Establishing and Maintaining an Etruscan Shrew Colony.

The Etruscan shrew (Suncus etruscus) is one of the smallest mammals on earth and is used in many fields of research, including physiology, behavioral science and neuroscience. However, establishing and maintaining a breeding colony of this species in the laboratory can be challenging, as it requires specific husbandry conditions that greatly differ from those of more common laboratory species such as mice or rats. Over the past 15 y, we have successfully established a long-term thriving colony of 150 to 200 animals originating from 36 founders. The colony shows longer life expectancy and larger litter sizes than wild conspecifics. Breeding occurs year-round, independent of seasons, and a breeding pair can regularly produce 2 to 6 offspring with an average life expectancy of more than 3 y. The shrews are housed in glass or plastic enclosures on a specific soil-sand-mixture bedding and are provided with hideouts and nesting material consisting of moss, wood, or bark. Due to their high basal metabolic rate, the shrews require food intake greater than their body weight per day, can hunt arthropods as large as themselves, and cannot survive more than a few hours without food. Live feed such as crickets or mealworms is crucial and must be provided daily or, at the very least, every 2 d. Although our husbandry practices have constantly been adapted and refined, shrew husbandry remains challenging, and great care is necessary to meet the specific needs of this species. Here, we describe the establishment of a long-term stable colony of Etruscan shrews in a research animal facility and the specific husbandry requirements for animal wellbeing.

PO2 of the metathoracic ganglion in response to progressive hypoxia in an insect.

Mammals regulate their brain tissue PO2 tightly, and only small changes in brain PO2 are required to elicit compensatory ventilation. However, unlike the flow-through cardiovascular system of vertebrates, insect tissues exchange gases through blind-ended tracheoles, which may involve a more prominent role for diffusive gas exchange. We tested the effect of progressive hypoxia on ventilation and the PO2 of the metathoracic ganglion (neural site of control of ventilation) using microelectrodes in the American locust, Schistocerca americana. In normal air (21 kPa), PO2 of the metathoracic ganglion was 12 kPa. The PO2 of the ganglion dropped as air PO2 dropped, with ventilatory responses occurring when ganglion PO2 reached 3 kPa. Unlike vertebrates, insects tolerate relatively high resting tissue PO2 levels and allow tissue PO2 to drop during hypoxia, activity and discontinuous gas exchange before activating convective or spiracular gas exchange. Tracheated animals, and possibly pancrustaceans in general, seem likely to generally experience wide spatial and temporal variation in tissue PO2 compared with vertebrates, with important implications for physiological function and the evolution of oxygen-using proteins.

[The effect of husbandry shortcomings on oxygen supply in pet fish tanks]. / Einfluss von Haltungsfehlern auf den Sauerstoffhaushalt in Standardaquarien.

OBJECTIVE: In order to investigate the suitability of standard fish tank setups for permanent keeping of ornamental pet fish, oxygen consumption and exchange rates were measured in a group of standard aquariums with a volume of 54 litres. MATERIALS AND METHODS: The effects of defined disturbances on oxygen partial pressure in fish tanks were measured. These simulated typical beginners' errors such as a high stocking density, excessive feeding, insufficient filter cleaning, lack of water movement, and plant coverage of the water surface. Quantitative changes in oxygen partial pressure were measured in the tank as well as in a simplified model tank. RESULTS: Oxygen uptake rate of the tank (substrate, aquatic plants, bacteria, reduced substances) was not quantifiable in the experiment. The metabolism of the fish, which increased sharply with the feeding dose, exhibited the greatest effect on oxygen consumption in the fish tank. Oxidative conversion of nitrogen from ammonia to nitrate also caused a decrease in oxygen content, however to a lesser extent. Oxygen uptake from the atmosphere was significantly modulated by water flow rate and size of the diffusion area of the water surface. CONCLUSION AND CLINICAL RELEVANCE: These results allow quantitative predictions concerning the interaction of fish stocking density and oxygen balance in standard commercial aquarium setups. Even under conditions of high stocking density, poor filter hygiene and excessive feeding, all tested tanks demon strated their suitability for permanent keeping of ornamental fish. Care is warranted, however, that water flow is maintained and its surface is not covered by plants. Ideally, the fish should be fed several small portions during daytime rather than a single large ration.

The effects of temperature, activity and convection on the plastron PO2 of the aquatic bug Aphelocheirus aestivalis (Hemiptera; Aphelocheiridae).

The aquatic bug Aphelocheirus aestivalis (Fabricius 1794) utilises a plastron, a thin bubble layer on the surface of its body to extract O2 from the water. Millions of tiny hairs keep the bubble from collapsing, enabling the bug to remain submerged indefinitely. The development of fibre optic O2-probes has allowed measurements of O2 pressure (PO2) surrounding the plastron, and within the plastron although only for short periods. Here we developed methods to continuously measure plastron PO2, and investigate how it is affected by temperature (15, 20, 25°C), activity, and water circulation. We also made measurements of water PO2, temperature and velocity in the field and swimming velocity at the treatment temperatures. Results show that plastron PO2 is inversely related to temperature, associated with differences in metabolic demand, and that small bouts of activity or changes in water convection result in rapid changes in plastron PO2. A model was developed to calculate the conditions under which Aphelocheirus would exist without becoming O2-limited in relation to water temperature, PO2 and boundary layer thickness. This suggests that Aphelocheirus at one of two field sites may have a reduced metabolic scope even in well convected water in association with low PO2 and moderate temperature, and that in well convected, air-saturated water, bugs may have a reduced metabolic scope where water temperatures are between 20 and 25°C. If exposed to 5kPa PO2, Aphelocheirus cannot sustain resting metabolic rate even in well-convected water and would die at temperatures above approximately 25°C.

Determination of Henry's constant, the dissociation constant, and the buffer capacity of the bicarbonate system in ruminal fluid.

Despite the clinical importance of ruminal acidosis, ruminal buffering continues to be poorly understood. In particular, the constants for the dissociation of H2CO3 and the solubility of CO2 (Henry's constant) have never been stringently determined for ruminal fluid. The pH was measured in parallel directly in the rumen and the reticulum in vivo, and in samples obtained via aspiration from 10 fistulated cows on hay- or concentrate-based diets. The equilibrium constants of the bicarbonate system were measured at 38°C both using the Astrup technique and a newly developed method with titration at 2 levels of partial pressure of CO2 (pCO2; 4.75 and 94.98 kPa), yielding mean values of 0.234 ± 0.005 mmol ∙ L(-1) ∙ kPa(-1) and 6.11 ± 0.02 for Henry's constant and the dissociation constant, respectively (n/n = 31/10). Both reticular pH and the pH of samples measured after removal were more alkalic than those measured in vivo in the rumen (by ΔpH = 0.87 ± 0.04 and 0.26 ± 0.04). The amount of acid or base required to shift the pH of ruminal samples to 6.4 or 5.8 (base excess) differed between the 2 feeding groups. Experimental results are compared with the mathematical predictions of an open 2-buffer Henderson-Hasselbalch equilibrium model. Because pCO2 has pronounced effects on ruminal pH and can decrease rapidly in samples removed from the rumen, introduction of a generally accepted protocol for determining the acid-base status of ruminal fluid with standard levels of pCO2 and measurement of base excess in addition to pH should be considered.

Respiratory function of the plastron in the aquatic bug Aphelocheirus aestivalis (Hemiptera, Aphelocheiridae).

The river bug Aphelocheirus aestivalis is a 40 mg aquatic insect that, as an adult, relies totally on an incompressible physical gill to exchange respiratory gases with the water. The gill (called a 'plastron') consists of a stationary layer of air held in place on the body surface by millions of tiny hairs that support a permanent air-water interface, so that the insect never has to renew the gas at the water's surface. The volume of air in the plastron is extremely small (0.14 mm(3)), under slightly negative pressure and connected to the gas-filled tracheal system through spiracles on the cuticle. Here, we measure PO2 of the water and within the plastron gas with O2-sensing fibre optics to understand the effectiveness and limitations of the gas exchanger. The difference in PO2 is highest in stagnant water and decreases with increasing convection over the surface. Respiration of bugs in water-filled vials varies between 33 and 296 pmol O2 s(-1), depending on swimming activity. The effective thickness of the boundary layer around the plastron was calculated from respiration rate, PO2 difference and plastron surface area, according to the Fick diffusion equation and verified by direct measurements with the fibre-optic probes. In stagnant water, the boundary layer is approximately 500 µm thick, which nevertheless can satisfy the demands of resting bugs, even if the PO2 of the free water decreases to half that of air saturation. Active bugs require thinner boundary layers (∼ 100 µm), which are achieved by living in moving water or by swimming.

Respiratory dynamics of discontinuous gas exchange in the tracheal system of the desert locust, Schistocerca gregaria.

Gas exchange dynamics in insects is of fundamental importance to understanding evolved variation in breathing patterns, such as discontinuous gas exchange cycles (DGCs). Most insects do not rely solely on diffusion for the exchange of respiratory gases but may also make use of respiratory movements (active ventilation) to supplement gas exchange at rest. However, their temporal dynamics have not been widely investigated. Here, intratracheal pressure, V(CO2) and body movements of the desert locust Schistocerca gregaria were measured simultaneously during the DGC and revealed several important aspects of gas exchange dynamics. First, S. gregaria employs two different ventilatory strategies, one involving dorso-ventral contractions and the other longitudinal telescoping movements. Second, although a true spiracular closed (C)-phase of the DGC could be identified by means of subatmospheric intratracheal pressure recordings, some CO(2) continued to be released. Third, strong pumping actions do not necessarily lead to CO(2) release and could be used to ensure mixing of gases in the closed tracheal system, or enhance water vapour reabsorption into the haemolymph from fluid-filled tracheole tips by increasing the hydrostatic pressure or forcing fluid into the haemocoel. Finally, this work showed that the C-phase of the DGC can occur at any pressure. These results provide further insights into the mechanistic basis of insect gas exchange.

Assessing honeybee and wasp thermoregulation and energetics-New insights by combination of flow-through respirometry with infrared thermography.

Endothermic insects like honeybees and some wasps have to cope with an enormous heat loss during foraging because of their small body size in comparison to endotherms like mammals and birds. The enormous costs of thermoregulation call for optimisation. Honeybees and wasps differ in their critical thermal maximum, which enables the bees to kill the wasps by heat. We here demonstrate the benefits of a combined use of body temperature measurement with infrared thermography, and respiratory measurements of energy turnover (O(2) consumption or CO(2) production via flow-through respirometry) to answer questions of insect ecophysiological research, and we describe calibrations to receive accurate results.To assess the question of what foraging honeybees optimise, their body temperature was compared with their energy turnover. Honeybees foraging from an artificial flower with unlimited sucrose flow increased body surface temperature and energy turnover with profitability of foraging (sucrose content of the food; 0.5 or 1.5 mol/L). Costs of thermoregulation, however, were rather independent of ambient temperature (13-30 °C). External heat gain by solar radiation was used to increase body temperature. This optimised foraging energetics by increasing suction speed.In determinations of insect respiratory critical thermal limits, the combined use of respiratory measurements and thermography made possible a more conclusive interpretation of respiratory traces.

Reactive oxygen species production and discontinuous gas exchange in insects.

While biochemical mechanisms are typically used by animals to reduce oxidative damage, insects are suspected to employ a higher organizational level, discontinuous gas exchange mechanism to do so. Using a combination of real-time, flow-through respirometry and live-cell fluorescence microscopy, we show that spiracular control associated with the discontinuous gas exchange cycle (DGC) in Samia cynthia pupae is related to reactive oxygen species (ROS). Hyperoxia fails to increase mean ROS production, although minima are elevated above normoxic levels. Furthermore, a negative relationship between mean and mean ROS production indicates that higher ROS production is generally associated with lower . Our results, therefore, suggest a possible signalling role for ROS in DGC, rather than supporting the idea that DGC acts to reduce oxidative damage by regulating ROS production.

The diving bell and the spider: the physical gill of Argyroneta aquatica.

Argyroneta aquatica is a unique air-breathing spider that lives virtually its entire life under freshwater. It creates a dome-shaped web between aquatic plants and fills the diving bell with air carried from the surface. The bell can take up dissolved O(2) from the water, acting as a 'physical gill'. By measuring bell volume and O(2) partial pressure (P(O(2))) with tiny O(2)-sensitive optodes, this study showed that the spiders produce physical gills capable of satisfying at least their resting requirements for O(2) under the most extreme conditions of warm stagnant water. Larger spiders produced larger bells of higher O(2) conductance (G(O(2))). G(O(2)) depended on surface area only; effective boundary layer thickness was constant. Bells, with and without spiders, were used as respirometers by measuring G(O(2)) and the rate of change in P(O(2)). Metabolic rates were also measured with flow-through respirometry. The water-air P(O(2)) difference was generally less than 10 kPa, and spiders voluntarily tolerated low internal P(O(2)) approximately 1-4 kPa before renewal with air from the surface. The low P(O(2)) in the bell enhanced N(2) loss from the bell, but spiders could remain inside for more than a day without renewal. Spiders appeared to enlarge the bells in response to higher O(2) demands and lower aquatic P(O(2)).

Modulation of cyclic CO(2) 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 CO(2) (interburst phase) was the only parameter of cyclic CO(2) gas exchange patterns that was adjusted to changing metabolic rates. The volume of CO(2) released in a burst was kept constant, suggesting a regulation for accumulation and release of a fixed amount of CO(2) 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 CO(2) release patterns at low metabolic rates was very likely caused by single defective non-occluding spiracles.

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 P(O(2)) and P(CO(2)) 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 P(O(2)) and P(CO(2)) 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 P(CO(2)) the minimum P(O(2)) resulting in open spiracles was linearly dependent upon P(CO(2)). Above a threshold of 1-1.5 kPa CO(2) the spiracles were open irrespective of P(O(2)). 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.

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 CO(2)-peaks on a continuous CO(2) 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 CO(2) 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.

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 ((cyc)GE) 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.

Cationic composition and acid-base state of the extracellular fluid, and specific buffer value of hemoglobin from the branchiopod crustacean Triops cancriformis.

Recent insights into the allosteric control of oxygen binding in the extracellular hemoglobin (Hb) of the tadpole shrimp Triops cancriformis raised the question about the physico-chemical properties of the protein's native environment. This study determined the cationic composition and acid-base state of the animal's extracellular fluid. The physiological concentrations of potential cationic effectors (calcium, magnesium) were more than one order of magnitude below the level effective to increase Hb oxygen affinity. The extracellular fluid in the pericardial space had a typical bicarbonate concentration of 7.6 mM but a remarkably high CO(2) partial pressure of 1.36 kPa at pH 7.52 and 20 degrees C. The discrepancy between this high CO(2) partial pressure and the comparably low values for water-breathing decapods could not solely be explained by the hemolymph-sampling procedure but may additionally arise from differences in cardiovascular complexity and efficiency. T. cancriformis hemolymph had a non-bicarbonate buffer value of 2.1 meq L(-1) pH(-1). Hb covered 40-60% of the non-bicarbonate buffering power. The specific buffer value of Hb of 1.1 meq (mmol heme)(-1) pH(-1) suggested a minimum requirement of two titratable histidines per heme-binding domain, which is supported by available information from N-terminal sequencing and expressed sequence tags.

Control of discontinuous gas exchange in Samia cynthia: effects of atmospheric oxygen, carbon dioxide and moisture.

The evolution of discontinuous gas exchange (DGE) in insects is highly controversial. Adaptive hypotheses which have obtained experimental support include a water savings mechanism for living in dry environments (hygric hypothesis), a reduction in oxidative damage due to a high-performance oxygen delivery system (oxidative damage hypothesis), and the need for steep intratracheal partial pressure gradients to exchange gases under the hypercapnic and/or hypoxic conditions potentially encountered in subterranean environments (chthonic hypothesis). However, few experimental studies have simultaneously assessed multiple competing hypotheses within a strong inference framework. Here, we present such a study at the species level for a diapausing moth pupa, Samia cynthia. Switching gas conditions from controlled normoxic, normocapnic and intermediate humidity to either high or low oxygen, high or low moisture, elevated carbon dioxide, or some combination of these, revealed that DGE was abandoned under all conditions except high oxygen, and high or low gas moisture levels. Thus, support is found for the oxidative damage hypothesis when scored as maintenance of DGE. Modulation of DGE under either dry or hyperoxic conditions suggested strong support for the oxidative damage hypothesis and some limited support for the hygric hypothesis. Therefore, this study demonstrates that the DGE can be maintained and modulated in response to several environmental variables. Further investigation is required using a strong-inference, experimental approach across a range of species from different habitats to determine how widespread the support for the oxidative damage hypothesis might be.

Bat breath reveals metabolic substrate use in free-ranging vampires.

We analysed the stable carbon isotope ratio in exhaled CO(2) (delta(13)C(breath)) of free-ranging vampires to assess the type of metabolized substrate (endogenous or exogenous substrate) and its origin, i.e. whether the carbon atoms came from a C(4) food web (grass and cattle) or the C(3) food web in which they were captured (a rainforest remnant and its mammals). For an improved understanding of factors influencing the delta(13)C(breath) of vampires, we conducted feeding experiments with captive animals. The mean delta(13)C(breath) of starved bats was depleted in (13)C in relation to the diet by 4.6 per thousand (n = 10). Once fed with blood, delta(13)C(breath )levelled off within a short time approximately 2.2 per thousand above the stable carbon isotope signature of the diet. The median time required to exchange 50% of the carbon atoms in exhaled CO(2) with carbon atoms from the ingested blood was 18.6 min (mean 29.5 +/- 19.0 min, n = 5). The average delta(13)C of wing membrane and fur in free-ranging vampire bats suggested that bats almost exclusively foraged for cattle blood during the past weeks. The delta(13)C(breath) of the same bats averaged -19.1 per thousand. Given that all free-ranging vampires were starving and that the delta(13)C of cattle was more in enriched in (13)C by 5-6 per thousand than the delta(13)C(breath) of vampires, we conclude that the vampire bats of our study metabolised fat that was predominantly built from carbon atoms originating from cattle blood. Since delta(13)C of wing membrane and fur integrates over weeks and months respectively and delta(13)C(breath) over hours and days, we also conclude that vampire bats of the studied population consistently ignored rainforest mammals and chose cattle as their prey during and prior to our study.

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.

Respiration of resting honeybees.

The relation between the respiratory activity of resting honeybees and ambient temperature (T(a)) was investigated in the range of 5-40 degrees C. Bees were kept in a temperature controlled flow through respirometer chamber where their locomotor and endothermic activity, as well as abdominal ventilatory movements was recorded by infrared thermography. Surprisingly, true resting bees were often weakly endothermic (thorax surface up to 2.8 degrees C warmer than abdomen) at a T(a) of 14-30 degrees C. Above 33 degrees C many bees cooled their body via evaporation from their mouthparts. A novel mathematical model allows description of the relationship of resting (standard) metabolic rate and temperature across the entire functional temperature range of bees. In chill coma (<11 degrees C) bees were ectothermic and CO(2) release was mostly continuous. CO(2) release rate (nls(-1)) decreased from 9.3 at 9.7 degrees C to 5.4 at 5 degrees C. At a T(a) of >11 degrees C CO(2) was released discontinuously. In the bees' active temperature range mean CO(2) production rate (nls(-1)) increased sigmoidally (10.6 at 14.1 degrees C, 24.1 at 26.5 degrees C, and 55.2 at 38.1 degrees C), coming to a halt towards the upper lethal temperature. This was primarily accomplished by an exponential increase in gas exchange frequency (0.54 and 3.1 breaths min(-1) at 14.1 and 38.1 degrees C) but not in released CO(2) volume per respiratory cycle (1487 and 1083 nl cycle(-1) at 14.1 and 38.1 degrees C). Emission of CO(2) bursts was mostly (98%) accompanied by abdominal ventilation movements even in small CO(2) bursts. Larger bursts coincided with a longer duration of active ventilation. An increased amount of CO(2) expelled per unit time of ventilation indicates a higher efficiency of ventilation at high ambient temperatures.