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. 2020 Apr 16;59:e12. doi: 10.6620/ZS.2020.59-12

Individual Heterozygosity Influences Arrival Times and Mating Success of Male Red-Breasted Flycatchers Ficedula parva

Cezary Mitrus 1,*, Joanna Mitrus 2, Robert Rutkowski 3
PMCID: PMC7396921  PMID: 32760458

Abstract

We examined the relationship between individual heterozygosity of male Red-breasted Flycatchers (Ficedula parva; a small long-distance migratory, socially monogamous bird species) and their mating success, arrival time and age. Using eight polymorphic microsatellite loci, we found that male heterozygosity is related to both mating success and arrival time, but not to age. Mated and earlier arriving males had higher heterozygosity than later arrivals and bachelors, but we did not find a relationship between age and individual heterozygosity of males. To our knowledge, this is the first study to provide evidence about the relationship between individual genetic diversity and arrival time, thus arrival time could be used as a signal of individual heterozygosity and quality.

Keywords: Multi-locus heterozygosity, Mate choice, Arrival time, Birds

BACKGROUND

Heterozygosity is considered as individual genetic diversity of individuals and can be used as a predictor of male quality (Brown 1997; Kempenaers 2007). Females should mate with males with high heterozygosity and such a choice should be dictated by the relationship between fitness and heterozygous advantage. Heterozygosity-fitness correlation (HFC) is explained by three main hypotheses (Kempenaers 2007): general effect, direct effect and local effect hypotheses. The general effect hypothesis states that heterozygosity in many loci is related to heterozygosity of the entire genome and marker loci and fitness loci are in identity disequilibrium (David 1998; Lynch and Walsh 1998; Hansson and Westerberg 2002). The direct effect hypothesis assumes that heterozygosity is more advantageous than homozygosity due to relative functional dominance (David 1998; Hansson and Westerberg 2002; Li et al. 2004). The third explains heterozygosity-fitness correlations as an effect of linkage between neutral loci and the loci affecting fitness (Lynch and Walsh 1998; Hansson and Westerberg 2002; Tiira et al. 2006, Malo and Coulson 2009).

Heterozygosity-fitness correlations are often reported (Szulkin et al. 2010) and are considered to influence various aspects of fitness including offspring number and survival, parasite resistance, reproductive success and characteristics of bird song (Daniels and Walters 2000; Kruuk et al. 2002; Slate et al. 2004; Whiteman et al. 2006; Reid et al. 2007; Cohas et al. 2009; Mainguy et al. 2009; Ryder et al. 2009; Harrison et al. 2011). On the other hand, there are a number of studies indicating weak or no correlation (Hoffman et al. 2006; Ortego et al. 2007 2010; Boerner et al. 2013; Zeng et al. 2013). Moreover, there is also the possibility that publication bias has falsely inflated the importance of HFCs in animals (Balloux et al. 2004; Chapman et al. 2009), as many negative results may not be published.

In migrating bird species, arrival time can be a good indicator of male quality. Patterns of arrival are often related to physical condition or age. Males that are higher quality, in better condition, and older usually arrive earlier (Stolt and Franson 1995; Møller 1994a; Arvidson and Neergaard 1991; Marra et al. 1998; Ninni et al. 2004; Mitrus 2007). To date, no reports about the relationship between the arrival time of migratory birds and individual heterozygosity have been published. To test for such a correlation, a long-distance migratory bird species can be used as a model.

The main aim of this study was to answer the question: is individual heterozygosity of male Red-breasted Flycatchers (Ficedula parva) related to their mating success, arrival time and/or age? We analysed a group of highly polymorphic microsatellite loci of a free-living population of this species breeding in natural conditions.

MATERIALS AND METHODS

Study area

The data were collected in the Białowieża Forest, northeast Poland (52°41'N, 23°52'E). The study was conducted over seven breeding seasons (2004–2011, excluding 2007) in three study plots (total area 79.5 ha) and along roads leading to those plots. The study plots were similar in floral composition and covered by old-growth lime-oak-hornbeam Tilio Querco-Carpinetum stands characterised mainly by Hornbeam Carpinus betulus, Small-leaved Lime Tilia cordata, Pendunculate Oak Quercus robur, Norway Maple Acer platanoides and Norway Spruce Picea abies. The Białowieża Forest is old-growth, deciduous temperate forest (Tomiałojć 1991; Wesołowski et al. 2003) characterised by a rich bird and mammal community (Wesołowski et al. 2003; Jędrzejewska and Jędrzejewski 1998). There are standing and fallen dead trees (Tomiałojć 1991; Wesołowski et al. 2003; Jaroszewicz et al. 2019).

Bird data

Data were collected for the Red-breasted Flycatcher, which is a small migratory, insectivorous bird. After arrival males try to attract females by singing and defending territories. The earliest males arrive to the Białowieża Forest in Poland from late April to early May (Mitrus et al. 2005). Based on the colour of the breast plumage, we estimated male age using two categories: old (after-2nd-year, with orange badge on the breast) and young (2nd-year, nestlings from the previous season, without orange breast feathers) (Svensson 1992). The old males arrive significantly earlier than younger ones (Mitrus 2007). In our studied population, old males are more numerous (2/3 of all males) and achieve higher mating success than young males (Mitrus 2006 2018). Most males are socially monogamous (only one case of polygyny has been reported to date), and low extra-pair paternity has been recorded (Mitrus and Soćko 2005; Mitrus et al. 2014). The Red-breasted Flycatcher is a rare species in Europe, and occurs at low density in the Białowieża Forest, i.e., in recent years, in optimum breeding habitat, up to 1.5 pairs/10 ha (Wesołowski et al. 2010 2015).

To determine the males’ arrival dates (usually from the end of April to the middle of May), up to four people searched for newly arriving males every day. In the statistical analyses, the arrival date each year was standardised by considering the arrival date of the first male as day 1. The active territories were checked every day to determine mating success. Unmated males of the Red-breasted Flycatcher sing intensely; therefore, if a male was seen to have stopped singing and was observed with a female, or was seen copulating, then it was defined as mated (Mitrus et al. 2012).

Males were captured using a concealed tape-recorder with a broadcasted conspecific song to lure birds into a mist net (mesh – 16 mm, dimensions – 6 × 25 m). Each male was ringed using a unique combination of aluminum and colour-coded rings. Additionally, three feathers from the lesser coverts were collected for DNA isolation and analysis of paternity.

Molecular data

The feathers were stored in disposable, sterile tubes in 96% alcohol in the fridge at 4°C. Before DNA isolation was performed, the alcohol was evaporated and the quills (source of DNA, Morin et al. 1994; Kawka et al. 2012) were cut into fragments (0.5–1 cm long) using a sterile scalpel. The DNA was extracted using the Qiagen® QIAamp DNA Investigator Kit.

The nine most polymorphic loci described for other species in the genus Ficedula were used: Fhy336, Fhy350, Fhy224, Fhy458, Fhy429, Fhy452, Fhy428 (Leder et al. 2008) and FhU, FhU4 (Primmer et al. 1996). Selected microsatellites were characterized by high allelic diversity, the number of alleles at a single locus ranged from 4 to 26 (mean = 15.4) (Mitrus et al. 2013). Amplification of locus Fhy350 failed in some individuals, which we assumed was due to a null allele. Many authors recommend avoiding the use of loci with null alleles (Dakin and Avise 2004; Selkoe and Toonen 2006) and this locus was not considered for heterozygosity analysis. Most of the loci (except Fhy350) were in Hardy-Weinberg equilibrium (Mitrus et al. 2013).

Statistical analysis

To characterise multi-locus individual heterozygosity the Excel macro Cernicalin was used (Aparicio et al. 2006). Cernicalin calculates observed homozygosity per individual (HO). We used standardized heterozygosity (HST) (Coltman et al. 1999), which takes into account the proportion of heterozygous loci divided by the mean observed heterozygosity. This measure of heterozygosity is highly conservative and performs better than other measures when there is allele dropout or when individuals are genotyped at different numbers of loci (Coulon 2010).

We first examined if there were differences in heterozygosity between paired and unpaired males. Most genotyped young and older males did not come back to the study area the following year and most adults were new individuals that came from other places. Therefore, in many cases an individual was observed for only one year and no repeated measurement was used in the analysed; in this case, the data for each individual were dealt with as independent data.

To determine factors affecting the heterozygosity of males General Linear Models (GLM) were constructed: heterozygosity was used as the dependent variable, age and matting success as categorical factors and arrival time as a continuous independent variable (covariate). Statistica for Windows v. 13.0 was used for statistical calculations

RESULTS

Individual standardised heterozygosity within males ranged from 0.26 to 1.03 (Me = 0.883) and was non-normal in distribution (Kolmogorov-Smirnov test; d = 0.19, p < 0.05). The lower quartile (Q1) was 0.773, the upper quartile (Q3) was 0.903 (Fig. 1). In the GLM (R2 = 0.21, F3.38 = 9.43, p < 0.01), mating success was related to the heterozygosity of males (Table 1) and the median heterozygosity of paired males was higher than that of unmated males (Fig. 2). The heterozygosity of males was related to their time of arrival (Table 1): and the earlier arriving males had a higher individual standardised heterozygosity than later arriving ones (Fig. 3). The age of males was not related to their individual heterozygosity (Table 1.)

Fig. 1.

Fig. 1.

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Distribution of individual heterozygosity of male Red-breasted Flycatchers.

Fig. 2.

Fig. 2.

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Individual heterozygosity in relation to the mating success of male Red-breasted Flycatchers.

Fig. 3.

Fig. 3.

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Relationship between arrival time and heterozygosity of male Red-breasted Flycatchers.

Table 1.

Results of General Linear Model testing the relationships between heterozygosity and arrival time, mating success and male age

Variable Degr. of freedom MS F p-value
Arrival time 1 0.286 14.873 < 0.01
Mating success 1 0.225 8.314 < 0.01
Age 1 0.013 0.503 0.48
Error 37 0.027
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DISCUSSION

We used a set of polymorphic microsatellite markers to estimate individual heterozygosity and then evaluated correlation between individual genetic diversity and fitness of Red-breasted Flycatcher males. Microsatellite markers are widely applied in studies of HFC (e.g., Wetzel et al. 2012; Forstmeier et al. 2012; Canal et al. 2014); however, their usefulness in such investigations remain a hotly debated issue (Väli et al. 2008; Ljungqvist et al. 2010). First, they are abundant in genomes, highly polymorphic and easy to analyse, allowing for a direct estimation of heterozygosity. On the other hand, complex mutational processes occurring within microsatellite loci lead to homoplasy which, in turn, can cause an underestimation of heterozygosity (Hansson and Westerberg 2002). Another problem is a connection between heterozygosity in microsatellite loci and genome-wide heterozygosity (Szulkin et al. 2010). As highly polymorphic microsatellites are usually desirable for population genetic studies, isolation and characterization of novel markers often lead to ascertainment bias and an upwardly skewed estimate of genome-wide diversity (Brandström and Ellegren 2008). However, some experimental and simulation studies indicated that microsatellites are good indicators of a genome-wide heterozygosity and are more informative than bi-allelic markers (e.g., SNPs) because they have a large number of variants per locus (Ljungqvist et al. 2010; Miller et al. 2014).

Another factor affecting the reliability of heterozygosity-fitness analyses is sample size and the number of markers used (David 1998). The basic assumption underlying the HFC concept is that the level of individual heterozygosity is associated with performance traits. This suggests that individuals with low heterozygosity are inbred, and thus present low genome-wide diversity. However, the idea that a handful of neutral markers reflects genome-wide heterozygosity has often been criticized (Balloux et al. 2004; Taylor et al. 2010). Undoubtedly, in our study both the sample size and the number of microsatellite loci are low. Low sample size generated weak correlations in HFC studies (David 1998). Similarly, it was shown that a large number of markers are needed to increase the strength of correlation between heterozygosity and inbreeding, whereas a number of markers below 20 should result in a weak correlation (Balloux et al. 2004; Taylor et al. 2010; Miller et al. 2014). Slate et al. (2004) showed that the strength of the correlation between heterozygosity and inbreeding is a function of inbreeding prevalence, as well as the number of loci used. Hence, relationships between heterozygosity and fitness should be hard to detect if there are a low number of markers employed or the sample size is low. However, a low number of markers will usually indicate a significant correlation in the most extreme cases, when associations should be especially strong, e.g., in highly inbred populations (Hedrick et al. 2001; Balloux et al. 2004; Slate et al. 2004). Hence, we can state that our results, despite having some flaws connected with sample size and number of markers, suggest that there are biologically important associations between individual heterozygosity and performance traits.

One finding of this study is that earlier arriving males of the Red-breasted Flycatcher had higher heterozygosity than later arriving males. To our knowledge, these results are the first documentation of a relationship between male heterozygosity and arrival time in a socially monogamous bird species. Protandry is often observed in migratory bird species, with males arriving before females (Kokko et al. 2006; Newton 2008; Harnos et al. 2014), and this is the case in Red-breasted Flycatchers. After arrival, males occupy and defend territories. The time that males arrive to the breeding areas can also be treated as an indicator of male quality. Arrival date is often correlated with size, condition and age of males (Mitrus 2007; Arvidson and Neergaard 1991; Marra et al. 1998; Forstmeier 2002; Stewart et al. 2002; Ninni et al. 2004; Risely et al. 2013). Individuals that arrive earlier may obtain higher quality territories (Potti and Montalvo 1991; Aebischer et al. 1996; Johnson 1997; Smith and Moore 2005) and can increase their probability of mating success (Møller 1994b; Lozano et al. 1996; Kokko 1999). Also, in the Red-breasted Flycatcher, earlier-arriving males are older, possess a larger plumage ornament and have higher mating success (Mitrus 2007; Mitrus et al. 2012), all of which suggest that arrival time can be used as a good indicator of male quality.

Our data show that mating success of Red-breasted Flycatcher males was related to their heterozygosity and that the mated males had higher individual heterozygosity than unmated ones. It is not clear how female preference for heterozygous mates could confer an indirect genetic benefit for her and/or her offspring. Heterozygosity is not a visible mark and females are unlikely to be able to distinguish it directly, but it is often related to condition-dependent phenotypic traits (Ditchkoff et al. 2001; Hansson and Westerberg 2002; Foerster et al. 2003; Roberts et al. 2005; Tiira et al. 2006; Malo and Coulson 2009). A male’s heterozygosity may also be correlated with characteristics related to male quality, such as song structure, reproductive and fledging success (Foerster et al. 2003; Seddon et al. 2004). Thus, female Red-breasted Flycatchers are predicted to value heterozygosity in their mates. The expression of vigour, condition-sensitive ornaments and symmetry in males may be a direct reflection not of “good genes” but of individual heterozygosity at key loci or at many loci.

The third finding of this study is that there were no differences in heterozygosity of males in relation to their age. Consistent with our finding, some other studies of wild populations of animals have also observed no differences in individual heterozygosity between various age classes (Diehl and Koehn 1985; Patarnello et al. 1991; Banks et al. 2010). However, a positive correlation between heterozygosity and survival is usually reported in various groups of animals, individuals with higher heterozygosity having a greater chance of survival (Coulson et al. 1998; Hansson et al. 2001; Lesbarrères et al. 2005; Acevedo-Whitehouse et al. 2003; Cohas et al. 2009). The difference in heterozygosity between younger and older males can be explained by higher selection pressure according to heterozygosity level early in life (Charlesworth and Hughes 1996; Keller et al. 2008) and differences in the rate of mortality and probability of survival among individuals in various stages of life. In birds, the highest mortality is observed in early life stages (Naef-Daenzer and Grüebler 2016), during which time individuals in bad condition and of low quality are likely to be eliminated (Newton 2008). In migratory birds, additional mortality occurs during the migration period, and may comprise 85% of apparent annual mortality and is at least 15 times higher than during the resident periods (Sillett and Holmes 2002). The Red-breasted Flycatcher is a long-distance migratory species; its one-way migration distance may reach 5000 km (Mitrus et al. 2005; Erciyas-Yavuz et al. 2015). Therefore, it should be expected that migration is an important factor causing high mortality in birds, especially young, first time migrating individuals (Owen and Black 1989; Newton 2008). Young birds that arrived on the breeding grounds have survived two long-distance migrations and their first winter, thus there is strong selection pressure for first time breeders to be high quality individuals. In a closely related species, the Pied Flycatcher Ficedula hypoleca⸺also a migratory species⸺the highest mortality is in young birds and fledgling recruitment is low, whereas mean survival probabilities of adult birds are much higher (Canal et al. 2014). In our studied population of the Red-breasted Flycatcher, older males arrived earlier (Mitrus et al. 2005 2012). Thus we expected that older, earlier-arriving birds would have higher heterozygosity than young males. However, within first arrivals, there were also young males that were in good condition (Mitrus 2006 2007) and had high heterozygosity. On the other hand, within older males we can distinguish only one class. However, this group includes birds in their third year of life that arrive later and this can influence the lower heterozygosity in this group of older males.

CONCLUSIONS

Our results show that individual heterozygosity in a migratory bird species is correlated with arrival time and mating success, but not to the age of males. Studies of heterozygosity-fitness correlations carried out under natural conditions in long distance migratory bird species are scarce. Migration is a very important part of bird life; it is also a factor influencing survival and applies strong selection pressure on individuals. In the future, more studies should be conducted on the relationships between heterozygosity, condition and survival during migration.

Acknowledgments

We extend our gratitude to Magdalena Sikora, Beata Jarkiewicz, Tomasz Wesołowski, Patryk Rowiński, and Willem van Manen for their help with fieldwork. The research was funded by The National Science Centre – grant no. N N304 372938. We would like to thank the Białowieża Geobotanical Station of Warsaw University for logistical support. We are also grateful to Shelley Hinsley for providing comments that improved the quality of the manuscript and for correcting the English.

Footnotes

Authors’ contributions: CM and JM performed the field work. CM and RR wrote the manuscript. All authors contributed to drafting and revising the manuscript. All authors read and approved the final manuscript.

Competing interests: All authors declare that they have no conflict of interests regarding the publication of this paper.

Availability of data and materials: Data and materials are available from the Department of Vertebrate Ecology and Paleontology Wrocław University of Environmental and in Life Sciences and in the Museum and Institute of Zoology Polish Academy of Sciences.

Consent for publication: Not applicable.

Ethics approval consent to participate: This study complies with current Polish laws.

References

  1. Aebischer A, Perrin M, Krieg M, Studer J, Meyer DR. 1996. The role of territory choice, mate choice and arrival date on breeding success in the Savi’s Warbler Locustella luscinioides. J Avian Biol 27:134–152. doi:10.2307/3677143.
  2. Aparicio JM, Ortego J, Cordero PJ. 2006. What should we weigh to estimate heterozygosity, alleles or loci? Mol Ecol 15:4659–4665. doi:10.1111/j.1365-294X.2006.03111.x. [DOI] [PubMed]
  3. Arvidson BL, Neergaard R. 1991. Mate choice in the Willow Warbler – a field experiment. Behav Ecol Sociobiol 29:225–229. doi:10.1007/BF00166406.
  4. Acevedo-Whitehouse K, Gulland F, Greig D, Amos W. 2003. Inbreeding: Disease susceptibility in California sea lions. Nature 422:35. doi:10.1038/422035a. [DOI] [PubMed]
  5. Balloux F, Amos W, Coulson T. 2004. Does heterozygosity estimate inbreeding in real populations? Mol Ecol 13:3021–3031. doi:10.1111/j.1365-294X.2004.02318.x. [DOI] [PubMed]
  6. Banks SC, Dubach J, Viggers KL, Lindenmayer DB. 2010. Adult survival and microsatellite diversity in possums: effects of major histocompatibility complex-linked microsatellite diversity but not multilocus inbreeding estimators. Oecologia 162:359. doi:10.1007/s00442-009-1464-0. [DOI] [PubMed]
  7. Boerner M, Hoffman JI, Amos W, Chakarov N, Kruger O. 2013. No correlation between multi-locus heterozygosity and fitness in the common buzzard despite heterozygote advantage for plumage colour. J Evol Biol 26:2233–2243. doi:10.1111/jeb.12221. [DOI] [PubMed]
  8. Brandström M, Ellegren H. 2008. Genome-wide analysis of microsatellite polymorphism in chicken circumventing the ascertainment bias. Genome Res 18:881–887. doi:10.1101/gr.075242.107. [DOI] [PMC free article] [PubMed]
  9. Brown JL. 1997. A theory of mate choice based on heterozygosity. Behav Ecol 8:60–65. doi:10.1093/beheco/8.1.60.
  10. Canal D, Serrano D, Potti J. 2014. Exploring heterozygosity-survival correlations in a wild songbird population: contrasting effects between juvenile and adult stages. PLoS ONE 9(8):e105020 doi:10.1371/journal.pone.0105020. [DOI] [PMC free article] [PubMed]
  11. Chapman JR, Nakagawa S, Coltman DW, Slate J, Sheldon BC. 2009. A quantitative review of heterozygosity-fitness correlations in animal populations. Mol Ecol 18:2746–2765. doi:10.1111/j.1365-294X.2009.04247.x. [DOI] [PubMed]
  12. Charlesworth B, Hughes KA. 1996. Age-specific inbreeding depression and components of genetic variance in relation to the evolution of senescence. Proc Nat Acad Sci USA 93:6140–6145. doi:10.1073/pnas.93.12.6140. [DOI] [PMC free article] [PubMed]
  13. Cohas A, Bonenfant C, Kempenaers B, Allaine D. 2009. Age-specific effect of heterozygosity on survival in alpine marmots Marmota marmot. Mol Ecol 18:1491–1503. doi:10.1111/j.1365-294X.2009.04116.x. [DOI] [PubMed]
  14. Coltman DW, Pilkington JG, Smith JA, Pemberton JM. 1999. Parasite-mediated selection against inbred Soay Sheep in a free-living, island population. Evolution 53:1259–1267. doi:10.1111/j.1558-5646.1999.tb04538.x. [DOI] [PubMed]
  15. Coulson TN, Pemberton JM, Albon SD, Beaumont M, Marshall TC, Slate J, Guinness EE, Clutton-Brock TH. 1998. Microsatellites reveal heterosis in red deer. Proc R Soc Lond B Biol Sci 265:489–495. doi:10.1098/rspb.1998.0321. [DOI] [PMC free article] [PubMed]
  16. Coulon A. 2010. GENHET: An easy-to-use R function to estimate individual heterozygosity. Mol Ecol Res 10:167–169. doi:10.1111/j.1755-0998.2009.02731.x. [DOI] [PubMed]
  17. Daniels SJ, Walters JR. 2000. Inbreeding depression and its effects on natal dispersal in red-cockaded woodpeckers. Condor 102:482– 491. doi:10.2307/1369779.
  18. Dakin EE, Avise JC. 2004. Microsatellite null alleles in parentage analysis. Heredity 93:504–509. doi:10.1038/sj.hdy.6800545. [DOI] [PubMed]
  19. David P. 1998. Heterozygosity-fitness correlations: new perspectives on old problems. Heredity 80:531–537. doi:10.1046/j.1365-2540.1998.00393.x. [DOI] [PubMed]
  20. Diehl WJ, Koehn RK. 1985. Multiple-locus heterozygosity, mortality, and growth in a cohort of Mytilus edulis. Mar Biol 88:265–271. doi:10.1007/BF00392588.
  21. Ditchkoff SS, Lochmiller RL, Masters RE, Hoofer SR, van den Bussche RA. 2001. Major-histocompatibility-complex-associated variation in secondary sexual traits of white-tailed deer Odocoileus virginianus. Evidence for good-genes advertisement. Evolution 55:616–625. doi:10.1111/j.0014-3820.2001.tb00794.x. [DOI] [PubMed]
  22. Erciyas-Yavuz K, Zduniak P, Bariş YS. 2015. Spring and autumn migration of the red-breasted flycatcher through the Kizilirmak delta, Turkey. Curr Zool 61:412–420. doi:10.1093/czoolo/61.3.412.
  23. Foerster K, Delhey K, Johnson A, Lifjeld JT, Kempenaers B. 2003. Females increase offspring heterozygosity and fitness through extra-pair matings. Nature 425:714–717. doi:10.1038/nature01969. [DOI] [PubMed]
  24. Forstmeier W. 2002. Benefits of early arrival at breeding grounds vary between males. J Anim Ecol 71:1–9. doi:10.1046/j.0021-8790.2001.00569.x.
  25. Forstmeier W, Schielzeth H, Mueller JC, Ellegren H, Kempenaers B. 2012. Heterozygosity-fitness correlations in zebra finches: microsatellite markers can be better than their reputation. Mol Ecol 21:3237–3249. doi:10.1111/j.1365-294X.2012.05593.x. [DOI] [PubMed]
  26. Hansson B, Bensch S, Hasselquist D, Akesson M. 2001. Microsatellite diversity predicts recruitment of sibling great reed warblers. Proc R Soc B Biol Sci 268:1287–1291. doi:10.1098/rspb.2001.1640. [DOI] [PMC free article] [PubMed]
  27. Hansson B, Westerberg L. 2002. On the correlation between heterozygosity and fitness in natural populations. Mol Ecol 11:2467–2474. doi:10.1046/j.1365-294x.2002.01644.x. [DOI] [PubMed]
  28. Harrison XA, Bearhop S, Inger R, Colhoun K, Gudmundsson GA, Hodgson D, McElwaine G, Tregenza T. 2011. Heterozygosity– fitness correlations in a migratory bird: an analysis of inbreeding and single-locus effects. Mol Ecol 20:4786–4795. doi:10.1111/j.1365-294X.2011.05283.x. [DOI] [PubMed]
  29. Harnos A, Nora Á, Kovács Sz, Lang Zs, Csörgő T. 2014. Increasing protandry in the spring migration of the Pied Flycatcher (Ficedula hypoleuca) in Central Europe. J Ornithol 156:543–546. doi:10.1007/s10336-014-1148-3.
  30. Hedrick P, Fredrickson R, Ellegren H. 2001. Evaluation of d2, a microsatellite measure of inbreeding and outbreeding, in wolves with a known pedigree. Evolution 55:1256–1260. doi:10.1111/j.0014-3820.2001.tb00646.x. [DOI] [PubMed]
  31. Hoffman JI, Forcada J, Amos W. 2006. No relationship between microsatellite variation and neonatal fitness in Antarctic fur seals, Arctocephalus gazella. Mol Ecol 15:1995–2005. doi:10.1111/j.1365-294X.2006.02894.x. [DOI] [PubMed]
  32. Johnson MS. 1997. The effect of age on nest concealment and its complementary effect on production of Wood Thrush. Wilson Bull 109:68–73.
  33. Jaroszewicz B, Cholewiński O, Gutowski JJ, Samojlik T, Zimny M, Latałow M. 2019. Białowieża Forest -A relic of the high naturalness of European forests. Forests 10:849. doi:10.3390/f10100849.
  34. Jędrzejewska B, Jędrzejewski W. 1998. Predation in vertebrate communities The Białowieża Primeval Forest as a case study. Ecological Studies 135, Springer-Verlag, Berlin, Germany.
  35. Kawka M, Parada R, Jaszczak K, Horbańczuk JO. 2012. The use of microsatellite polymorphism in genetic mapping of the ostrich Struthio camelus. Mol Biol Rep 39:3369–3374. doi:10.1007/s11033-011-1107-5. [DOI] [PMC free article] [PubMed]
  36. Kempenaers B. 2007. Mate choice and genetic quality: a review of the heterozygosity theory. Adv Study Behav 37:189–278. doi:10.1016/S0065-3454(07)37005-8.
  37. Keller L, Reid J, Arcese P. 2008. Testing evolutionary models of senescence in a natural population: age and inbreeding effects on fitness components in song sparrows. Proc R Soc Lond B Biol Sci 275:597–604. doi:10.1098/rspb.2007.0961. [DOI] [PMC free article] [PubMed]
  38. Kokko H. 1999. Competition for early arrival in migratory birds. J Anim Ecol 68:940–950.
  39. Kokko H, Gunnarsson TG, Morrell LJ, Gill JA. 2006. Why do female migratory birds arrive later than males? J Anim Ecol 75:1293– 1303. doi:10.1111/j.1365-2656.2006.01151.x. [DOI] [PubMed]
  40. Kruuk LEB, Sheldon BC, Merila J. 2002. Severe inbreeding depression in collared flycatchers (Ficedula albicolis). Proc R Soc Lond B Biol Sci 269:1581–1589. doi:10.1098/rspb.2002.2049. [DOI] [PMC free article] [PubMed]
  41. Leder EH, Karaiskou N, Primmer CR. 2008. Seventy new microsatellites for the pied flycatcher, Ficedula hypoleuca and amplification in other passerine birds. Mol Ecol Res 8:874–880. doi:10.1111/j.1755-0998.2008.02096.x. [DOI] [PubMed]
  42. Lesbarrères D, Primmer CR, Laurila A, Merila J. 2005. Environmental and population dependency of genetic variability-fitness correlations in Rana temporaria. Mol Ecol 14:311–323. doi:10.1111/j.1365-294X.2004.02394.x. [DOI] [PubMed]
  43. Li YC, Korol AB, Fahima T, Nevo E. 2004. Microsatellites within genes: Structure, function, and evolution. Mol Biol Evol 21:991– 1007. doi:10.1093/molbev/msh073. [DOI] [PubMed]
  44. Ljungqvist M, ÅKesson M, Hansson B. 2010. Do microsatellites reflect genome-wide genetic diversity in natural populations? A comment on Väli et al. (2008). Mol Ecol 19:851–855. doi:10.1111/j.1365-294X.2010.04522.x. [DOI] [PubMed]
  45. Lozano GA, Perreault S, Lemon RE. 1996. Age, arrival date and reproductive success of male American Redstarts Setophaga ruticilla. J Avian Biol 27:164–170. doi:10.2307/3677146.
  46. Lynch M, Walsh B. 1998. Genetics and Analysis of Quantitative Traits. Sinauer Associates, editor Sunderland, Massachusetts, USA: Sinauer Associates, Inc.
  47. Mainguy J, Coté SD, Festa-Bianchet M, Coltman DW. 2009. Father-offspring phenotypic correlations suggest intralocus sexual conflict for a fitness-linked trait in a wild sexually dimorphic mammal. Proc R Soc Lond B Biol Sci 276:4067–4075. doi:10.1098/rspb.2009.1231. [DOI] [PMC free article] [PubMed]
  48. Malo AF, Coulson T. 2009. Heterozygosity-fitness correlations and associative overdominance: new detection method and proof of principle in the Iberian wild boar. Mol Ecol 18:2741–2747. doi:10.1111/j.1365-294X.2009.04219.x. [DOI] [PubMed]
  49. Marra PP, Hobson KA, Holmes RT. 1998. Linking winter and summer events in a migratory bird by using stable-carbon isotopes. Science 282:1884–1886. doi:10.1126/science.282.5395.1884. [DOI] [PubMed]
  50. Miller JM, Malenfant RM, David P, Davis CS, Poissant J, Hogg JT, Coltman DW. 2014. Estimating genome-wide heterozygosity: effects of demographic history and marker type. Heredity 112:240–247. doi:10.1038/hdy.2013.99. [DOI] [PMC free article] [PubMed]
  51. Mitrus C. 2006. The influence of male age and phenology on reproductive success of the red-breasted flycatcher (Ficedula parva Bechst.). Ann Zool Fenn 43:358–365.
  52. Mitrus C. 2007. Is the later arrival of young male red-breasted flycatchers (Ficedula parva) related to their physical condition? J Ornithol 148:53–58. doi:10.1007/s10336-006-0099-8.
  53. Mitrus C. 2018. Early arriving males wait longer for a mate than later arrivals: the case of a migratory monogamous passerine bird species. J Ethol 36:93–98. doi:10.1007/s10164-017-0531-y. [DOI] [PMC free article] [PubMed]
  54. Mitrus C, Soćko B. 2005. Polyterritoriality and the first record of polygyny in the Red-breasted Flycatcher Ficedula parva in a primeval forest. Acta Ornithol 40:170–172.
  55. Mitrus C, Sparks TH, Tryjanowski P. 2005. First evidence of phenological change in a transcontinental migrant overwintering in the Indian sub-continent: the Red-breasted flycatcher Ficedula parva. Ornis Fenn 82:13–19.
  56. Mitrus C, Mitrus J, Sikora M. 2012. Badge size and arrival time predict mating success of red-breasted flycatcher Ficedula parva males. Zool Sci 29:795–799. doi:10.2108/zsj.29.795. [DOI] [PubMed]
  57. Mitrus J, Mitrus C, Rutkowski R, Sikora M. 2014. Extra-pair paternity in relation to age of the Red-breasted Flycatcher Ficedula parva males. Avian Biol Res 7:111–116. doi:10.3184/17581551 4X13948188185179.
  58. Mitrus J, Mitrus C, Rutkowski R, Sikora M, Suchecka E. 2013. Characterisation of cross-amplified microsatellite markers in the red-breasted flycatcher Ficedula parva. Ann Zool 63:517–523. doi:10.3161/000345413X676731.
  59. Morin PA, Messier J, Woodruff DS. 1994. DNA extraction, amplification, and direct sequencing from hornbill feathers. J Sci Soc Thailand 20:31–41.
  60. Møller AP. 1994a. Phenotype-dependent arrival time and its consequences in a migratory bird. Behav Ecol Sociobiol 35:115– 122.
  61. Møller AP. 1994b. Sexual selection and the Barn Swallow. Oxford: Oxford University Press.
  62. Naef-Daenzer B, Grüebler M. 2016. Post-fledging survival of altricial birds: ecological determinants and adaptation. J Field Ornithol 87:227–250. doi:10.1111/jofo.12157.
  63. Ninni P, De Lope F, Saino N, Haussy C, Møller AP. 2004. Antioxidants and condition dependence of arrival date in a migratory passerine Oikos 105:55–64.
  64. Newton I. 2008. The Ecology of Bird Migration. London: Academic Press.
  65. Ortego J, Cordero PJ, Aparicio JM, Calabuig G. 2007. No relationship between individual genetic diversity and prevalence of avian malaria in a migratory kestrel. Mol Ecol 16:4858–4866. doi:10.1111/j.0030-1299.2004.12516.x. [DOI] [PubMed]
  66. Ortego J, Cordero PJ, Aparicio JM, Calabuig G. 2010. Parental genetic characteristics and hatching success in a recovering population of Lesser Kestrels. J Ornithol 151:155–162. doi:10.1007/s10336-009-0438-7.
  67. Owen M, Black JM. 1989. Factors affecting the survival of barnacle geese on migration from the breeding grounds. J Anim Ecol 58:603–617. doi:10.2307/4851.
  68. Patarnello T, Guinez R, Battaglia B. 1991. Effects of pollution on heterozygosity in the barnacle Balanus amphitrite (Cirripedia: Thoracica). Mar Ecol Progr 70:237–243. doi:10.3354/meps070237.
  69. Potti J, Montalvo S. 1991. Male arrival and female mate choice in Pied Flycatchers Ficedula hypoleuca. in central Spain. Ornis Scand 22:45–54. doi:10.2307/3676620.
  70. Primmer CR, Anders M, Ellegren H. 1996. New microsatellites from the pied flycatcher Ficedula hypoleuca and the swallow Hirundo rustica genomes. Hereditas 124:281–283. doi:10.1111/j.1601-5223.1996.00281.x. [DOI] [PubMed]
  71. Reid JM, Arcese P, Keller LF, Elliot KH, Sampson L, Hasselquist D. 2007. Inbreeding effects on immune response in free-living song sparrow (Melospiza melodia). Proc R Soc Lond B Biol Sci 274:697–706. doi:10.1098/rspb.2006.0092. [DOI] [PMC free article] [PubMed]
  72. Risely A, Nightingale J, Richardson DS, Barr I. 2013. Wing length and age, but not tarsus or mass, independently determine spring arrival at breeding territories in a long-distance migrant the Common Whitethroat, Sylvia communis. Bird Study 60:539–546. doi:10.1080/00063657.2013.846293.
  73. Roberts SC, Little AC, Gosling LM, Perrett DI, Carter V, Jones BC, Penton-Voak I, Petrie M. 2005. MHC-heterozygosity and human facial attractiveness. Evol Hum Behav 26:213–226. doi:10.1016/j.evolhumbehav.2004.09.002.
  74. Ryder TB, Tori WP, Blake JG, Loiselle BA, Parker PG. 2009. Mate choice for genetic quality: a test of the heterozygosity and compatibility hypotheses in a lek breeding bird. Behav Ecol 21:203–210. doi:10.1093/beheco/arp176.
  75. Seddon N, Amos W, Mulder RA, Tobias JA. 2004. Male heterozygosity predicts territory size, song structure and reproductive success in a cooperatively breeding bird. Proc R Soc Lond B Biol Sci 271:1823–1829. doi:10.1098/rspb.2004.2805. [DOI] [PMC free article] [PubMed]
  76. Selkoe KA, Toonen RJ. 2006. Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers. Ecol Lett 9:615–629. doi:10.1111/j.1461-0248.2006.00889.x. [DOI] [PubMed]
  77. Sillett TS, Holmes RT. 2002. Variation in survivorship of a migratory songbird throughout its annual cycle. J Anim Ecol 71:296–308. doi:10.1046/j.1365-2656.2002.00599.x.
  78. Slate J, David P, Dodds K, Veenvliet BA, Glass BC, Broad TE, McEwan JC. 2004. Understanding the relationship between the inbreeding coefficient and multilocus heterozygosity: theoretical expectations and empirical data. Heredity 93:255–265. doi:10.1038/sj.hdy.6800485. [DOI] [PubMed]
  79. Smith RJ, Moore FR. 2005. Arrival timing and seasonal reproductive performance in a long-distance migratory landbird. Behav Ecol Sociobiol 57:231–239. doi:10.1007/s00265-004-0855-9.
  80. Stewart RLM, Francis CM, Massey C. 2002. Age-related differential timing of spring migration within sexes in passerines. Wilson Bull 114:264–271. doi:10.1676/0043-5643(2002)114[0264:ARD TOS]2.0.CO;2.
  81. Stolt BO, Fransson T. 1995. Body mass, wing length and spring arrival of the Ortolan Bunting Emberiza hortulana. Ornis Fenn 72:14–18.
  82. Szulkin M, Bierne N, David P. 2010. Heterozygosity-fitness correlations: A time for reappraisal. Evolution 64:1202–1217. doi:10.1111/j.1558-5646.2010.00966.x. [DOI] [PubMed]
  83. Svensson L. 1992. Identification guide to European passerines. 4th edit Naturhistoriska. Riksmuseet, Stockholm, Sweden.
  84. Taylor SS, Sardell RJ, Reid JM, Bucher T, Taylor NG, Arcese P, Keller LF. 2010. Inbreeding coefficient and heterozygosity-fitness correlations in unhatched and hatched song sparrow nestmates. Mol Ecol 19:4454–4461. doi:10.1111/j.1365-294X.2010.04824.x. [DOI] [PubMed]
  85. Tiira K, Laurila A, Enberg K, Piironen, J, Aikio S, Ranta E, Primmer CR. 2006. Do dominants have higher heterozygosity? Social status and genetic variation in brown trout, Salmo trutta. Behav Ecol Soc 59:657–665. doi:10.1007/s00265-005-0094-8.
  86. Tomiałojć L. 1991. Characteristics of old growth in the Bialowieża Forest, Poland. Nat Areas J 11:7–18.
  87. Väli Ü, Einarsson A, Waits L, Ellegren H. 2008. To what extent do microsatellite markers reflect genome-wide genetic diversity in natural populations. Mol Ecol 17:3808–3817. doi:10.1111/j.1365-294X.2008.03876.x. [DOI] [PubMed]
  88. Wesołowski T, Czeszczewik D, Mitrus C, Rowiński P. 2003. Birds of the Białowieża National Park. Not Orn 44:1–31.
  89. Wesołowski T, Mitrus C, Czeszczewik D, Rowiński P. 2010. Breeding bird dynamics in a primeval temperate forest over thirty-five years: variation and stability in the changing world. Acta Ornithol 45:209–232. doi:10.3161/000164510X551354.
  90. Wesołowski T, Czeszczewik D, Hebda G, Maziarz M, Mitrus C, Rowiński P. 2015. 40 years of breeding bird community dynamics in a primeval temperate forest (Białowieża National Park, Poland). Acta Ornithol 50:95–120. doi:10.3161/00016454 AO2015.50.1.010.
  91. Wetzel DP, Stewart IR, Westneat DF. 2012. Heterozygosity predicts clutch and egg size but not plasticity in a house sparrow population with no evidence of inbreeding. Mol Ecol 21:406– 420. doi:10.1111/j.1365-294X.2011.05380.x. [DOI] [PubMed]
  92. Whiteman NK, Matson KD, Bollmer JL, Parker PG. 2006. Disease ecology in the Galapagos Hawk Buteo galapagoensis: host genetic diversity, parasites, and natural antibodies. Proc R Soc Lond B Biol Sci 273:797–804. doi:10.1098/rspb.2005.3396. [DOI] [PMC free article] [PubMed]
  93. Zeng Y, Li C, Zhang L, Zhong Z, Jiang Z. 2013. No correlation between neonatal fitness and heterozygosity in a reintroduced population of Père David’s deer. Curr Zool 59:249–256. doi:10.1093/czoolo/59.2.249.

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