Wintering Snow Buntings Elevate Cold Hardiness to Extreme Levels but Show No Changes in Maintenance Costs.

AbstractResident temperate passerines adjust their phenotypes to cope with winter constraints, with peak performance in metabolic traits typically occurring during the coldest months. However, it is sparsely known whether cold-adapted northern species make similar adjustments when faced with variable seasonal environments. Life in near-constant cold could be associated with limited flexibility in traits underlying cold endurance. We investigated this by tracking individual physiological changes over five consecutive winters in snow buntings (Plectrophenax nivalis), an Arctic-breeding migratory passerine typically confronted with nearly constant cold. Buntings were held in an outdoor aviary and exposed to seasonal temperature variation typical of temperate zone climates. We measured phenotypic changes in body composition (body, fat, and lean mass, pectoralis muscle thickness), oxygen transport capacity (hematocrit), metabolic performance (basal metabolic rate [BMR] and summit metabolic rate [Msum]), thermogenic endurance (time to reach Msum), and cold tolerance (temperature at Msum). Snow buntings showed flexibility in functions underlying thermogenic capacity and cold endurance comparable to that observed in temperate resident passerines wintering at similar latitudes. Specifically, they increased body mass (13%), fat mass (246%), hematocrit (23%), pectoralis muscle thickness (8%), and Msum (27%). We also found remarkable cold tolerance in these birds, with individuals reaching Msum in helox at temperatures equivalent to less than -90°C in air. However, in contrast with resident temperate passerines, lean mass decreased by 12%, and there was no clear increase in maintenance costs (BMR). Our results show that the flexibility of traits underlying thermal acclimatization in a cold-adapted northern species is comparable to that of temperate resident species living at lower latitudes and is therefore not limited by life in near-constant cold.

Body temperature responses to handling stress in wintering Black-capped Chickadees (Poecile atricapillus L.).

Body temperature variation in response to acute stress is typically characterized by peripheral vasoconstriction and a concomitant increase in core body temperature (stress-induced hyperthermia). It is poorly understood how this response differs between species and within individuals of the same species, and how it is affected by the environment. We therefore investigated stress-induced body temperature changes in a non-model species, the Black-capped Chickadee, in two environmental conditions: outdoors in low ambient temperature (mean: -6.6°C), and indoors, in milder ambient temperature close to thermoneutrality (mean: 18.7°C). Our results show that the change in body temperature in response to the same handling stressor differs in these conditions. In cold environments, we noted a significant decrease in core body temperature (-2.9°C), whereas the response in mild indoor conditions was weak and non-significant (-0.6°C). Heat loss in outdoor birds was exacerbated when birds were handled for longer time. This may highlight the role of behavioral thermoregulation and heat substitution from activity to body temperature maintenance in harsh condition. Importantly, our work also indicates that changes in the physical properties of the bird during handling (conductive cooling from cold hands, decreased insulation from compression of plumage and prevention of ptiloerection) may have large consequences for thermoregulation. This might explain why females, the smaller sex, lost more heat than males in the experiment. Because physiological and physical changes during handling may carry over to affect predation risk and maintenance of energy balance during short winter days, we advice caution when designing experimental protocols entailing prolonged handling of small birds in cold conditions.

Chickadees Faced with Unpredictable Food Increase Fat Reserves but Certain Components of Their Immune Function Decline.

In winter, temperate resident birds are often faced with periodic low natural food availability. This reduction or unpredictability in resource availability might then have a negative impact on immune function, given that immune system support is highly resource dependent. We investigated the balance between energetic and immune management in captive black-capped chickadees (Poecile atricapilus) by manipulating the predictability of resources. The control group received food ad lib. every day, while the experimental group received a reduced amount of food on random days and food ad lib. on all other days. We measured two key metrics of energetic management (body and fat mass) as well as a suite of immune system components. Compared with control birds, experimental birds maintained significantly higher total body and fat mass, had lower acute phase protein concentrations, and had decreased body temperature and lost more body mass during the fever response following injection with lipopolysaccharides. Interestingly, birds in both groups had similar levels of complement lysis, delayed-type hypersensitivity response (phytohemagglutinin), and primary antibody production (keyhole limpet hemocyanin). This experiment demonstrates that black-capped chickadees strategically increase their fat mass in response to decreased food availability and that this might allow the birds to maintain most of the immune system unaltered, except some of the most costly immune components.

Increasing Winter Maximal Metabolic Rate Improves Intrawinter Survival in Small Birds.

Small resident bird species living at northern latitudes increase their metabolism in winter, and this is widely assumed to improve their chances of survival. However, the relationship between winter metabolic performance and survival has yet to be demonstrated. Using capture-mark-recapture, we followed a population of free-living black-capped chickadees (Poecile atricapillus) over 3 yr and evaluated their survival probability within and among winters. We also measured the size-independent body mass (Ms), hematocrit (Hct), basal metabolic rate (BMR), and maximal thermogenic capacity (Msum) and investigated how these parameters influenced survival within and among winters. Results showed that survival probability was high and constant both within (0.92) and among (0.96) winters. They also showed that while Ms, Hct, and BMR had no significant influence, survival was positively related to Msum-following a sigmoid relationship-within but not among winter. Birds expressing an Msum below 1.26 W (i.e., similar to summer levels) had a <50% chance of survival, while birds with an Msum above 1.35 W had at least a 90% chance of surviving through the winter. Our data therefore suggest that black-capped chickadees that are either too slow or unable to adjust their phenotype from summer to winter have little chances of survival and thus that seasonal upregulation of metabolic performance is highly beneficial. This study is the first to document in an avian system the relationship between thermogenic capacity and winter survival, a proxy of fitness.

Estimation of Muscle Mass by Ultrasonography Differs between Observers and Life States of Models in Small Birds.

Ultrasonography has proven to be a valuable noninvasive method of measure of muscle size in birds, but validation of its use in birds as small as black-capped chickadees (Poecile atricapillus; 11 g) is scarce. The effect of observers and life state (dead or alive) of models used for calibration on measurement quality is also poorly documented. Using 31 dead and 22 live chickadees, linear regressions between ultrasound and dissection measurements of pectoral and thigh muscles were fitted and compared between five different observers. R(2) values varied greatly between observers and were generally weaker in live birds, ranging between 0.02 and 0.59, despite high repeatability of measurement. Using equations calculated from dead birds to estimate muscle mass of live birds yielded much higher measurement errors (9%-18%) than when using equations calculated from live birds (5%-8%). Our results suggest that with careful training and using only calibration from live birds, ultrasonography can be a useful but limited tool to estimate muscle size of birds as small as the black-capped chickadee.

Individual inconsistencies in basal and summit metabolic rate highlight flexibility of metabolic performance in a wintering passerine.

Resident passerines inhabiting high latitude environments are faced with strong seasonal changes in thermal conditions and energy availability. Summit metabolic rate (maximal metabolic rate elicited by shivering during cold exposure: M(sum)) and basal metabolic rate (BMR) vary in parallel among seasons and increase in winter due to cold acclimatization, and these adjustments are thought to be critical for survival. Wintering individuals expressing consistently higher M(sum) and BMR could therefore be seen as better performers with higher chances of winter survival than those exhibiting lower metabolic performance. In this study, we calculated repeatability to evaluate temporal consistency of body mass, BMR and M(sum) within and across three consecutives winters in black-capped chickadees (Poecile atricapillus). We found that body mass was significantly repeatable both within and across winters (R 0.51-0.90). BMR (R 0.29-0.47) was only repeatable within winter while M(sum) was repeatable both among (R 0.33-0.49) and within winters (R 0.33-0.49) with the magnitude and significance of repeatability in both variables depending on the year and whether they were corrected for body mass or body size. The patterns of repeatability observed among years also differed between the two variables. Our findings suggest that the relative ranking of individuals in winter metabolic performance is affected by local ecological conditions and can change within relatively short periods of time.

Reaction norms in natural conditions: how does metabolic performance respond to weather variations in a small endotherm facing cold environments?

Reaction norms reflect an organisms' capacity to adjust its phenotype to the environment and allows for identifying trait values associated with physiological limits. However, reaction norms of physiological parameters are mostly unknown for endotherms living in natural conditions. Black-capped chickadees (Poecile atricapillus) increase their metabolic performance during winter acclimatization and are thus good model to measure reaction norms in the wild. We repeatedly measured basal (BMR) and summit (Msum) metabolism in chickadees to characterize, for the first time in a free-living endotherm, reaction norms of these parameters across the natural range of weather variation. BMR varied between individuals and was weakly and negatively related to minimal temperature. Msum varied with minimal temperature following a Z-shape curve, increasing linearly between 24°C and -10°C, and changed with absolute humidity following a U-shape relationship. These results suggest that thermal exchanges with the environment have minimal effects on maintenance costs, which may be individual-dependent, while thermogenic capacity is responding to body heat loss. Our results suggest also that BMR and Msum respond to different and likely independent constraints.

Lipid metabolites as markers of fattening rate in a non-migratory passerine: effects of ambient temperature and individual variation.

Plasma lipid metabolites triglycerides (TRIG) and glycerol (GLY) are used as indicators of fattening rate and nutritional condition in migratory birds. Requiring only one blood sample, they could also be used for studying daily and seasonal fattening rates in relation with habitat quality or weather variations in species wintering in cold climates. Using black-capped chickadees exposed to three experimental temperatures (0 °C, 15 °C, and 30 °C), the goal of this experiment was to determine the relationship between plasma levels of TRIG and GLY and fattening rate measured over periods from a few hours to the previous two days. Results showed that birds maintained in the cold had metabolite levels 39-81% higher than those at thermoneutrality, likely reflecting the size of their fat reserves, and that TRIG and total GLY were highly correlated across treatments. Fattening rate was also higher at 0 °C (+35%) and 30 °C (+24%) relative to that measured at 15 °C and, as expected, was positively correlated with metabolite levels across treatments. However, despite fattening rates similar to that observed at the other temperatures, the relationships were uncoupled at 30 °C, implying that the technique may not be easily applicable at temperatures within or close to thermoneutrality. We also found a strong individual effect in the relationships between fattening rate and TRIG levels, suggesting high individual consistency in these parameters in conditions of unrestricted food access such as in captivity. Our study confirms that plasma TRIG and GLY levels can be used as relative indexes of condition and fattening rates in wintering passerines.

How does flexibility in body composition relate to seasonal changes in metabolic performance in a small passerine wintering at northern latitude?

Abstract Small avian species wintering at northern latitudes typically show increases in basal metabolic rate (BMR) and maximal thermogenic capacity (Msum). Those are widely assumed to reflect changes in body composition, with enlargement of digestive and excretory organs resulting in elevated winter BMR and larger body muscles driving the increase in Msum. Using free-living black-capped chickadees (Poecile atricapillus) as our model species, we investigated seasonal changes in body composition and tested for relationships between mass variations of body organs and variability of both BMR and Msum. Our results confirmed the expected winter increase in mass of body muscles and cardiopulmonary organs (heart + lungs) and showed that 64% of the observed Msum variations throughout the year were explained by changes in these organs. In contrast, we found little support for an effect of the digestive organs (gizzard + intestines) on BMR seasonal changes. Instead, this variable was mainly influenced by variations in mass of body muscles and excretory organs (liver + kidney), explaining up to 35% of its variability.

Phenotype manipulations confirm the role of pectoral muscles and haematocrit in avian maximal thermogenic capacity.

In small resident bird species living at northern latitudes, winter cold acclimatization is associated with an increase in pectoral muscle size and haematocrit level, and this is thought to drive the seasonal increase in summit metabolic rate (Msum, a measure of maximal shivering thermogenic capacity). However, evidence suggesting that pectoral muscle size influences Msum is correlational and the link between haematrocrit level and Msum remains to be demonstrated. We experimentally tested the relationship between pectoral muscle size and Msum by manipulating muscle size using a feather clipping protocol in free-living wintering black-capped chickadees (Poecile atricapillus). This also allowed us to investigate the link between haematocrit and thermogenic capacity. After a first series of measures on all birds, we cut half of the flight feathers of experimental individuals (N=14) and compared their fat and pectoral muscle scores, Msum and haematocrit level at recapture with their previous measures and with those of control birds (N=17) that were captured and recaptured at comparable times. Results showed that: (1) experimental birds developed larger pectoral muscles than control individuals and (2) mass-independent Msum was up to 16% higher in birds expressing large pectoral muscles. Msum was also positively correlated with haematocrit, which was not affected by the experimental manipulation. These findings demonstrate that, for a given body mass, large pectoral muscles are associated with a higher Msum in black-capped chickadees and that oxygen carrying capacity likely supports thermogenesis in this species.

Intra-seasonal flexibility in avian metabolic performance highlights the uncoupling of basal metabolic rate and thermogenic capacity.

Stochastic winter weather events are predicted to increase in occurrence and amplitude at northern latitudes and organisms are expected to cope through phenotypic flexibility. Small avian species wintering in these environments show acclimatization where basal metabolic rate (BMR) and maximal thermogenic capacity (MSUM) are typically elevated. However, little is known on intra-seasonal variation in metabolic performance and on how population trends truly reflect individual flexibility. Here we report intra-seasonal variation in metabolic parameters measured at the population and individual levels in black-capped chickadees (Poecileatricapillus). Results confirmed that population patterns indeed reflect flexibility at the individual level. They showed the expected increase in BMR (6%) and MSUM (34%) in winter relative to summer but also, and most importantly, that these parameters changed differently through time. BMR began its seasonal increase in November, while MSUM had already achieved more than 20% of its inter-seasonal increase by October, and declined to its starting level by March, while MSUM remained high. Although both parameters co-vary on a yearly scale, this mismatch in the timing of variation in winter BMR and MSUM likely reflects different constraints acting on different physiological components and therefore suggests a lack of functional link between these parameters.

Dominant black-capped chickadees pay no maintenance energy costs for their wintering status and are not better at enduring cold than subordinate individuals.

Winter requires physiological adjustments in northern resident passerines. Cold acclimatization is generally associated with an increase in physiological maintenance costs, measured as basal metabolic rate (BMR), and cold endurance, reflected by summit metabolic rate (M(sum)). However, several northern species also form social groups in winter and a bird's hierarchical position may influence the size of its metabolically active organs as well as its BMR. Winter metabolic performance in these species may therefore reflect a complex set of adjustments to both seasonal climatic variations and social environment. We studied the effect of social status on parameters of cold acclimatization (body mass, size of fat reserves and pectoral muscles, BMR and M(sum)) in free-living black-capped chickadees (Poecile atricapillus). Birds that were structurally large and heavy for their body size, mostly dominant individuals, carried more fat reserves and had larger pectoral muscles. However, social status had little effect on metabolic performance in the cold. Indeed, M(sum) was independent of social rank while mass-corrected BMR was slightly lower in dominant individuals, likely due to a statistical dilution effect caused by large metabolically inactive fat reserves. BMR and M(sum), whether considered in terms of whole-animal values, corrected for body mass or body size were nevertheless correlated, suggesting a functional link between these metabolic components. Our results therefore indicate that the energy cost of social dominance is not a generalized phenomenon in small wintering birds.

Ambient temperature does not affect fuelling rate in absence of digestive constraints in long-distance migrant shorebird fuelling up in captivity.

Pre-flight fuelling rates in free-living red knots Calidris canutus, a specialized long-distance migrating shorebird species, are positively correlated with latitude and negatively with temperature. The single published hypothesis to explain these relationships is the heat load hypothesis that states that in warm climates red knots may overheat during fuelling. To limit endogenous heat production (measurable as basal metabolic rate BMR), birds would minimize the growth of digestive organs at a time they need. This hypothesis makes the implicit assumption that BMR is mainly driven by digestive organ size variation during pre-flight fuelling. To test the validity of this assumption, we fed captive knots with trout pellet food, a diet previously shown to quickly lead to atrophied digestive organs, during a fuelling episode. Birds were exposed to two thermal treatments (6 and 24 degrees C) previously shown to generate different fuelling rates in knots. We made two predictions. First, easily digested trout pellet food rather than hard-shelled prey removes the heat contribution of the gut and would therefore eliminate an ambient temperature effect on fuelling rate. Second, if digestive organs were the main contributors to variations in BMR but did not change in size during fuelling, we would expect no or little change in BMR in birds fed ad libitum with trout pellets. We show that cold-acclimated birds maintained higher body mass and food intake (8 and 51%) than warm-acclimated birds. Air temperature had no effect on fuelling rate, timing of fuelling, timing of peak body mass or BMR. During fuelling, average body mass increased by 32% while average BMR increased by 15% at peak of mass and 26% by the end of the experiment. Our results show that the small digestive organs characteristic of a trout pellet diet did not prevent BMR from increasing during premigratory fuelling. Our results are not consistent with the heat load hypothesis as currently formulated.

Limited access to food and physiological trade-offs in a long-distance migrant shorebird. I. Energy metabolism, behavior, and body-mass regulation.

Previous experiments showed reduction of basal metabolic rate (BMR) in birds facing energetic challenges. We alternately exposed two groups of red knots (Calidris canutus) to either 6 h or 22 h of food availability for periods of 22 d. Six h of access to food led to a 6%-10% loss of body mass over the first 8 d, with nearly all of the birds' daily energy expenditures supported by body nutrient stores during the first 2 d. Birds responded by increasing feeding behavior and food intake, but the response was slow. There were no gains of mass before day 15, which suggests a digestive bottleneck and a period of physiological adjustment. Food-restricted birds exhibited decreases in pectoral-muscle thickness and BMR in association with a loss of body mass. Although a decrease in BMR saves energy, savings represented only 2%-7% of the daily energy spent in excess of that acquired during the deficit period. Red knots did not downregulate mass-independent BMR. On the bases of recent independent findings and the pattern of mass gain observed when food access was switched from 6 h to 22 h, we suggest that these birds routinely maintain nutrient stores as a buffer against periods of energy shortages, thereby precluding the need for downregulation of mass-independent BMR.

Limited access to food and physiological trade-offs in a long-distance migrant shorebird. II. Constitutive immune function and the acute-phase response.

In response to unbalanced energy budgets, animals must allocate resources among competing physiological systems to maximize fitness. Constraints can be imposed on energy availability or energy expenditure, and adjustments can be made via changes in metabolism or trade-offs with competing demands such as body-mass maintenance and immune function. This study investigates changes in constitutive immune function and the acute-phase response in shorebirds (red knots) faced with limited access time to food. We separated birds into two experimental groups receiving either 6 h or 22 h of food access and measured constitutive immune function. After 3 wk, we induced an acute-phase response, and after 1 wk of recovery, we switched the groups to the opposite food treatment and measured constitutive immune function again. We found little effect of food treatment on constitutive immune function, which suggests that even under resource limitation, a baseline level of immune function is maintained. However, birds enduring limited access to food suppressed aspects of the acute-phase response (decreased feeding and mass loss) to maintain energy intake, and they downregulated thermoregulatory adjustments to food treatment to maintain body temperature during simulated infection. Thus, under resource-limited conditions, birds save energy on the most costly aspects of immune defense.