Sándor Zsebők

Among-year variation in the repeatability, within- and between-individual, and phenotypic correlations of behaviors in a natural population

When mean behaviors correlate among individuals, they form behavioral syndromes. One way to understand the evolution of such a group-level phenomenon is to compare horizontally patterns of correlations among populations (or species) or follow longitudinally the same population over years in the light of parallel differences in the environment. We applied the longitudinal approach to 8-year field data and analyzed phenotypic correlations, and their within- and between-individual components, among three behaviors (novelty avoidance, aggression, and risk-taking) in male collared flycatchers, Ficedula albicollis, in a meta-analytic framework. The phenotypic correlation between novelty avoidance and aggression varied heterogeneously (it was positive in some years, while it was negative in other years), while the other pair-wise correlations were consistently positive over the study period. We investigated four potential socio-ecological factors, and found evidence that the among-year alterations in the demographic structure of the population (density, age composition) can be responsible for the heterogeneous effect sizes. Comparing within- and between-individual correlations across pairs of traits, we found that the correlation between aggression and risk-taking at the among-individual level was the strongest suggesting that this relationship has the highest potential to form a behavioral syndrome. Within-year repeatabilities varied among traits, but were systematically higher than between-year repeatabilities. Our study highlights on an empirical basis that there can be several biological and statistical reasons behind detecting a phenotypic correlation in a study, but only few of these imply that fixed behavioral syndromes are maintained in a natural population. In fact, some correlations seem to be shaped by environmental fluctuations.

The relationship between DRD4 polymorphisms and phenotypic correlations of behaviors in the collared flycatcher

There is increasing evidence that the genetic architecture of exploration behavior includes the dopamine receptor D4 gene (DRD4). Such a link implies that the within-individual consistency in the same behavior has a genetic basis. Behavioral consistency is also prevalent in the form of between-individual correlation of functionally different behaviors; thus, the relationship between DRD4 polymorphism and exploration may also be manifested for other behaviors. Here, in a Hungarian population of the collared flycatcher, Ficedula albicollis, we investigate how males with distinct DRD4 genotypes differ in the consistent elements of their behavioral displays during the courtship period. In completely natural conditions, we assayed novelty avoidance, aggression and risk-taking, traits that were previously shown repeatable over time and correlate with each other, suggesting that they could have a common mechanistic basis. We identified two single-nucleotide polymorphisms (SNP554 and SNP764) in the exon 3 of the DRD4 gene by sequencing a subsample, then we screened 202 individuals of both sexes for these SNPs. Focusing on the genotypic variation in courting males, we found that “AC” heterozygote individuals at the SNP764 take lower risk than the most common “AA” homozygotes (the “CC” homozygotes were not represented in our subsample of males). We also found a considerable effect size for the relationship between SNP554 polymorphism and novelty avoidance. Therefore, in addition to exploration, DRD4 polymorphisms may also be associated with the regulation of behaviors that may incur fear or stress. Moreover, polymorphisms at the two SNPs were not independent indicating a potential role for genetic constraints or another functional link, which may partially explain behavioral correlations.

Sex-dependent role of vesicular glutamate transporter 3 in stress-regulation and related anxiety phenotype during the early postnatal period

Abstract Stress and related disorders are in the focus of interest and glutamate is one of the most important neurotransmitters that can affect these processes. Glutamatergic neurons are characterized by vesicular glutamate transporters (VGluT1-3) among which vGluT3 is unique contributing to the non-canonical, neuromodulatory effect of glutamate. We aimed to study the role of vGluT3 in stress axis regulation and related anxiety during the early postnatal period using knockout (KO) mice with special focus on sex differences. Anxiety was explored on postnatal day (PND) 7-8 by maternal separation-induced ultrasonic vocalization (USV). Stress-hormone levels were detected 60 min after intraperitoneal lipopolysaccharide (LPS) injection 7 days later. Both genotypes gained weight, but on PND 14-15 KO mice pups had smaller body weight compared to wild type (WT). vGluT3 KO mice reacted to an immune stressor with enhanced adrenocorticotropin (ACTH) and corticosterone secretion compared to WT. Although there was a tendency for enhanced anxiety measured by more emitted USV, this did not reach the level of significance. The only sex-related effect was the enhanced corticosterone reactivity in male pups. For the HPA axis regulation in neonates vGluT3 expression seems to be dispensable under basal conditions, but is required for optimal response to immune stressors, most probably through an interaction with other neurotransmitters. Disturbance of the fine balance between these systems may result in a borderline enhanced anxiety-like behavior in vGluT3 KO pups.

Minimum spanning tree as a new, robust repertoire size comparison method: simulation and test on birdsong

The comparison of acoustic complexity across individuals is often essential for understanding the evolution of acoustic signals. In many animal taxa, as a proxy of acoustic complexity, repertoire size is intensively studied; however, its estimation is challenging in species with large repertoires, as this process is time-consuming and may involve considerable subjectivity for the classification of signal elements. Here, we propose a novel application of the minimum spanning tree (MST) method for comparing individuals’ signal complexity, an approach that does not require classification process. We suggest that the differences in the MST length predict the differences in the repertoire sizes between individuals. To evaluate these proposals, first, we performed a simulation study investigating the effect of the practically important variables (repertoire size, number of acoustic parameters, sample size, distribution of element types and within-group variance) on the MST length. Second, we compared repertoire size estimates from the same song data from male collared flycatchers obtained using the fully manual, computer-aided manual and MST methods. In our simulation study, we found that the repertoire size strongly correlated with MST length. We also found significant effects of sample size, number of parameters and within-group variance, as well as how uniformly the samples were distributed between the groups, on the MST length. Our empirical data also revealed a strong correlation between the computer-aided manual estimation of repertoire sizes and MST length, which was comparable to the correlation between the estimations of repertoire size obtained using the two different manual methods. Therefore, we suggest using the MST method to compare the acoustic complexity among individuals in birds and other animals, with the practical restrictions suggested by our simulation results.

MHC-mediated sexual selection on birdsong: Generic polymorphism, particular alleles and acoustic signals

Several hypotheses predict that the major histocompatibility complex (MHC) drives mating preference in females. Olfactory, colour or morphological traits are often found as reliable signals of the MHC profile, but the role of avian song mediating MHC‐based female choice remains largely unexplored. We investigated the relationship between several MHC and acoustic features in the collared flycatcher (Ficedula albicollis), a European passerine with complex songs. We screened a fragment of the class IIB second exon of the MHC molecule, of which individuals harbour 4–15 alleles, while considerable sequence diversity is maintained at the population level. To make statistical inferences from a large number of comparisons, we adopted both null‐hypothesis testing and effect size framework in combination with randomization procedures. After controlling for potential confounding factors, neither MHC allelic diversity nor the presence of particular alleles was associated remarkably with the investigated qualitative and quantitative song traits. Furthermore, genetic similarity among males based on MHC sequences was not reflected by the similarity in their song based on syllable content. Overall, these results suggest that the relationship between features of song and the allelic composition and diversity of MHC is not strong in the studied species. However, a biologically motivated analysis revealed that individuals that harbour an MHC allele that impairs survival perform songs with broader frequency range. This finding suggests that certain aspects of the song may bear reliable information concerning the MHC profile of the individuals, which can be used by females to optimize mate choice.

“Ficedula”: an open-source MATLAB toolbox for cutting, segmenting and computer-aided clustering of bird song

AbstractQualitative and quantitative assessments of bird song repertoires are important in studies related to song learning, sexual selection and cultural evolution. Despite methods for automatic analysis, it is still necessary to engage in manual cutting, segmenting and clustering of bird song elements in many cases. Here, we describe a program, the Ficedula Toolbox, which has been made available for free to the bird song research community and has recently come into extensive use. The main advantages of this package are the opportunity to conduct all processing steps in one framework and the option of carrying out computer-aided manual clustering. Output files are ready for further analyses, such as estimation of repertoire size, sequential analysis and repertoire overlap calculation. With this program, findings based on empirical data from the Collared Flycatcher (Ficedula albicollis) song show high inter-observer similarity, and thus, reproducible results. The toolbox may be especially applicable to the analysis of song in species with moderately high repertoires.Zusammenfassung“Ficedula”—eine open-source MATLAB toolbox für Schnitt, Segmentierung und computerunterstütztes Clustering von Vogelgesang Die qualitative und quantitative Bewertung von Gesangsrepertoires von Vögeln ist wesentlich in Studien im Zusammenhang mit Themen wie Gesangslernen, sexuelle Selektion und kulturelle Evolution. Trotz automatischer Analysemöglichkeiten ist es in vielen Fällen immer noch nötig, die Gesänge manuell zu Schneiden, zu Segmentierung und zu Clustern. Hier beschreiben wir das Programm „Ficedula Toolbox“, welches für Vogelgesangsforscher frei verfügbar gemacht wurde und das neuerdings in größerem Umfang genutzt wird. Die größten Vorteile dieses Paketes sind zum einen die Möglichkeit, alle Verarbeitungsschritte in einem System vorzunehmen und zum anderen die Option der Durchführung von computerunterstütztem manuellen Clustern. Die Ausgabedateien sind bereit für weitere Auswertungen, wie beispielsweise die Bestimmung des Repertoireumfangs, sequentielle Analysen und die Berechnung von Repertoireüberlappungen. Mit Hilfe dieses Programms zeigen die Ergebnisse, basierend auf empirischen Daten zum Halsbandschnäpper-Gesang (Ficedula albicollis) eine große Ähnlichkeit zwischen den Beobachtern und daher reproduzierbare Resultate. Die Nutzung dieser Toolbox könnte insbesondere für Arten mit einem moderat hohen Gesangsrepertoire von Vorteil sein.

A test on within-individual changes in risk-taking behaviour due to experience to predation in the Collared Flycatcher (Ficedula albicollis)

Summary Different experiences from the past may have influence on individual’s behaviour through feedback mechanisms that can weaken or preserve the within-individual consistency of behavioural traits. Here, we aimed to find evidence for such feedback mechanisms that may operate on risk-taking behaviour via the effect of former experience to potential predation events in male Collared Flycatchers (Ficedula albicollis). We predicted that risk-taking of males would decrease after experiencing a predator’s attack in previous breeding seasons (negative feedback). We assessed risk-taking by flight initiation distance (FID) that is the distance at which an individual flees from an approaching predator, which was estimated for 234 individuals from different breeding seasons. Information on predation experience (i.e. occurrence of nest predation, the incidence of capture by human observers) was available from our long-term database on individual life histories. In a horizontal approach, we found no difference in FID when comparing males with former experience to predation with males naive to predators. A longitudinal approach relying on the repeated tests of the same individuals from different years yielded analogous results, we could not show a significant change in the risk-taking behaviour of the males as a consequence of experience to predation in past years. However, we found that individuals systematically took less risk over the years, which might be a consequence of acquiring general experience with age.