas beetles given access to food and water showed a form of continuous CO₂ release. These data give support to the respiratory water conservation hypothesis for the discontinuous gas exchange cycle.     

Hold your breath beetle—Mites!

Respiratory gas exchange in insects occurs via a branching tracheal system. The entrances to the air‐filled tracheae are the spiracles, which are gate‐like structures in the exoskeleton. The open or closed state of spiracles defines the three possible gas exchange patterns of insects. In resting insects, spiracles may open and close over time in a repeatable fashion that results in a discontinuous gas exchange (DGE) pattern characterized by periods of zero organism‐to‐environment gas exchange. Several adaptive hypotheses have been proposed to explain why insects engage in DGE, but none have attracted overwhelming support. We provide support for a previously untested hypothesis that posits that DGE minimizes the risk of infestation of the tracheal system by mites and other agents. Here, we analyze the respiratory patterns of 15 species of ground beetle (Carabidae), of which more than 40% of individuals harbored external mites. Compared with mite‐free individuals, infested one's engaged significantly more often in DGE. Mite‐free individuals predominantly employed a cyclic or continuous gas exchange pattern, which did not include complete spiracle closure. Complete spiracle closure may prevent parasites from invading, clogging, or transferring pathogens to the tracheal system or from foraging on tissue not protected by thick chitinous layers.     

Tradeoffs between metabolic rate and spiracular conductance in discontinuous gas exchange of Samia cynthia (Lepidoptera, Saturniidae)

The insect tracheal system is a unique respiratory system, designed for maximum oxygen delivery at high metabolic demands, e.g. during activity and at high ambient temperatures. Therefore, large safety margins are required for tracheal and spiracular conductance. Spiracles are the entry to the tracheal system and play an important role in controlling discontinuous gas exchange (DGC) between tracheal system and atmosphere in moth pupae. We investigated the effect of modulated metabolic rate (by changing ambient temperature) and modulated spiracular conductance (by blocking all except one spiracles) on gas exchange patterns in Samia pupae. Both, spiracle blocking and metabolic rates, affected respiratory behavior in Samia cynthia pupae. While animals showed discontinuous gas exchange cycles at lower temperatures with unblocked spiracles, the respiratory patterns were cyclic at higher temperatures, with partly blocked spiracles or a combination of these two factors. The threshold for the transition from a discontinuous (DGC) to a cyclic gas exchange (^cycGE) was significantly higher in animals with unblocked spiracles (18.7 nmol g⁻¹ min⁻¹ vs. 7.9 nmol g⁻¹ min⁻¹). These findings indicate an important influence of spiracle conductance on the DGC, which may occur mostly in insects showing high spiracular conductances and low metabolic rates.     

The use of the anaesthetic, enflurane, for determination of metabolic rates and respiratory parameters in insects, using the ant, Camponotus maculatus (Fabricius) as the model

This study investigated the effects of the anaesthetic, enflurane, on metabolic rates and ventilation patterns in the spotted sugar ant, Camponotus maculatus, using flow-through respirometry. The standard metabolic rate was not affected by the anaesthetic. While the ants were anaesthetised they exhibited a similar discontinuous gas exchange cycle to that observed when they were voluntarily motionless, but their spiracles remained open for a longer time during the open or burst phase even though the amount of CO(2) emitted during this phase remained constant. We discuss this finding in the context of the central nervous system control of the spiracle muscle. For both the determination of standard metabolic rate and ventilation patterns the individual ant has to be motionless. From this study we recommend the use of enflurane to ensure immobility in ants, and other small active insects, during the determination of standard metabolic rates, but the anaesthetic cannot be used to quantify the respiration pattern.     

昆虫不连续气体交换

许多昆虫呼吸时气体交换是不连续的循环式进行的。根据气门开闭,一个典型的不连续气体交换循环（discontinuous gas exchangecycle, DGC）可以明显分为3个阶段: 关闭阶段,极少或没有气体交换;颤动阶段,气门迅速微开和关闭,O₂进入气管,少量CO₂释放;最后是开放阶段,大量的CO₂释放。该文综述了DGC特征及昆虫活动、温度、体重对DGC的影响,并讨论了DGC与呼吸失水、缺氧或高CO₂浓度环境有关的进化适应意义。     

The respiratory basis of locomotion in Drosophila

The respiratory system of insects has evolved to satisfy the oxygen supply during rest and energetically demanding processes such as locomotion. Flapping flight in particular is considered a key trait in insect evolution and requires an increase in metabolic activity of 10-15-fold the resting metabolism. Two major trade-offs are associated with the extensive development of the tracheal system and the function of spiracles in insects: the risk of desiccation because body water may leave the tracheal system when spiracles open for gas exchange and the risk of toxic tracheal oxygen levels at low metabolic activity. In resting animals there is an ongoing debate on the function and evolution of spiracle opening behavior, focusing mainly on discontinuous gas exchange patterns. During locomotion, large insects typically satisfy the increased respiratory requirements by various forms of ventilation, whereas in small insects such as Drosophila diffusive processes are thought to be sufficient. Recent data, however, have shown that during flight even small insects employ ventilatory mechanisms, potentially helping to balance respiratory currents inside the tracheal system. This review broadly summarizes our current knowledge on breathing strategies and spiracle function in the genus Drosophila, highlighting the gas exchange strategies in resting, running and flying animals.     

Gas exchange patterns of Pterostichus niger (Carabidae) in dry and moist air

Gas exchange patterns of adult male Pterostichus niger Schaller after hydration (i.e. given access to food and water) are compared in dry air [5–7% relative humidity (RH)] and moist air (90–97% RH) by means of flow‐through CO₂ respirometry combined with infrared probe actography. Of thirty beetles examined, slightly more than 50% showed continuous gas exchange and are not considered further. Of the remaining beetles, the majority (approximately 71%) display a pattern of cyclic gas exchange in both dry and moist air (i.e. CO₂ gas is released in bursts, with a low level of CO₂ release during the interburst periods). A minority of the beetles (four out of 30) are found to exhibit discontinuous gas exchange in both dry and moist air; this is characterized by three clearly separated states of the spiracles: closed (C), flutter (F) and open (O) phases. The pattern of cyclic gas exchange is associated with weak abdominal pulsations. After switching from moist to dry air, a small modulation of the discontinuous gas exchange cycles (maximum mean CO₂ production rate) occurs, providing no clear support for the hygric theory of discontinuous gas exchange in this species (i.e. that it serves to restrict respiratory water loss).     

Spiracle activity in moth pupae—The role of oxygen and carbon dioxide revisited

After decades of intensive research, the actual mechanism behind discontinuous gas exchange in insects has not been fully understood. One open question concerns the actual way (closed, flutter, and open) of how spiracles respond to tracheal gas concentrations. As the results of a classic paper [Burkett, B.N., Schneiderman, H.A., 1974. Roles of oxygen and carbon dioxide in the control of spiracular function in cecropia pupae. Biological Bulletin 147, 274-293] allow ambiguous interpretation, we thus reexamined the behavior of the spiracles in response to fixed, controlled endotracheal gas concentrations.The tracheal system of diapausing pupae of Attacus atlas (Saturniidae, Lepidoptera) was flushed with gas mixtures varying in PO₂ and PCO₂ while the behavior of the spiracles was monitored using changes in the pressure signal. This novel pressure based technique proved to be superior to classic visual observation of single spiracles. A two-dimensional map of the spiracle behavior in response to endotracheal PO₂ and PCO₂ was established. Typically, it contained two distinct regions only, corresponding to “closed” and “open” spiracles. A separate “flutter” region was missing. Because fluttering is commonly observed in moth pupae, we suggest that the intermittent spiracle opening during a flutter phase is an effect of non-steady-state conditions within the tracheal system. For low PCO₂ the minimum PO₂ resulting in open spiracles was linearly dependent upon PCO₂. Above a threshold of 1-1.5 kPa CO₂ the spiracles were open irrespective of PO₂. We propose a hypothetical spiracular control model, which is simple and explains the time course of endotracheal partial pressures during all phases of discontinuous gas exchange.     

Discontinuous gas exchange, water loss, and metabolism in Protaetia cretica (Cetoniinae, Scarabaeidae)

Insects are at high risk of desiccation because of their small size, high surface-area-to-volume ratio, and air-filled tracheal system that ramifies throughout their bodies to transport O(2) and CO(2) to and from respiring cells. Although the tracheal system offers a high-conductance pathway for the movement of respiratory gases, it has the unintended consequence of allowing respiratory transpiration to the atmosphere. When resting, many species exchange respiratory gases discontinuously, and an early hypothesis for the origin of these discontinuous gas exchange cycles (DGCs) is that they serve to reduce respiratory water loss. In this study, we test this "hygric" hypothesis by comparing rates of CO(2) exchange and water loss among flower beetles Protaetia cretica (Cetoniinae, Scarabaeidae) breathing either continuously or discontinuously. We show that, consistent with the expectations of the hygric hypothesis, rates of total water loss are higher during continuous gas exchange than during discontinuous gas exchange and that the ratio of respiratory water loss to CO(2) exchange is lower during discontinuous gas exchange. This conclusion is in agreement with other studies of beetles and cockroaches that also support the hygric hypothesis. However, this result does not exclude other adaptive hypotheses supported by work on ants and moth pupae. This ambiguity may arise because there are multiple independent evolutionary origins of DGCs and no single adaptive function underlying their genesis. Alternatively, the observed reduction in water loss during DGCs may be a side effect of a nonadaptive gas exchange pattern that is elicited during periods of inactivity.     

Pump out the volume—The effect of tracheal and subelytral pressure pulses on convective gas exchange in a dung beetle, Circellium bacchus (Fabricus)

Many flightless beetles like the large apterous dung beetle Circellium bacchus, possess a subelytral cavity (SEC) providing an extra air space below the elytra which connects to the tracheal system (TS) via metathoracic and abdominal spiracles. By measuring subelytral and intratracheal pressure as well as body movements and gas exchange simultaneously in a flow-through setup, we investigated the contribution of convection on Circellium respiratory gas exchange.No constriction phase was observed. TS and SEC pressures were always around atmospheric values. During interburst phase open abdominal spiracles and a leaky SEC led to small CO₂-peaks on a continuous CO₂ baseline, driven by intermittent positive tracheal pressure peaks in anti-phase with small negative subelytral pressure peaks caused by dorso-ventral tergite action.Spiracle opening was accompanied by two types of body movements. Higher frequency telescoping body movements at the beginning of opening resulted in high amplitude SEC and TS pressure peaks. High frequency tergite movements caused subelytral pressure peaks and led to a saw tooth like CO₂ release pattern in a burst. We propose that during the burst open mesothoracic spiracles increase the compliance of the subelytral cavity allowing big volumes of tracheal air being pulled out by convection.     

Spiracular closing mechanisms in the firebrat, Thermobia domestica (packard) (Thysanura)

Mechanisms for regulating the degree of opening of its spiracles are present in Thermobia. That of the mesothoracic spiracle is of the external type with a flap-like hood guarding the spiracular aperture. Contraction of muscles open the spiracle by raising the hood. Closure is brought about by muscular relaxation and elastic cuticular recoil. Opening is either partial, with small-scale oscillatory movements ('fluttering'), or complete ('wide-opening'). Wide-opening follows bouts of muscular activity. Carbon dioxide anaesthesia relaxes the opener muscles causing the spiracles to close by elastic recoil. This explains continued low tracheal water loss during anaesthesia, and also in death. The control mechanisms of the metathoracic and 8 pairs of abdominal spiracles are of the internal type, with a crypt-like atrium leading into the slit-like neck region of the spiracular pit, one side of which has an elastic cuticular rod running along it. Muscles inserted on the opposite side widen the aperture. As with the mesothoracic spiracle, closure is brought about by muscular relaxation and elastic cuticular recoil.     

Modulation of cyclic CO2 release in response to endogenous changes of metabolism during pupal development of Zophobas rugipes (Coleoptera: Tenebrionidae)

Understanding the mechanisms of gas exchange regulation in insects currently is a hot topic of insect physiology. Endogenous variation of metabolism during pupal development offers a great opportunity to study the regulation of respiratory patterns in insects. Here we show that metabolic rates during pupal development of the tenebrionid beetle Zophobas rugipes reveal a typical U-shaped curve and that, with the exception of 9-day-old pupae, the time between two bursts of CO₂ (interburst phase) was the only parameter of cyclic CO₂ gas exchange patterns that was adjusted to changing metabolic rates. The volume of CO₂ released in a burst was kept constant, suggesting a regulation for accumulation and release of a fixed amount of CO₂ throughout pupal development. We detected a variety of discontinuous and cyclic gas exchange patterns, which were not correlated with any periods of pupal development, suggesting a high among individual variability. An occasional occurrence of continuous CO₂ release patterns at low metabolic rates was very likely caused by single defective non-occluding spiracles.     

Feeding and respiratory gas exchange in the American dog tick,Dermacentor variabilis

This study investigated the effect of blood feeding on respiratory gas exchange in the dog tick Dermacentor variabilis. Adult male and female ticks were fed on bovine hosts from 1 to 11days. Females fed slowly for the first 6days and then rapidly engorged on blood 2-3days prior to dropping from the host. Ticks were removed at daily intervals during feeding, weighed and CO(2) emission measured at 25 degrees C using flow-through respirometry. During feeding, females (N=39) showed a 100-fold gain in mass from 5.78+/-1.05mg to 541.15+/-18.60mg while standard metabolic rate (Vdot;co(2)) increased from 0.179+/-0.030&mgr;lh(-1) in unfed ticks to 87.32+/-5.72&mgr;lh(-1) in fully engorged ticks. CO(2) release prior to feeding was highly discontinuous with discrete spiracular bursts of CO(2) emission approximately every 30min. For CO(2) emission measured in detached partially or completely fed ticks, burst frequency became more and more rapid as feeding progressed and changed to continuous sustained CO(2) output during rapid engorgement. In contrast to females, male ticks (N=20) showed little change in mass and maintained discontinuous CO(2) throughout the 11day attachment period on the host. The switch from discontinuous to continuous CO(2) release and presumed increase in respiratory water loss in female ticks is correlated to an increase in metabolic expenditure associated with blood meal digestion rather than any factor relating directly to maintenance of water balance.     

Discontinuous gas exchange and water loss in the keratin beetle Omorgus radula: further evidence against the water conservation hypothesis?

Discontinuous gas exchange cycles are demonstrated in Omorgus radula (Erichson) (Coleoptera, Trogidae) for the first time, thus extending evidence for such cycles to another family of beetles. The closed, flutter and open phases of the cycle were clearly distinguishable in this species, and the duration of these phases was 221 ± 28, 1403 ± 148 and 755 ± 43 s (mean ± SE), respectively. No evidence for significant intraspecific mass scaling of VCO₂ or any of the components of the cycle was found. Although the prolonged F‐phase recorded here is unusual for many insects, it has previously been found in other scarabaeoid beetles, especially those from xeric environments. It has been suggested that such modulation of the discontinuous gas exchange cycle may result in a reduced VCO₂ and, consequently, reduced water loss. In O. radula VCO₂ (15.25 ± 1.49 μl/h) was considerably lower than that predicted from its body mass (0.207 ± 0.006 g). However, the small relative contribution of respiratory transpiration (6.5%) to total water loss indicated that reduced VCO₂ has little to do with water economy. Rather, it may be a consequence of generally low activity levels of these beetles. The low respiratory water loss, but distinct subterranean component in the adult life of O. radula, lend some credence to the hypothesis suggesting that regular use of subterranean habitats might have been responsible for the evolution of discontinuous gas exchange cycles. However, non‐adaptive hypotheses can still not be discounted.     

Gas exchange characteristics, metabolic rate and water loss of the Heelwalker, Karoophasma biedouwensis (Mantophasmatodea: Austrophasmatidae)

This study presents the first physiological information for a member of the wingless Mantophasmatodea, or Heelwalkers. This species shows cyclic gas exchange with no evidence of a Flutter period (more typical of discontinuous gas exchange in insects) and no indication that the spiracles are fully occluded during quiescent metabolism. Standard metabolic rate at 20 degrees C was 21.32+/-2.73 microl CO(2)h(-1) (mean+/-S.E.), with a Q(10) (10-25 degrees C) of 1.7. Increases in V()CO(2) associated with variation in mass and with trial temperature were modulated by an increase in burst period volume and a decline in cycle frequency. Total water loss rate, determined by infrared gas analysis, was 0.876+/-0.08 mg H(2)Oh(-1) (range 0.602-1.577, n=11) whilst cuticular water loss rate, estimated by linear regression of total water loss rate and metabolic rate, was 0.618+/-0.09 mg H(2)Oh(-1) (range 0.341-1.363, n=11). Respiratory water loss rate was therefore no more than 29% of the total rate of water loss. Both total water loss rate and estimated cuticular water loss rate were significantly repeatable, with intraclass correlation coefficients of 0.745 and 0.553, respectively.     

Discontinuous gas exchange in the Pseudoscorpion Garypus californicus is regulated by hypoxia,not hypercapnia

The discontinuous gas exchange cycle (DGC) of the pseudoscorpion Garypus californicus is characterized by periodic bursts of CO(2) emission and by high rates of interburst CO(2) emission. We investigated the mechanism that triggers the burst phase by manipulating ambient oxygen partial pressures (Po(2)). The ventilatory trigger in most land animals is hypercapnia; in insects, for example, the burst phase is triggered when endotracheal Pco(2) reaches about 4 kPa. In insects with a DGC, hypoxia induces prolonged interburst phases because spiracular conductance is elevated to supply oxygen to the tissues, thus delaying the onset of the hypercapnia-triggered burst phase because CO(2) accumulates more slowly. In G. californicus, hypoxia induced a decrease in interburst phase length, while hyperoxia increased its duration relative to normoxia. This is opposite to the condition in insects. In addition, CO(2) emission fell during the interburst phase as ambient Po(2) rose, also opposite to the condition in insects. Thus, the burst phase is triggered in G. californicus (and presumably in other pseudoscorpions) not by hypercapnia but by hypoxia, a situation that is seldom encountered in terrestrial animals.