Dark respiration of the stipe of Ecklonia cava (Laminariales, Phaeophyta) in relation to temperature

Sporophytes of Ecklonia cava Kjellman (Laminariales, Phaeophyta) with a stipe length of 22–102 cm were collected at 6–9 m depth in Nabeta Bay, Shimoda, central Japan by scuba diving in February (winter) and in August (summer) 1998. Dark respiration of the intact stipe of E. cava was measured at various water temperatures ranging from 15 to 27.5°C in winter and 15–30°C in summer in a closed system by using a dissolved oxygen meter. The stipe respiration was compared on whole stipe, length, surface area, volume, wet weight and dry weight bases. On each basis, the stipe respiration always increased with a rise in water temperature within the temperature range investigated. The stipes showed similar respiration rates on each basis of length, surface area, volume, wet weight and dry weight at each temperature, irrespective of the stipe length. The mean respiration rates in winter (at 15–27.5°C) were: length, 16.7–32.5 μL O₂ cm^?1 h^?1; surface area, 3.2–6.2 μL O₂ cm^?2 h^?1; volume, 7.6–15.0 μL O₂ cm^?3 h^?1; wet weight, 6.2–12.2 μL O₂ g (wet weight)^?1 h^?1; and dry weight, 43.8–88.0 μL O₂ g (dry weight)^?1 h^?1. Those for summer (at 15–30°C) were: length, 17.1–32.0 μL O₂ cm^?1 h^?1; surface area, 3.6–6.8 μL O₂ cm^?2 h^?1; volume, 9.7–18.7 μL O₂ cm^?3 h^?1; wet weight, 7.6–14.6 μL O₂ g (wet weight)^?1 h^?1; and dry weight, 49.4–95.8 μL O₂ g (dry weight)^?1 h^?1. This is the first report of the intact stipe respiration of E. cava at various temperatures.     

Foraging in male bumblebees (Bombus lucorum L.): maximizing energy or minimizing water load?

Summary The O₂, CO₂, and H₂O exchange of single flying male bumblebees (Bombus lucorum and B. terrestris) were measured simultaneously. A respiratory quotient RQ=1 was found for all activities investigated (torpor-flight). The dependence of respiratory CO₂ production in flight on body-weight was measured: for a 220-mg male bumblebee it amounts to 24.5 mg CO₂/h (=56.4 ml O₂/g·h). The corresponding evaporative water loss amounts to 6 mg H₂O/h. Males tranferred to a climatic test chamber and conditioned to artificial flower feeders started to fly, after a few days of acclimatization, in typical scent-marked flight-paths. The daily pattern of flight activity was recorded: the mean total time in flight amounts to 244 min, and the corresponding daily flight length is about 17 km. At 20°C and 50% relative humidity (RH) a daily uptake of 180 l ( 220mg) of 50% sugar solution was measured, equal to the mean body weight of the male bumblebees. Since the body weight remains constant on consecutive days a 24-h energy- and water-budget could be calculated. The energy-budget is balanced; the activities observed can be fuelled with the sugar available. About 70% of the energy is used for the 4 h of flight activity. With the daily nectar volume 110 mg of water is ingested; in the oxidation of 110 mg sugar, 66 mg of metabolic water is produced and 40 mg water is dissipated by the evaporative water-loss. Thus, to have a balanced water-budget, 136 mg of water must be voided in 24 h, which equals the total body-water of the bumblebees. Nectar is a nutrient of high water content which not only provides the sugar necessary for activity but also, in most circumstances, an excess of water. The effect of this high water load in limiting daily activity is discussed and compared with the water- and osmoregulation of hummingbirds. The strategy of maximizing energy for a male bumblebee must be one of minimizing water load.     

The effects of temperature and dissolved oxygen on antioxidant defences and oxidative damage in the fathead minnow Pimephales promelas

Fathead minnows Pimephales promelas maintained at 25° C for 6 h had significantly higher superoxide dismutase (SOD) activity than fish maintained at 7 or 32° C, but hypoxic conditions (3 mg l^?1 O₂) over the same time period did not affect SOD activity. Fish in better body condition (length‐adjusted mass) had higher SOD activity. In a separate experiment, P. promelas maintained at three water temperatures (7, 23 and 32° C) for 31 days did not differ in liver acrolein, a biomarker of oxidative stress.     

Hypoxia tolerance of the mummichog: the role of access to the water surface

Low dissolved oxygen (DO) had a significant effect on specific growth rate (G_S), length increment (I_L) and haematocrit (Hct) of the mummichog Fundulus heteroclitus. Regardless of access to the water surface, F. heteroclitus maintained high growth rates (G_S and I_L) at DO concentrations as low as 3 mg O₂ l^?1. With access to the water surface, both G_S and I_L of F. heteroclitus decreased by c. 60% at 1·0 mg O₂ l^?1 compared to all higher DO treatments. When denied access to the water surface, a further decrease in G_S (c. 90%) and I_L (c. 75%) was observed at 1 mg O₂ l^?1. There was no effect of diel‐cycling DO (1–11 mg O₂ l^?1) with or without surface access on G_S, I_L or Hct of F. heteroclitus. Similar trends between G_S and faecal production across DO treatments suggest that decreased feeding contributed significantly to the observed decrease in growth rate. Haematocrit was significantly elevated at 1 mg O₂ l^?1 for fish with and without access to the water surface. Increased Hct, however, was not sufficient to maintain high G_S or I_L at severely low DO. When permitted to respire in the surface layer, however, F. heteroclitus was capable of maintaining moderate growth rates at DO concentrations of 1 mg O₂ l^?1 (c. 15% saturation). Although aquatic surface respiration (ASR) was not quantified in this study, F. heteroclitus routinely swam in contact with the water surface and performed ASR at DO concentrations ≤3 mg O₂ l^?1. No hypoxia‐related mortality was observed in any DO or surface access treatment for as long as 9 days. This study demonstrates that surface access, and thus potential for ASR, plays an important role in providing F. heteroclitus substantial independence of growth rate over a wide range of low DO conditions commonly encountered in shallow estuarine environments.     

Utilization of metabolic scope in relation to feeding and activity by individual and grouped zebrafish, Brachydanio rerio (Hamilton-Buchanan)

Metabolic scope and its utilization in relation to feeding and activity were measured in individual and grouped zebrafish (weight range, 430–551 mg) at 24° C by respirometry. Mean maximum metabolic rate, induced by swimming to exhaustion, R_max(i), was 1223 (s.d. , 157) mg O₂, kg^?1 h^?1 for individuals. Standard metabolic rate, R_s. was 364 mg O₂ kg^?1 h^?1, as estimated by extrapolating to zero activity from measurements of unfed, spontaneously active individuals. Mean routine metabolic rate, R_rout, of individuals was 421 (s.d. , 58) mg O₂, kg_-1 h_-1. The mean voluntary maximum metabolic rate, R_max(v), following transfer of minimally exercised fish to the respirometer, was 1110 (s.d. , 83) mg O₂ kg ^?1 h^?1 for groups of six fish, and was not significantly different from the value measured for individuals, 1066 (s.d. , 122) mg O₂, kg^?1 h^?1. Grouped fish acclimated to the respirometer more slowly than individual fish and exhibited significantly higher R_rout, apparently a result of greater social interaction and activity in groups. Mean R_rout for groups was 560 (s.d. , 78) mg O₂, kg^?1 h^?1. While groups of zebrafish fed a ration of 5% wet body weight day^?1 exhibited consistently higher metabolic rates than fish fed rations of 2.5% wet body weight day^?1 the high ration group still used only a maximum of 77% of the metabolic scope. Zebrafish of the size studied do not appear to demonstrate a high degree of conflict in utilization of metabolic scope by different respiratory components. The metabolic rates measured for zebrafish are among the highest yet measured for fish of similar size and at similar temperatures.     

Effect of water-borne copper on respiratory and cardiac function during the early ontogeny of the brine shrimp,Artemia franciscana Kellogg 1908 (Branchiopoda: Anostraca)

There was no direct effect of copper on the ontogeny or function of the heart of the brine shrimp Artemia franciscana in sea water (salinity= 36 mg·ml^-1, 25°C). There was, however, an indirect effect as an increase in copper concentration resulted in a reduced growth rate. There was no difference between the critical O₂ tensions of newly hatched (stage 0/1) nauplii of control and treated (<0.32 and 10.11 mol·l^-1 copper, respectively) individuals. However by developmental stages 4–6, when both the heart and thoracic gills are in the process of differentiating, respiratory performance had improved (i.e. critical O₂ tension decreased from 6.27±0.45 to 4.69±0.24 kPa) in control but not in copper-treated individuals. It is suggested that respiratory impairment of stages 4–6 individuals is unlikely to be related to differences in cardiac performance or cellular respiration. Instead it may be related to metal-related damage to newly differentiating gill tissue and/or by copper in some way compromising the normal ontogenic shift in haemoglobin O₂ affinity. Copper-related respiratory impairment develops at a critical point in brine shrimp organogenesis when a good supply of O₂ is essential for normal development and if compromised may reduce the ability of this species to survive copper exposure.Abbreviations BL body length - BW body weight - HR heart rate - HM heavy metals - SW sea water - P _c critical oxygen tension     

Standard metabolic rate models for Carmine Shiner (Notropis percobromus) and Common Shiner (Luxilus cornutus) across different temperature regimes

Standard metabolic rates (SMR) were measured empirically for carmine shiner Notropis percobromus and common shiner Luxilus cornutus to develop SMR models that predict metabolic responses of each species under thermal conditions observed in the wild. SMR increased significantly with body mass and rising water temperature, ranging from 0.05 mg O₂ h⁻¹ at 10°C to 0.89 mg O₂ h⁻¹ at 20°C for N. percobromus weighing 0.6–2.5 g and from 0.11 mg O₂ h⁻¹ at 10°C to 0.98 mg O₂ h⁻¹ at 20°C for L. cornutus weighing 0.8–6.6 g. SMR models significantly differed between sympatric species on account of differences in model intercepts (RA) and temperature coefficients (RQ), however, the allometric relationships between mass and SMR did not significantly differ between species. Known distribution of N. percobromus and L. cornutus includes the Birch River located in Manitoba, Canada, where N. percobromus is listed as Endangered. Little is known about the physiology of N. percobromus or the species' ability to acclimate or adapt to different environmental conditions. While size differences between species contributed, in part, to differences in SMR predictions for Birch River populations, SMR trends (< 2 mg O₂ h⁻¹) for individuals weighing 1 g were similar for both species across daily temperatures. Respirometry experiments contributed to developing species-specific SMR models and inform on the effect of natural and anthropogenic stressors, namely water temperature, on the conservation of N. percobromus in this ecosystem.     

Blood oxygen dissociation curve of the frogsRana catesbeiana andRana brevipoda

Summary The blood O₂ dissociation curve was determined with a microphotometric reaction apparatus in two frog species,Rana catesbeiana andRana brevipoda, and formulated in terms of a modified Hill equation. The O₂ dissociation curve was in reasonable agreement with determinations of O₂ content, O₂ capacity, and pH (or ) in blood samples taken from various blood vessels and heart cavities. The blood O₂ affinity inRana catesbeiana (mean body weight 230 g, kept at 25°C) was low:P ₅₀=42 Torr (at 25°C and at ventricular blood pH=7.79). The blood of the smaller frog,Rana brevipoda (mean body weight 16 g), had even lower O₂ affinity,P ₅₀ being 52 Torr (at 25°C, pH=7.72). The decrease ofP ₅₀ with increasing body mass, resulting from comparison of these two frog species, is in accordance with literature data on mammals and birds.     

Effect of meal size on postprandial metabolic response in Chinese catfish (Silurus asotus Linnaeus)

The effect of relative meal size (0.5–24% body mass) on specific dynamic action (SDA) was assessed in Chinese catfish (Silurus asotus Linnaeus) (30.90±1.30 g) at 25.0°C; the cutlets of freshly killed loach without viscera, head and tail were used as a test meal. There was no significant difference in either SDA duration or peak oxygen consumption (VO₂) among low meal size ranges. But both increased linearly as meal size increased from 2 to 24% without reaching a plateau. Factorial metabolic scope was 5.92 in fish fed with 24% body mass, the highest documented feeding metabolic scope value in fish till now. The Peak VO₂ of satiated meal size groups (175.85±10.55 mg O₂ h⁻¹) was above 80% of maximum metabolic rate during locomotion recovery process (215.48±7.07 mg O₂ h⁻¹). The relationship between energy expended on SDA (E) and energy ingested (I) was described as: E=0.0000432I ²+0.140I+2.12. The lowest value of SDA coefficient appeared at 2% body mass group.     

High-dose chromium(III) supplementation has no effects on body mass and composition while altering plasma hormone and triglycerides concentrations

Chromium is generally believed to be an essential element and is often claimed to have value as a weight loss or muscle building agent. Recent studies in humans and rats have failed to demonstrate effects on body composition, although recent studies with pharmacological doses of the cation [Cr(III)₃O(O₂CCH₂CH₃)6(H₂O)₃]⁺ (or Cr3) (≤1 mg Cr/kg body mass) in rats have noted a trend toward body mass loss and fat mass loss. Thus, the effects of large gavage doses of Cr3 (1–10 mg Cr/kg) on body mass, organ mass, food intake, and blood plasma variables (insulin, glucose, leptin, cholesterol, and triglycerides) were examined over a 10-wk period using male Sprague-Dawley rats. No effects on body composition were noted, although Cr3 administration lowered (p<0.05) plasma insulin, leptin, and triglycerides concentrations. As Cr3 is absorbed greater than 10-fold better than commercially available nutritional supplements, the lack of an effect of the Cr(III) compound at these levels of administration clearly indicates that Cr(III) supplements do not have an effect on body composition at any reasonable dosage.     

O2 uptake through water during early life of Heteropneustes fossilis (Bloch)

The O₂ uptake through water has been measured in case of Heteropneustes fossilis during development and growth and its relationship to body size established. A higher rate of O₂ uptake during the early phase of ontogenesis is related to intense growth of the respiratory surface area and increasing metabolic demand of the fish.The logarithmic plot of data for O₂ uptake in relation to body size shows a statistically significant two-component curve; one related to the fish when it is a fully aquatic breather and the other when it changes to bimodal gas exchange. The onset of the air breathing habit brings about a 40% drop in O₂ uptake through water, which is made good through the newly developed air breathing organ.     

Responses to hypoxia of plaice,Pleuronectes platessa,and dab,Limanda limanda,in the south-east Kattegat: distribution and growth

Synopsis Plaice, Pleuronectes platessa, and dab, Limanda limanda, were sampled with a Glommen lobster trawl at 25 to 40 m depth in the SE Kattegat during spring and autumn of 1984 to 1990. During autumn, hypoxia (O₂-concentration < 3 mg I^–1) occurred in the bottom water below the halocline for four to ten weeks every year, except in 1984 when moderate hypoxia (O₂-concentration 3–5 mg 1^–1) occurred. Biomass of both species was shown to be negatively correlated with oxygen concentration during autumn. Further, a decrease in population mean total length was observed during the study period in both spring and autumn samples. Laboratory studies of growth of juvenile plaice and dab, at 15° C and 30–34%, showed that growth is reduced at 50 and 30% O₂-saturation for both species during a 20 d period. There was some adaptation to hypoxia resulting in less reduction of growth during the second half of the experiment. The frequency of fish eating was reduced in plaice at 30% O₂-saturation. Reduced mean total length of the plaice and dab population of the SE Kattegat is discussed in view of sublethal effects of oxygen deficiency.     

Protective effect of Zizyphus lotus jujube fruits against cypermethrin-induced oxidative stress and neurotoxicity in mice

Context: Cypermethrin (CYP) is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control. The toxicity of CYP is well studied in many organisms.

Objective: The aim of present study was to investigate the protective effect of Zizyphus lotus (Zizyp) fruit against neurotoxicity and oxidative stress induced by CYP in mice.

Materials and methods: Mice were divided into four groups of six each: groups I and II were used as control and CYP control (20?mg/kg body weight). While, groups III was orally treated with Zizyphus lotus fruit (5?g/kg body weight) plus CYP (20?mg/kg body weight) for 18?days. Furthermore, HPLC–ESI–MS–MS (Q-Tof) and GC–MS were used to identify the compounds fraction.

Results: Antioxidant enzyme catalase (CAT), neurotoxicity enzyme acetylcholinesterase (AChE) activities and hydrogen peroxide (H₂O₂)_, malondialdehyde (MDA) levels were determined in the liver, kidney and heart. CYP caused decreased CAT activity, inhibition of AChE activity and increased the levels of H₂O₂ and MDA in heart, liver and kidney.

Conclusion: Our results indicate that Zizyp fruit is markedly effective in protecting mice against CYP-induced biochemical changes. This protection may be due to its antioxidant property and scavenging ability against active free radicals.     

Oxygen consumption by ammocoetes of the lampreyGeotria australis in air

 When covered by moistened lint-free gauze, the larvae (ammocoetes) of the lamprey Geotria australis survived, without apparent discomfort, for 4 days in water-saturated air at 10, 15 and 20 °C. In air, the mean standard rates of O₂ consumption of medium to large ammocoetes of G. australis (xˉ=0.52 g) at 10, 15 and 20 °C were 14.5, 35.7 and 52.1 μl⋅g^-1⋅h^-1, respectively. At 15 °C, the slope of the relationship between log O₂ consumption (μl O₂⋅h^-1) and log body weight for ammocoetes over a wide range in body weight was 0.987. The Q ₁₀s for rate of O₂ consumption between 10 and 15 °C, 15 and 20 °C and 10 and 20 °C were 4.9, 2.9 and 3.6, respectively. Our results and observations of the ammocoetes suggest that, when out of water, larval G. australis derives most of its O₂ requirements from cutaneous respiration, particularly at lower temperatures. This would be facilitated by the small size and elongate shape (and thus a relatively high surface-to-volume ratio), low metabolic rate, thin dermis, extensive subdermal capillary network and high haemoglobin concentration of larval G. australis. Accepted: 28 March 1996     

Influence of daylength on metabolic rate and daily water loss in the male prosimian primate Microcebus murinus

In its natural habitat, Microcebus murinus, a small malagasy prosimian primate, is exposed to seasonal shortage of water and resources. During the winter dry season, animals enter a pronounced fattening period with concurrent decrease in behavioural/physiological activities, whereas the breeding season is restricted to the rainy summer months. To determine the role of daylength on metabolic rate and water loss in this nocturnal primate, we measured body mass, oxygen consumption at 25°C (RMR), circadian water loss through urine output (UO) and evaporation (EWL) in eight males exposed to either short days (8L:16D SD) or long days (14L:10D LD), under controlled captive conditions. Exposure to SD led to a ponderal increase (maximal body mass: 125±4 g, N=8), and to significant changes in RMR and water loss, both reaching lowest values after 3 months under SD (0.84±0.04 ml O₂ h⁻¹ g⁻¹ and 38±0.3 mg H₂O g⁻¹ day⁻¹, respectively). Following exposure to LD, body mass decreased to 77±3 g (N=8), whereas both RMR and water loss, mainly through EWL, significantly increased (P<0.001), the highest value occurring after 2 months (1.51±0.08 ml O₂ h^.−1 g⁻¹ and 87±7 mgH₂O g⁻¹ day⁻¹, respectively). Moreover, independent of daylength, circadian changes in EWL were characterized by significantly reduced values during the diurnal rest. The results demonstrate that daylength variations affect the physiology of this tropical primate, allowing anticipatory adaptation to seasonal environmental constraints.     

Feeding dynamics and respiration of the bottom-dwelling caridean shrimp Nauticaris marionis Bate, 1888 (Crustacea: Decapoda) in the vicinity of Marion Island (Southern Ocean)

The feeding dynamics and oxygen uptake of the bottom-dwelling caridean shrimp Nauticaris marionis were studied during the April/May 1984, 1996 and 1997 cruises to Marion Island (Prince Edward Islands, Southern Ocean). N. marionis is thought to have an opportunistic feeding mode. Prey composition varied considerably between the years and sites investigated. Overall, benthic (mainly hydrozoans and bottom-dwelling polychaetes) and, at times, pelagic (largely euphausiids and copepods) prey items dominated in the stomachs of N. marionis both by occurrence and by volume. Generally, pelagic prey contributed more to the diets of smaller shrimps, while benthic prey was a more important component in the guts of larger specimens. Wet, dry and ash-free dry weight were determined for specimens used in respiration experiments. The respiration rates of N. marionis females with carapace length 6.6–11.1 mm ranged from 80 to 250 μl O₂ individual⁻¹ · h⁻¹, or from 0.588 to 2.756 μl O₂ · mg⁻¹ dry weight h⁻¹. Regression analyses showed highly significant correlations between oxygen consumption and carapace length for N. marionis. Daily ingestion rates estimated using an in situ gut content analysis technique (4.4% of body dry weight) and an energy budget approach (average 4.7% of body dry weight, range 2.0–7.5%) showed good agreement with each other. Accepted: 29 July 1998     

Oxygen uptake in tobacco budworm larvae (Heliothis virescens) infected with cytoplasmic polyhedrosis virus

We measured oxygen uptake in 3- to 13-day-old Heliothis virescens (tobacco budworm) larvae. The “test” group was infected with cytoplasmic polyhedrosis virus (CPV) while the “control” group was not. The healthy budworms had an average oxygen uptake of 7.64 μl of O₂/mg body weight, while those infected with CPV had an average uptake of 47.11 μl of O₂/mg body weight/hr. The weights of larvae from the two groups were likewise recorded. Budworms from the “control” group showed an average weight of 186.9 mg, while larvae infected with CPV had an average weight of 18.3 mg.     

Cytochrome c oxidase: the mechanistic significance of structural H+ in energy transduction

Changes in the bulk-phase concentration of O₂ and H⁺ associated with the reduction of O₂ to water are simultaneously determined in reactions catalyzed by fully reduced cytochrome c oxidase both isolated and embedded in liposomes. Consistent with the polyphasic kinetics of electron transfer through the oxidase, the time course of O₂ consumption and H⁺ translocation exhibit the following novel characteristics: (1) The uptake of scalar protons (H_m ⁺), the ejection of vectorial protons (H⁺ _v), and the consumption of O₂, all proceed in a kinetically polyphasic process. (2) During the first phase of the reaction the rates of O₂ uptake and H⁺ transfer are extremely fast and compatible with the rates of electron flow through the oxidase. (3) The K_m of the oxidase for O₂ is close to 75 M, the same for O₂ consumption and scalar H⁺ uptake. The V_max of O₂ reduction to water in reactions catalyzed by the isolated enzyme is, at least, 0.5 × 10⁴ s^–1. (4) The extent of vectorial H⁺ ejection by cytochrome c oxidase embedded in liposomes is an exponential function dependent on both enzyme concentration and extent of O₂ consumption. (5) The H⁺/O stoichiometry of H⁺ ejection is a variable that may reach a maximum value of 4.0 only when the enzyme undergoes net oxidation at extremely high enzyme/O₂ molar ratios. It is postulated that the generation of useful energy at the level of cytochrome c oxidase depends not only on the number of molecules of O₂ reduced to water but also on the extent and state of reduction and/or protonation of the enzyme.     

Hypoglycemic activity of the fungi Cordyceps militaris, Cordyceps sinensis, Tricholoma mongolicum, and Omphalia lapidescens in streptozotocin-induced diabetic rats

Crude extracts were prepared from fruiting bodies and mycelia of the medicinal fungus Cordyceps militaris, and a polysaccharide-enriched fraction was obtained after extraction with hot water and ethanol precipitation. Polysaccharide-enriched fractions were similarly prepared from Cordyceps sinensis, Omphalia lapidescens, and Tricholoma mongolicum. The various aforementioned preparations were orally administered into different groups of adult rats 24 h before an intraperitoneal injection of streptozotocin (40 mg/kg body weight), and subsequently daily for another 4 days. The dosage used was 10 mg/kg body weight for polysaccharide-enriched preparations and 100 mg/kg body weight for crude extracts. Control rats received distilled water instead of crude extract or polysaccharide-enriched preparation. It was found in the control rats that plasma glucose level rose from about 90 mg/dl before streptozotocin injection to levels that were maintained at about 300 mg/dl postinjection. All preparations produced hypoglycemic effects. C. militaris polysaccharide-enriched fraction displayed a more prominent effect than that of C. sinensis polysaccharide-enriched fraction which in turn was more potent than that of O. lapidescens and T. mongolicum polysaccharide-enriched fractions. The hypoglycemic effect of C. militaris polysaccharide-enriched fraction was dose-dependent.     

Water balance of over-wintering beetles in relation to strategies for cold tolerance

The vapour pressure of the haemolymph of a supercooled insect is higher than the vapour pressure of the haemolymph of a frozen insect at the same temperature. The aim of the study was to see whether this may affect the water loss of freeze-avoiding and freezetolerant, over-wintering beetles. The rates of water loss were determined on freeze-tolerant Pytho depressus larvae and Upis ceramboides adults. Within each species one group was kept supercooled whereas another group was frozen. All groups were incubated at-5°C. Both species displayed significantly lower rates of water loss when they were frozen than when they were supercooled. Values of respiratory rate and water loss of freeze-avoiding and freeze-tolerant species were compared to corresponding values of desert beetles. The results indicate that the freeze-avoiding species have lower rates of cuticular water loss than freeze-tolerant species. This indicates that the freeze-avoiding species have developed more efficient water-saving mechanisms than freeze-tolerant species. The reason for this may be that the haemolymph in frozen animals will be in vapour pressure equlibrium with the ice in the hibernaculum and thus there is no danger of desiccation during winter. The supercooled insects will have a vapour pressure of the haemolymph that is higher than the vapour pressure of water in the surrounding air and will thus lose water.Abbreviations BW body weight - BW_i initial body weight - BW_t body weight at time t - P vapour pressure difference between the water in the haemolymph and the water in the air - DWL_t dry weight loss at time t - M _w rate of metabolic water production - MF_w mol fraction of water, in the haemolymph - MO₂ rate of oxygen consumption - Osm osmolality of the haemolymph - P _a vapour pressure of water in the air - P _h vapour pressure of water in the haemolymph - P _w vapour pressure of pure water - Q a constant (2,02 1 oxygen per g fat metabolized) relating oxygen consumption to dry weight loss when fat is metabolized - R a constant (1,89 1 oxygen per g water produced) relating metabolic water production to oxygen consumption when fat is metabolized - R _dwl rate of dry weight loss - RH relative humidity of the air - RWC_i initial relative water content measured by weighing - RWC_t relative water content at time t - STP standard temperature and pressure