The Relationship between Cutaneous Water Loss and Thermoregulatory State in Kuhl's Pipistrelle Pipistrellus kuhlii, a Vespertillionid Bat

Abstract Total evaporative water loss is the sum of respiratory water loss (RWL) and cutaneous water loss (CWL) and constitutes the main avenue of water loss in bats. Because bats fly and have large surface-to-volume ratios, they potentially have high rates of RWL and CWL. Most species of small insectivorous bats have the ability to reduce their body temperature (T(b)) at rest, which substantially reduces energy expenditure and water loss. We hypothesized that bats reduce evaporative water loss during bouts of deep hypothermia (torpor) by decreasing RWL and CWL. We measured T(b), RWL, CWL, and resting metabolic rate (RMR) in Kuhl's pipistrelle Pipistrellus kuhlii, a small insectivorous bat. In support of our hypothesis, we found that RWL decreased with decreasing RMR. We found that CWL was lower in torpid individuals than in normothermic bats; however, bats in deep torpor had similar or higher CWL than bats in shallow torpor, suggesting that they exert a less effective physiological control over CWL when in deep torpor. Because insectivorous bats spend most of their lives in torpor or hibernation, the regulation of CWL in different heterothermic states has relevant ecological and evolutionary consequences.     

Respiratory and cuticular water loss in insects with continuous gas exchange: comparison across five ant species

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.     

Cutaneous and respiratory water loss in larks from arid and mesic environments

Birds from deserts generally have lower total evaporative water loss (TEWL), the sum of cutaneous (CWL) and respiratory water loss (RWL), than species from mesic areas. We investigated the role of CWL and RWL as a function of air temperature (T(a)) in hoopoe larks (Alaemon alaudipes) and Dunn's larks (Eremalauda dunni) from the Arabian Desert and skylarks (Alauda arvensis) and woodlarks (Lullula arborea) from temperate mesic grasslands. The proportional contribution of CWL to TEWL in all larks at moderate T(a) ranged from 50% to 70%. At high T(a) (40 degrees -45 degrees C), larks enhanced CWL by only 45%-78% and relied on an increase in RWL by 676%-2,733% for evaporative cooling. Surface-specific CWL at 25 degrees C was 29% lower in the arid-zone species than in the mesic larks. When acclimated to constant T(a), 15 degrees C-acclimated hoopoe larks increased CWL by 22% compared with 35 degrees C-acclimated birds, but the other species did not change CWL. This study is consistent with the hypothesis that larks from deserts have a reduced CWL at moderate and low T(a) but provided no support for the hypothesis that at high T(a) larks from arid regions rely more on CWL than larks from mesic environments. Interspecific differences in CWL cannot be attributed to acclimation to environmental temperature and are possibly the result of genetic differences due to natural selection or of phenotypically plastic responses to divergent environments during ontogeny.     

The relative importance of respiratory water loss in scorpions is correlated with species habitat type and activity pattern

Scorpions exhibit some of the lowest recorded water loss rates compared with those of other terrestrial arthropods of similar body size. Evaporative water loss (EWL) includes cuticular transpiration and respiratory water loss (RWL) from gas exchange surfaces, that is, book lung lamellae. Estimated fractions of cuticular and respiratory losses currently available from the literature show considerable variation, at least partly as a result of differences in methodology. This study reports RWL rates and their relative importance in scorpions from two families (Buthidae and Scorpionidae), including both xeric and mesic species (or subspecies). Two of the included Buthidae were surface-dwelling species, and another inhabits empty burrows of other terrestrial arthropods. This experimental design enabled correlating RWL importance with scorpion phylogeny, habitat type, and/or homing behavior. Buthidae species exhibited significantly lower EWL rates compared with those of Scorpionidae, whereas effects of habitat type and homing behavior were not significant. Resting RWL rates were not significantly affected by scorpion phylogeny, but rates for the xeric species (totaling ~10% of EWL rates at 30°C) were significantly lower compared with those of mesic species. These lower RWL values were correlated with significantly lower H(2)O/CO(2) emission rates in xeric species. The experimental setup and ~24-h duration of each individual recording allowed estimating the effect of interspecific variation in activity on RWL proportions. The high respiratory losses in active hydrated Scorpio maurus fuscus, totaling 30% of EWL, suggest that behavioral discretion in this species is a more likely mechanism for body water conservation under stressful conditions when compared with the responses of other studied species.     

Effects of phosphine on the neural regulation of gas exchange in Periplaneta americana

Phosphine is used for fumigating stored commodities, however an understanding of the physiological response to phosphine in insects is limited. Here we show how the central pattern generator for ventilation in the central nervous system (CNS) responds to phosphine and influences normal resting gas exchange. Using the American cockroach, Periplaneta americana, that perform discontinuous gas exchange (DGE) at rest, we simultaneously measure ventilatory nervous output from the intact CNS, VCO(2) and water loss from live specimens. Exposure to 800 ppm phosphine at 25 degrees C for 2 h (n=13) during recording did not cause any mortality or obvious sub-lethal effects. Within 60 s of introducing phosphine into the air flow, all animals showed a distinct CNS response accompanied by a burst release of CO(2). The initial ventilatory response to phosphine displaced DGE and was typically followed by low, stable and continuous CO(2) output. CNS output was highest and most orderly under normoxic conditions during DGE. Phosphine caused a series of ventilatory CNS spikes preceding almost complete cessation of CNS output. Minimal CNS output was maintained during the 2 h normoxic recovery period and DGE was not reinstated. VCO(2) was slightly reduced and water loss significantly lower during the recovery period compared with those rates prior to phosphine exposure. A phosphine narcosis effect is rejected based on animals remaining alert at all times during exposure.     

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.     

Discontinuous gas exchange in insects: a clarification of hypotheses and approaches

Many adult and diapausing pupal insects exchange respiratory gases discontinuously in a three-phase discontinuous gas exchange cycle (DGC). We summarize the known biophysical characteristics of the DGC and describe current research on the role of convection and diffusion in the DGC, emphasizing control of respiratory water loss. We summarize the main theories for the evolutionary genesis (or, alternatively, nonadaptive genesis) of the DGC: reduction in respiratory water loss (the hygric hypothesis), optimizing gas exchange in hypoxic and hypercapnic environments (the chthonic hypothesis), the hybrid of these two (the chthonic-hygric hypothesis), reducing the toxic properties of oxygen (the oxidative damage hypothesis), the outcome of interactions between O(2) and CO(2) control set points (the emergent property hypothesis), and protection against parasitic invaders (the strolling arthropods hypothesis). We describe specific techniques that are being employed to measure respiratory water loss in the presence or absence of the DGC in an attempt to test the hygric hypothesis, such as the hyperoxic switch and H(2)O/CO(2) regression, and summarize specific areas of the field that are likely to be profitable directions for future research.     

Neural regulation of discontinuous gas exchange in Periplaneta americana

Patterns of gas exchange among terrestrial arthropods are highly variable from continuous to discontinuous with discretely partitioned phases. The underlying initiation and co-ordination of these patterns is relatively poorly understood. Here we present a novel method for the simultaneous measurement of central nervous system (CNS) activity of the metathoracic ganglion and VCO(2) in medium to large sized live terrestrial arthropods. Using Periplaneta americana at four oxygen levels (40%, 21%, 10% and 2% at 25 degrees C; n=6 per treatment), we present minimally invasive visualization of nervous output relative to typical resting discontinuous gas exchange (DGE) data for the first time. DGE was maintained when cockroaches were exposed to hyperoxia or moderate hypoxia, but was lost in severe hypoxia. CNS activity was manifested in three signal types: large CNS output coinciding with peak CO(2) production during a burst, moderate CNS output coinciding with CO(2) sawtoothing and fluttering, and minimal CNS activity during the closed phase of DGE in normoxia. Large and moderate CNS outputs were associated with observed abdominal pumping and congruent CO(2) peaks. At 10% oxygen, VCO(2) was significantly elevated during the inter-burst period in association with almost constant moderate CNS output between the periodic large CNS output. At 2% oxygen, DGE and large CNS output are lost to continuous CO(2) release and largely continuous moderate CNS output. As previously reported for this species, a central pattern generator for ventilation in the metathoracic ganglion is supported and we infer the presence of localized oxygen chemoreceptors based on clear CNS response to a change in oxygen tension.     

Effect of mating stage on water balance, cuticular hydrocarbons and metabolism in the desert harvester ant, Pogonomyrmex barbatus

Water-loss rates increase after mating in queens of the harvester ant Pogonomyrmex barbatus (Formicidae: Myrmicinae), then increase again after the mated queens excavate an incipient nest. We determined the mechanistic basis for these increased water-loss rates by examining cuticular permeability, respiratory water loss, metabolic rates, and cuticular hydrocarbons for queens at three stages in the mating sequence: unmated alate queens, newly mated dealate queens, and mated queens excavated from their incipient nest. Both total water loss and cuticular transpiration increased significantly following mating, with cuticular transpiration accounting for 97% of the increased water loss. In contrast, metabolic rate and respiratory water loss were unaffected by mating stage. The total quantity of cuticular hydrocarbons did not vary by mating stage. However, relative amounts of four of the most abundant cuticular hydrocarbons did vary by mating stage, as did quantities of n-alkanes and methylalkanes. The general pattern was that percent composition of n-alkanes decreased through the mating sequence, while percent composition of methylalkanes increased over the same sequence. We discuss three mechanisms that might cause these post-mating increases in cuticular permeability. Our data support the hypothesis that part of this increase results from soil particles abrading the cuticle during the process of nest excavation.     

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.     

beta-Glucocerebrosidase activity in the stratum corneum of house sparrows following acclimation to high and low humidity

Skin is an important avenue of water loss in terrestrial birds, so environmental conditions that necessitate water conservation should favor physiological mechanisms that reduce cutaneous water loss (CWL). Skin resistance to CWL is conferred by a barrier of lipid molecules located in the stratum corneum (SC), the outer layer of the epidermis. In mammals, SC barrier function depends on the conversion of cerebrosides to ceramides by the enzyme beta -glucocerebrosidase ( beta -GlcCer'ase). Avian SC contains both cerebrosides and ceramides, suggesting that observed plasticity in CWL may be mediated by changes in beta -GlcCer'ase activity and resultant SC lipid composition. We tested the hypothesis that changes in ambient humidity would alter beta -GlcCer'ase activity by acclimating house sparrows (Passer domesticus) to either dry (6.5 g H(2)O m(-3) absolute humidity) or humid (31 g H(2)O m(-3)) conditions for 5 and 21 d at 30 degrees C and then measuring beta -GlcCer'ase activity from SC homogenates. Our results provide the first characterization of beta -GlcCer'ase activity in any nonmammalian vertebrate. Relative to nonacclimated controls, both dry- and humid-acclimated sparrows had significantly elevated beta -GlcCer'ase activity at 21 d postacclimation. Across individuals, we observed negative correlations between beta -GlcCer'ase activity and both CWL and SC ceramide content. Although dry- and humid-acclimated sparrows did not differ in beta -GlcCer'ase activity, these results are consistent with our findings that both humidity treatments caused a reduction in CWL and similar changes in SC lipid composition. Our results demonstrate physiological plasticity in CWL and provide tentative support for a role of beta -GlcCer'ase in mediating this response.     

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.     

Effects of hyperoxic gas mixtures on energy metabolism during prolonged work.

These experiments were designed to study selected respiratory and metabolic responses to exercise in hyperoxia. Four subjects were examined during 30-min bicycle ergometer rides at both 40% and 80% of their aerobic maximum. The VO2 was significantly increased at both work levels breathing 60% O2 versus 21% O2, while VCO2 showed no significant change during the 40% exercise tests but was significantly decreased during the 80% intensity rides. The average increase in the volume of O2 taken up during 30 min of hyperoxic exercise, compared with normoxia, was 3.3 liters at the 40% exercise level and 5.6 liters at the 80% level. Neither the magnitude of the O2 nor the CO2 storage calculated for the exercise bouts could explain these increases. Steady-state criteria for the gas stores were established by the stable values of PETCO2, VO2, VCO2, and VI from minute 6 through 30 at both work levels. R values decreased during the hyperoxic tests suggesting the possibility of a shift toward increased fatty acid metabolism.     

Development of skin structure and cutaneous water loss in nestling desert house sparrows from Saudi Arabia

The outer layer of the epidermis, the stratum corneum (SC), contains lipids and corneocytes, which together form layers that limit cutaneous water loss (CWL). We examined the development of structure of the SC and CWL in nestling House Sparrows (Passer domesticus) from Saudi Arabia. We measured CWL of nestlings, and characterized development of their epidermis using electron microscopy. We tested two antagonistic hypotheses, that CWL decreases as nestlings age, a response to increased thickness of SC, and an opposite idea that CWL increases as nestlings age even though the number of layers of the SC remains constant. CWL of nestling House Sparrows varied with developmental stages, in a non-linear fashion, but not significantly so. CWL of nestlings averaged 7.31+/-1.5 g H(2)O/(m(2) h), whereas for adults it was 4.95 g/(m(2) h); adult CWL was 67.7% that of nestlings. We found that morphology of the SC did not change linearly with age, but seemed to vary with developmental stage. CWL decreased as the SC thickness increased and as the total thickness of the corneocytes increased. Further, we found that CWL decreased as the thickness of the extracellular space increased, number of corneocytes increased, and proportion of the SC that is extracellular space increased.     

The length of discontinuous gas exchange cycles in lepidopteran pupae may serve as a mechanism for natural selection

Gas exchange is studied in diapausing pupae of Mamestra brassicae L., whose larvae are reared under identical conditions. The release of CO₂ gas is recorded with infrared gaseous analyzers. Oxygen convective uptake into the tracheae and oxygen consumption rates are recorded by means of a constant‐volume coulometric respirometer. Outputs from both of these respirometry systems are combined with infrared actographs. All 3‐month‐old pupae of M. brassicae display a pattern of discontinuous gas exchange (DGE) cycles of CO₂ gas release by bursts, although the lengths of these cycles varies between individuals. Some pupae exhibit long DGE cycles of at least 20 h in duration, with negligible CO₂ gas release during interburst periods, and there is presumed to be a convective gas exchange at this time. As a result of a partial vacuum inside the tracheae, a large oxygen convective uptake always occurs at the start of the spiracular opening phase. Other pupae have short DGE cycles of less than 3 h in duration, with elevated CO₂ gas release during the interburst period, when gas exchange is predominantly diffusive. The spiracular open phase in these pupae consists of frequent separate convective bursts of CO₂ gas release, with the opening–closing rhythms of the spiracles, which are considered as O phase fluttering. The pupae with long DGE cycles exhibit extremely low metabolic rates and very low total water loss rates, whereas those with short DGE cycles have higher metabolic and total water loss rates. The pupae with long DGE cycles live approximately twice as long as those with short cycles; thus, the present study demonstrates that long DGE cycles confer a fitness benefit on pupae as a result of a lower metabolic rate associated with water economy, conferring on them a longer life.     

Simple computer measurement of pulmonary VCO2 per breath.

Measurements of the volume of CO2 exhaled per breath (VCO2/br) are preferable to end-tidal PCO2, when the exhaled flow and CO2 waveforms may be changing during unsteady states, such as during alterations in positive end-expiratory pressure or alterations in cardiac output. We describe computer algorithms that determine VCO2/br from digital measurements of exhaled flow (including discontinuous signals common in anesthesia circuits) and CO2 concentration at the airway opening. Fractional concentration of CO2 is normally corrected for dynamic response and transport delay (TD), measured in a separate procedure. Instead, we determine an on-line adjusted TD during baseline ventilation. In six anesthetized dogs, we compared the determination of VCO2/br with a value measured in a simultaneous collection of expired gas. Over a wide range of tidal volume (180-700 ml), respiratory rate (3-30 min-1), and positive end-expiratory pressure (0-14 cmH2O), VCO2/br was more accurate with use of the adjusted TD than the measured TD (P less than 0.05).     

Cutaneous water loss and the development of the stratum corneum of nestling house sparrows (Passer domesticus) from desert and mesic environments

Evaporation through the skin contributes to more than half of the total water loss in birds. Therefore, we expect the regulation of cutaneous water loss (CWL) to be crucial for birds, especially those that live in deserts, to maintain a normal state of hydration. Previous studies in adult birds showed that modifications of the lipid composition of the stratum corneum (SC), the outer layer of the epidermis, were associated with changes in rates of CWL. However, few studies have examined the ontogeny of CWL and the lipids of the SC in nestling birds. In this study, we measured CWL and the lipid composition of the SC during development of nestlings from two populations of house sparrows, one from the deserts of Saudi Arabia and the other from mesic Ohio. We found that desert and mesic nestlings followed different developmental trajectories for CWL. Desert nestlings seemed to make a more frugal use of water than did mesic nestlings. To regulate CWL, nestlings appeared to modify the lipid composition of the SC during ontogeny. Our results also suggest a tighter regulation of CWL in desert nestlings, presumably as a result of the stronger selection pressures to which nestlings are exposed in deserts.     

Cutaneous evaporimetry. IX. Cutaneous water loss in old age

The cutaneous water loss (CWL) were investigated in 15 old human subjects by Evaporimeter Ep-1. The mean value of the CWL resulted 11,9 +/- 4,3 g/h. This result is significantly lower compared with the value of normal people equal to 17, 5 +/- 4,0 g/h as reported in foregoing notes.     

Measurement of cutaneous evaporation. 5. Cutaneous water loss in peripheral vascular disease

The purpose of this research is the measurement of the cutaneous water loss in 19 angiopathie patients by the Evaporimeter. Ep I in order to compare it with the values of normal people. We found a statistical significant increase of the CWL without difference between the normal and pathologic limb.     

Phenotypic flexibility in cutaneous water loss and lipids of the stratum corneum in house sparrows (Passer domesticus) following acclimation to high and low humidity

Resistance to water-vapor diffusion through the skin is thought to be conferred by lipids in the stratum corneum (SC), the outer layer of the epidermis. We tested the effect of ambient humidity on cutaneous water loss (CWL) and lipid composition of the SC by acclimating house sparrows (Passer domesticus) to either a dry (6.5 g/m(3) absolute humidity) or a humid (31 g/m(3)) environment for 3 wk at a thermoneutral temperature (30 degrees C). Sparrows in the dry-acclimated group reduced CWL by 36% compared with those in the humid environment. Relative to initial values, both groups of sparrows decreased CWL, 45% in the dry-acclimated group and 23% in the humid group, suggesting that temperature is also an important stimulus for CWL apart from humidity. Both groups of acclimated sparrows decreased quantities of cholesterol, free fatty acids, and cerebrosides and increased the proportion of ceramides in their SC. Lipid amounts or proportions in the SC did not differ between dry- and humid-acclimated sparrows, but the free fatty acid : ceramide ratio was significantly lower in dry-acclimated birds. Also, lipid composition was only correlated with CWL in dry-acclimated sparrows, suggesting that structural changes to SC lipids are more tightly linked to CWL regulation in response to low humidity. Our results demonstrate phenotypic flexibility in CWL and lipid composition of the SC and provide support for a functional relationship between these traits.