MC1R-dependent, melanin-based colour polymorphism is associated with cell-mediated response in the Eleonora's falcon.

Colour polymorphism in vertebrates is usually under genetic control and may be associated with variation in physiological traits. The melanocortin 1 receptor (Mc1r) has been involved repeatedly in melanin-based pigmentation but it was thought to have few other physiological effects. However, recent pharmacological studies suggest that MC1R could regulate the aspects of immunity. We investigated whether variation at Mc1r underpins plumage colouration in the Eleonora's falcon. We also examined whether nestlings of the different morphs differed in their inflammatory response induced by phytohemagglutinin (PHA). Variation in colouration was due to a deletion of four amino acids at the Mc1r gene. Cellular immune response was morph specific. In males, but not in females, dark nestling mounted a lower PHA response than pale ones. Although correlative, our results raise the neglected possibility that MC1R has pleiotropic effects, suggesting a potential role of immune capacity and pathogen pressure on the maintenance of colour polymorphism in this species.

Physiological stress links parasites to carotenoid-based colour signals.

Vertebrates commonly use carotenoid-based traits as social signals. These can reliably advertise current nutritional status and health because carotenoids must be acquired through the diet and their allocation to ornaments is traded-off against other self-maintenance needs. We propose that the coloration more generally reveals an individual's ability to cope with stressful conditions. We tested this idea by manipulating the nematode parasite infection in free-living red grouse (Lagopus lagopus scoticus) and examining the effects on body mass, carotenoid-based coloration of a main social signal and the amount of corticosterone deposited in feathers grown during the experiment. We show that parasites increase stress and reduce carotenoid-based coloration, and that the impact of parasites on coloration was associated with changes in corticosterone, more than changes in body mass. Carotenoid-based coloration appears linked to physiological stress and could therefore reveal an individual's ability to cope with stressors.

Stress response during development predicts fitness in a wild, long lived vertebrate.

Short-term elevation of circulating glucocorticosteroids (GCs) in vertebrates facilitates the adoption of a distinct emergency life history state, which allows individuals to cope with perturbations and recover homeostasis at the expense of temporarily suppressing nonessential activities. Although GC responses are viewed as a major evolutionary mechanism to maximize fitness through stress management, phenotypic variability exists within animal populations, and it remains unclear whether interindividual differences in stress physiology can explain variance in unequivocal components of fitness. We show that the magnitude of the adrenocortical response to a standardized perturbation during development is negatively related to survival and recruitment in a wild population of long lived birds. Our results provide empirical evidence for a link between stress response, not exposure to stressors, and fitness in a vertebrate under natural conditions. Recent studies suggest that variability in the adrenocortical response to stress may be maintained if high and low GC responders represent alternative coping strategies, with differential adaptive value depending on environmental conditions. Increased fitness among low GC responders, having a proactive personality, is predicted under elevated population density and availability of food resources, conditions that characterize our study population.

Testosterone increases bioavailability of carotenoids: insights into the honesty of sexual signaling.

Androgens and carotenoids play a fundamental role in the expression of secondary sex traits in animals that communicate information on individual quality. In birds, androgens regulate song, aggression, and a variety of sexual ornaments and displays, whereas carotenoids are responsible for the red, yellow, and orange colors of the integument. Parallel, but independent, research lines suggest that the evolutionary stability of each signaling system stems from tradeoffs with immune function: androgens can be immunosuppressive, and carotenoids diverted to coloration prevent their use as immunostimulants. Despite strong similarities in the patterns of sex, age and seasonal variation, social function, and proximate control, there has been little success at integrating potential links between the two signaling systems. These parallel patterns led us to hypothesize that testosterone increases the bioavailability of circulating carotenoids. To test this hypothesis, we manipulated testosterone levels of red-legged partridges Alectoris rufa while monitoring carotenoids, color, and immune function. Testosterone treatment increased the concentration of carotenoids in plasma and liver by >20%. Plasma carotenoids were in turn responsible for individual differences in coloration and immune response. Our results provide experimental evidence for a link between testosterone levels and immunoenhancing carotenoids that (i) reconciles conflicting evidence for the immunosuppressive nature of androgens, (ii) provides physiological grounds for a connection between two of the main signaling systems in animals, (iii) explains how these signaling systems can be evolutionary stable and honest, and (iv) may explain the high prevalence of sexual dimorphism in carotenoid-based coloration in animals.

Organochlorine contamination and physiological responses in nestling tree swallows in Point Pelee National Park, Canada.

Point Pelee National Park in southwestern Ontario, Canada--a major migratory route and vital breeding area for many birds--has localized areas of organochlorine (OC) contamination from the 1950s and 1960s. During 2002, we investigated the effect of tissue OC contaminant levels on the physiology and growth of nestling tree swallows (Tachycineta bicolor). Polychlorinated biphenyl (PCB) contamination in the nestlings positively correlated with hepatic ethoxy-, benzyloxy-, and pentoxyresorufin-o-dealkylase enzyme activities and liver size. Despite detectable physiologic changes associated with body burdens of PCBs, reproductive success of breeding pairs was not affected. Hatching date was negatively correlated with PCB levels, alkoxyresorufin-O-dealkylase (alkROD) activities, liver size, and serum protein levels. alkROD activities were largely dependent on hatching date because insect prey contaminated with PCBs were only abundant during a limited period of time during the breeding season.

Thyroid hormone suppression and cell-mediated immunomodulation in American kestrels (Falco sparverius) exposed to PCBs.

Exposure to environmental contaminants can induce physiological changes in animals through various mechanisms. One manifestation of subclinical toxicity from polychlorinated biphenyl (PCB) exposure is the disruption of normal immune function described in numerous species, including American kestrels (Falco sparverius). In 1998, 152 mature male and female kestrels were fed either a mixture of Aroclor 1248:1254:1260 (approximately 7 mg/kg kestrel/day) through their food items, or control diets. Offspring produced by 50 breeding pairs (thus, half received in ovo PCB exposure only) were also studied. Total and differential white blood cell counts, the phytohemagglutinin (PHA) skin response, as well as thyroid hormone levels were tested in vivo in nonbreeding adults (1998 only) and nestlings (1998 and 1999). In 1999, nestlings came from three parental groups; adults exposed in 1998, birds produced by PCB-exposed parents, and unexposed birds. In 1998, directly exposed males but not females had increased total white blood cell counts driven by lymphocytosis, plus a decreased heterophil-to-lymphocyte ratio relative to controls. PCB-exposed birds had a significantly greater response to PHA than did controls, with sex as a significant factor and plasma triiodothyonine (T(3)) as a significant covariate. Levels of T(3) were significantly depressed in PCB-exposed birds of both sexes. The 1999 nestlings (F1 generation with respect to PCB exposure) did not show any effect of parental treatment group on the PHA skin response, yet T(3) remained as a significant covariate. Immunological effects are discussed in light of the antibody-mediated immunotoxicity found in the same birds and reported previously.

Bioaccumulation and toxicokinetics of 42 polychlorinated biphenyl congeners in American kestrels (Falco sparverius).

The bioaccumulation and toxicokinetics of 42 polychlorinated biphenyls (PCBs) was determined in male American kestrels exposed to an Aroclor-contaminated diet for 120 d followed by a 348-d depuration period. The birds were housed under ambient outdoor temperatures to permit normal fluctuations in body weight during the study. Whole body PCB clearance, plasma/fat distribution coefficients, and plasma PCB clearance constants were determined for individual PCBs to calibrate a two-compartment rate constant model in order to describe PCB elimination in the birds. Plasma/fat partition coefficients (K(PF)) averaged 0.0060 +/- 0.0001 for all congeners of study, were not dependent on chemical hydrophobicity, and did not change in summer versus winter sacrificed animals. Plasma clearance constants (k'pc) for PCBs were observed to be dependent on both chlorine substitution patterns and congener hydrophobicity. Polychlorinated biphenyl congeners categorized as readily cleared congeners contained vicinal meta-para hydrogen substituents on at least one phenyl ring, while slowly cleared congeners were chlorine hindered at these positions. A general equation was derived to predict plasma clearance constants for all tri- to octachlorobiphenyls based on the presence of an open meta-para site on one of the phenyl rings and from the n-octanol-water partition coefficient of the chemical. The equation was validated by comparing predicted versus measured relative biomagnification factors of PCBs determined in birds at the end of the dosing period. The two-compartment model calibrated for PCB elimination in American kestrels may be used to describe PCB toxicokinetics in wild birds provided that seasonal fluctuations in the fat content of the modeled population is known.

Hemograms for and nutritional condition of migrant bald eagles tested for exposure to lead.

Plasma proteins, hematocrit, differential blood counts were examined and nutritional condition was estimated for bald eagles (Haliaeetus leucocephalus) trapped (n = 66) during antumn migration, 1994-95 at Galloway Bay (Saskatchewan, Canada), for the purposes of estimating prevalence of exposure to lead. Sex and age differences in hematocrit and plasma proteins were not observed; however, female eagles exhibited larger median absolute heterophil counts than males. Hematologic values were similar to those previously reported from eagles in captivity. Departures from expected hematological values from a healthy population of eagles were not observed in birds with elevated levels of blood lead (> or =0.200 microg/ml). Similarly, nutritional condition was not related to blood-lead concentrations. Therefore, it appears that lead exposure in this population was below a threshold required to indicate toxicological alteration in the hematological values and index of nutritional condition that we measured.

In ovo exposure to polychlorinated biphenyls: reproductive effects on second-generation American kestrels.

The reproductive success of wild birds has been affected by exposure to multiple contaminants. Reproduction of captive American kestrels (Falco sparverius) was suppressed when adult birds were exposed to dietary polychlorinated biphenyls (PCBs). In this study, the reproductive effects of in ovo exposure to PCBs is explored, along with determining effects on reproduction in second-generation birds indirectly exposed to PCBs. Reproductive changes in this subsequent generation are examined separately in male and female birds. Captive American kestrels (F. sparerius) were hatched from clutches with eggs containing environmentally relevant levels of total PCBs (34.0 microg/g whole egg WW versus 0 microg/g controls); parent birds had been fed PCB-spiked (Aroclor 1248:1254:1260) food (7 mg/kg BW day(-1)) for 100 days until their eggs hatched. In 1999, the second-generation PCB birds were paired with unexposed kestrels having reproductive experience. In ovo PCB exposure suppressed egg laying completely in 25% of PCB females and resulted in delays in clutch initiation and smaller clutch sizes for PCB male and female pairs. There was no evidence in this study of in ovo PCB treatment effects on fertility or hatching success. The decline in reproductive success was also reflected in the reduced fledging success and higher incidence of complete brood mortality of PCB nestlings. Differences between in ovo-exposed PCB males and females but not between controls were evident in reproductive success. In ovo PCB exposure appears to have had greater effects on female kestrels until clutch completion, with a greater time lag between pairing and egg laying, reduced numbers of pairs laying eggs, and smaller clutches being laid. In ovo PCB exposure has greater effects later in the breeding season on male kestrels, which had poorer hatching and fledging success relative to the PCB females. Possible behavioral and physiological mechanisms involved in these reproductive changes are discussed.

Courtship behavior of captive American kestrels (Falco sparverius) exposed to polychlorinated biphenyls.

Polychlorinated biphenyls (PCBs) adversely affect reproduction in birds. Captive adult male and female American kestrels (Falco sparverius) were studied to investigate the potential behavioral and hormonal alterations during the courtship period resulting from clinical exposure to PCBs. American kestrels ingested 7 mg/kg/body weight/bird/day of a 1:1:1 mixture of Aroclors 1248, 1254, and 1260 through their diet of day-old cockerels. The dietary dosage of Aroclors resulted in environmentally relevant total PCB residues in the eggs, averaging 34.1 microg/g wet weight (geometric mean). There was no difference between treatment and control birds in the circulating levels of total androgens (p = 0.44) or in 17 beta-estradiol (p = 0.29), one week following pairing. Male kestrels exposed to dietary PCBs exhibited significantly more sexual behaviors (p = 0.034) and flight behaviors (p = 0.026) than the control males. Sexual behaviors of male kestrels included; nest-box inspections, solicitation of copulation, the offer of food to the female, and giving the female food. The flight behaviors of the male included; flying from one perch to another and aerial display. In addition, the frequency of male sexual behaviors were correlated (r = 0.605, p = 0.001) with total PCB residues in the eggs of their mates. A concurrent study found that these same PCB-exposed kestrels experienced a delay in clutch initiation as well as a greater number of completely infertile clutches.

Reproduction success of American kestrels exposed to dietary polychlorinated biphenyls.

While reproduction of wild birds is adversely affected by multiple environmental contaminants, we determined that polychlorinated biphenyls (PCBs) alone alter reproduction. Captive American kestrels (Falco sparverius), fed PCB-spiked (Aroclor 1248:1254:1260) food (7 mg/kg body weight/d) prior to and during the first breeding season only (100 d) laid eggs with environmentally relevant levels of total PCBs (34.0 microg/g whole egg wet wt vs 0 microg/g for controls). Reproduction changed during, not after, PCB exposure in this two-year study. The PCB-exposed pairs laid smaller clutches later in the season and laid more totally infertile clutches. Hatching success was reduced in PCB-exposed pairs, and 50% of PCB nestlings died within 3 d of hatching. Nearly 60% of PCB-exposed pairs with hatchlings failed to produce fledglings. Higher levels of total PCB residues and congeners were associated with later clutch initiation and fewer fertile eggs, hatchlings, and fledglings. We suggest that nonpersistent PCB congeners have a greater influence on reproduction than do persistent congeners.

Carotenoid discrimination by the avian embryo: a lesson from wild birds.

The concentrations (microg/g wet yolk) of total carotenoids in eggs of the common moorhen (Gallinula chloropus), American coot (Fulica americana) and lesser black-backed gull (Larus fuscus), collected in the wild, were 47.5, 131.0 and 71.6, respectively. In contrast to data for eggs of the domestic chicken, beta-carotene was a significant component in the yolks of these three wild species, forming 25-29% by wt. of the total carotenoids present. The concentration of total carotenoids in the livers of the newly-hatched chicks was 5-10 times higher than in the other tissues and beta-carotene was again a major component, forming 37-58% of the hepatic carotenoids. In the newly-hatched gull, the proportions of both lutein and zeaxanthin were very low in the liver but high in the heart and muscle when compared with the yolk. By contrast canthaxanthin, echinenone and beta-carotene were very minor constituents of heart and muscle when compared with their proportions in the yolk of the gull. The proportions of lutein and zeaxanthin in the liver of the newly-hatched coot and moorhen were also far lower than in the yolk whereas the liver was relatively enriched with beta-cryptoxanthin, beta-carotene and (in the moorhen) echinenone. The results indicate that avian embryos discriminate between different carotenoids during their distribution from the yolk to the various tissues.

Exposure of migrant bald eagles to lead in prairie Canada.

The prevalence of elevated exposure to lead was assessed in a migrant population of bald eagles (Haliaeetus leucocephalus) at a waterfowl staging area in the southern portion of the Canadian prairies, from September to November, 1992-1995. Of 103 eagles, 8% exhibited blood lead (PbB) concentrations suggestive of elevated exposure to lead (> or = 0.200 microgram ml-1 wet wt.). PbB concentrations in eagles from the study area ranged from < 0.01 to 0.585 microgram ml-1, while those of nestling eagles from a reference site indicated normal or background exposure (< 0.01 microgram ml-1). No differences in the prevalence of elevated exposure were detected among genders or age classes (0.5- and > or = 1.5-year-old birds) (P > 0.05). The prevalence of elevated exposure was significantly greater in November than in October (21.7 vs. 3.8%) (all years: chi 2Y = 5.75, P = 0.017). Eagles with shotshell pellets in the digestive tract did not have accompanying high PbB concentrations. The prevalence of elevated lead exposure in this study was low in comparison to other areas in North America. Potential biases in the trapping technique as they relate to interpreting the results are addressed.

Captivity diets alter egg yolk lipids of a bird of prey (the American kestrel) and of a galliforme (the red-legged partridge).

The salient feature of the fatty acid profile of kestrel eggs collected in the wild was the very high proportion of arachidonic acid (15.2%+/-0.7% of fatty acid mass, n=5) in the phospholipid fraction of the yolk. Kestrels in captivity fed on day-old chickens produced eggs that differed from those of the wild birds in a number of compositional features: the proportion of linoleic acid was increased in all the lipid fractions; the proportion of arachidonic acid was increased in yolk phospholipid and cholesteryl ester; the proportion of alpha-linolenic acid was decreased in all lipid classes, and that of docosahexaenoic acid was decreased in phospholipid and cholesteryl ester. Partridge eggs from the wild contained linoleic acid as the main polyunsaturate of all the yolk lipid fractions. Captive partridges maintained on a formulated diet very rich in linoleic acid produced eggs with increased levels of linoleic, arachidonic, and n-6 docosapentaenoic acids in the phospholipid fraction; reduced proportions of alpha-linolenic acid were observed in all lipid classes, and the proportion of docosahexaenoic acid was markedly reduced in the phospholipid fraction. Thus, captive breeding of both the kestrel and the partridge increases the n-6/n-3 polyunsaturate ratio of the yolk lipids.

Antibody-mediated immunotoxicity in American kestrels (Falco sparverius) exposed to polychlorinated biphenyls.

Antibody-mediated immune function in adult and recently fledged (30 to 33 d old) American kestrels (Falco sparverius) was examined in birds exposed directly, or only in ovo, to polychlorinated biphenyls (PCBs). In 1998, 9 mature male and 9 female kestrels were fed PCBs, whereas 9 females and 10 males served as controls. A mixture of Aroclors 1248:1254:1260 suspended in safflower oil was injected into the kestrels' food items, while in control diets only the same volume of oil was added. The dosage of PCBs was approximately 7 mg/kg kestrel/d, beginning in March 1998 and continuing for 120 d. In 1998, the antibody-mediated immune response was stimulated by immunization and booster vaccinations of the kestrels using a nonpathogenic antigen, dinitrophenol-keyhole limpet hemocyanin (DNP-KLH). In 1999, offspring from three treatment groups based upon maternal exposure to PCBs were similarly tested for their antibody response. None of these mothers was vaccinated with DNP-KLH the previous year. The maternal groups were: (1) exposed to PCBs in 1998 for 120 d, (2) exposed in ovo in 1998 (i.e., mothers were produced by PCB-exposed parents), or (3) unexposed to PCBs. Serum antibody levels were determined using an enzyme-linked immunosorbent assay (ELISA). In 1998, PCB-exposed adult females had a significantly higher antibody response than did controls, whereas adult males exposed to PCBs had significantly suppressed antibody production. For the nestlings produced in 1999, maternal treatment significantly affected antibody response. Generally, the antibody response in the nestlings was much lower than that seen in adult kestrels. Yet both male and female offspring from mothers that had been fed PCBs the previous year had significantly higher postbooster anti-DNP-KLH titers than control and in ovo-exposed maternal groups, thus mimicking the response seen in the adult females the previous year. These sex-specific responses in PCB-exposed birds provide further evidence of the endocrine-disrupting behavior of PCBs. Both suppression and stimulation of the antibody response are undesirable because this indicates that the immune system is not able to respond normally to challenges by infectious or other disease-causing agents.

Genetics, local environment and health as factors influencing plasma carotenoids in wild American kestrels (Falco sparverius).

Carotenoids are important as pigments for bright coloration of animals, and as physiologically active compounds with a wide array of health-related functions. Carotenoid-dependent coloration may have evolved as a signal to conspecifics; however, factors that may limit availability of carotenoids are poorly known. We investigated how the acquisition of carotenoids may be constrained by availability in the environment, diet, genetic make-up and health status of wild American kestrels (Falco sparverius). Plasma concentrations of siblings at the time of fledging showed a high degree of resemblance; however, a cross-fostering experiment revealed that variance was largely explained by nest of rearing, rather than nest of origin, thus indicating a low genetic component. A multivariate analysis of attributes of nestlings (sex, size, plasma proteins, immune function), parental reproduction (laying date, clutch size) and rearing conditions (brood size, size hierarchy, nestling mortality) showed only a small significant effect of leucocyte differentials on carotenoid concentrations of nestlings. A strong environmental effect on plasma carotenoids was demonstrated by levels of adult kestrels being correlated within mated pairs, and having a significant association with the abundance of voles, the primary prey species, per territory.

Changes in egg composition of American kestrels exposed to dietary polychlorinated biphenyls.

Changes in the quality of eggs of birds exposed to polychlorinated biphenyls (PCBs) have been described, but have never been directly attributed to PCBs. Polychlorinated biphenyl residues in eggs have been associated with reduced reproductive success and embryonic deformities in wild birds. Egg size and composition, specifically the amount of albumen, yolk, and water in an egg, also influence the growth and viability of embryos and hatchlings, and consequently the reproductive success of birds. To deter mine whether PCB exposure of adult birds affected the size and composition of their eggs, 25 pairs of captive American kestrels (Falco sparverius) were fed a mixture of PCB-spiked (1248:1254:1260) food to give an approximate exposure of 7 mg/kg body weight/d, beginning 1 mo prior to pairing, and continuing throughout the courtship, egg-laying, and incubation periods. This dietary level in the adult female kestrels resulted in mean total PCB residues in the eggs of 34.1 microg/g wet weight (geometric mean), which is environmentally relevant. PCB residues in eggs increased with the time of female exposure to the contaminated diet and laying date. Variation in egg size within PCB clutches was significantly greater than within control clutches, although absolute egg mass and volume did not differ markedly by treatment. Only infertile eggs and only one egg per clutch were used for egg composition analysis. Yolks in the PCB-contaminated eggs were heavier, with less wet and dry albumen relative to control eggs. Water content and eggshell thickness were not significantly affected by PCB exposure. These results suggest that eggs from the PCB treatment have relatively more lipid and less protein available for embryonic development. Changes in egg composition were not associated with egg size, lay date, ambient temperature, humidity, or precipitation, which are factors known to affect these variables in bird eggs. The PCB-induced changes in egg composition described here provide insight into possible mechanisms contributing to reduced reproductive performance in wild birds exposed to PCBs.

The T-cell-mediated immune response and return rate of fledgling American kestrels are positively correlated with parental clutch size.

Life-history theory predicts that parents face a trade-off between the number and viability of the progeny they produce. We found evidence for an apparent trade-off in a free-living population of American kestrels (Falco sparverius), as larger clutches produced more but lighter fledglings. However, while the body mass of fledglings has traditionally been used as a measure of survival prospect, offspring immunocompetence should also play an important role. We thus measured the T-cell-mediated immune response of fledgling kestrels in relation to brood traits and nest-rearing conditions through a cross-fostering experiment. The immune response was positively correlated with the body condition of fledglings, but was also higher in those hatched from five-egg than four-egg clutches. These results were not influenced by other brood traits, nor by current exposure to stressors and infectious agents, as measured by serological variables. Such ability to resist pathogens may account for why the probability of offspring returning to the study area in subsequent years, when controlling for brood size, was higher for five-egg than four-egg clutches. These results suggest an optimal clutch size through maternal effects on offspring immunocompetence rather than a trade-off between the number and quality of the offspring.

Environmental and genetic variation in T-cell-mediated immune response of fledgling American kestrels.

We investigated genetic and environmental components of variance in avian T-cell-mediated immune response (CMI) through a cross-fostering experiment conducted on wild American kestrels (Falco sparverius). CMI was evaluated in vivo by an experimental challenge with phytohaemagglutinin, a T-cell mitogen, injected intradermally in fledglings. Additionally, we assessed two measures of nutritional condition (body mass and circulating plasma proteins) which could influence the variance components of CMI. A two-way nested ANOVA indicated that CMI of fledgling kestrels was explained more by the nest where the bird was reared (33% of the explained variance) than by the nest of origin (12%). Body mass was explained equally by familial and environmental components, while plasma proteins were only related to the rearing environment. CMI of fledglings was not related to their circulating plasma proteins, but was positively correlated with their body mass. Fledgling body mass seemed to be influenced by pre-hatching or post-hatching maternal effects prior to manipulation since resemblance in body mass of sibships at the age of manipulation was high (h 2≤0.58), and body mass at this age predicted body mass at fledging. Therefore, pre-manipulation parental effects on body mass, such as investment in egg size, could have inflated the familial effects on body mass of fledglings and then on its correlated CMI. When controlling for body mass, most of the variation in CMI of fledglings was explained by the nest where the bird was reared (36.6%), while the variance explained by the nest of origin (4%) was not significant. This means that environmental influences are major determinants of offspring CMI. The low proportion of variance explained by the familial component may have been due to the high correlation of CMI to fitness.