Parus Major Research Articles

First person – Elisa Thoral

ABSTRACT First Person is a series of interviews with the first authors of a selection of papers published in Biology Open, helping researchers promote themselves alongside their papers. Elisa Thoral is first author on ‘ The relationship between mitochondrial respiration, resting metabolic rate and blood cell count in great tits’, published in BiO. Elisa is a postdoctoral researcher in the lab of Dr Andreas Nord at Lund University, Lund, Sweden, investigating I am interested in the effects of environmental parameters, such as temperature and diet, on animals’ performance at both the organismal and cellular level.

Early developmental carry-over effects on exploratory behaviour and DNA methylation in wild great tits (Parus major).

Adverse, postnatal conditions experienced during development are known to induce lingering effects on morphology, behaviour, reproduction and survival. Despite the importance of early developmental stress for shaping the adult phenotype, it is largely unknown which molecular mechanisms allow for the induction and maintenance of such phenotypic effects once the early environmental conditions are released. Here we aimed to investigate whether lasting early developmental phenotypic changes are associated with post-developmental DNA methylation changes. We used a cross-foster and brood size experiment in great tit (Parus major) nestlings, which induced post-fledging effects on biometric measures and exploratory behaviour, a validated personality trait. We investigated whether these post-fledging effects are associated with DNA methylation levels of CpG sites in erythrocyte DNA. Individuals raised in enlarged broods caught up on their developmental delay after reaching independence and became more explorative as days since fledging passed, while the exploratory scores of individuals that were raised in reduced broods remained stable. Although we previously found that brood enlargement hardly affected the pre-fledging methylation levels, we found 420 CpG sites that were differentially methylated between fledged individuals that were raised in small versus large sized broods. A considerable number of the affected CpG sites were located in or near genes involved in metabolism, growth, behaviour and cognition. Since the biological functions of these genes line up with the observed post-fledging phenotypic effects of brood size, our results suggest that DNA methylation provides organisms the opportunity to modulate their condition once the environmental conditions allow it. In conclusion, this study shows that nutritional stress imposed by enlarged brood size during early development associates with variation in DNA methylation later in life. We propose that treatment-associated DNA methylation differences may arise in relation to pre- or post-fledging phenotypic changes, rather than that they are directly induced by the environment during early development.

Exploring the Impact of Urbanization on Avian Malaria Dynamics in Great Tits: Insights from a Study Across Urban and Non-Urban Environments

Exploring the Impact of Urbanization on Avian Malaria Dynamics in Great Tits: Insights from a Study Across Urban and Non-Urban Environments

The relationship between mitochondrial respiration, resting metabolic rate and blood cell count in great tits.

Although mitochondrial respiration is believed to explain a substantial part of the variation in resting metabolic rate (RMR), few studies have empirically studied the relationship between organismal and cellular metabolism. We therefore investigated the relationship between RMR and mitochondrial respiration of permeabilized blood cells in wild great tits (Parus major L.). We also studied the correlation between mitochondrial respiration traits and blood cell count, as normalizing mitochondrial respiration by the cell count is a method commonly used to study blood metabolism. In contrast to previous studies, our results show that there was no relationship between RMR and mitochondrial respiration in intact blood cells (i.e. with the ROUTINE respiration). However, when cells were permeabilized and interrelation re-assessed under saturating substrate availability, we found that RMR was positively related to phosphorylating respiration rates through complexes I and II (i.e. OXPHOS respiration) and to the mitochondrial efficiency to produce energy (i.e. net phosphorylation efficiency), though variation explained by the models was low (i.e. linear model: R2=0.14 to 0.21). However, unlike studies in mammals, LEAK respiration without [i.e. L(n)] and with [i.e. L(Omy)] adenylates was not significantly related to RMR. These results suggest that phosphorylating respiration in blood cells can potentially be used to predict RMR in wild birds, but that this relationship may have to be addressed in standardized conditions (permeabilized cells) and that the prediction risks being imprecise. We also showed that, in our conditions, there was no relationship between any mitochondrial respiration trait and blood cell count. Hence, we caution against normalising respiration rates using this parameter as is sometimes done. Future work should address the functional explanations for the observed relationships, and determine why these appear labile across space, time, taxon, and physiological state.

The ontogeny of social networks in wild great tits (Parus major).

Sociality impacts many biological processes and can be tightly linked to an individual's fitness. To maximize the advantages of group living, many social animals prefer to associate with individuals that provide the most benefits, such as kin, familiar individuals, or those of similar phenotypes. Such social strategies are not necessarily stable over time but can vary with changing selection pressures. In particular, young individuals transitioning to independence should continuously adjust their social behavior in light of developmental changes. However, social strategies exhibited during adolescence in animals are understudied, and the factors underlying social network formation during ontogeny remain elusive. Here, we tracked associations of wild great tits (Parus major) during the transition to independence and across their first year of life. Both spatial and social factors predicted dyadic associations. During the transition to independence in spring, fledglings initially preferred to associate with siblings and peers over non-parent adults. We found no evidence for preferred associations among juveniles of similar age or fledge weight during that time but weak evidence for some potential inheritance of the parental social network. By autumn, after juveniles had reached full independence, they exhibited social strategies similar to those of adults by establishing stable social ties based on familiarity that persisted through winter into the next spring. Overall, this research demonstrates dynamic changes in social networks during ontogeny in a species with a fast life history and limited parental care, which likely reflect changes in selective pressures. It further highlights the importance of long-term social bonds based on familiarity in this species.

Metabolic adjustments to winter severity in two geographically separated great tit (Parus major) populations.

Understanding the potential limits placed on organisms by their ecophysiology is crucial for predicting their responses to varying environmental conditions. A main hypothesis for explaining avian thermoregulatory mechanisms is the aerobic capacity model, which posits a positive correlation between basal (basal metabolic rate [BMR]) and summit (Msum) metabolism. Most evidence for this hypothesis, however, comes from interspecific comparisons, and the ecophysiological underpinnings of avian thermoregulatory capacities hence remain controversial. Indeed, studies have traditionally relied on between-species comparisons, although, recently, there has been a growing recognition of the importance of intraspecific variation in ecophysiological responses. Therefore, here, we focused on great tits (Parus major), measuring BMR and Msum during winter in two populations from two different climates: maritime-temperate (Gontrode, Belgium) and continental (Zvenigorod, Russia). We tested for the presence of intraspecific geographical variation in metabolic rates and assessed the predictions following the aerobic capacity model. We found that birds from the maritime-temperate climate (Gontrode) showed higher BMR, whereas conversely, great tits from Zvenigorod showed higher levels of Msum. Within each population, our data did not fully support the aerobic capacity model's predictions. We argued that the decoupling of BMR and Msum observed may be caused by different selective forces acting on these metabolic rates, with birds from the continental-climate Zvenigorod population facing the need to conserve energy for surviving long winter nights (by keeping their BMR at low levels) while simultaneously being able to generate more heat (i.e., a high Msum) to withstand cold spells.

Short-time exposure to light at night affects incubation patterns and correlates with subsequent body weight in great tits (Parus major).

Artificial light at night (ALAN) widely affects wildlife by blurring light-dark differences, including transitions such as sunrise and sunset, thereby affecting regulation of diel rhythms. As a result, activity onsets in many wild diurnal songbirds advance under ALAN. From chronobiological studies, it is known that the direction and strength of the response to light depends on when during the night exposure takes place. However, these experiments are mostly done under continuous light conditions, when animals have free-running rhythms. It remains unclear whether phase-dependence also holds in entrained, wild songbirds; i.e., does the effect of ALAN on activity patterns differ between exposure in the morning compared to the evening? This information is essential to assess the effects of mitigation measures by limiting ALAN to selected times of the night. We exposed incubating great tits (Parus major) inside the nest-box to 4 h of dim light, of which 1 h overlapped with dawn before sunrise or dusk after sunset. We found a small advancing effect of morning-light on activity onset and of evening-light on offset compared to dark controls but not vice versa. Breeding success and chick condition were unaffected by the light treatments. However, light-treated females had lower weights 9-18 days after the end of the treatment compared to the controls, independent of whether ALAN occurred in the morning or the evening, indicating possible costs of ALAN. Despite the weak behavioral response, ALAN might have affected the females' circadian clock or physiology resulting in lower body condition.

Divergence in evolutionary potential of life history traits among wild populations is predicted by differences in climatic conditions.

Short-term adaptive evolution represents one of the primary mechanisms allowing species to persist in the face of global change. Predicting the adaptive response at the species level requires reliable estimates of the evolutionary potential of traits involved in adaptive responses, as well as understanding how evolutionary potential varies across a species' range. Theory suggests that spatial variation in the fitness landscape due to environmental variation will directly impact the evolutionary potential of traits. However, empirical evidence on the link between environmental variation and evolutionary potential across a species range in the wild is lacking. In this study, we estimate multivariate evolutionary potential (via the genetic variance-covariance matrix, or G-matrix) for six morphological and life history traits in 10 wild populations of great tits (Parus major) distributed across Europe. The G-matrix significantly varies in size, shape, and orientation across populations for both types of traits. For life history traits, the differences in G-matrix are larger when populations are more distant in their climatic niche. This suggests that local climates contribute to shaping the evolutionary potential of phenotypic traits that are strongly related to fitness. However, we found no difference in the overall evolutionary potential (i.e., G-matrix size) between populations closer to the core or the edge of the distribution area. This large-scale comparison of G-matrices across wild populations emphasizes that integrating variation in multivariate evolutionary potential is important to understand and predict species' adaptive responses to new selective pressures.

Nestling Diet of Two Sympatric Insectivorous Passerines in Different Habitats—A Metabarcoding Study

Increasing landscape transformations and urbanisation affect insectivorous bird populations in various ways such as food availability, breeding phenology, or reproductive success. Especially during the breeding season, many passerine birds rely on the availability of caterpillars as the main prey for their nestlings. Previous studies suggested that similar diet preferences of sympatric species may result in interspecific competition, as demonstrated for Blue and Great Tits in forest habitats. However, nestling diet and prey preferences in other habitats are not fully understood. Prey availability, especially caterpillars, is lower in cities than in forests, thus influencing prey choice and interspecific competition. Here we used faecal DNA metabarcoding to investigate if nestling diet composition of the two sympatric species Blue Cyanistes caeruleus and Great Tits Parus major varied among species and different habitats (forest, traditional orchards, and urban parks). Furthermore, we examined food availability by DNA barcoding of the arboreal arthropod communities among habitats and compared them to the nestling diet to infer parental prey selectivity. The study was carried out in central Germany from 2018 to 2019. Blue and Great Tits showed a diverse diet which was dominated by Lepidoptera in all habitats. Lepidopteran diet components were most similar between forest and orchard sites, as were the components with other arthropods between orchard and urban sites. Both tit species showed selectivity for the lepidopteran families Geometridae and Tortricidae in all habitats, and for Noctuidae (Lepidoptera), Tenthredinidae and Braconidae (Hymenoptera) in forest and orchard sites. As the tits showed preferences for mainly families of Lepidoptera, and Hymenoptera, our approach provides a baseline to support monitoring of these groups.

Plumage coloration differs between offspring raised in natural cavities and nestboxes

Abstract Most of our knowledge of secondary hole-nesting birds comes from populations breeding in human-provided nestboxes, yet these might differ from natural cavities in a number of parameters, including internal dimensions or microclimate, leading to differences in reproductive ecology. Here, we demonstrate differences in plumage coloration, an important visual signal of individual quality, in Blue Tit (Cyanistes caeruleus) and Great Tit (Parus major) nestlings raised in natural cavities and nestboxes. For this study, we collected feather samples over two breeding seasons and applied reflectance spectrophotometry to calculate color metrics. Carotenoid chroma was higher in nestbox-reared Blue Tits, whereas brightness was higher in nestbox-reared Great Tits (with a similar tendency for Blue Tits). The latter result might be explained by the better nutritional condition of Great Tit nestlings raised in nestboxes. Furthermore, we found no evidence for preference of adults expressing more elaborate plumage towards a specific cavity type in either species. Consequently, we assume that differences in nestling plumage reported here are driven by rearing conditions (nestboxes vs. natural cavities) and not by differences in plumage-based parental quality. Our study adds to the growing body of evidence confirming that anthropogenic environmental modifiers, such as nestboxes, might influence avian physiology and the resulting phenotype.

Variation in the use of anthropogenic materials in tit nests: influence of human activities and pandemic restrictions

As urbanization expands, so does the presence of anthropogenic materials within bird’s nests. However, our understanding of this phenomenon remains limited, particularly in terrestrial birds. This study describes and compares the use of anthropogenic materials in great tit (Parus major) and blue tit (Cyanistes caeruleus) nests in relation to the intensity of anthropogenic activities, distance to contamination sources and reproductive metrics, in nest boxes in a suburban wood in Coimbra, Portugal, during the breeding seasons of 2020 and 2021. Nests were collected after fledging and sorted to remove anthropogenic materials. From a total of 9.71% anthropogenic materials in great tit nests we observed an abundance of uncoloured tangles of mixed fibres, representing 78% of the total weight of the anthropogenic materials in great tit nests in 2021. The chemical composition of these mixtures was determined by spectroscopy analysis, using Fourier transform infrared spectroscopy (FTIR), and 56% of the nests (16/25) contained fibres from synthetic origin. The percentage of anthropogenic materials was higher in great tit than in blue tit nests and increased in 2021, comparatively to 2020. The proximity to potential contamination sources influenced positively the percentage of anthropogenic materials in nests, as those closer to areas with more intense anthropogenic activities (the bar/tennis court and the wastewater treatment plant) had a higher percentage of these materials. No effects on productivity were detected. Our results suggest that the use of these materials is probably related to the intensity of anthropogenic activities.

Responses of avian predators to the polymorphic harlequin ladybird (Harmonia axyridis)

Abstract The harlequin ladybird (Harmonia axyridis) shows a high level of colour polymorphism. Particular forms differ in their colour combination, pattern and abundance. Two species of wild‐caught passerines native to Central Europe were offered various forms of ladybirds, differing in their colour pattern and abundance in nature. We predicted that those forms that are more abundant in the wild are better protected, as they are more familiar to predators. Forms novemdecimsignata and spectabilis, which represent 97% of individuals in the wild population, significantly differ in their visual appearance (mostly orange vs. mostly black). The form axyridis is very rare in the wild, and suturalis and aninkae do not occur in the wild and were derived from laboratory breedings As predators, we used great tits (Parus major), which are very aversive towards ladybirds, and tree sparrows (Passer montanus), which are quite willing to attack and even eat ladybirds. We compared bird responses to particular ladybird colour forms and included a brown‐painted control to test the effect of conspicuous colours. We showed that both species of birds attacked all conspicuous forms of the ladybird equally and usually very rarely. The brown‐painted novemdecimsignata form was attacked more frequently compared with the conspicuous forms, showing that the visual appearance prevents birds from attacking any conspicuous colour combination. Sparrows tended to eat the attacked ladybirds of forms novemdecimsignata, spectabilis and brown painted. The rare axyridis form and laboratory forms were very well protected from the attack, very likely due to neophobia. We may conclude that despite the outstanding polymorphism of H. axyridis, its protection against avian predators is very effective as long as the red‐and‐black pattern is preserved.

Hard life for sons in the nest? Sex-dependent offspring mortality in Great Tits in urban and forest areas

Sex-biased mortality can occur in birds during development, for example due to sexual differences in energy requirement and/or environmental sensitivity, or the effects of sex hormones or sex differences in the expression of mutations linked to sex chromosomes. The extent of sex-bias in mortality may also be related to environmental conditions that influence offspring development and survival. Urban areas often provide poorer conditions for nestling development resulting in higher offspring mortality compared to natural areas, which may accelerate sex differences in offspring mortality in cities. To test this hypothesis, we examined the sex ratio of dead offspring in Great Tits (Parus major), using 427 samples of unhatched eggs and dead nestlings collected in two urban and two forest sites between 2013 and 2019. The ratio of males in the whole sample of dead offspring (56.9%) was significantly higher than expected by an 1:1 ratio, and the strongest sex biases were detected in urban areas (57.6% males) and in young nestlings (<14 days old, 59.0% males). However, the sex ratios of dead offspring did not differ significantly among study sites and between offspring developmental stages. 29.3% of unhatched eggs contained a visible embryo, and the proportion of embryo-containing unhatched eggs did not differ significantly between urban and forest study sites. These results suggest male-biased offspring mortality in Great Tits, and highlight the need of large datasets to detect subtle differences between habitats and developmental stages.

Effects of Ecological Factors on Occupancy of Artificial Nest Boxes by the Great Tit (Parus major): A Comprehensive Analysis

Effects of Ecological Factors on Occupancy of Artificial Nest Boxes by the Great Tit (Parus major): A Comprehensive Analysis

Breeding success of passerine birds in the forest steppe of European Russia

In the period from 1985 to 2023, the breeding success of 15 bird species was studied in the Voronezh Nature Reserve (50°21ʹ-52°02ʹ N, 39°21ʹ-39°47ʹ E), where there has been no economic activity for 100 years. The plant cover consists of pine (32.3% of the total area), oak (29.3%), aspen (19.3%), birch (5.7%) and alder (5.2%) forests. 2004 nests were monitored. Low breeding success (25.4-27.9%, determined by the modified Mayfield method) was found in the Blackbird Turdus merula , the Song Thrush T. philomelos , the Finch Fringilla coelebs and the Greenfinch Chloris chloris. From 1.6 to 1.8 fledglings fly out for one breeding attempt. The average level of breeding success (38.7-50.4%) was found in the Common Chiffchaff Phylloscopus collybita , the Red-backed Shrike Lanius collurio , Redwing Turdus iliacus , Eurasian Blackcap Sylvia atricapilla , Garden Warbler S. borin and Hawfinch Coccothraustes coccothraustes . From 2.1 to3.4 fledglings fly out for one breeding attempt. A high level of breeding success (60.6-82.5%) was found in the Great Tit Parus major , the Common Redstart Phoenicurus phoenicurus , the Pied Flycatcher Ficedula hypoleuca , the Collared Flycatcher F. albicollis and the Spotted Flycatcher Muscicapa striata . From 3.7 to 7 fledglings fly out for one breeding attempt. Due to the increased pressure of nest predators, the breeding success of birds in the Voronezh Nature Reserve is often lower than in other areas. The greatest breeding success is typical for birds that place nests in hollows and other shelters. Breeding success is significantly lower in openly nesting species, especially in birds that begin nesting before the leaves bloom on trees and shrubs.

Аналіз активності угрупувань птахів у міських парках: приклад з містом Дніпро (Україна)

The article presents the results of the analysis of bird communities’ activity in the parks of the right-bank part of Dnipro City during the breeding season of 2022–2023. It was established that 51 bird species consistently exhibit activity in the city parks. Among them, more than half (68.6 %) belong to the order Passeriformes. The analysis of the obtained data indicates that common species such as the house sparrow, chiffchaff, great tit, and white wagtail demonstrate the highest activity in all investigated parks and squares. The widespread presence of these species suggests their flexible use of spatial resources in urban parks, indicating their adaptability to anthropogenic environments. Applying the Rodgers–Shannon overlap index allowed determining the similarity degree of bird community parameters. The results indicate significant similarity among communities in several parks in the right-bank part of Dnipro, associated with their small area, proximity to residential buildings, and common tree crown pruning stages. The correlation between bird diversity and the size and characteristics of park territories was investigated. The correlation matrix revealed a clear positive relationship between park area and the number of bird species exhibiting activity. This is attributed to the greater environmental heterogeneity and the presence of resources for survival, which, in turn, enables representatives of different ecological complexes to realise their ecological and spatial niches. This is explained by the greater heterogeneity and the availability of resources for survival, facilitating the realisation of various ecological and spatial niches for different species. Some parks in the right-bank part of the city exemplify a clear connection between park area and bird species diversity. It was found that with an increase in park area by an average of 2.9556 hectares, the number of bird species increases by one. Additionally, a correlation was observed between the number of tree species in the territory and the bird communities’ biodiversity of the park or square, emphasising the importance of the plant cover for preserving and attracting bird diversity.

Nearly (?) sterile avian egg in a passerine bird.

During early ontogeny, microbiome affects development of the gastrointestinal tract, immunity, and survival in vertebrates. Bird eggs are thought to be (1) initially sterile (sterile egg hypothesis) and (2) colonized after oviposition through horizontal trans-shell migration, or (3) initially seeded with bacteria by vertical transfer from mother oviduct. To date, however, little empirical data illuminate the contribution of these mechanisms to gut microbiota formation in avian embryos. We investigated microbiome of the egg content (day 0; E0-egg), embryonic gut at day 13 (E13) and female faeces in a free-living passerine, the great tit (Parus major), using a methodologically advanced procedure combining 16S rRNA gene sequencing and microbe-specific qPCR assays. Our metabarcoding revealed that the avian egg is (nearly) sterile, but acquires a slightly richer microbiome during the embryonic development. Of the three potentially pathogenic bacteria targeted by qPCR, only Dietzia was found in E0-egg (yet also in negative controls), E13 gut and female samples, which might indicate possible vertical transfer. Unlike in poultry, we have shown that major bacterial colonization of the gut in passerines does not occur before hatching. We emphasize that protocols that carefully check for environmental contamination are critical in studies with low-bacterial biomass samples.

Measuring body temperature in birds - the effects of sensor type and placement on estimated temperature and metabolic rate.

Several methods are routinely used to measure avian body temperature, but different methods vary in invasiveness. This may cause stress-induced increases in temperature and/or metabolic rate and, hence, overestimation of both parameters. Choosing an adequate temperature measurement method is therefore key to accurately characterizing an animal's thermal and metabolic phenotype. Using great tits (Parus major) and four common methods with different levels of invasiveness (intraperitoneal, cloacal, subcutaneous, cutaneous), we evaluated the preciseness of body temperature measurements and effects on resting metabolic rate (RMR) over a 40°C range of ambient temperatures. None of the methods caused overestimation or underestimation of RMR compared with un-instrumented birds, and body or skin temperature estimates did not differ between methods in thermoneutrality. However, skin temperature was lower compared with all other methods below thermoneutrality. These results provide empirical guidance for future research that aims to measure body temperature and metabolic rate in small bird models.

Ecological value of wood energy plantations in the support of some animal groups

Today, some environmental problems have reached such severe proportions that it is no longer enough to recognise them, but environmentally friendly solutions must be used to reduce them. The reduction in the area of natural forests of native species is causing problems in several ways. This research aimed to highlight how environmental, conservation and economic interests can be reconciled. In addition to natural forests, wood energy plantations are becoming increasingly important. Energy import dependency is a problem for most countries, for which wood energy plantations can partly offer an alternative. Native forests can be protected, and their area increased where possible. Meanwhile, energy plantations can be established in areas with low agricultural productivity. In this experiment, I studied a plantation of Pedunculate Oak (Quercus robur), a Black Locust (Robinia pseudoacacia) and a Paulownia (Paulownia Shan Tong). I selected burrowing birds and ground-dwelling arthropods as indicator groups. I did this by establishing a nesting colony and soil trapping. I wanted to demonstrate that, in addition to natural forests, wood energy plantations have a role not only in economic terms but also in maintaining certain animal groups. Soil trapping tests were carried out in all three tree plantations. The obtained results showed that in the Paulownia plantation, the occupancy rate of nest boxes was almost 100%, while in the Black Locust plantation it was around 30%. Among the species that occupied the nesting sites, the Common Starling (Sturnus vulgaris) and the Eurasian Tree Sparrow (Passer montanus) were more abundant, while Great Tit (Parus major) was present in the Black Locust plantation. These are opportunistic species for which nesting opportunity is the most important factor, since their feeding area (in the case of the Common Starling and the Eurasian Tree Sparrow) is not typical of the nesting area. The soil trap investigations show that there are no significant differences in the composition of the arthropod group (beetles, spiders) in the study areas.

Great tits (Parus major) in a west European temperate forest show little seasonal variation in metabolic energy requirements

Understanding how birds annually allocate energy to cope with changing environmental conditions and physiological states is a crucial question in avian ecology. There are several hypotheses to explain species' energy allocation. One prominent hypothesis suggests higher energy expenditure in winter due to increased thermoregulatory costs. The "reallocation" hypothesis suggests no net difference in seasonal energy requirements, while the "increased demand" hypothesis predicts higher energy requirements during the breeding season. Birds are expected to adjust their mass and/or metabolic intensity in ways that are consistent with their energy requirements. Here, we look for metabolic signatures of seasonal variation in energy requirements of a resident passerine of a temperate-zone (great tit, Parus major). To do so, we measured whole-body and mass-independent basal (BMR), summit (Msum), and field (FMR) metabolic rates during late winter and during breeding in Belgian great tits. During the breeding season, birds had on average 10% higher whole-body BMR and FMR compared to winter, while their Msum decreased by 7% from winter to breeding. Mass-independent metabolic rates showed a 10% increase in BMR and a 7% decrease in Msum from winter to breeding. Whole-body BMR was correlated with Msum, but this relationship did not hold for mass-independent metabolic rates. The modest seasonal change we observed suggests that great tits in our temperature study area maintain a largely stable energy budget throughout the year, which appears mostly consistent with the reallocation hypothesis.