Detailed knowledge of habitat utilization by animals is integral in understanding their ability to adjust resource exploitation to the conditions of heterogeneous and constantly changing habitats. To identify the most important micro- and macrohabitat landscape characteristics that are important factors for birds when choosing foraging patches in their territory, we used a radio-telemetric approach with nestling-feeding great tit (Parus major) females in a heterogeneous habitat mosaic. We showed that birds preferred to forage in areas with low or high but not mid-range normalized difference vegetation index (NDVI) values, similarly in coniferous and deciduous forests. At the same time, foraging birds avoided areas with low or high but not mid-range canopy cover only in the dense deciduous forest, whereas in the more open coniferous forest, females used areas, which vary widely in the amount of canopy coverage. Females employing this foraging behaviour raised larger broods on average. Our results suggest that foraging microhabitat selection in forest passerines is a complicated process subject to diverse and interacting effects of habitat characteristics other than simply a habitat's position on a food-richness axis. Moreover, the study demonstrates how using a micro-scale approach can reveal subtle, but significant, habitat selection mechanisms that remain hidden at a macro-scale level.

Although avian basal metabolic rate (BMR) has been widely used in interspecific comparative studies, the sources of within-species variation in this parameter are still relatively poorly understood. Individually differing levels of activity and stress responsiveness have been proposed as potential sources for such variation in BMR. Here, I used an open flow-through respirometry system to examine the possible correlates of the time it takes individual Laughing Doves Spilopelia senegalensis to reach lowest oxygen consumption level in a metabolic chamber ('BMR time'). Also, the association between individual response to handling stress and BMR was studied. Breath rate, measured while holding the bird in hand, was used as a measure of stress response to handling. It was found that BMR time' was not related to BMR or breath rate. However, its values were positively predicted by individuals' hematocrit. This indicates the potential importance of 'BMR time' as an indicator of activity. It was also found that breath rate was individually repeatable when two measurements were taken 12 hours apart. Breath rate was also positively related to BMR; however, the effect disappeared when mass-specific residuals of BMR were used. These results suggest that individual differences in response to standard handling stress probably do not affect BMR measurements.

In vertebrates, stress experienced by mothers during the early stages of reproduction is an important source of epigenetic modifications in their offspring. Birds represent excellent models to test such effects as their maternal investment can be quantified in terms of egg quality. Recently, it has been demonstrated that corticosterone (CORT) can be transmitted from a female bird into its eggs. However, there is little published evidence about maternal effects that are mediated by acute stress. In this study, we demonstrated that female great tits Parus major facing an aerial predator during egg formation increased CORT concentration in eggs that were laid the morning after the treatment. By presenting a predator model to each experimental nest twice a day, we found that maternal stress influences corticosterone content in eggs during a time period from albumen production in the magnum until the initial phase of shell secretion, when additional water is added to the egg in the shell gland. We also found a positive correlation between the duration of parental alarm calls and CORT concentration in eggs. In conclusion, the response of female passerines to predatory cues brings about a maternal effect that may have negative consequences for offspring performance. We also suggest that the total duration of the behavioral response to a stressor is an important determinant of CORT levels in the albumen of bird eggs.

Plumage bacteria may play an important role in shaping the life histories of birds. However, few studies have examined natural patterns of variation in plumage bacteria. We have previously shown that plumage bacterial load is higher during the pre-laying period than during the brood rearing period in female Great Tits (Parus major). Here, we examined whether the pre-laying bacterial peak in female Great Tits develops during nest-building, where the females come into increased contact with the ground and nest materials, or dates back to an earlier period. Females were captured during three stages of nest-building (nest initiated, nest half ready, and nest completed). The density of plumage bacteria and the species richness of feather-degrading bacterial assemblages were studied using flow cytometry and ribosomal intergenic spacer analysis (RISA), respectively. The density of attached bacteria on feathers increased significantly between nest initiation and nest completion. No effect of nest-building stage on feather-degrading bacterial species richness was found. Our results indicate that the density of attached bacteria in the plumage of free-living birds can change rapidly during nest-building, providing one potential cost of nest-building for individual birds.

Microorganisms are known to play an important role in shaping the life histories of animals. Recent studies have proposed that the coloration of birds' plumage could reflect individual quality through associations with feather-degrading bacteria. However, few studies have explored such relationships. We studied breeding female Great Tits (Parus major) during nest building and chick rearing to explore associations between bacteria inhabiting their yellow chest feathers and feather coloration. Specifically, we used flow cytometry and ribosomal intergenic spacer analysis (RISA), respectively, to study the densities of all free-living and attached bacteria and the phylotypic richness of feather-degrading bacterial assemblages. We used chroma (color saturation) as a measure of feather coloration. During chick rearing but not during nest building, the female's chroma was negatively related to the phylotypic richness of feather-degrading bacteria. Also, a seasonal change in the density of attached bacteria associating with individual birds was negatively associated with change in chroma over the same period. These findings suggest that conspicuous coloration of female Great Tits may reflect the numbers and character of bacteria inhabiting feathers.

Plumage bacteria may play an important role in shaping the life histories of birds. However, to design suitable experiments to examine causal relationships between plumage bacteria and the fitness of host birds, natural variation in plumage bacterial communities needs to be better understood. We examined within-individual consistency of plumage bacterial contamination in Great Tits (Parus major), comparing different body regions (ventral vs. dorsal) and comparing contamination between years. Numbers of free-living and attached bacteria and the species richness of feather-degrading bacterial assemblages were studied using flow cytometry and ribosomal intergenic spacer analysis (RISA). Numbers of both types of bacteria were higher on dorsal than on ventral feathers. Numbers of free-living, but not attached, bacteria on the two body regions were highly positively correlated. There was also a strong within-individual correlation between numbers of attached bacteria during the same breeding stages in different years. These results suggest that, despite variation in absolute levels of feather bacterial loads between years and different body regions, sampling individual birds can provide reliable estimates of relative levels of bacterial contamination, as long as sampling time and body region are carefully standardized.

Plumage bacteria might influence the trade-off between parental and self-preening efforts in birds, therefore affecting breeding success. However, too little is known about natural variation patterns in plumage bacterial communities for these hypotheses to be thoroughly assessed. We studied the density and phylotypic richness of plumage bacterial assemblages in wild breeding populations of Pied Flycatchers Ficedula hypoleuca and Great Tits Parus major in the same area and breeding season, using flow cytometry and ribosomal intergenic spacer analysis (RISA). The density of plumage bacteria was higher in Tits than in Flycatchers, providing evidence that bacterial microflora differs even between co-occurring hosts that share habitat, nest site and foraging preferences. It is concurrent with the finding that migratory birds might have lower bacterial loads than sedentary birds. In both species bacterial loads were higher in females than in males, which along with two earlier studies, indicates the generality of this sex pattern. A negative correlation between parental body mass and the richness of feather-degrading bacterial phylotypes was found in Pied Flycatchers. In Great Tits, higher bacterial densities in the plumage of parent birds were associated with the production of fewer fledglings. However, the causality of these associations remains to be tested experimentally.

Microorganisms have been shown to play an important role in shaping the life histories of animals, and it has recently been suggested that feather-degrading bacteria influence the trade-off between parental effort and self-preening behavior in birds. We studied a wild breeding population of great tits (Parus major) to explore habitat-, seasonal-, and sex-related variation in feather-degrading and free-living bacteria inhabiting the birds' yellow ventral feathers and to investigate associations with body condition. The density and species richness of bacterial assemblages was studied using flow cytometry and ribosomal intergenic spacer analysis. The density of studied bacteria declined between the nest-building period and the first brood. The number of bacterial phylotypes per bird was higher in coniferous habitat, while bacterial densities were higher in deciduous habitat. Free-living bacterial density was positively correlated with female mass; conversely, there was a negative correlation between attached bacterial density and female mass during the period of peak reproductive effort. Bacterial species richness was sex dependent, with more diverse bacterial assemblages present on males than females. Thus, this study revealed that bacterial assemblages on the feathers of breeding birds are affected both by life history and ecological factors and are related to body condition.

Differences in competitive abilities of siblings in birds can be caused by a combination of hatching asynchrony and intra-clutch variation in egg quality. However, very little is known how within-brood hierarchies affect the allocation of resources between different functions of the body. We examined the effects of within-brood hierarchy on growth of morphological parameters, blood plasma antioxidant protection and immune function of free-living great tit Parus major nestlings. To assure that competitive hierarchies occur, we experimentally delayed the start of incubation of the last two eggs in the clutch. At pre-fledging stage (day 13 post-hatch), late-hatched nestlings were smaller in body mass and wing length when compared to early-hatched nestlings, but no differences between siblings were found in tarsus length, plasma antioxidant potential, uric acid concentration, residual antioxidant potential (from regression with uric acid), hematocrit and response to phytohaemagglutinin injection. In early-hatched nestlings, the antioxidant potential and residual antioxidant potential measured in the middle of nestling period (day 6 post-hatch) were negatively related to body mass growth at early nestling stage, indicating that fast initial growth could reduce antioxidant properties of blood plasma.

Environmental factors during early development may have profound effects on subsequent life-history traits in many bird species. In wild birds, sex-specific effects of early ontogeny on natal dispersal and future reproduction are not well understood. The objective of this work was to determine whether hatching date and pre-fledging mass and condition of free-living Great Tits Parus major have any subsequent effect on individuals' natal dispersal and reproductive performance at first breeding. Both males and females dispersed longer distances in coniferous than in deciduous forests, while dispersal was condition-dependent only in males (heavier as nestlings dispersed farther). In females, mass and condition at pre-fledging stage correlated significantly with clutch size, but not with subsequent reproductive performance as measured by fledging success or offspring quality. In contrast, heavier males as nestlings had higher future fledging success and heavier offspring in their broods compared with those in worse condition as nestlings. The hatching date of female as well as male parents was the only parental parameter related to the number of eggs hatched at first breeding. These results indicate that pre-fledging mass and condition predict subsequent fitness components in this bird species. We suggest that sex-specific relationships between a disperser's condition and its selectivity with respect to breeding habitat and subsequent performance need to be considered in future models of life-history evolution.

Habitat-related variation has been revealed in the seasonal reproductive patterns of great tits inhabiting mosaics of deciduous woodland fragments and managed coniferous forests. The number and quality of offspring tend to be higher in the non-preferred coniferous habitat compared with the preferred deciduous habitat. We explored whether these patterns can be explained by variation in food abundance and/or parental provisioning ability. Frass fall seemed not to be a reliable measure of the seasonal dynamics of nestling feeding conditions, although it is widely used for this purpose. No habitat-related differences were found in the tendency of parents to increase provisioning frequency in response to increased hunger levels of nestlings, suggesting that provisioning frequency as such is not a limiting factor for nestling growth. Higher feeding rates in deciduous habitat were associated with lower proportions of high-quality food items among prey delivered to offspring. These findings confirm that relatively high nestling feeding rates in birds may reflect the low quality of available food rather than the quality of parental care or an abundance of food in the environment. The results also indicate that deciduous forest habitat, though preferred by tits, may sometimes provide poorer brood-rearing conditions than the non-preferred, coniferous habitat. We suggest that the great tit's preference for deciduous habitat, which presumably evolved in more southern regions, may be maladaptive in northern regions where deciduous woods are mainly young, secondary stands of alder and birch.

Recent studies have suggested that bone alkaline phosphatase (ALP) may be a valuable indicator of skeletal development in wild birds. However, the information about age-related dynamics of ALP isoforms in passerines is very scanty. We examined age-related changes in the activity of bone ALP and liver ALP and tested the applicability of these isoenzymes as indicators of chick maturity in randomly selected nestlings of a small passerine bird, the Great Tit (Parus major). Bone ALP activity was elevated in the middle of the nestling period (day 8), when skeletal growth is assumed to be most rapid, and declined significantly during the prefledging stage (day 15 posthatch). Bone ALP activity at this age was positively and highly significantly related to the overall duration of nestling period and negatively and less significantly related to wing length and body mass. All three morphological traits of the 15-day-old nestlings were nearly significantly negatively correlated with the duration of the nestling period. Liver ALP activity neither changed with nestling age nor was related to nestling morphology. We suggest that prefledging activity of bone ALP is a more reliable indicator of nestling maturity than traditionally used morphological measurements.

Little is known about the physiological causes and consequences of habitat choice decisions in birds. We compared size-corrected body mass, hematocrit, plasma albumin, beta- and gamma-globulin and triglyceride concentrations, as well as the albumin/globulin ratio of female Great Tits breeding in deciduous and coniferous forests in Estonia. Females were sampled during incubation and during the chick-rearing period. Whereas the nest box occupation rate and clutch size were higher in the deciduous habitat, the female nutritional condition during incubation was better in the coniferous habitat. This habitat difference in condition was not explained by the differences in the start of egg-laying or clutch size. Females lost more mass from incubation to the chick-rearing stage in the coniferous than in the deciduous forests. Although the values of most blood parameters changed from incubation to the chick-rearing stage, no habitat-related change was observed. There were no habitat differences in the number and quality of fledglings or in female condition during the chick-rearing period. Our results indicate that incubating female Great Tits are in a worse nutritional state in the preferred deciduous habitat fragments than those in the non-preferred coniferous habitat. However, habitat-related differences in condition during incubation seem to be unrelated to habitat-specific reproductive decisions.

It has been suggested that a bird's clutch size is not limited by the amount of resources available at the time of laying but that differences in the availability of food for nestlings is the ultimate underlying factor determining spatio-temporal variations in clutch size. However, habitat-related variations in egg production ability has yet to be investigated explicitly. We studied the breeding of Great Tits Parus major in deciduous and coniferous forests in the same area. The sizes of both the clutches and the eggs were, on average, larger in the former habitat than in the latter. A number of females were induced to lay more eggs than usual by removing four eggs from designated experimental clutches early in the laying period. These manipulated females laid approximately one egg more than control females, with the number of additional eggs laid not differing between the habitats. However, in both study years the relative size of the extra eggs – relative to the mean size of earlier laid eggs of the same clutch – was smaller in the coniferous habitat than in the deciduous habitat, while there was no habitat-related difference in the relative size of the last-laid eggs of control clutches. This result indicates that some form of proximate limitation during egg-laying period can contribute to the relatively small clutches and eggs in the coniferous habitat. Our results emphasize the need to take egg production costs into account when attempting to account for spatial variation in the reproductive behaviour of birds.

Recent studies have suggested that a biochemical marker, plasma alkaline phosphatase ( ALP), can be used as a general indicator of skeletal development in vertebrate animals. In birds, age-related variation in ALP activity, presumably due to bone formation processes, has been demonstrated, but to date, a direct connection between bone mineralization and enzyme activity has been elusive. In this study, we show that the activity of a bone isoform of ALP ( bone ALP) is closely related to the overall rate of skeletal mineralization in nestlings of a small passerine bird, the great tit (Parus major L). Moreover, bone ALP activity predicted the rate of mineralization of leg and wing bones but not that of the skull. Liver isoform of ALP was only marginally related to the overall rate of skeletal mineralization, while no association with the mineralization of long bones was found. We conclude that bone ALP activity in the blood plasma is a reliable biomarker for skeletal mineralization in birds. This marker enables detection of subtle developmental differences between chicks of similar structural size, potentially facilitating the prediction of offspring mid- and long-term survival.

Carotenoids are biologically active pigments, which are important for animals due to their dual role in health maintenance and ornamental signalling. In adult birds, immunostimulatory properties of carotenoids have been repeatedly demonstrated while much less is known about the importance of carotenoids as antioxidants. We studied the relationships between plasma carotenoid levels, as well as total antioxidant protection, and various hemato-serological health state indices in female great tits (Parus major L.), incubating their second clutches in two contrasting (coniferous and deciduous) habitats in southwest Estonia. To manipulate reproductive effort, four eggs were removed from half of the clutches during laying to stimulate females to lay additional eggs. However, egg removal had no effect on the final number of eggs laid. Plasma carotenoid levels increased seasonally in parallel with caterpillar food availability. However, no between-habitat differences in carotenoid levels, total antioxidant capacity, or indices of health state could be found despite the apparently better feeding conditions in the coniferous habitat. No correlation was detected between plasma carotenoid levels and measures of total antioxidant capacity, which suggests that at least for the adult birds feeding on naturally carotenoid-rich diet, antioxidant function of carotenoids is not of primary importance. A strong non-linear association between the measures of antioxidant protection and leukocytic markers of inflammation was found, which suggests that measures of total antioxidant capacity deserve further attention in ecophysiological studies as potential indicators of immunopathology.

Very little is known about the causes and correlates for variation of individual condition in the wild. However, such knowledge is essential for understanding the mechanisms that mediate environmental effects to populations. We studied the variation of several hematological condition indices (hematocrit, albumin, globulin and triglyceride concentrations, albumin/globulin ratio, lymphocyte and heterophile concentrations and heterophile/lymphocyte ratio) and body mass in brood-rearing great tits (Parus major) in relation to habitat, multiple breeding and gender. Although great tits prefer deciduous forest to coniferous forests, individuals breeding in coniferous forests tended to be in a superior health state than those breeding in deciduous habitat. We suggest that this difference in adult condition can be caused by differences in breeding densities between habitats. Although there was some variation in condition indices between breeding attempts, none of these parameters measured at the end of the first breeding attempt predicted the probability of double breeding. We also found that females were in poorer condition and probably more stressed than males, both during the first and the second breeding attempt. These findings demonstrate that hematological parameters can be used to assess spatial and temporal variation of individual condition in the wild.

In birds, it has been shown that reproductive effort may impair parental condition, while the relation of different condition indices to subsequent survival is still poorly understood. In this study, we measured body mass and various hematological condition indices in breeding great tits in relation to local survival. Number and quality of nestlings and the occurrence of second broods, potentially reflecting parents' breeding effort, were also considered in analyses. The great tits, both male and female, that returned the following year had had a higher albumin/globulin ratio, lower plasma globulin concentration, and a lower heterophile/lymphocyte ratio during breeding in the preceding year, compared to those who did not return. Surviving males (but not females) also had had a higher level of circulating lymphocytes, compared to nonsurvivors. There was no correlation between breeding effort and survival. We conclude that better immunological state and lower stress in great tits during breeding were positively related to their survival probability.