Sympatric divergence and clinal variation in multiple coloration traits of Ficedula flycatchers.

Geographic variation in phenotypes plays a key role in fundamental evolutionary processes such as local adaptation, population differentiation and speciation, but the selective forces behind it are rarely known. We found support for the hypothesis that geographic variation in plumage traits of the pied flycatcher Ficedula hypoleuca is explained by character displacement with the collared flycatcher Ficedula albicollis in the contact zone. The plumage traits of the pied flycatcher differed strongly from the more conspicuous collared flycatcher in a sympatric area but increased in conspicuousness with increasing distance to there. Phenotypic differentiation (PST ) was higher than that in neutral genetic markers (FST ), and the effect of geographic distance remained when statistically controlling for neutral genetic differentiation. This suggests that a cline created by character displacement and gene flow explains phenotypic variation across the distribution of this species. The different plumage traits of the pied flycatcher are strongly to moderately correlated, indicating that they evolve non-independently from each other. The flycatchers provide an example of plumage patterns diverging in two species that differ in several aspects of appearance. The divergence in sympatry and convergence in allopatry in these birds provide a possibility to study the evolutionary mechanisms behind the highly divergent avian plumage patterns.

[Cytophotometric determination of genome size in two species of Cyclops Lake Baikal (Crustacea: Copepoda: Cyclopoina) in ontogenetic development].

Genome size of Cyclops in cells at early stages of cleavage (up to the 5th division) and in somatic cells were estimated by a static digital Feulgen cytophotometry in order to study the quantitative changes in DNA content during chromatin diminution. Our realization of the cytophotometric method was approbeted on five different digital-imaging systems in blood cells of four vertebrate species. In all cases, we observed a direct correlation of the obtained and known from the literature data on the genome size and a high reproducibility, which allows to use these systems in future work. We also optimized the conditions for DNA hydrolysis of both blood smears, and for two species of Cyclops from the Moscow population, as 30 min in 5 N HCl at 24 degrees C. Here we first revealed chromatin diminution in two endemic Baikal species of Cyclopoida: Acanthocyclops incolotaenia and Diacyclops galbinus estimated the extent ofchromatin diminution in Diacyclops galbinus as 95.5-96.2 %. Cytometric analysis of the third species, Mesocyclops leuckarti, did not reveal obvious chromatin diminution. We also optimized the conditions for DNA hydrolysis of both blood smear preparations, and for two species of copepods from the Moscow population, as 30 min in 5N HCl at 24 degrees C.

[Specific immunity and polymorphism of breeding plumage in pied flycatcher (Ficedula hypoleuca) males (Aves: passeriformes)].

The relationship between the type of melanin-based plumage coloration and the strength of experimentally induced immune response was studied using as an example a pied flycatcher population from the Moscow Region. The plumage of pied flycatcher males exhibits the full spectrum of transitions from contrasting black-and-white to cryptic brownish, the latter being very similar to the coloration of females. In spite of numerous studies, the nature of this polymorphism still remains vague. Unlike many other avian species with monocyclic breeding, a considerable fraction of pied flycatchers combines two energy-consuming productive processes, breeding and molt. During the main experimental treatment we activated the humoral immunity of free-living males in chick-rearing period by injection of nonpathogenic multigenic antigen (sheep red blood cells, SRBC) and estimated the strength of the immune responses after repeated captures in 6-8 days. In addition, after each capture we estimated the numbers of leucocytes (WBC), heterophil to lymphocyte ratios (H/L) and measured night time basal metabolic rates (BMR). Non-molting males of different color types showed the same immune responses. Among molting birds, the strength of the immune response was significantly higher in pale males (morphs 4-7 by Drost's scale) than in bright males with rich melanin-based coloration (morphs 2-3). This difference resulted from two opposite processes. During molting, pale males heightened the antibody titer after immunization, while bright males tended to reduce the strength of immune response. Possibly such an asymmetry in immunocompetence at the first stage of molt reflects the different life strategies of pied flycatcher males - conspicuous birds less commonly combine breeding with molt than cryptic ones.

[DNA contents in nuclei of Cyclops kolensis and C. insignis (Crustacea, Copepoda)].

Chromatin diminution (CD) in two Cyclopoida species, Cyclops kolensis and C. insignis, was studied by static digital Feulgen cytophotometry. DNA content (pg/cell) was evaluated by standard curves builded up using blood cells of five organisms with known DNA content, which ranged from 1.25 to 14.70 pg. According to data obtained, diploid genome of C. kolensis has about 40 pg DNA before CD and 1.8-2.0 pg DNA after CD. These values are similar for both Moscow and Baikal populations of C. kolensis and 6-10 times exceed estimates made earlier (Grishanin, 2008), Our data confirm that CD in C. kolensis is 94-96% of DNA. In mitotic dividing cells of C. insignis, DNA content was about 7.5 pg both in early and late embryos, and CD was not revealed for this species. The data obtained show that, among Cyclopoida studied, the genome of C. kolensis before CD has a maximum content of DNA.

[Estimation of heritability and repeatability of resting metabolic rate in birds, with free-living pied flycatchers Ficedula hypoleuca (Aves: Passeriformes) as an example].

Estimates of a trait heritability and repeatability can get at an idea of its usefulness for being an individual characteristic and its ability to change under selection pressure. Heritability and repeatability of energetic parameters still poorly studied in birds. The most important physiological characteristic of homoiotherms is resting metabolic rate (RMR), which, in the absence of productive processes, does not exceed basal metabolic rate (BMR). We estimated BMR repeatability in free-living pied flycatchers in Moscow Region (55 degrees 44' N, 36 degrees 51' E; 1992-2008) and Tomsk (56 degrees 20' N, 84 degrees 56' E; 2008-2009) populations over intervals from 40 days to 3 years. In Moscow Region population, BMR repeatability amounted to tau = 0.34 +/- 0.10 (n=80) if measured over 1 year interval, tau = 0.60 +/- 0.15 (n=19) if measured over 2 years interval, and tau = 0.85 +/- 0.13 (n=6) if measured over 3 years interval providing that consecutive BMR measurements were done in the same period of reproductive season. In Tomsk population, BMR repeatability, measured over 1 year interval, amounted to tau = 0.49 +/- 0.11 (n=50). Repeatability is a measure of a trait constancy and sets the upper limit of its heritability. To estimate RMR heritability, cross-fostering experiments have been conducted in 2003-2005 with flycatchers of Moscow Region population. RMR of chicks positively correlated with BMR of their biological fathers, whereas such correlation in metabolic rates between chicks and their foster fathers was absent. The RMR heritability estimate turned out to be h2 = 0.43 +/- 0.17 (n=210). The obtained estimates of heritability and repeatability of fundamental energetic traits are rather high for physiological features. This suggests the existence of a potential for direct selection on BMR and evolutionary stable diversity of avian populations with regard to basal metabolic rate.

[Structural and functional aspects of variation of the breeding plumage ornamentation in the male Pied Flycatcher Ficedula hypoleuca (Aves: passeriformes)].

Inter- and intraindividual variation of the breeding plumage ornamentation in male Pied Flycatcher Ficedula hypoleuca from the Moscow province population (55 degrees 44' N 36 degrees 51' E) was studied using long-term data (1996-2005). Eumelanin prevailing colour type (CT) scored according to the seven-step Dorst scale (Dorst, 1936), age, and reproductive status during the current breeding season were analyzed as potential sources of variation of the white ornamentation on the forehead, the wings, and the tail in males. Male CT strongly varied, from almost black to cryptic brownish (2-7, average 4.4, N = 1403). On average CT became I1degree darker with age and tended to be stable after the age of two years. Initial and final CT strongly correlated (r = 0.76). Wing ornamentation and forehead patch (FP) were associated with both initial and final CT, whereas correlation between tail ornamentation and CT was observed only in aged males. Ornamentation was most variable in males with cryptic (pale) CT. Such males had, on average, less intense forehead and wing ornamentation than conspicuous males, compensating this difference by having more intense ornamentation next year. These data suggest the weakening of morphogenetic relations between prevailing colour and ornamentation in males with cryptic phenotype. The return rate (rate of conspicuous males returning to their previous nesting territory) was not related to the success of their previous breeding attempts. Among pale males, the return rate was higher in breeders than in non-breeders. The relation between FP shape and reproductive status was pronounced only in pale males. Among pale males, previous breeders and non-breeders displayed no difference in the degree of prevailing colour darkening next year. In conspicuous males, previous breeding led to the weakening of dorsal eumelanin pigmentation of the new plumage, which suggests the existence of tradeoffs between reproductive efforts and the development of rich eumelanin coloration. Among pale males, previous breeders displayed a higher rate of FP widening next year than previous non-breeders. Diffrerent patterns of relations between male ornamentation and reproductive experience suggest that the females can use plural clues to evaluate the quality of their potential mates.

Immune challenge affects basal metabolic activity in wintering great tits.

The costs of exploiting an organism's immune function are expected to form the basis of many life-history trade-offs. However, there has been debate about whether such costs can be paid in energetic and nutritional terms. We addressed this question in a study of wintering, free-living, male great tits by injecting them with a novel, non-pathogenic antigen (sheep red blood cells) and measuring the changes in their basal metabolic rates and various condition indices subsequent to immune challenge. The experiment showed that activation of the immune system altered the metabolic activity and profile of immune cells in birds during the week subsequent to antigen injection: individuals mounting an immune response had nearly 9% higher basal metabolic rates, 8% lower plasma albumin levels and 37% higher heterophile-to-lymphocyte ratios (leucocytic stress indices) than sham-injected control birds. They also lost nearly 3% (0.5 g) of their body mass subsequent to the immune challenge. Individuals that mounted stronger antibody responses lost more mass during the immune challenge. These results suggest that energetic expenditures to immune response may have a non-trivial impact upon an individual's condition.