Testing the shared-pathway hypothesis in the carotenoid-based coloration of red crossbills

The mechanisms involved in the production of red carotenoid-based ornaments of vertebrates are still poorly understood. These colorations often depend on enzymatic transformations (ketolation) of dietary yellow carotenoids, which could occur in the inner mitochondrial membrane (IMM). Thus, carotenoid ketolation and cell respiration could share biochemical pathways, favoring the evolution of ketocarotenoid-based ornaments as reliable indices of individual quality under sexual selection. Captive male red crossbills (Loxia curvirostra Linnaeus) were exposed to redox-active compounds designed to penetrate and act in the IMM: an ubiquinone (mitoQ) or a superoxide dismutase mimetic (mitoTEMPO). MitoQ can act as an antioxidant but also distort the IMM structure, increasing mitochondrial free radical production. MitoQ decreased yellow carotenoids and tocopherol levels in blood, perhaps by being consumed as antioxidants. Contrarily, mitoTEMPO-treated birds rose circulating levels of the second most abundant ketocarotenoid in crossbills (i.e., canthaxanthin). It also increased feather total red ketocarotenoid concentration and redness, but only among those birds exhibiting a redder plumage at the start of the study, that is, supposedly high-quality individuals. The fact that mitoTEMPO effects depended on original plumage color suggests that the red-ketocarotenoid-based ornaments indicate individual quality as mitochondrial function efficiency. The findings would thus support the shared pathway hypothesis.     

Relationships between hair melanization, glutathione levels, and senescence in wild boars

The synthesis of melanins, which are the most common animal pigments, is influenced by glutathione (GSH), a key intracellular antioxidant. At high GSH levels, pheomelanin (the lightest melanin form) is produced, whereas production of eumelanin (the darkest melanin form) does not require GSH. Oxidative damage typically increases with age, and age-related decreases in GSH have accordingly been found in diverse organisms. Therefore, there should be positive associations between the capacity to produce eumelanic traits, GSH levels, and senescence, whereas there should be negative associations between the capacity to produce pheomelanic traits, GSH levels, and senescence. We explored this hypothesis in a free-ranging population of wild boars Sus scrofa of different ages. As expected from the fact that pheomelanogenesis consumes GSH, levels of this antioxidant in muscle tended to be negatively related to pheomelanization and positively related to eumelanization in pelage, and the degree of pelage pheomelanization was positively related to oxidative damage as reflected by levels of thiobarbituric-acid-reactive substances (TBARS), which is consistent with the hypothesis that pheomelanin synthesis has physiological costs. In our cross-sectional sample, GSH levels did not show senescence effects, and we did not detect senescence effects in pelage melanization. Prime body condition and low TBARS levels were also associated with hair graying, which is attributable to a loss of melanin produced by oxidative stress, thus raising the possibility that hair graying constitutes a signal of resistance to oxidative stress in wild boars. Our results suggest that the degree of melanization is linked to GSH levels in wild boars and that their antioxidant damage shows senescence effects.     

Variation in plasma biochemical parameters in captive adult red-legged partridges (Alectoris rufa) during daylight hours

Blood biochemical parameters may provide useful information about the physical condition of the individual, making them a useful tool in ecological studies. However, to avoid biases, factors affecting circulating levels of plasma metabolites must be investigated. In this paper, we analyse the effect of daytime sampling hour on seven plasma metabolites in adult red-legged partridges (Alectoris rufa) with free access to food during the breeding season. We found that sampling hour affected circulating levels of glucose and triglycerides but not those of cholesterol, total protein, uric acid, urea or albumin. A sex effect was found only for glucose, uric acid, urea and triglycerides. Repeated sampling affected all the parameters studied. These results suggest that the effect of sex, sampling hour and repeated sampling should be carefully controlled for (methodologically or by statistical procedures) to avoid undesirable sources of error at least in some blood parameters.     

Avoiding bad genes: oxidatively damaged DNA in germ line and mate choice

August Weismann proposed that genetic changes in somatic cells cannot pass to germ cells and hence to next generations. Nevertheless, evidence is accumulating that some environmental effects can promote heritable changes in the DNA of germ cells, which implies that some somatic influence on germ line is possible. This influence is mostly detrimental and related to the presence of oxidative stress, which induces mutations and epigenetic changes. This effect should be stronger in males due to the particular characteristics of sperm. Here, we propose the hypothesis that females are able to avoid males with oxidatively damaged DNA in the germ line by using oxidative-dependent (pre- and post-mating) signals. This new hypothesis may shed light on unsolved questions in evolutionary biology, such as the benefits of polyandry, the lek paradox, or the role of sexual selection on the evolution of aging.     

A biochemical study of fasting, subfeeding, and recovery processes in yellow-legged gulls

An investigation of the effects of fasting, subfeeding, and refeeding on plasma biochemistry was carried out on 22 captive yellow-legged gulls Larus cachinnans Pallas. These birds showed the same fasting endurance model described in other species, but with an important decrease in glucose plasma concentration and very great differences between individuals when reaching the deterioration limit, suggesting a moderate physiological adaptation to long periods of fasting. A different model was proposed in subfed gulls in relation to fasted gulls, based on lipid and protein use, which could be reflected by changes in nitrogen wastes and triglyceride levels in this experiment. Thus, the subfed gulls might use protein directly from the diet as an energy source, thereby reducing the use of fat stores. The gulls quickly recovered body mass during the refeeding period, but while some plasma substances quickly reached their initial values, others showed many changes before the end of the experiment, which could reflect a process of metabolic restabilization. These results contribute to a better knowledge of fasting, subfeeding, and refeeding processes in birds and can be added to a recent study about fasting in gulls.     

Testosterone and oxidative stress: the oxidation handicap hypothesis

Secondary sexual traits (SST) are usually thought to have evolved as honest signals of individual quality during mate choice. Honesty of SST is guaranteed by the cost of producing/maintaining them. In males, the expression of many SST is testosterone-dependent. The immunocompetence handicap hypothesis has been proposed as a possible mechanism ensuring honesty of SST on the basis that testosterone, in addition to its effect on sexual signals, also has an immunosuppressive effect. The immunocompetence handicap hypothesis has received mixed support. However, the cost of testosterone-based signalling is not limited to immunosuppression and might involve other physiological functions such as the antioxidant machinery. Here, we tested the hypothesis that testosterone depresses resistance to oxidative stress in a species with a testosterone-dependent sexual signal, the zebra finch. Male zebra finches received subcutaneous implants filled with flutamide (an anti-androgen) or testosterone, or kept empty (control). In agreement with the prediction, we found that red blood cell resistance to a free radical attack was the highest in males implanted with flutamide and the lowest in males implanted with testosterone. We also found that cell-mediated immune response was depressed in testosterone-treated birds, supporting the immunocompetence handicap hypothesis. The recent finding that red blood cell resistance to free radicals is negatively associated with mortality in this species suggests that benefits of sexual signalling might trade against the costs derived from oxidation.     

Repeated sampling but not sampling hour affects plasma carotenoid levels

Carotenoid pigments have become a central subject of research on animal signaling systems during the past decade. Thus, measurement of plasma carotenoid levels is widespread in the literature. Many plasma biochemical parameters tend to vary with the hour of sampling, which may be an important source of sampling error. However, little is known about this kind of variation for circulating carotenoids. With a sampling protocol that allowed us to separately analyze the effect of sampling hour and repeated blood extraction at the within-individual level, we evaluated the effect of these two parameters on plasma carotenoid concentration in captive red-legged partridges (Alectoris rufa). The hour of sampling did not show a significant influence on carotenoid variability. Nevertheless, carotenoid levels significantly decreased as a result of repeated sampling. In fact, carotenoid variability was positively related to hematocrit values, which also decreased throughout the experiment. Furthermore, the effect was evident from the second sampling event. These results suggest that blood samples for carotenoid analysis may be obtained at random during the daytime with no serious risk of adding variance to data obtained. However, the effect of hemodilution associated with repeated blood extraction should be carefully considered in those studies involving repeated sampling.     

The effects of testosterone manipulation on the body condition of captive male yellow-legged gulls

Persistently high testosterone levels are believed to be costly to males due to their negative effect on body condition. However, this assumption could not be validated when we analysed birds isolated from all social interactions. The hypothesis was tested on birds kept in isolation in order to analyse the effects of testosterone per se, and thereby exclude the influence of social interactions. Adult male yellow-legged gulls (Larus cachinnans) were captured, and after a period of adjustment, some individuals were subcutaneously implanted with testosterone, while the rest were used as controls. The gulls received ad libitum food for 10 days and were then fasted for 4 days. Thyroid hormones, body-mass change, daily food intake, hematocrit and several plasma biochemical parameters were analysed. Treated (T)-males maintained constant levels of plasma total protein throughout the experiment, whilst control (C)-males showed a decrease. We did not find any other differences between groups for the other variables analysed. Since the implanted birds sustained high testosterone levels for a number of days, any cost to body condition would have been revealed if these costs levels were actually important. Our results do not support the hypothesis that a reduction in body condition can be directly produced by plasma testosterone, although total protein changes do suggest different anabolic patterns in testosterone-treated gulls.     

Ultraviolet reflectance affects male-male interactions in the blue tit (Parus caeruleus ultramarinus)

Several animal species have been shown to use phenotypic traitsto assess the competitive ability of opponents and adjust theiraggressiveness depending on the likelihood to win the contest.In birds, these phenotypic traits usually involve patches ofcolored feathers. The benefit to harbor honest signals of malequality is the avoidance of wasteful aggressive interactions.Recent work has shown that ultraviolet (UV) plumage reflectanceis an important signal used by females during mate choice. Surprisingly,however, the role of UV signaling on intrasexual selection hasbeen neglected. In the present study, we aimed to test whetherUV reflectance of crown feathers was used as a signal of malecompetitive ability during male-male interactions. Breedingmale blue tits (Parus caeruleus ultramarinus) were exposed duringthe female egg-laying period to blue tit taxidermic mounts witheither control or reduced UV reflectance of crown feathers.In agreement with the prediction that UV reflectance advertisesmale quality, we found that breeding blue tits behaved lessaggressively toward the UV-reduced decoy. To our knowledge,this is the first experimental evidence suggesting a role forUV signaling on intrasexual selection.     

Female body condition and brood sex ratio in Yellow-legged Gulls Larus cachinnans

In the Yellow-legged Gull Larus cachinnans , males are the larger sex, and show more reproductive variance than females. We predicted that the proportion of male chicks in a brood should increase with female body condition. We investigated brood sex ratio by using DNA markers taken from samples of hatchlings or dead embryos, and female body condition using plasma cholesterol concentration as a reliable indicator. The brood sex ratio of females in good condition was male biased and the sex ratio of females in poor condition was female biased. This relationship was also significant in those nests where all the eggs laid were sexed. Thus, manipulation of embryo mortality cannot explain the biases reported in this study, suggesting that the sex ratio of the eggs was biased prior to laying. These results confirm that sex-ratio manipulation in gulls operates under natural conditions, and supports earlier experimental findings.