Gustavo Tomás

Corticosterone metabolites in blue tit and pied flycatcher droppings : Effects of brood size, ectoparasites and temperature

The hypothalamic-pituitary-adrenal (HPA) axis of birds induces the secretion of corticosterone (CORT) as a response to different ecological variables. In this study we tested experimentally if manipulations of brood size or ectoparasitism led to subsequent differences in the concentration of excreted CORT metabolites of adult and nestling blue tits (Cyanistes caeruleus). No significant effect of the manipulation of brood size was detected in adults or nestlings. No significant effect of ectoparasitism was detected in males or nestlings, although females from uninfested nests showed lower concentrations of excreted CORT metabolites. In addition, we analysed if weather conditions had an influence on the concentration of excreted CORT metabolites of blue tits and pied flycatchers (Ficedula hypoleuca) breeding in the same forest. We detected no effect of weather conditions on adults, but nestlings of both species showed a negative correlation between their excreted CORT metabolites and the average mean temperatures they were subjected to during their growth. This effect was not found in blue tits in a colder year, suggesting that the sensitivity of the HPA axis to ambient temperature may be subjected to interannual variation. Moreover, we found a positive effect of the maximum temperature on the day of sampling on the concentration of CORT metabolites of blue tit nestlings in one of the years. These results suggest that weather conditions may act as environmental stressors to which the HPA axis of blue tit and pied flycatcher nestlings may be sensitive.

NEST WEIGHT AND FEMALE HEALTH IN THE BLUE TIT (CYANISTES CAERULEUS)

Abstract We explored two hypotheses that may explain intraspecific variability in nest size. The “thermoregulatory” hypothesis states that species adjust nest size to maintain egg temperature and minimize temperature fluctuations in the nest. Recently, the suggestion has been made that nest size may reflect the health status or phenotypic quality of the builder, potentially making it a sexually selected trait (“sexual selection” hypothesis). For two years, we weighed nests of Blue Tit (Cyanistes caeruleus) at initiation of reproduction in a nest-box breeding population in central Spain. We recorded laying date, clutch size, incubation period, and hatching success. We measured and took blood samples of adult females when nestlings reached three days of age. General regression models controlling for potential variables that could affect nest weight revealed that prevalence of Trypanosoma avium and immunoglobulin levels in females were significantly related to nest weight in only one of the study years. Females not infected with Trypanosoma avium built heavier nests than infected ones, whereas female immunoglobulin levels were negatively associated with nest weight. Hatching success and duration of the incubation period were not related to nest weight in either year. Thus, our results do not support the thermoregulatory hypothesis and show that certain measures of female health are related to nest-building effort in some years, probably depending on environmental conditions. Peso del Nido y Estado de Salud de la Hembra en el Cyanistes caeruleus

Sex recognition by odour and variation in the uropygial gland secretion in starlings

1. Although a growing body of evidence supports that olfaction based on chemical compounds emitted by birds may play a role in individual recognition, the possible role of chemical cues in sexual selection of birds has been only preliminarily studied. 2. We investigated for the first time whether a passerine bird, the spotless starling Sturnus unicolor, was able to discriminate the sex of conspecifics by using olfactory cues and whether the size and secretion composition of the uropygial gland convey information on sex, age and reproductive status in this species. 3. We performed a blind choice experiment during mating, and we found that starlings were able to discriminate the sex of conspecifics by using chemical cues alone. Both male and female starlings preferred male scents. Furthermore, the analysis of the chemical composition of the uropygial gland secretion by using gas chromatography-mass spectrometry (GC-MS) revealed differences between sexes, ages and reproductive status. 4. In conclusion, our study reveals for first time that a passerine species can discriminate the sex of conspecifics by relying on chemical cues and suggests that the uropygial gland secretion may potentially function as a chemical signal used in mate choice and/or intrasexual competition in this species.

Carotenoid-based plumage colouration is associated with blood parasite richness and stress protein levels in blue tits (Cyanistes caeruleus).

Carotenoids are molecules that birds are not able to synthesize and therefore, must be acquired through their diet. These pigments, besides their function of giving birds red and yellow colouration when deposited in feathers, seem to act as immune-stimulators and antioxidants in the organism. Hence, only the healthiest individuals would be able to express carotenoid-based ornaments to a larger extent without compromising the physiological functions of carotenoids. Various studies have reported that birds infected by parasites are paler than those uninfected, but, to our knowledge, none of them has assessed the possible effect of multiple infections by blood parasites on plumage colour. By comparing the yellow colour in the breast plumage of blue tits, Cyanistes caeruleus, between birds infected by different numbers of blood parasite genera, we found that those birds infected by more than one genus were paler than those parasitized just by one. In addition, we examined the potential role of carotenoid-based plumage colour of blue tits as a long-term indicator of other parameters of health status, such as body condition and immunoglobulin and heat shock protein (HSP) levels. Our results indicate that more brightly coloured birds had lower HSP70 levels than paler birds, but we did not find any significant association between colour and body condition or immunoglobulin levels. In addition, we found a positive significant association between Haemoproteus density of infection and HSP60 levels. Overall, these results support the role of carotenoid-based colours as indicators of health status in blue tits and show detrimental effects of parasitism on this character.

Determinants of abundance and effects of blood-sucking flying insects in the nest of a hole-nesting bird

Compared to non-flying nest-dwelling ectoparasites, the biology of most species of flying ectoparasites and its potential impact on avian hosts is poorly known and rarely, if ever, reported. In this study we explore for the first time the factors that may affect biting midge (Diptera: Ceratopogonidae) and black fly (Diptera: Simuliidae) abundances in the nest cavity of a bird, the hole-nesting blue tit Cyanistes caeruleus, and report their effects on adults and nestlings during reproduction. The abundance of biting midges was positively associated with nest mass, parental provisioning effort and abundance of blowflies and black flies, while negatively associated with nestling condition. Furthermore, a medication treatment to reduce blood parasitaemias in adult birds revealed that biting midges were more abundant in nests of females whose blood parasitaemias were experimentally reduced. This finding would be in accordance with these insect vectors attacking preferentially uninfected or less infected hosts to increase their own survival. The abundance of black flies in the population was lower than that of biting midges and increased in nests with later hatching dates. No significant effect of black fly abundance on adult or nestling condition was detected. Blood-sucking flying insects may impose specific, particular selection pressures on their hosts and more research is needed to better understand these host–parasite associations.

BENEFICIAL EFFECTS OF CLOACAL BACTERIA ON GROWTH AND FLEDGING SIZE IN NESTLING PIED FLYCATCHERS (FICEDULA HYPOLEUCA) IN SPAIN

Abstract Effects of bacteria on avian hosts in the wild have received little attention until recently. Whereas the pathogenic effects of bacteria are well known, positive effects of symbiotic bacteria are more rarely considered. Nestling growth has important repercussions for offspring fitness in avian populations and may be affected by microbial colonization of the gut. Enterococcus faecalis is a common opportunistic pathogen, whereas E. faecium has been used as a growth promoter because it interacts competitively with pathogenic bacteria, E. faecalis included. We followed the growth in tarsus length and mass of 18 Pied Flycatcher (Ficedula hypoleuca) broods. Chicks were weighed and measured on days 4, 8, 10, and 13 after hatching. On day 13, wing length was also measured and cloacal swabs were taken of two chicks in each brood for detection of enterococci. In all, the methods used allowed us to detect six species of bacteria among a possibly much richer community. Most chicks had E. faecalis, whereas E. faecium was less prevalent. There was a negative association between scores for E. faecalis and for the rest of the species pooled. The presence of E. faecalis showed no detectable association with nestling mass or size at any age, whereas the presence of the other species showed significantly positive associations with mass and size on day 13, but not before. Presence of E. faecium on its own was positively associated with nestling mass and size shortly before fledging. E. faecium may act as a growth promoter in the wild through its competitive interactions with facultative pathogenic bacteria. The presence of some microbes are critically important in avian growth and development.

Nest Construction Rate and Stress in Female Pied Flycatchers Ficedula hypoleuca

Abstract. Nest building effort has received scant attention in the literature although it may involve costs which can be detected as physiological stress. We prolonged nest construction effort in a population of Spanish Pied Flycatchers by removing nests from nest-boxes and forcing females to build a second nest. In comparison with control nests, the experimental females had to work for longer periods and accumulate more nest material, but nest construction rates (g of nest material per day of construction) were not affected. There was a positive association of clutch mass with nest construction rate. To measure physiological stress, we captured females shortly after laying to obtain blood samples for heat-shock protein quantification. Heat-shock proteins quantify stress at cell level. The level of HSP60 in peripheral blood was positively associated with total nest construction rate (including second nests for experimental females), but not with laying date, clutch mass or experimental treatment. A third of the variation in the HSP60 level was explained by the nest construction rate. Fast nest builders are physiologically stressed, suggesting that the nest construction rate may constitute an index of female physiological performance.

Evidence for differential maternal allocation to eggs in relation to manipulated male attractiveness in the pied flycatcher (Ficedula hypoleuca)

We present evidence of differential maternal allocation to eggs in response to manipulated male attractiveness in the migratory pied flycatcher (Ficedula hypoleuca). We manipulated the size of a male secondary sexual trait, the white forehead patch, right after male arrival to the breeding area and before female arrival. Patch size was (1) enlarged to the maximum observed in the population, (2) reduced by 40% or (3) kept constant by painting with indelible felt markers over the natural feather patch. Male behaviour was affected by the experimental manipulation, as individuals with enlarged patches performed more approaches to the nestbox in response to song playback during the territory occupation and nest-site presentation phases. Females paired with males with reduced forehead patches laid significantly smaller eggs than those paired with males in the control and enlarged-patch treatments. Laying date and clutch size did not differ among the experimental groups. We discuss that manipulations of ornaments designed to study differential allocation at laying should reduce as well as enlarge their expression.

Collection and analysis of blood samples to detect stress proteins in wild birds

Abstract The blood stress protein response can be used to evaluate physiological or environmental stress in individuals and populations. It is important that sample conservation and transport are correct to avoid confounding effects on protein analyses. We show that Great Tit (Parus major) blood samples can be centrifuged and frozen any time within eight hours of collection, as during this period there are no significant changes in stress protein HSP70 and HSP60 levels. These findings simplify future research on the stress protein response in wild birds because samples can be safely kept in a cool box for a relatively long time.

Factors affecting Culicoides species composition and abundance in avian nests

Mechanisms affecting patterns of vector distribution among host individuals may influence the population and evolutionary dynamics of vectors, hosts and the parasites transmitted. We studied the role of different factors affecting the species composition and abundance of Culicoides found in nests of the blue tit (Cyanistes caeruleus). We identified 1531 females and 2 males of 7 different Culicoides species in nests, with C. simulator being the most abundant species, followed by C. kibunensis, C. festivipennis, C. segnis, C. truncorum, C. pictipennis and C. circumscriptus. We conducted a medicationxfumigation experiment randomly assigning birds nests to different treatments, thereby generating groups of medicated and control pairs breeding in fumigated and control nests. Medicated pairs were injected with the anti-malarial drug Primaquine diluted in saline solution while control pairs were injected with saline solution. The fumigation treatment was carried out using insecticide solution or water for fumigated and control nests respectively. Brood size was the main factor associated with the abundance of biting midges probably because more nestlings may produce higher quantities of vector attractants. In addition, birds medicated against haemoparasites breeding in non-fumigated nests supported a higher abundance of C. festivipennis than the rest of the groups. Also, we found that the fumigation treatment reduced the abundance of engorged Culicoides in both medicated and control nests, thus indicating a reduction of feeding success produced by the insecticide. These results represent the first evidence for the role of different factors in affecting the Culicoides infracommunity in wild avian nests.