Validating accelerometry-derived proxies of energy expenditure using the doubly labelled water method in the smallest penguin species.

Understanding energy use is central to understanding an animal's physiological and behavioural ecology. However, directly measuring energy expenditure in free-ranging animals is inherently difficult. The doubly labelled water (DLW) method is widely used to investigate energy expenditure in a range of taxa. Although reliable, DLW data collection and analysis is both financially costly and time consuming. Dynamic body acceleration (e.g. VeDBA) calculated from animal-borne accelerometers has been used to determine behavioural patterns, and is increasingly being used as a proxy for energy expenditure. Still its performance as a proxy for energy expenditure in free-ranging animals is not well established and requires validation against established methods. In the present study, the relationship between VeDBA and the at-sea metabolic rate calculated from DLW was investigated in little penguins (Eudyptula minor) using three approaches. Both in a simple correlation and activity-specific approaches were shown to be good predictors of at-sea metabolic rate. The third approach using activity-specific energy expenditure values obtained from literature did not accurately calculate the energy expended by individuals. However, all three approaches were significantly strengthened by the addition of mean horizontal travel speed. These results provide validation for the use of accelerometry as a proxy for energy expenditure and show how energy expenditure may be influenced by both individual behaviour and environmental conditions.

Daily energy expenditure and water turnover in two breeds of laying hens kept in floor housing.

Laying hens are increasingly kept in barn or free-range systems, which not only allows birds to move freely but also potentially entails higher energy expenditures due to higher locomotor activity. Therefore, the aim of our study was to quantify the daily energy expenditure (DEE) and water turnover in freely moving laying hens. For that purpose, 10 Lohmann Selected Leghorn (LSL) and 10 Lohmann Brown (LB) hens were obtained from a conventional breeding company at 17 weeks of age. The trial started when birds reached an age of 34 weeks. All 20 birds were kept together in the same littered floor pen (12.1 m2). The pen was equipped with perches, a nest box, feeding and nipple drinkers. The DEE was determined individually for all experimental birds (n = 20) for a total of nine days using the doubly labelled water (DLW) method. Lohmann Brown hens were heavier than LSL hens, but laying rate did not differ between the two breeds, that is, one egg per hen and day during the study period. Average egg mass was 63.1 ±â€¯0.20 g in LB and 61.7 ±â€¯0.12 g in LSL hens, which converted to an egg energy content of 420 and 410 kJ/egg, respectively. Dilution spaces for oxygen and hydrogen differed between the breeds but not the respective turnover rates. Total body water as a percentage of body mass (LB: 54.4%, LSL: 53.8%; SEM = 0.7, F1,18 = 0.41, P = 0.513) and total water intake (TWI) per day (LB: 275 ml/day, LSL: 276 ml/day; SEM = 20, F1,17 = 0, P = 0.994) did not differ between LB and LSL hens. Individual DEE increased with body mass in LB but not in LSL hens. Average DEE did not differ between the two breeds (LB: 1501 kJ/day; LSL: 1520 kJ/day; SEM = 32.1, F1,17 = 2.54, P = 0.131). However, when comparing the DEE on a metabolic mass basis, LSL hens expended with 984 kJ/kg0.75 on average significantly more energy per day than LB hens (895 kJ/kg0.75; SEM = 20.3, F1,18 = 10.1, P = 0.005). Our results suggest that the DLW technique is a viable method to measure the energy expenditure and water turnover over several days in laying hens. Furthermore, we show that laying hens kept in floor pens fit into the general pattern of DEE among wild birds.

Oxidative costs of reproduction: Oxidative stress in mice fed standard and low antioxidant diets.

Lactation is one of the most energetically expensive behaviours, and trade-offs may exist between the energy devoted to it and somatic maintenance, including protection against oxidative damage. However, conflicting data exist for the effects of reproduction on oxidative stress. In the wild, a positive relationship is often observed, but in laboratory studies oxidative damage is often lower in lactating than in non-breeding animals. We hypothesised that this discrepancy may exist because during lactation food intake increases many-fold resulting in a large increase in the intake of dietary antioxidants which are typically high in laboratory rodent chow where they are added as a preservative. We supplied lactating and non-breeding control mice with either a standard or low antioxidant diet and studied how this affected the activity of endogenous antioxidants (catalase, superoxide dismutase; SOD, and glutathione peroxidise; GPx) and oxidative damage to proteins (protein carbonyls, PC) in liver and brain tissue. The low antioxidant diet did not significantly affect activities of antioxidant enzymes in brain or liver, and generally did not result in increased protein damage, except in livers of control mice on low antioxidant diet. Catalase activity, but not GPx or SOD, was decreased in both control and lactating mice on the low antioxidant diet. Lactating mice had significantly reduced oxidative damage to both liver and brain compared to control mice, independent of the diet they were given. In conclusion, antioxidant content of the diet did not affect oxidative stress in control or reproductive mice, and cannot explain the previously observed reduction in oxidative stress in lactating mammals studied in the laboratory. The reduced oxidative stress in the livers of lactating mice even under low antioxidant diet treatment was consistent with the 'shielding' hypothesis.

Effects of age and body mass on development of diving capabilities of gray seal pups: costs and benefits of the postweaning fast.

Development of adequate diving capabilities is crucial for survival of seal pups and may depend on age and body size. We tracked the diving behavior of 20 gray seal pups during their first 3 mo at sea using satellite relay data loggers. We employed quantile analysis to track upper limits of dive duration and percentage time spent diving, and lower limits of surface intervals. When pups first left the breeding colony, extreme (ninety-fifth percentile) dive duration and percentage time spent diving were positively correlated with age, but not mass, at departure. Extreme dive durations and percentage time spent diving peaked at [Formula: see text] d of age at values comparable with those of adults, but were not sustained. Greater peaks in extreme percentage time spent diving occurred in pups that had higher initial values, were older at their peak, and were heavier at departure. Pups that were smaller and less capable divers when they left the colony improved extreme dive durations and percentage time spent diving more rapidly, once they were at sea. Minimum survival time correlated positively with departure mass. Pups that were heavier at weaning thus benefitted from being both larger and older at departure, but smaller pups faced a trade-off. While age at departure had a positive effect on early dive performance, departure mass impacted on peak percentage time spent diving and longer-term survival. We speculate that once small pups have attained a minimum degree of physiological development to support diving, they would benefit by leaving the colony when younger but larger to maximize limited fuel reserves, rather than undergoing further maturation on land away from potential food resources, because poor divers may be able to "catch up" once at sea.

Physical activity energy expenditure has not declined since the 1980s and matches energy expenditures of wild mammals.

OBJECTIVE: Obesity results from protracted energy imbalance. Whether this comprises excessive energy intake, lowered physical activity or both, remains disputed. DESIGN: Physical activity energy expenditure, evaluated in three different ways from daily energy expenditure (DEE) measured using doubly labelled water, was examined for trends over time. Data included subjects in Europe (Maastricht, the Netherlands) and North America extending back to the 1980s. These data were compared with measures from the third world, and measures made on wild terrestrial mammals. RESULTS: Physical activity expenditure in Europe (residual of the regression of DEE on basal energy expenditure (BEE)) has slightly but significantly increased since the 1980s. There was no trend over time in physical activity level (PAL=DEE/BEE), or in the residual variance in DEE once mass, sex and age were accounted for. This latter index of physical activity expenditure also significantly increased over time in North America. DEE of individuals in Europe and North America was not significantly different from individuals measured in the third world. In wild terrestrial mammals, DEE mostly depended on body mass and ambient temperature. Predicted DEE for a 78 kg mammal living at 20 degrees C was 9.2 MJ per day (95% CI: 7.9-12.9 MJ per day), not significantly different from the measured DEE of modern humans (around 10.2-12.6 MJ per day). CONCLUSION: As physical activity expenditure has not declined over the same period that obesity rates have increased dramatically, and daily energy expenditure of modern man is in line with energy expenditure in wild mammals, it is unlikely that decreased expenditure has fuelled the obesity epidemic.

Energetic costs of parasitism in the Cape ground squirrel Xerus inauris.

Parasites have been suggested to influence many aspects of host behaviour. Some of these effects may be mediated via their impact on host energy budgets. This impact may include effects on both energy intake and absorption as well as components of expenditure, including resting metabolic rate (RMR) and activity (e.g. grooming). Despite their potential importance, the energy costs of parasitism have seldom been directly quantified in a field setting. Here we pharmacologically treated female Cape ground squirrels (Xerus inauris) with anti-parasite drugs and measured the change in body composition, the daily energy expenditure (DEE) using doubly labelled water, the RMR by respirometry and the proportions of time spent looking for food, feeding, moving and grooming. Post-treatment animals gained an average 19g of fat or approximately 25kJd-1. DEE averaged 382kJd-1 prior to and 375kJd-1 post treatment (p>0.05). RMR averaged 174kJd-1 prior to and 217kJd-1 post treatment (p<0.009). Post-treatment animals spent less time looking for food and grooming, but more time on feeding. A primary impact of infection by parasites could be suppression of feeding behaviour and, hence, total available energy resources. The significant elevation of RMR after treatment was unexpected. One explanation might be that parasites produce metabolic by-products that suppress RMR. Overall, these findings suggest that impacts of parasites on host energy budgets are complex and are not easily explained by simple effects such as stimulation of a costly immune response. There is currently no broadly generalizable framework available for predicting the energetic consequences of parasitic infection.

The energy costs of sexual dimorphism in mole-rats are morphological not behavioural.

Different reproductive strategies of males and females may lead to the evolution of differences in their energetic costs of reproduction, overall energetic requirements and physiological performances. Sexual dimorphism is often associated with costly behaviours (e.g. large males might have a competitive advantage in fighting, which is energetically expensive). However, few studies of mammals have directly compared the energy costs of reproductive activities between sexes. We compared the daily energy expenditure (DEE) and resting metabolic rate (RMR) of males and females of two species of mole-rat, Bathyergus janetta and Georychus capensis (the former is sexually dimorphic in body size and the latter is not) during a period of intense digging when males seek females. We hypothesized that large body size might be indicative of greater digging or fighting capabilities, and hence greater mass-independent DEE values in males of the sexually dimorphic species. In contrast to this prediction, although absolute values of DEE were greater in B. janetta males, mass-independent values were not. No differences were apparent between sexes in G. capensis. By comparison, although RMR values were greater in B. janetta than G. capensis, no differences were apparent between the sexes for either species. The energy cost of dimorphism is most likely to be the cost of maintenance of a large body size, and not the cost of behaviours performed when an individual is large.

Associations between BMI, social strata and the estimated energy content of foods.

OBJECTIVE: Many studies have shown that the prevalence of obesity is greater in lower social classes. The reasons for this effect however are unclear. Since there is also a link between education and social class, and an association between education and prevalence of obesity, one hypothesis is that lack of education about energy contents of foods may contribute to the effects of social class on obesity. SUBJECTS: We tested the hypothesis that knowledge of food energy contents is associated with differences in body mass index (BMI) in a sample of 346 people of both genders, aged between 18 and 45 y, of variable body mass index and drawn from different social strata. RESULTS: Estimates of food energy contents were on average well correlated with the actual energy contents, but individual estimates were very poor in all subpopulations of this sample. We found that subjects of different BMIs did not differentially estimate the energy contents of foods high in carbohydrate, but low in fat and protein (fruit and bread). However, foods that contained high fat contents, independent of the other macronutrients present, were generally perceived to have significantly lower energy contents by obese people than nonobese subjects (although this was not observed for all high-fat foods). Overall, this difference interacted with social class, such that the difference between the BMI groups was exaggerated in the lower social stratum but abolished in the higher social class. Binary logistic regressions revealed that the probability of being obese (BMI>30 kg/m2) in the lower social class group was significantly negatively associated with the estimated food energy content of most high-fat foods. Such an association was not found in the higher social class group. In the lower social class group, overall food knowledge appeared superior in the leaner subject group (BMI<30 kg/m2), but obese subjects were actually better at estimating the energy contents of snacks and alcoholic beverages. The leaner group significantly overestimated the energy contents of these items. CONCLUSION: Differences between individuals in estimates of food energy contents may contribute to the development of obesity in lower social strata. Whether this is driven by a protective effect in lean subjects of overestimating the energy contents of certain foods (snacks and alcoholic beverages) or a susceptibility in the obese because they underestimate the energy contents of other foods is not certain. Knowing which of these alternatives is true is important and may help design public health education programmes directed at these people to help alleviate the obesity epidemic.

Comparison of the cost of short flights in a nectarivorous and a non-nectarivorous bird.

Although most birds are accustomed to making short flights, particularly during foraging, the flight patterns during these short periods of activity differ between species. Nectarivorous birds, in particular, often spend time hovering, while non-nectarivorous birds do not. The cost of short flights is likely therefore to differ between nectarivorous and non-nectarivorous birds because of the different energetic contributions of different flight types to the behaviour. The 13C-labelled bicarbonate technique was used to measure the energy cost of short flights in the nectarivorous Palestine sunbird Nectarinia osea (mean mass 6.17+/-0.16 g, N=8) and the non-nectarivorous starling Sturnus vulgaris (mean mass 70.11+/-1.11 g, N=9). The technique was initially calibrated in five individuals for each species at temperatures ranging from 1 to 35 degrees C, by comparing the isotope elimination rate to the metabolic rate measured simultaneously by indirect calorimetry. The cost for short intermittent flight was then measured by encouraging birds to fly between two perches at either end of a narrow corridor (perch distance for sunbirds, 6 m; for starlings, 5 m), and measuring the amount of isotope eliminated during the flight. The isotope elimination rate was interpolated onto the calibration equation to predict flight cost, as a direct calibration could not be performed during flight. Mean energy expenditure during flight was 1.64+/-0.32 W in sunbirds, while in starlings the flight costs averaged 20.6+/-0.78 W. Energy cost of flight relative to basal metabolic rate was substantially greater in the starling than the sunbird. Phylogenetic analysis of different modes of flight in these and additional species suggests that differences in flight behaviour may cause these elevated costs in slow flying non-nectarivores such as starlings, compared to birds that are more prone to short intermittent flights like the sunbirds.

The energy cost of loaded flight is substantially lower than expected due to alterations in flight kinematics.

The effect of experimentally increased wing loading on the energy cost of flight was examined in cockatiels Nyphicus hollandicus. Five individuals were flown for periods of approximately 2 min, while carrying additional payload mass amounting to between 5 and 20% of unloaded body mass. The energy cost of flight was measured using the 13C-labelled bicarbonate technique, which was also calibrated in a separate experiment on resting birds, by comparing the elimination rate of 13C in breath with a simultaneous measurement of oxygen consumption by indirect calorimetry. It was not possible to perform a similar calibration during flight when energy costs were higher, so we extrapolated the relationship from the resting calibration to predict flight cost. Flight cost in the pre-manipulated individuals averaged 16.7+/-1.8 W. Flight cost in the pre-manipulated birds was significantly related to the interaction between downstroke duration and flight speed. There was no significant increase in flight cost with increases in payload mass. The birds responded to payload masses between 5 and 15% of their unloaded body mass by decreasing flight speed relative to unloaded birds, while maintaining wing beat frequency (Fb). At a payload mass equivalent to 20% of body mass, however, the birds flew at higher speeds than unloaded controls, and had a significantly higher Fb, generated by a reduction in both the upstroke and downstroke durations. Wing amplitude was unaffected by the increase in loading. Using the measured flight parameters, the effect of loading was not significantly different than predicted using aerodynamic models.

The energetic cost of variations in wing span and wing asymmetry in the zebra finch Taeniopygia guttata.

Asymmetry is a difference in the sizes of bilaterally paired structures. Wing asymmetry may have an effect on the kinematics of flight, with knock-on effects for the energetic cost of flying. In this study the 13C-labelled bicarbonate technique was used to measure the energy expended during the flight of zebra finches Taeniopygia guttata, prior to and after experimental manipulation to generate asymmetry and a change in wing span by trimming the primary feathers. In addition, simultaneous high-speed video footage enabled differences in flight kinematics such as flight speed, wing amplitude, up- and downstroke duration and wing beat frequency to be examined. In 10 individuals, the primary feathers on the right wing were trimmed first, by 0.5 cm, and then by an additional 0.5 cm in six of these individuals. In a separate 'control' group (N=7), approximately 0.25 cm was trimmed off the primary feathers of both wings, to produce the same reduction in wing span as 0.5 cm trimmed from one wing, while maintaining symmetry. When birds were manipulated to become asymmetric they maintained flight speed. They also increased the left wing amplitude and decreased the right up- and downstroke durations to counteract the changes in wing shape, which meant that they had an increase in wing beat frequency. When the wing area was reduced while maintaining symmetry, birds flew with slower flight speed. In this case wing amplitude did not change and wing upstroke slightly decreased, causing an increased wing beat frequency. The mean flight cost in the pre-manipulated birds was 1.90+/-0.1 W. There was a slight increase in flight cost with both of the asymmetry manipulations (0.5 cm, increase of 0.04 W; 1.0 cm, increase of 0.12 W), neither of which reached statistical significance. There was, however, a significantly increased flight cost when the wing span was reduced without causing asymmetry (increase of 0.45 W; paired t-test T=2.3, P=0.03).

Resting and daily energy expenditures of free-living field voles are positively correlated but reflect extrinsic rather than intrinsic effects.

Resting metabolic rates at thermoneutral (RMRts) are unexpectedly variable. One explanation is that high RMRts intrinsically potentiate a greater total daily energy expenditure (DEE), but recent work has suggested that DEE is extrinsically defined by the environment, which independently affects RMRt. This extrinsic effect could occur because expenditure is forced upwards in poor habitats or enabled to rise in good habitats. We provide here an intraspecific test for an association between RMRt and DEE that separates intrinsic from extrinsic effects and forcing from enabling effects. We measured the DEE and RMRt of 75 free-living short-tailed field voles at two time points in late winter. Across all sites, there was a positive link between individual variation in RMRt and DEE. This correlation, however, emerged only because of an effect across sites, rather than because of an intrinsic association within sites. We defined site quality from the survivorship of voles at the sites and the time at which they commenced breeding in spring. The associations between DEE/RMRt and site quality suggested that in February voles in poorer sites had higher energy demands, indicating that DEE was forced upwards, but in March the opposite was true, with higher demands in good sites, indicating that high expenditure was enabled. These data show that daily energy demands are extrinsically defined, with a link to RMRt that is secondary or independent. Both forcing and enabling effects of the environment may pertain at different times of year.

Cost of flight in the zebra finch ( Taenopygia guttata): a novel approach based on elimination of (13)C labelled bicarbonate.

On three separate occasions, five zebra finches ( Taenopygia guttata) were injected intraperitoneally with 0.2 ml 0.29 M NaH(13)CO(3)solution and placed immediately into respirometry chambers to explore the link between (13)C elimination and both O(2) consumption (VO(2)) and CO(2) production (VCO(2)). Isotope elimination was best modelled by a mono-exponential decay. The elimination rate (k(c)) of the (13)C isotope in breath was compared to VO(2) (ml O(2)/min) and VCO(2) (ml CO(2)/min) over sequential 5-min time intervals following administration of the isotope. Elimination rates measured 15-20 min after injection gave the closest relationships to VO(2) ( r(2) =0.82) and VCO(2) ( r(2)=0.63). Adding the bicarbonate pool size (N(c)) into the prediction did not improve the fit. A second group of birds ( n=11) were flown for 2 min (three times in ten birds and twice in one) between 15 min and 20 min following an injection of 0.2 ml of the same NaH(13)CO(3) solution. Breath samples, collected before and after flight, were used to calculate k(c) over the flight period, which was converted to VO(2) and VCO(2) using the equation generated in the validation experiment for the corresponding time period. The energy expenditure (watts) during flight was calculated from these values using the average RQ measured during flight of 0.79. The average flight cost measured using the bicarbonate technique was 2.24+/-0.11 W (mean+/-SE). This average flight cost did not differ significantly from predictions generated by an allometric equation formulated by Masman and Klaassen (1987 Auk 104:603-616). It was however substantially higher than the predictions based on the aerodynamic model of Pennycuick (1989 Oxford University Press), which assumes an efficiency of 0.23 for flight. The flight efficiency in these birds was 0.11 using this model. Flight cost was not related to within-individual variation [general linear model (GLM) F(1,31)=1.16, P=0.29] or across-individual variations in body mass (GLM F(1,31)=0.26, P=0.61), wingspan (regression F(1,10)=0.01, P=0.94) or wing loading (regression F(1, 31)=0.001, P=0.99) in this sample of birds.

Energetic and fitness costs of mismatching resource supply and demand in seasonally breeding birds.

By advancing spring leaf flush and ensuing food availability, climatic warming results in a mismatch between the timing of peak food supply and nestling demand, shifting the optimal time for reproduction in birds. Two populations of blue tits (Parus caeruleus) that breed at different dates in similar, but spatially distinct, habitat types in Corsica and southern France provide a unique opportunity to quantify the energetic and fitness consequences when breeding is mismatched with local productivity. As food supply and demand become progressively mismatched, the increased cost of rearing young pushes the metabolic effort of adults beyond their apparent sustainable limit, drastically reducing the persistence of adults in the breeding population. We provide evidence that the economics of parental foraging and limits to sustainable metabolic effort are key selective forces underlying synchronized seasonal breeding and long-term shifts in breeding date in response to climatic change.

Daily energy expenditure of the grey mouse lemur (Microcebus murinus): a small primate that uses torpor.

We aimed to investigate the pattern of utilisation of torpor and its impact on energy budgets in free-living grey mouse lemurs (Microcebus murinus), a small nocturnal primate endemic to Madagascar. We measured daily energy expenditure (DEE) and water turnover using doubly labelled water, and we used temperature-sensitive radio collars to measure skin temperature (Tsk) and home range. Our results showed that male and female mouse lemurs in the wild enter torpor spontaneously over a wide range of ambient temperatures (Ta) during the dry season, but not during the rainy season. Mouse lemurs remained torpid between 1.7-8.9 h with a daily mean of 3.4 h, and their Tsk s fell to a minimum of 18.8 degrees C. Mean home ranges of mouse lemurs which remained normothermic were similar in the rainy and dry season. During the dry season, the mean home range of mouse lemurs showing daily torpor was significantly smaller than that of animals remaining normothermic. The DEE of M. murinus remaining normothermic in the rainy season (122 +/- 65.4 kJ x day(-1)) was about the same of that of normothermic mouse lemurs in the dry season (115.5 +/- 27.3 kJ x day(-1)). During the dry season, the mean DEE of M. murinus that utilised daily torpor was 103.4 +/- 32.7 kJ x day(-1) which is not significantly different from the mean DEE of animals remaining normothermic. We found that the DEE of mouse lemurs using daily torpor was significantly correlated with the mean temperature difference between Tsk and Ta (r2 = 0.37) and with torpor bout length (r2 = 0.46), while none of these factors explained significant amounts of variation in the DEE of the mouse lemurs remaining normothermic. The mean water flux rate of mouse lemurs using daily torpor (13.0 +/- 4.1 ml x day(-1)) was significantly lower than that of mouse lemurs remaining normothermic (19.4 +/- 3.8 ml x day(-1)), suggesting the lemurs conserve water by entering torpor. Thus, this first study on the energy budget of free-ranging M. murinus demonstrates that torpor may not only reflect its impact on the daily energy demands, but involve wider adaptive implications such as water requirements.

Validation of the labeled bicarbonate technique for measurement of short-term energy expenditure in the mouse.

The energy expenditure of free-living animals has been studied extensively by the doubly-labeled water (DLW) technique. This method provides a reasonably accurate estimate of daily energy needs. However, there is considerable interest in the energy demands of animals over much shorter timescales, for which the DLW technique is less useful. We examined the possibility of measuring the expenditure of small animals over these shorter timescales from the washout kinetics of a bolus dose of 13C labeled bicarbonate. The study involved 19 laboratory mice which were injected either i.p. or s.c. with 0.2 ml of 13C labeled bicarbonate in water. Mice were placed in a standard respirometry system, maintained at different temperatures to precipitate a 3 fold variation in metabolism. Samples of breath were collected from the chamber into vacutainers at one minute intervals for approximately 40 minutes to an hour. Samples were analyzed by admission to a mass spectrometer (VG Optima) via a GC interface which identified and admitted the CO2 peak. The log converted isotope elimination was linear (r2 > 98% in all cases) indicating a single pool was involved. We evaluated the pool size from a dilution series of the injectate in equilibrium with CO2 gas. Conventional compartmental analysis produced an estimate which on average across the 19 individuals provided a reasonable estimate of the CO2 production. Individual estimates were however imprecise and the overall correlation between isotope and calorimeter estimates had an r2 of only 15%. Reasons for this discrepancy are unclear. Nevertheless an empirical model, using the elimination gradient, pool size and route of isotope administration as predictors explained 86% of the variation in CO2 production. Elimination of a bolus dose of 13C labeled bicarbonate provides a useful tool for estimating the energy metabolism of mice over intervals between 15 and 40 minutes.

No cost of echolocation for bats in flight.

Echolocation has evolved in relatively few animal species. One constraint may be the high cost of producing pulses, the echoes of which can be detected over useful distances. The energy cost of echolocation in a small (6 g) insectivorous bat, when hanging at rest, was recently measured at 0.067 Joules per pulse, implying a mean cost for echolocation in flight of 9.5 x basal metabolic rate (range 7 to 12x). Because flight is very costly, whether the costs of echolocation and flying are additive is an important question. We measured the energy costs of flight in two species of small echolocating Microchiroptera using a novel combination of respirometry and doubly-labelled water. Flight energy expenditure (adjusted for body mass) was not significantly different between echolocating bats and non-echolocating bats and birds. The low cost of echolocation for flying vertebrates may have been a significant factor favouring its evolution in these groups.

Principles, problems and a paradox with the measurement of energy expenditure of free-living subjects using doubly-labelled water.

Many aspects of the study of human nutrition would benefit from the ability to measure the energy which is expended by subjects as they go about their normal activities. The doubly-labelled water technique is a method which allows such measures to be made. The technique depends on the principle that a measure of carbon dioxide production is possible from the difference in the rates at which isotopic labels of hydrogen and oxygen are eliminated from the body. This simple explanation however conceals a host of assumptions. Several issues have emerged as important in the application of the technique to humans. These issues include first, the use of two samples (an initial and final sample) or multiple samples to estimate the rates of isotope elimination, and secondly the appropriate use of dilution spaces. This paper reviews the current status of the debates concerning these two issues. Paradoxically, improvements in our understanding of the technique, in the methods for calculating carbon dioxide production and in the accuracy of mass spectrometry, have not led to improvements in the accuracy of the technique. The mean deviation of estimates of carbon dioxide production using the technique from estimates made by conventional methods averages 7 per cent.