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The increase of urbanization and agricultural activities is causing a dramatic reduction of natural environments. As a consequence, animals need to physiologically adjust to these novel environments, in order to exploit them for foraging and breeding. The aim of this work was to compare the physiological status among nestling common kestrels (Falco tinnunculus) that were raised in nest-boxes located in more natural, rural, or urban areas in a landscape with a mosaic of land uses around Rome in Central Italy. A blood-based multi-biomarker approach was applied to evaluate physiological responses at multiple levels, including antioxidant concentrations, immunological functions, genotoxicity, and neurotoxicity. We found lower concentrations of glutathione and GSH:GSSG ratio values and higher proportions of monocytes in urban birds compared to the other areas. We also found higher DNA damage in rural compared to urban and natural krestels and inhibition of butyrylcholinesterase activity in urban and natural birds compared to rural area. Finally, we found similar values among study areas for respiratory burst, complement system, bactericidal capacity, and plasma non-enzymatic antioxidant capacity. These results suggest that (i) city life does not necessarily cause physiological alterations in kestrels compared to life in other habitats, and (ii) environmental pressures are likely to differ in typology and intensity across habitats requiring specific responses that a multi-biomarker approach can help to detect. Further studies are needed to assess which factors are responsible for the physiological differences among city, rural, and natural birds, and whether these differences are consistent across time and space., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Urbanisation is one of the main anthropogenic forms of land cover affecting an ever-increasing number of wild animals and their habitats. Physiological plasticity represents an important process through which animals can adjust to the novel conditions of anthropogenic environments. Relying on the analysis of gene expression, it is possible to identify the molecular responses to the habitat conditions and infer possible environmental factors that affect the organismal physiology. We have quantified for the first time the blood transcriptome of common kestrel (Falco tinnunculus) nestlings living in urban sites and compared it to the transcriptome of kestrel nestlings inhabiting rural and natural environments. We found mild differences in the expression of genes among sites, indicating adaptability or acclimation of the birds to the urban habitat. We identified 58 differentially expressed genes between urban and natural kestrels, and 12 differentially expressed genes between urban and rural kestrels. The most striking differences among sites involved inflammatory-immunological, metabolic, apoptosis, DNA repair and development genes. In particular, we found that (i) urban kestrel nestlings had higher expression of genes linked to inflammation, repair of DNA damage, or apoptosis than natural kestrel nestlings, and (ii) natural and rural kestrel nestlings had higher expression of genes linked to the development and activation of immune cells, type I interferon response, or major histocompatibility complex than urban kestrel nestlings. Finally, the KEGG enrichment analysis identified the insulin signalling as the main pathway that differed between natural and urban kestrel nestlings. This is one of a limited number of studies on vertebrates that revealed habitat-associated differences in the transcriptome. It paves the way for further in-depth studies on the links between physiological variation and habitat structure at different spatial and temporal scales., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Many large birds rely on thermal soaring flight to travel cross-country. As such, they are under selective pressure to minimise the time spent gaining altitude in thermal updrafts. Birds should be able to maximise their climb rates by maintaining a position close to the thermal core through careful selection of bank angle and airspeed; however, there have been few direct measurements of either parameter. Here, we apply a novel methodology to quantify the bank angles selected by soaring birds using on-board magnetometers. We couple these data with airspeed measurements to parameterise the soaring envelope of two species of Gyps vulture, from which it is possible to predict 'optimal' bank angles. Our results show that these large birds respond to the challenges of gaining altitude in the initial phase of the climb, where thermal updrafts are weak and narrow, by adopting relatively high, and conserved, bank angles (25-35 deg). The bank angle decreased with increasing altitude, in a manner that was broadly consistent with a strategy of maximising the rate of climb. However, the lift coefficients estimated in our study were lower than those predicted by theoretical models and wind-tunnel studies. Overall, our results highlight how the relevant currency for soaring performance changes within individual climbs: when thermal radius is limiting, birds vary bank angle and maintain a constant airspeed, but speed increases later in the climb in order to respond to decreasing air density., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

Carotenoid-based skin colorations vary seasonally in many bird species and are thought to be honest sexually selected signals. In order to provide more insight in the potential signal function and underlying mechanisms of such colorations we here quantified patterns of variation of leg coloration in adult male and female Eurasian kestrels (Falco tinnunculus tinnunculus) over the breeding season, and evaluated the relationship between coloration and levels of carotenoids, androgens and estrogens, oxidative damage and plasma non-enzymatic antioxidant capacity. We studied both reproducing wild and non-reproducing captive birds to test for the effect of diet and breeding effort. Males were more colored than females only during mating, and independently of diet, suggesting that leg-color is a sexually selected trait. Seasonal variation in leg color was associated with circulating carotenoids, but concentrations of these molecules were not related to antioxidant capacity, body condition or oxidative damage. These results indicate that carotenoid-based colorations may not be an honest signal of health status in this species. Production of carotenoid rich eggs coincided with low levels of circulating carotenoids in females, indicating that carotenoids might be a limited resource for laying female kestrels. Finally, young rearing males had higher levels of oxidative damage than females, and wild birds of both sexes had higher levels of these parameters than captive birds. These results may indicate that parental effort and physical activity are costly, independently from hormonal status. Since androgens did not explain carotenoid variation we suggest that multiple interacting factors can regulate carotenoid levels along the season., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Nesting on high voltage transmission line towers exposes birds to electric and magnetic fields for long periods. Nestlings are exposed from their development in ovo until fledging. This is a critical period for them because the quality of the developmental environment may affect their fitness at adulthood. We carried out a field study on Eurasian kestrels, Falco tinnunculus, to compare chicks from pairs nesting on high voltage power lines vs. those nesting in control sites in similar habitats. The magnetic field (MF) was measured in each nest-box and analysed in relation to growth curves, melatonin levels, leukocyte counts, and fledging success. None of the variables differed between exposed and control nestlings. Wing length (proxy of age) showed a negative covariation with serum melatonin concentration. Our findings suggest that exposure to MFs produced by high voltage power lines during the embryonic and post-hatching period (until fledging) does not have significant short-term physiological effects on kestrel nestlings.

Fail-to-hatch kestrel (Falco tinnunculus) eggs collected at the end of the 1999 and 2005 breeding seasons from nest boxes in and around the city of Rome, Italy, were analyzed by gas chromatography with electron capture detection for their PCB content and for the presence of DDT derivatives and other organochlorines. Among the various PCBs, congeners 153 and 180 were detected in all the eggs and showed the highest concentrations. Eggs collected from the same nest from a polluted location in Rome during 2 different years showed similar type and number of PCB congeners. These data and the fact that eggs from another nest near a sulphate mine had, atypically, low-chlorinated congeners support the conclusion that eggs of this species, whose adults in the Mediterranean and continental Europe perform only short or no migration movements, might be indicative of local pollution. When multiple eggs in the same clutch were analyzed, the PCBs were similar in type but their concentration decreased within clutch, likely in parallel to the laying order.

Birds control body homeostasis through the secretion of corticosterone. This hormone is the end-product of the hypothalamic-pituitary-adrenal (HPA) axis response to stressors. High levels of corticosterone may be associated with low individual fitness and may affect balance between pro-oxidants and antioxidants. Given these points, chronic stress modulated by hormones could undermine individual fitness by increasing oxidative tissue damage. In this study, we administered corticosteroids by diet (20 mg/kg of diet) to captive adult kestrels (Falco tinnunculus) over a 14-day period to evaluate the effects of a simulated chronic stress modulated by corticosteroids. We found that dietary administration of corticosterone caused a 32% increase of reactive oxygen metabolites, but did not impair total serum antioxidant capacity, serum carotenoids or body mass. Oxidative stress had a 64% increase in treated birds compared to 30% in controls. The two groups did not differ in the total serum antioxidant capacity, which showed a significant decrease over the study period. In contrast, circulating carotenoids and body mass increased in both groups. These results suggest that stress hormones, such as corticosterone, may also act as modulators of oxidative stress in birds.

Carotenoids are considered a limited resource for animals because they are not synthesised by the body. Birds use carotenoids, mainly xanthophylls, for physiological functions, such as anti-oxidant activity, and for colour expression; hence, they need to shunt carotenoids between competitive demands. Recent studies suggest that the anti-oxidant role of xanthophylls might not be as important as previously thought and that at high concentrations they may, in fact, acquire pro-oxidant properties. In this work, we studied the effects of a moderate xanthophyll supplementation (115 mg of carotenoids/kg diet/day; 4 weeks) on serum carotenoids, serum concentration of reactive oxygen metabolites (ROMs), serum anti-oxidant capacity (OXY), the degree of oxidative stress (OS; ROMs/OXY x 1,000), body mass, and skin colour, in rehabilitated captive adult Eurasian kestrels (Falco tinnunculus). The supplementation caused increased levels of serum carotenoids (approximately 90%), ROMs (approximately 82%), OS (approximately 115%) and an immediate loss of body mass (approximately 6.2%), but it did not affect OXY and tarsi skin hue. The red (approximately 16%) and yellow (approximately 15%) colorimetric components were increased after the first week of supplementation and the effect persisted during the rest of the experiment. Two months after the end of supplementation, serum carotenoids, OS and ROMs returned to baseline levels, however the body mass did not. Our findings suggest that, above a certain physiological threshold, carotenoids can cause detrimental effects. This is relevant for the trade-off between expression of sexual signals and the costs of maintaining/producing them.

Pylons of utility lines are commonly used by breeding birds as structures for supporting their nests. Nesting near power lines, however, exposes adult birds and their offspring to the electric and magnetic fields (EMFs) produced by the current. Therefore, we searched for possible relationships between the magnetic field (MF) magnitude experienced by wild kestrel (Falco tinnunculus) nestlings grown on pylons and different health-related variables: body condition, serum concentration of carotenoids, reactive oxygen metabolites (ROMs; marker of early oxidative damage), serum anti-oxidant capacity (OXY), and the ratio between ROMs and OXY (index of oxidative stress). No significant relationships were found between the MF magnitude or squared MFs and any of the variables considered. Comparisons with values recorded in nestlings from non-exposed nests seem to confirm the absence of any effect of exposure to MFs produced by power lines on the variables considered.

It is recognized that carotenoids are useful anti-oxidants in embryo and hatchling avian models. However, recent evidence suggests that the anti-oxidant role of carotenoids in nestling or adult birds may not be as important as previously thought. The aim of the present work was to investigate if supplemental carotenoids decreased the level of oxidative damage (by reactive oxygen metabolites, ROMs) and increased the serum anti-oxidant capacity (OXY) in nestling Eurasian kestrels Falco tinnunculus. Circulating carotenoids in supplemented nestlings increased about 1.5-fold compared to the control and pre-treatment levels at the end of the supplementation period. There was no effect on ROMs, OXY or the level of oxidative stress (ratio between ROMs and OXY), however, or on body mass or body condition of nestlings. ROMs and OXY decreased with age, but this pattern varied across the nests. Our results show that (i) in general, younger nestlings actually have to cope with a high free radical production, and (ii) the ability of wild nestling kestrels to cope with oxidative stress is not affected by carotenoid availability.

We evaluated the oxidative cost paid by birds when coping with an immune challenge. We used the phytohaemagglutinin skin test (PHA) to assess the effects of the T-cell-mediated immune response on the concentration of reactive oxygen metabolites (ROMs), total antioxidant barrier (OXY) and total serum carotenoid concentration in wild nestlings of the Eurasian kestrel (Falco tinnunculus). Immunostimulation caused increased levels of ROMs, decreased OXY and increased circulating levels of carotenoids. These results suggest that an immune challenge can increase avian oxidative stress, and that carotenoids were remobilised from other tissues likely because their circulating levels were not sufficiently high to sustain an effective immune response.