Proximity to roads, but not exposure to metal pollution, is associated with accelerated developmental telomere shortening in nestling great tits.

Comprehensively understanding the factors affecting physiology and fitness in urban wildlife requires concurrently considering multiple stressors. To this end, we simultaneously assessed how metal pollution and proximity to roads affect body condition and telomere shortening between days 8 and 15 of age in nestling great tits (Parus major), a common urban bird. We employed a repeated-measures sampling design to compare telomere shortening and body condition between nestlings from four urban study sites south of Antwerp, Belgium, which are located at different distances from a metal pollution point source. In addition, we explored associations between metal exposure and telomere dynamics on the individual level by measuring blood concentrations of five metals/metalloids, of which lead, copper and zinc were present at concentrations above the limit of detection. To assess whether roadway-associated stressors (e.g. noise and air pollution) might affect nestling condition and telomere shortening, we measured the proximity of nest boxes to roads. Metal exposure was not associated with nestling telomere length or body condition, despite elevated blood lead concentrations close to the metal pollution source (mean ±â€¯SE = 0.270 ±â€¯0.095 µg/g wet weight at the most polluted study site), suggesting that nestlings may have some capacity to detoxify metals. However, nestlings from nest boxes near roads exhibited more telomere shortening between days 8 and 15 of age, and shorter telomeres at day 15. Nestlings in poorer condition also had shorter telomeres, but proximity to the road was unrelated to body condition. Thus, nutritional stress is unlikely to mediate the relationship between proximity to roads and telomere length. Rather, proximity to roads could have affected telomere shortening by exposing nestlings to air or noise pollution. Our study highlights that traffic-related pollution, which is implicated in human health problems, might also affect urban wildlife.

Accumulation, tissue-specific distribution and debromination of decabromodiphenyl ether (BDE 209) in European starlings (Sturnus vulgaris).

In this study we investigated the accumulation, tissue-specific distribution and possible debromination of BDE 209 in a terrestrial songbird species, the European starling, using silastic implants as a method of exposure. BDE 209 accumulated in the blood of the exposed starlings to a mean peak concentration of 16+/-4.1 ng/ml on day 10. After this peak, there was a decline to 3.3+/-0.4 ng/ml blood at the end of the exposure period of 76 days, which suggests elimination of BDE 209. In the exposed group, the muscle concentrations (461 ng/g lipid weight [lw], 430 ng/g lw) were about twofold those in liver (269 ng/g lw, 237 ng/g lw). In addition to BDE 209, other PBDE congeners, particularly octa- and nonaBDEs, were also present in the muscle and liver, suggesting bioformation from BDE 209. To our knowledge, these results are the first indications for the debromination of BDE 209 in birds.

Experimental evaluation of the usefulness of feathers as a non-destructive biomonitor for polychlorinated biphenyls (PCBs) using silastic implants as a novel method of exposure.

Risk assessment of pollutants requires both monitoring studies in the field and experimental exposure studies. In this study, we evaluated silastic implants as an alternative method of exposure for use in toxicological studies and at the same time evaluated the usefulness of feathers as a non-destructive biomonitor for PCBs. European starlings (Sturnus vulgaris) were exposed to different doses (including a control group) of environmentally relevant concentrations of PCB 153 during a 15-week period using silastic implants with both ends/only one end sealed. After implantation, there was a rapid and significant increase in PCB 153 blood concentration in the exposed groups. The significant differences in blood concentrations among the treatment groups show that silastic implants are useful as a method of exposure. Moreover, the ratio between the tissue concentrations of two treatment groups reflected the difference in implantation doses between these groups. There was also a clear difference in tissue concentrations among the treatment groups, although we could not test this statistically due to the small sample sizes. The slow release kinetics for a prolonged period and the relatively stable blood concentrations during the 15-week period render silastic tubes very interesting to study the effects of chronic exposure to pollutants. Our results also revealed that sealing both ends of the implant instead of only one did not significantly affect the exposure. There were strong, significant positive correlations between the blood and the tissues, which confirm the use of blood to monitor PCBs. To evaluate the usefulness of feathers as a non-destructive biomonitor for PCBs, we plucked the original and newly grown wing and tail feathers. We observed strong, significant positive correlations between the concentrations in the newly grown feathers and concentrations in the muscle, liver, brain and blood. PCB 153 concentrations in the newly grown feathers differed among the treatment groups. To our knowledge, our results provide the first experimental evidence that feathers are useful as a non-destructive biomonitoring tool for PCBs.

Within- and among-clutch variation of organohalogenated contaminants in eggs of great tits (Parus major).

In this study we investigated the variation of organohalogenated compounds, such as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and organochlorine pesticides (OCPs), within and among clutches of a small insectivorous songbird, the great tit (Parus major), and determined whether egg laying order affects the concentrations of these compounds. PCBs were the major organohalogenated contaminants in the eggs of great tits (mean 4778 ng/g lipid weight), while PBDEs (mean 204 ng/g lipid weight) and DDTs (mean 601 ng/g lipid weight) were found at much lower concentrations. Within-clutch variability of PCBs, PBDEs, and DDTs residues in the egg samples was small (7%, 3%, and 22%, respectively) compared to among-clutch variability (93%, 97%, and 78%, respectively). The small within-clutch variability may be in part related to the small home range of great tits and low spatial heterogeneity of the contaminants. We found no laying order effects on the concentrations of PCBs, PBDEs, and DDTs. Our results show that random egg samples of great tits are suitable as a biomonitoring tool for contamination with persistent organohalogenated pollutants in terrestrial environments.

Effects of heavy metal exposure on aggressive behavior in a small territorial songbird.

Several studies have shown that elevated levels of certain heavy metals may affect the behavior of birds and mammals. However, most of these studies were carried out under controlled laboratory conditions; results from free-living populations are scarce. In this study we examined the possible effects of exposure to high heavy metal concentrations on the aggressive behavior of resident great tit (Parus major) males by means of simulated territorial intrusion experiments during the egg-laying and incubation period at a highly polluted and a reference site. We also examined the effects of heavy metal exposure on the breeding performance of the tested couples, and we examined the plasma testosterone concentrations of great tit males in both study sites. We found no significant differences in the aggressive behavior between great tits at the polluted and the reference site during the egg-laying period. During the incubation period, responding male great tits at the polluted site performed significantly more aggressive flights around the decoy than did males at the reference site. During this period, there was also a tendency that more responding males were out of sight of the observer for some time during the experiment at the reference site than at the polluted site and this also for a longer period of time. Other behavioral parameters did not differ significantly between the two sites. Although plasma testosterone levels were higher at the polluted site, they did not differ significantly between the two sites. The tested couples at the polluted site interrupted their laying sequence significantly more and hatching success was significantly reduced compared to the tested birds at the reference site. We found no significant difference in clutch size, the length of the laying interruptions, fledging success, and fledgling age between the two sites. Although our results showed only minor alterations in the territorial aggressive behavior of male great tits exposed to heavy metal pollution, these small changes may have an important influence on the breeding and survival of these birds.

Variation of heavy metals within and among feathers of birds of prey: effects of molt and external contamination.

In this study we examined the effect of external contamination on the heavy metal (Ag, Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn) concentration in feathers. We compared the heavy metal content among the 10 primary wing feathers of sparrowhawks (Accipiter nisus), little owls (Athene nocta) and barn owls (Tyto alba) and the variation within the outermost tail feather of sparrowhawks and tawny owls (Strix aluco). The concentration of Hg was significantly higher in feathers molted first, suggesting that levels in feathers reflect levels in the blood during formation. For some other elements (Al, Co, Ni, Pb, Zn) on the other hand, there are strong indications that external contamination may have an important impact on the levels detected in the feathers. This should be taken into account in future monitoring studies.

Seasonal variation in androgen-metabolizing enzymes in the diencephalon and telencephalon of the male European starling (Sturnus vulgaris).

In seasonally breeding songbirds, seasonal fluctuations occur in serum testosterone (T) concentrations and reproductive behaviours. Many T-dependent behaviours are regulated by the activity of androgenic and oestrogenic metabolites within specific brain regions. Male European starlings breed in spring when circulating T concentrations peak. T and its metabolites act within portions of the diencephalon to regulate the pituitary-gonadal axis and to activate courtship and copulation. Song in male starlings is critical for mate attraction during the breeding season and is regulated by steroid-sensitive nuclei in the telencephalon and diencephalon. Outside the breeding season, T is undetectable, however, males continue to sing at high levels. This suggests that singing outside of the breeding season might not be T-dependent as it appears to be in the spring. Alternatively, singing when T is low might continue to be regulated by T due to increased sensitivity of the brain to the action of the steroid. This increased sensitivity could be mediated by changes in intracellular T metabolism leading to increased production of active or decreased production of inactive metabolites. To explore the relationship between T-metabolism and reproductive behaviour, we analysed seasonal changes in the activity of four brain T-metabolizing enzymes: aromatase, 17beta-hydroxysteroid dehydrogenase (17beta-HSDH), 5alpha-reductase (all three convert T into active metabolites) and 5beta-reductase (converts T into an inactive metabolite) in the diencephalon and telencephalon. In the anterior and posterior diencephalon, the highest aromatase was observed in spring when this region is critical for courtship and copulation. In the telencephalon, aromatase was highest and 5beta-reductase was lowest throughout the winter months well prior to the reproductive season and these enzymes presumably maximize T-activity within this region. Although these data do not indicate whether the metabolic changes occur specifically within song nuclei, these findings are compatible with the idea that singing in male starlings outside the breeding season may be regulated by steroids despite the presence of low serum T concentrations. Overall, seasonal changes in T-metabolizing enzymes appear to play a significant role in seasonal changes in behaviour and reproductive physiology.

Seasonal changes in courtship song and the medial preoptic area in male European starlings (Sturnus vulgaris).

In male starlings (Sturnus vulgaris) courtship song plays a critical role in mate attraction. During the breeding season courtship song occurs prior to copulation and appears to reflect male sexual arousal. Outside the breeding season starlings sing, but song appears unrelated to reproduction. The aromatization of testosterone (T), likely within the medial preoptic nucleus (POM), is critical for the expression of male sexual arousal. The present study was performed to determine whether seasonal changes in the POM might relate to seasonal changes in courtship singing behavior in male starlings. T concentrations, the volume of the POM, and aromatase within the POM were examined both during and outside of the breeding season in male starlings. Song was also recorded at these times both with and without a female present. The POM was largest and contained dense aromatase immunostaining only during the spring breeding season, when T concentrations were highest and males responded to a female with an increase in courtship song. Outside the breeding season the volume of the POM was small, T concentrations were low, and males displayed no changes in song expression in response to female conspecifics. Song bout length was positively related to POM volume, and males sang longer songs in spring. Only males with nestboxes in spring responded to a female, and the POM tended to be larger in these males, suggesting that nestbox possession might influence neuroplasticity within the POM. Overall, the findings suggest that T-dependent plasticity and aromatase activity within the POM might regulate courtship singing in a wild songbird.

Sex-role reversal in vertebrates: behavioural and endocrinological accounts.

Sex-role reversal occurs when females compete more intensely than males for access to mates. In this paper, we survey the occurrence of sex-role reversal in vertebrates: we focus on behavioural aspects of sex-role reversal and we examine possible endocrinological correlates of this phenomenon. The best documented cases among vertebrates of sex-role reversal occur in fish and birds. In nearly all sex-role reversed species or populations, females have higher potential reproductive rates than males. Some species in which females were previously thought to be the predominant competitors for mates (for instance seahorses and a dendrobatid frog), appear not to be sex-role reversed according to recent studies. The endocrinology of sex-role reversal has been studied in only a few species and therefore remains poorly understood. In birds, which probably have been studied the most in this respect, steroid hormones appear to follow the typical ancestral conditions (for instance no reversal of testosterone levels) in sex-role reversed species, whereas prolactin, a principal regulator of the onset and maintenance of incubation, departs from the usual avian pattern in that it is higher in males than in females. The study of sex-role reversed behaviour offers unique opportunities not only to test sexual selection theory, but also to enhance our understanding of the neuroendocrine mechanisms mediating behavioural sex differences.

Can excrement and feathers of nestling songbirds be used as biomonitors for heavy metal pollution?

Although birds have been frequently used as indicators of heavy metal pollution, few studies have examined pollutant levels in nestling passerines. In this paper we determined the levels of two essential (zinc and copper) and three nonessential heavy metals (lead, cadmium, and arsenic) in the excrement and feathers of great (Parus major) and blue tit (Parus caeruleus) nestlings at a polluted site (near a metallurgic factory) and a reference site (4 km farther east). The excrement of both great and blue tit nestlings contained significantly higher concentrations of arsenic, cadmium, copper, and lead at the polluted site. Zinc concentrations did not differ significantly between sites for both species. The feathers of great and blue tit nestlings accumulated significantly higher concentrations of lead at the polluted site than at the reference site. Zinc levels in the feathers of great tit nestlings were significantly higher at the reference site than at the polluted site. For all other elements considered, concentrations did not differ significantly between the two sites. There were no interspecific differences in metal levels between great and blue tits in both excrement and feathers. There was a significant positive correlation between the lead concentration in the excrement and feathers for both great and blue tits. We therefore conclude that excrement of great and blue tit nestlings can be used as a biomonitor for heavy metals (lead, cadmium, arsenic, and copper), whereas feathers appear only to be suitable as a biomonitor for lead pollution.

Shield characteristics are testosterone-dependent in both male and female moorhens.

The immunocompetence handicap hypothesis proposes that the expression of secondary sexual characteristics is positively related to testosterone levels, but that elevated testosterone levels also impose costs from immune suppression. Hence, testosterone-dependent characteristics should accurately reflect male quality because only high-quality males are able to invest in large sexual characteristics without detrimental effects upon their own immune system. Most studies to date have focused on the role of testosterone in the expression of male ornaments and on the possible immunosuppressant effects of androgens in males. In the moorhen (Gallinula chloropus), a sexually monomorphic monogamous bird species showing a partial sex-role reversal, both sexes have a prominent frontal shield. We implanted both sexes with testosterone-filled implants to examine the effects of testosterone on shield characteristics and immune function. Shield size, thickness, and color were significantly increased by an experimental increase in testosterone concentrations in both males and females. Likewise, removal of the implants led to a rapid decrease in shield size and thickness in both males and females, suggesting that both sexes responded quickly to an increase or a decrease in testosterone. Moorhens implanted with testosterone had higher intensities of ectoparasite infestations than control birds, but other indirect measures of immunocompetence did not differ significantly between the two categories of birds.

Great and blue tits as indicators of heavy metal contamination in terrestrial ecosystems.

Although there are abundant data on heavy metal contamination in the feathers of seabird species and birds of prey, relatively few studies have addressed the use of passerine birds as indicators of point-source contamination. Concentrations of three essential elements (Al, Zn, and Cu) and two toxic metals (Cd and Pb) were determined in the outer tail feathers of great and blue tits (Parus major and Parus caeruleus), two small insectivorous passerines, at a presumed polluted site (in an urban area close to household waste incinerators) and at a reference site (in a nature reserve). For the great tit, both Cd and Cu levels were significantly higher at the polluted site than in the nature reserve. For the blue tit, Cd, Pb, and Cu levels were significantly higher at the polluted site than at the reference site. There were striking differences in metal levels between blue and great tits inhabiting the same polluted site: lead, zinc, and copper were significantly higher in the smaller blue tit than in the great tit. We suggest that tit (Parus) species, which have been important models in behavioral and ecological research, may be very useful as biomonitors for local contamination.

Copulation and mate-guarding patterns in polygynous European starlings

We recorded the timing and frequency of copulation and mate-guarding behaviour during the fertile period in relation to day of first ovulation and time of day in the facultatively polygynous European starling, Sturnus vulgarisAll within-pair copulations were female-solicited. Females solicited them at a high rate during both the pre-ovulatory and ovulatory period, but there were differences in solicitation rates between monogamous, primary and secondary females. Extra-pair copulation attempts were all male-initiated and females rarely engaged in them. Attempts by males were most frequent during the ovulatory period, when the probability of extra-pair fertilization was highest. Males intensively guarded their mate during both the pre-ovulatory and ovulatory period. In contrast to most other passerines, female starlings lay their eggs not at dawn but usually between 0900 and 1100 hours, implying that they may be most fertile in the late morning. During the ovulatory period, extra-pair copulation attempts were more frequent in the late morning than in the early morning, suggesting that males try to take advantage of this putative 'insemination window'. Accordingly, there was some indication that mate guarding during the ovulatory period was more intense in the late than early morning. Females solicited within-pair copulations at a higher rate in the early morning than in the late morning, but copulation rate did not differ between early and late morning. The proportion of monogamous and polygynous males attempting extra-pair copulations did not differ significantly, but mate-guarding intensity differed significantly between female categories. Primary females were guarded less intensively than monogamous females suggesting that males trade mate guarding against mate attraction/courting additional females.