Telomeres in ecology and evolution: A review and classification of hypotheses.

Research on telomeres in the fields of ecology and evolution has been rapidly expanding over the last two decades. This has resulted in the formulation of a multitude of, often name-given, hypotheses related to the associations between telomeres and life-history traits or fitness-facilitating processes (and the mechanisms underlying them). However, the differences (or similarities) between the various hypotheses, which can originate from different research fields, are often not obvious. Our aim here is therefore to give an overview of the hypotheses that are of interest in ecology and evolution and to provide two frameworks that help discriminate among them. We group the hypotheses (i) based on their association with different research questions, and (ii) using a hierarchical approach that builds on the assumptions they make, such as about causality of telomere length/shortening and/or the proposed functional consequences of telomere shortening on organism performance. Both our frameworks show that there exist parallel lines of thoughts in different research fields. Moreover, they also clearly illustrate that there are in many cases competing hypotheses within clusters, and that some of these even have contradictory assumptions and/or predictions. We also touch upon two topics in telomere research that would benefit from further conceptualization. This review should help researchers, both those familiar with and those new to the subject, to identify future avenues of research.

Autumn migration direction of juvenile willow warblers (Phylloscopus t. trochilus and P. t. acredula) and their hybrids assessed by qPCR SNP genotyping.

BACKGROUNDS: Geographic regions, where two closely related taxa with different migration routes come into contact, are known as migratory divides. Hybrids originating from migratory divides are hypothesized to migrate intermediately relative to the parental populations. Few studies have tested this hypothesis in wild birds, and only in hybrids that have completed the migration back to the breeding grounds. Here, we make use of the well-established migration routes of willow warblers (Phylloscopus trochilus), for which the subspecies trochilus and acredula have migration-associated genetic markers on chromosomes 1 and 5. The genetic approach enabled us to analyze the geographic distribution of juveniles during their first autumn migration, predicting that hybrids should be more frequent in the central flyway over Italy than along the typical SW routes of trochilus and SE routes of acredula. METHODS: Blood and feather samples were collected from wintering birds in Africa (n = 69), and from juveniles during autumn migration in Portugal (n = 33), Italy (n = 38) and Bulgaria (n = 32). Genotyping was carried out by qPCR SNP assays, on one SNP each on chromosome 1 (SNP 65) and chromosome 5 (SNP 285). Both these SNPs have alternative alleles that are highly fixed (> 97%) in each of the subspecies. RESULTS: The observed combined genotypes of the two SNPs were associated with the known migration routes and wintering distributions of trochilus and acredula, respectively. We found hybrids (HH) among the juveniles in Italy (5/38) and in Portugal (2/33). The proportion of hybrids in Italy was significantly higher than expected from a background rate of hybrid genotypes (1.5%) in allopatric populations of the subspecies. CONCLUSIONS: Our genetic approach to assign individuals to subspecies and hybrids allowed us to investigate migration direction in juvenile birds on their first migration, which should better reflect the innate migratory direction than studies restricted to successful migrants. The excess of hybrids in Italy, suggests that they employ an intermediate route relative to the parental populations. Our qPCR SNP genotyping method is efficient for processing large sample sizes, and will therefore be useful in migration research of species with known population genetic structure.

Geographic patterns of mtDNA and Z-linked sequence variation in the Common Chiffchaff and the 'chiffchaff complex'.

The Common Chiffchaff Phylloscopus collybita is an abundant, polytypic Palearctic bird. Validity of some of its subspecies is controversial and birds from some parts of the species range remain unclassified taxonomically. The relationships among populations from different geographic areas have not been sufficiently explored with molecular data. In this study we analyzed the relationships among the four species in the 'chiffchaff complex' (Common Chiffchaff, Iberian Chiffchaff P. ibericus, Canary Islands Chiffchaff P. canariensis and Mountain Chiffchaff P. sindianus), and the patterns of intraspecific geographic variation in the mtDNA ND2 gene and intron 9 of the Z-linked aconitase gene (ACO1I9) across the Common Chiffchaff range, including a recently discovered population breeding on Mt. Hermon (Anti-Lebanon mountains). Our data supported the monophyly of the chiffchaff complex and its current systematics at the species level. Within the Common Chiffchaff, the Siberian race P. c. tristis was the most differentiated subspecies and may represent a separate or incipient species. Other Common Chiffchaff subspecies also were differentiated in their mtDNA, however, lineages of neighboring subspecies formed wide zones of introgression. The Mt. Hermon population was of mixed genetic origin but contained some birds with novel unique lineage that could not be assigned to known subspecies. All Common Chiffchaff lineages diverged at the end of the Ionian stage of Pleistocene. Lineage sorting of ACO1I9 alleles was not as complete as that of mtDNA. Chiffchaff species were mostly distinct at ACO1I9, except the Common and Canary Islands Chiffchaffs that shared multiple alleles. An AMOVA identified geographic structure in Common Chiffchaff ACO1I9 variation that was broadly consistent with that of mtDNA ND2 gene. The genetic and other data suggest the chiffchaff complex to be a group of evolutionarily young taxa that represent a paradigm of 'species evolution in action' from intergrading subspecies through to apparently complete biological speciation.

Differential timing and latitudinal variation in sex ratio of Aquatic Warblers during the autumn migration.

Differential migration has been extensively reported in spring, but less so in autumn, particularly in relation to sex in monomorphic bird species. Here, we analysed the autumn passage of a monomorphic, globally threatened passerine, the Aquatic Warbler Acrocephalus paludicola throughout Western Europe, with regard to age and sex. We showed that, overall, adults migrated earlier than first-year birds, and males migrated earlier than females during the autumn migration. This may be caused by an overall social dominance of adults over immatures, and differentiated migration strategy of males and females. In addition, we found male-skewed sex proportions, with a tendency to an equalised ratio in more southern stopover sites. This may indicate a male bias in the global population or different migration strategies of the sexes. Differential migration may cause the age and sex classes to be exposed differently to various threats affecting demographic structure of the species.

Provenance does matter: links between winter trophic segregation and the migratory origins of European robins.

Amongst migratory species, it is common to find individuals from different populations or geographical origins sharing staging or wintering areas. Given their differing life histories, ecological theory would predict that the different groups of individuals should exhibit some level of niche segregation. This has rarely been investigated because of the difficulty in assigning migrating individuals to breeding areas. Here, we start by documenting a broad geographical gradient of hydrogen isotopes (δ 2H) in robin Erithacus rubecula feathers across Europe. We then use δ 2H, as well as wing-tip shape, as surrogates for broad migratory origin of birds wintering in Iberia, to investigate the ecological segregation of populations. Wintering robins of different sexes, ages and body sizes are known to segregate between habitats in Iberia. This has been attributed to the despotic exclusion of inferior competitors from the best patches by dominant individuals. We find no segregation between habitats in relation to δ 2H in feathers, or to wing-tip shape, which suggests that no major asymmetries in competitive ability exist between migrant robins of different origins. Trophic level (inferred from nitrogen isotopes in blood) correlated both with δ 2H in feathers and with wing-tip shape, showing that individuals from different geographic origins display a degree of ecological segregation in shared winter quarters. Isotopic mixing models indicate that wintering birds originating from more northerly populations consume more invertebrates. Our multi-scale study suggests that trophic-niche segregation may result from specializations (arising in the population-specific breeding areas) that are transported by the migrants into the shared wintering grounds.

Phenotypic divergence among west European populations of Reed Bunting Emberiza schoeniclus: the effects of migratory and foraging behaviours.

Divergent selection and local adaptation are responsible for many phenotypic differences between populations, potentially leading to speciation through the evolution of reproductive barriers. Here we evaluated the morphometric divergence among west European populations of Reed Bunting in order to determine the extent of local adaptation relative to two important selection pressures often associated with speciation in birds: migration and diet. We show that, as expected by theory, migratory E. s. schoeniclus had longer and more pointed wings and a slightly smaller body mass than the resident subspecies, with the exception of E. s. lusitanica, which despite having rounder wings was the smallest of all subspecies. Tail length, however, did not vary according to the expectation (shorter tails in migrants) probably because it is strongly correlated with wing length and might take longer to evolve. E. s. witherbyi, which feed on insects hiding inside reed stems during the winter, had a very thick, stubby bill. In contrast, northern populations, which feed on seeds, had thinner bills. Despite being much smaller, the southern E. s. lusitanica had a significantly thicker, longer bill than migratory E. s. schoeniclus, whereas birds from the UK population had significantly shorter, thinner bills. Geometric morphometric analyses revealed that the southern subspecies have a more convex culmen than E. s. schoeniclus, and E. s. lusitanica differs from the nominate subspecies in bill shape to a greater extent than in linear bill measurements, especially in males. Birds with a more convex culmen are thought to exert a greater strength at the bill tip, which is in agreement with their feeding technique. Overall, the three subspecies occurring in Western Europe differ in a variety of traits following the patterns predicted from their migratory and foraging behaviours, strongly suggesting that these birds have became locally adapted through natural selection.

Phylogeography of a habitat specialist with high dispersal capability: the Savi's Warbler Locustella luscinioides.

In order to describe the influence of Pleistocene glaciations on the genetic structure and demography of a highly mobile, but specialized, passerine, the Savi's Warbler (Locustella luscinioides), mitochondrial DNA sequences (ND2) and microsatellites were analysed in c.330 individuals of 17 breeding and two wintering populations. Phylogenetic, population genetics and coalescent methods were used to describe the genetic structure, determine the timing of the major splits and model the demography of populations. Savi's Warblers split from its sister species c.8 million years ago and have two major haplotype groups that diverged in the early/middle Pleistocene. One of these clades originated in the Balkans and is currently widespread, showing strong evidence for population expansion; whereas the other is restricted to Iberia and remained stable. Microsatellites agreed with a genetic break around the Pyrenees, but showed considerable introgression and a weaker genetic structure. Both genetic markers showed an isolation-by-distance pattern associated with the population expansion of the eastern clade. Breeding populations seem to be segregated at the wintering sites, but results on migratory connectivity are preliminary. Savi's Warbler is the only known migratory bird species in which Iberian birds did not expand beyond the Pyrenees after the last glaciation. Despite the long period of independent evolution of western and eastern populations, complete introgression occurred when these groups met in Iberia. Mitochondrial sequences indicated the existence of refugia-within-refugia in the Iberian Peninsula during the last glacial period, which is surprising given the high dispersal capacity of this species. Plumage differences of eastern subspecies seemed to have evolved recently through natural selection, in agreement with the glacial expansion hypothesis. This study supports the great importance of the Iberian Peninsula and its role for the conservation of genetic variation.

Concentrations of Fe, K, Na, Ca, P, Zn and Mg in maternal colostrum and mature milk.

The objective of this study was to determine the concentrations of Fe, K, Na, Ca, P, Zn and Mg in colostrum and mature milk (2 months) from nursing mothers, and the correlation between the concentration of these micronutrients and newborn weight, maternal age, gestational age GA, inter-pregnancy interval, and gestational weight gain. Thirty one women donated milk on the 2nd postpartum day and on the 2nd month of lactation. Micronutrients were analysed by atomic emission spectrophotometry. The concentrations (mean+/-SD) of Fe (0.12+/-0.08; 0.09+/-0.5 mg/100 ml), Zn (0.93+/-0.36; 0.15+/-0.06 mg/100 ml), K (62.8+/-11.5; 46.2+/-0.84 mg/100 ml) and Na (33.3+/-12.3; 20.5+/-15.6 mg/100 ml) decreased from colostrum to mature milk, whereas Ca (21.4+/-5.8; 25.0+/-3.1 mg/100 ml) and P (11.12+/-2.8; 13.7+/-2.0 mg/100 ml) increased. Correlations were observed between Zn and Ca concentrations and GA (r=-0.41, p=0.042; r=0.48, p=0.014, respectively), between Ca and K and inter-pregnancy interval (r=0.56, p=0.001; r=0.38, p=0.033, respectively), and between Mg and P and maternal age (r=0.49, p=0.005; r=0.37, p=0.042, respectively). The Zn concentration in colostrum showed a negative correlation with gestational weight gain (r=-0.49, p=0.006), and Na concentration showed a negative correlation with newborn weight (r=-0.38, p=0.036). The concentration of micronutrients in colostrum and mature milk (2 months) suffers alterations, including a decrease in Fe, Zn, K and Na and an increase in Ca and P, probably in order to satisfy the requirements of the nursing infant. Micronutrients are influenced by birth weight, maternal age, gestational weight gain, GA and inter-pregnancy interval.