The Yellow-browed Warbler (Phylloscopus inornatus) as a model to understand vagrancy and its potential for the evolution of new migration routes.

Why and how new migration routes emerge remain fundamental questions in ecology, particularly in the context of current global changes. In its early stages, when few individuals are involved, the evolution of new migration routes can be easily confused with vagrancy, i.e. the occurrence of individuals outside their regular breeding, non-breeding or migratory distribution ranges. Yet, vagrancy can in theory generate new migration routes if vagrants survive, return to their breeding grounds and transfer their new migration route to their offspring, thus increasing a new migratory phenotype in the population. Here, we review the conceptual framework and empirical challenges of distinguishing regular migration from vagrancy in small obligate migratory passerines and explain how this can inform our understanding of migration evolution. For this purpose, we use the Yellow-browed Warbler (Phylloscopus inornatus) as a case study. This Siberian species normally winters in southern Asia and its recent increase in occurrence in Western Europe has become a prominent evolutionary puzzle. We first review and discuss available evidence suggesting that the species is still mostly a vagrant in Western Europe but might be establishing a new migration route initiated by vagrants. We then list possible empirical approaches to check if some individuals really undertake regular migratory movements between Western Europe and Siberia, which would make this species an ideal model for studying the links between vagrancy and the emergence of new migratory routes.

Magnetoreceptory Function of European Robin Retina: Electrophysiological and Morphological Non-Homogeneity.

The avian magnetic compass allows orientation during migration and is shown to function properly under short-wavelength but not long-wavelength visible light. Therefore, the magnetoreceptive system is assumed to be light- and wavelength-dependent and localized in the retina of the eye. Putative candidates for the role of primary magnetosensory molecules are the cryptochromes that are known to be expressed in the avian retina and must be able to interact with phototransduction proteins. Previously, we reported that in migratory birds change in magnetic field direction induces significant effects on electroretinogram amplitude in response to blue flashes, and such an effect was observed only in the nasal quadrant of the retina. Here, we report new electroretinographic, microscopic and microspectrophotometric data on European robins, confirming the magnetosensitivity of the retinal nasal quadrant after applying the background illumination. We hypothesized that magnetoreceptive distinction of this region may be related to its morphology and analyzed the retinal distribution and optical properties of oil droplets, the filtering structures within cones. We found that the nasal quadrant contains double cones with the most intensely colorized oil droplets compared to the rest of the retina, which may be related to its magnetosensory function.

Access to the sky near the horizon and stars does not play a crucial role in compass calibration of European songbird migrants.

Migratory birds use different global cues including celestial and magnetic information to determine and maintain their seasonally appropriate migratory direction. A hierarchy among different compass systems in songbird migrants is still a matter for discussion owing to highly variable and apparently contradictory results obtained in various experimental studies. How birds decide whether and how they should calibrate their compasses before departure remains unclear. A recent 'extended unified theory' suggested that access to both a view of the sky near the horizon and stars during the cue-conflict exposure might be crucial for the results of cue-conflict experiments. In this study, we performed cue-conflict experiments in three European songbird species with different migratory strategies (garden warbler, Sylvia borin; pied flycatcher, Ficedula hypoleuca; and European robin, Erithacus rubecula; juveniles and adults; spring and autumn migrations) using a uniform experimental protocol. We exposed birds to the natural celestial cues in a shifted (120 deg clockwise/counterclockwise) magnetic field from sunset to the end of the nautical twilight and tested them in orientation cages immediately after cue-conflict treatments. None of the species (apart from adult robins) showed any sign of calibration even if they had access to a view of the sky and local surroundings near the horizon and stars during cue-conflict treatments. Based on results of our experiments and data from previous contradictory studies, we suggest that no uniform theory can explain why birds calibrate or do not calibrate their compass systems. Each species (and possibly even different populations) may choose its calibration strategy differently.

Geographical and Tick-Dependent Distribution of Flavi-Like Alongshan and Yanggou Tick Viruses in Russia.

The genus Flavivirus includes related, unclassified segmented flavi-like viruses, two segments of which have homology with flavivirus RNA-dependent RNA polymerase NS5 and RNA helicase-protease NS3. This group includes such viruses as Jingmen tick virus, Alongshan virus, Yanggou tick virus and others. We detected the Yanggou tick virus in Dermacentor nuttalli and Dermacentor marginatus ticks in two neighbouring regions of Russia. The virus prevalence ranged from 0.5% to 8.0%. We detected RNA of the Alongshan virus in 44 individuals or pools of various tick species in eight regions of Russia. The virus prevalence ranged from 0.6% to 7.8%. We demonstrated the successful replication of the Yanggou tick virus and Alongshan virus in IRE/CTVM19 and HAE/CTVM8 tick cell lines without a cytopathic effect. According to the phylogenetic analysis, we divided the Alongshan virus into two groups: an Ixodes persulcatus group and an Ixodes ricinus group. In addition, the I. persulcatus group can be divided into European and Asian subgroups. We found amino acid signatures specific to the I. ricinus and I. persulcatus groups and also distinguished between the European and Asian subgroups of the I. persulcatus group.

Navigation by extrapolation of geomagnetic cues in a migratory songbird.

Displacement experiments have demonstrated that experienced migratory birds translocated thousands of kilometers away from their migratory corridor can orient toward and ultimately reach their intended destinations.1 This implies that they are capable of "true navigation," commonly defined2-4 as the ability to return to a known destination after displacement to an unknown location without relying on familiar surroundings, cues that emanate from the destination, or information collected during the outward journey.5-13 In birds, true navigation appears to require previous migratory experience5-7,14,15 (but see Kishkinev et al.16 and Piersma et al.17). It is generally assumed that, to correct for displacements outside the familiar area, birds initially gather information within their year-round distribution range, learn predictable spatial gradients of environmental cues within it, and extrapolate from those to unfamiliar magnitudes-the gradient hypothesis.6,9,18-22 However, the nature of the cues and evidence for actual extrapolation remain elusive. Geomagnetic cues (inclination, declination, and total intensity) provide predictable spatial gradients across large parts of the globe and could serve for navigation. We tested the orientation of long-distance migrants, Eurasian reed warblers, exposing them to geomagnetic cues of unfamiliar magnitude encountered beyond their natural distribution range. The birds demonstrated re-orientation toward their migratory corridor as if they were translocated to the corresponding location but only when all naturally occurring magnetic cues were presented, not when declination was changed alone. This result represents direct evidence for migratory birds' ability to navigate using geomagnetic cues extrapolated beyond their previous experience.

Ontogeny of the star compass in birds: pied flycatchers (Ficedula hypoleuca) can establish the star compass in spring.

The star compass of birds, like the sun compass, is not innate. To possess either of them, birds have to observe the rotating sky and determine its centre of rotation (in the case of the star compass) or the sun's movement (for the sun compass). Young birds are believed to learn how to use the star compass before their first migration, even though the evidence of this is lacking. Here, we tested whether hand-raised Pied flycatchers (Ficedula hypoleuca) that had not established the star compass prior to their first autumn migration can gain it later in their ontogeny, in spring. We also attempted to examine whether the observation of diurnal celestial cues (the sun and polarized light) prior to autumn migration would affect the process of star compass learning in spring. When tested in the vertical magnetic field under the natural starry sky, the group of birds that observed the stars in spring as the first celestial cues were able to choose the migratory direction. In contrast, the birds that had never seen the stars were not able to use the nightly celestial cues in the vertical magnetic field. However, birds that had seen the daytime celestial cues till autumn and the stars at spring were disoriented, although this might be due to the small sample size. Our data suggest the possibility that the star compass may be learned in spring and emphasize the necessity for further research into the interaction of celestial compasses.

Electroretinographic study of the magnetic compass in European robins.

Migratory birds are known to be sensitive to external magnetic field (MF). Much indirect evidence suggests that the avian magnetic compass is localized in the retina. Previously, we showed that changes in the MF direction could modulate retinal responses in pigeons. In the present study, we performed similar experiments using the traditional model animal to study the magnetic compass, European robins. The photoresponses of isolated retina were recorded using ex vivo electroretinography (ERG). Blue- and red-light stimuli were applied under an MF with the natural intensity and two MF directions, when the angle between the plane of the retina and the field lines was 0° and 90°, respectively. The results were separately analysed for four quadrants of the retina. A comparison of the amplitudes of the a- and b-waves of the ERG responses to blue stimuli under the two MF directions revealed a small but significant difference in a- but not b-waves, and in only one (nasal) quadrant of the retina. The amplitudes of both the a- and b-waves of the ERG responses to red stimuli did not show significant effects of the MF direction. Thus, changes in the external MF modulate the European robin retinal responses to blue flashes, but not to red flashes. This result is in a good agreement with behavioural data showing the successful orientation of birds in an MF under blue, but not under red illumination.

Phlebovirus sequences detected in ticks collected in Russia: Novel phleboviruses, distinguishing criteria and high tick specificity.

Phlebovirus is an abundant and rather heterogeneous genus within the Phenuiviridae family (order Bunyavirales). The genus Phlebovirus is divided into two antigenic complexes, which also correspond to the main vector: sandflies/mosquitoes and ticks. Previously, only sandfly/mosquito-borne phleboviruses were associated with human disease, such as Rift Valley fever virus, Toscana virus, Sicilian and Naples Sandfly fever viruses and others. Until recently, tick-borne phleboviruses were not considered as human pathogens. After the discovery of severe fever with thrombocytopenia syndrome, interest to tick-borne phleboviruses has increased dramatically. In the last decade, many novel phleboviruses have been reported in different regions. Despite this, the diversity, ecology and pathogenicity of these viruses still remain obscure. The aim of this work was to study the diversity of phleboviruses in ticks collected in several regions of Russia. We used pan-phlebovirus RT-PCR assays based on multiple degenerate primers targeting the polymerase gene fragment. Arthropod specimens were collected from 2005 to 2018. A total of 5901 Ixodidae ticks combined into 1116 pools were screened. A total of 160 specific amplicons were produced. In three cases RT-PCR assays amplified two distinct viruses from same tick pools. Direct sequencing of amplicons and subsequent phylogenetic analysis revealed twelve representatives of divergent phlebovirus groups. Based on the distribution of pairwise nucleotide sequence identity values, a cut-off (88%) was suggested to distinguish tick-borne phleboviruses. According to this provisional criterion, two viruses found here could be termed novel, while ten viruses have been described in previous studies. Detected phleboviruses demonstrated almost perfect specificity to a tick species or, at least, a genus. The same pattern was observed for tick-borne phleboviruses found in different studies around the world. Viruses that grouped together on a phylogenetic tree and differed less than this sequence identity threshold suggested above were hosted by ticks from the same genus.

No evidence for the use of magnetic declination for migratory navigation in two songbird species.

Determining the East-West position was a classical problem in human sea navigation until accurate clocks were manufactured and sailors were able to measure the difference between local time and a fixed reference to determine longitude. Experienced night-migratory songbirds can correct for East-West physical and virtual magnetic displacements to unknown locations. Migratory birds do not appear to possess a time-different clock sense; therefore, they must solve the longitude problem in a different way. We showed earlier that experienced adult (but not juvenile) Eurasian reed warblers (Acrocephalus scirpaceus) can use magnetic declination (the difference in direction between geographic and magnetic North) to solve this problem when they were virtually displaced from Rybachy on the eastern Baltic coast to Scotland. In this study, we aimed to test how general this effect was. Adult and juvenile European robins (Erithacus rubecula) and adult garden warblers (Sylvia borin) under the same experimental conditions did not respond to this virtual magnetic displacement, suggesting significant variation in how navigational maps are organised in different songbird migrants.

Searching for magnetic compass mechanism in pigeon retinal photoreceptors.

Migratory birds can detect the direction of the Earth's magnetic field using the magnetic compass sense. However, the sensory basis of the magnetic compass still remains a puzzle. A large body of indirect evidence suggests that magnetic compass in birds is localized in the retina. To confirm this point, an evidence of visual signals modulation by magnetic field (MF) should be obtained. In a previous study we showed that MF inclination impacts the amplitude of ex vivo electroretinogram (ERG) recorded from isolated pigeon retina. Here we present the results of an analysis of putative MF effect on one component of ERG, the photoreceptor's response, isolated from the total ERG by adding sodium aspartate and barium chloride to the perfusion solution. Photoresponses were recorded from isolated retinae of domestic pigeons Columba livia. The retinal samples were placed in MF that was modulated by three pairs of orthogonal Helmholtz coils. Light stimuli (blue and red) were applied under two inclinations of MF, 0° and 90°. In all the experiments, preparations from two parts of retina were used, red field (with dominant red-sensitive cones) and yellow field (with relatively uniform distribution of cone color types). In contrast to the whole retinal ERG, we did not observe any effect of MF inclination on either amplitude or kinetics of pharmacologically isolated photoreceptor responses to blue or red half-saturating flashes. A possible explanations of these results could be that magnetic compass sense is localized in retinal cells other than photoreceptors, or that photoreceptors do participate in magnetoreception, but require some processing of compass information in other retinal layers, so that only whole retina signal can reflect the response to changing MF.

Magnetic compass of garden warblers is not affected by oscillating magnetic fields applied to their eyes.

The magnetic compass is an important element of the avian navigation system, which allows migratory birds to solve complex tasks of moving between distant breeding and wintering locations. The photochemical magnetoreception in the eye is believed to be the primary biophysical mechanism behind the magnetic sense of birds. It was shown previously that birds were disoriented in presence of weak oscillating magnetic fields (OMF) with frequencies in the megahertz range. The OMF effect was considered to be a fingerprint of the photochemical magnetoreception in the eye. In this work, we used miniaturized portable magnetic coils attached to the bird's head to specifically target the compass receptor. We performed behavioural experiments on orientation of long-distance migrants, garden warblers (Sylvia borin), in round arenas. The OMF with the amplitude of about 5 nT was applied locally to the birds' eyes. Surprisingly, the birds were not disoriented and showed the seasonally appropriate migratory direction. On the contrary, the same birds placed in a homogeneous 5 nT OMF generated by large stationary coils showed clear disorientation. On the basis of these findings, we suggest that the disruption of magnetic orientation of birds by oscillating magnetic fields is not related to photochemical magnetoreceptors in their eyes.

Magnetic map navigation in a migratory songbird requires trigeminal input.

Recently, virtual magnetic displacement experiments have shown that magnetic cues are indeed important for determining position in migratory birds; but which sensory system(s) do they use to detect the magnetic map cues? Here, we show that Eurasian reed warblers need trigeminal input to detect that they have been virtually magnetically displaced. Birds with bilaterally ablated ophthalmic branches of the trigeminal nerves were not able to re-orient towards their conspecific breeding grounds after a virtual magnetic displacement, exactly like they were not able to compensate for a real physical displacement. In contrast, sham-operated reed warblers re-oriented after the virtual displacement, like intact controls did in the past. Our results show that trigeminally mediated sensory information is necessary for the correct function of the reed warblers' magnetic positioning system.

Migratory Eurasian Reed Warblers Can Use Magnetic Declination to Solve the Longitude Problem.

The longitude problem (determining east-west position) is a classical problem in human sea navigation. Prior to the use of GPS satellites, extraordinarily accurate clocks measuring the difference between local time and a fixed reference (e.g., GMT) [1] were needed to determine longitude. Birds do not appear to possess a time-difference clock sense [2]. Nevertheless, experienced night-migratory songbirds can correct for east-west displacements to unknown locations [3-9]. Consequently, migratory birds must solve the longitude problem in a different way, but how they do so has remained a scientific mystery [10]. We suggest that experienced adult Eurasian reed warblers (Acrocephalus scirpaceus) can use magnetic declination to solve the longitude problem at least under some circumstances under clear skies. Experienced migrants tested during autumn migration in Rybachy, Russia, were exposed to an 8.5° change in declination while all other cues remained unchanged. This corresponds to a virtual magnetic displacement to Scotland if and only if magnetic declination is a part of their map. The adult migrants responded by changing their heading by 151° from WSW to ESE, consistent with compensation for the virtual magnetic displacement. Juvenile migrants that had not yet established a navigational map also oriented WSW at the capture site but became randomly oriented when the magnetic declination was shifted 8.5°. In combination with latitudinal cues, which birds are known to detect and use [10-12], magnetic declination could provide the mostly east-west component for a true bi-coordinate navigation system under clear skies for experienced migratory birds in some areas of the globe.

Very weak oscillating magnetic field disrupts the magnetic compass of songbird migrants.

Previously, it has been shown that long-distance migrants, garden warblers (Sylvia borin), were disoriented in the presence of narrow-band oscillating magnetic field (1.403 MHz OMF, 190 nT) during autumn migration. This agrees with the data of previous experiments with European robins (Erithacus rubecula). In this study, we report the results of experiments with garden warblers tested under a 1.403 MHz OMF with various amplitudes (∼0.4, 1, ∼2.4, 7 and 20 nT). We found that the ability of garden warblers to orient in round arenas using the magnetic compass could be disrupted by a very weak oscillating field, such as an approximate 2.4, 7 and 20 nT OMF, but not by an OMF with an approximate 0.4 nT amplitude. The results of the present study indicate that the sensitivity threshold of the magnetic compass to the OMF lies around 2-3 nT, while in experiments with European robins the birds were disoriented in a 15 nT OMF but could choose the appropriate migratory direction when a 5 nT OMF was added to the stationary magnetic field. The radical-pair model, one of the mainstream theories of avian magnetoreception, cannot explain the sensitivity to such a low-intensity OMF, and therefore, it needs further refinement.

Theoretically possible spatial accuracy of geomagnetic maps used by migrating animals.

Many migrating animals, belonging to different taxa, annually move across the globe and cover hundreds and thousands of kilometres. Many of them are able to show site fidelity, i.e. to return to relatively small migratory targets, from distant areas located beyond the possible range of direct sensory perception. One widely debated possibility of how they do it is the use of a magnetic map, based on the dependence of parameters of the geomagnetic field (total field intensity and inclination) on geographical coordinates. We analysed temporal fluctuations of the geomagnetic field intensity as recorded by three geomagnetic observatories located in Europe within the route of many avian migrants, to study the highest theoretically possible spatial resolution of the putative map. If migratory birds measure total field intensity perfectly and take the time of day into account, in northern Europe 81% of them may return to a strip of land of 43 km in width along one of coordinates, whereas in more southern areas such a strip may be narrower than 10 km. However, if measurements are performed with an error of 0.1%, the strip width is increased by approximately 40 km, so that in spring migrating birds are able to return to within 90 km of their intended goal. In this case, migrating birds would probably need another navigation system, e.g. an olfactory map, intermediate between the large-scale geomagnetic map and the local landscape cues, to locate their goal to within several kilometres.

Compass systems.

Three compass systems based on global cues known to exist in migrating birds are reviewed. Two of these systems are based on celestial cues, a time-dependent sun compass and time-independent, i.e. not involving the internal clock, star compass. The third system is the magnetic compass, based on a separate sensory modality, which currently attracts much attention from behavioural ecologists, physiologists and physicists. The complex pattern of hierarchy and interactions between these compass systems is briefly discussed. It is argued that rules of integration of information from different compass cues are likely dependent on ecological and geographic conditions the birds are facing during their journey, so it is likely that no single set of rules is shared by all migrating birds.

Magnetic orientation of garden warblers (Sylvia borin) under 1.4 MHz radiofrequency magnetic field.

We report on the experiments on orientation of a migratory songbird, the garden warbler (Sylvia borin), during the autumn migration period on the Courish Spit, Eastern Baltics. Birds in experimental cages, deprived of visual information, showed the seasonally appropriate direction of intended flight with respect to the magnetic meridian. Weak radiofrequency (RF) magnetic field (190 nT at 1.4 MHz) disrupted this orientation ability. These results may be considered as an independent replication of earlier experiments, performed by the group of R. and W. Wiltschko with European robins (Erithacus rubecula). Confirmed outstanding sensitivity of the birds' magnetic compass to RF fields in the lower megahertz range demands for a revision of one of the mainstream theories of magnetoreception, the radical-pair model of birds' magnetic compass.

Contrasting patterns of genetic differentiation among Blackcaps (Sylvia atricapilla) with divergent migratory orientations in Europe.

Migratory divides are thought to facilitate behavioral, ecological, and genetic divergence among populations with different migratory routes. However, it is currently contentious how much genetic divergence is needed to maintain distinct migratory behavior across migratory divides. Here we investigate patterns of neutral genetic differentiation among Blackcap (Sylvia atricapilla) populations with different migratory strategies across Europe. We compare the level of genetic divergence of populations migrating to southwestern (SW) or southeastern (SE) wintering areas with birds wintering in the British Isles following a recently established northwesterly (NW) migration route. The migratory divide between SW and SE wintering areas can be interpreted as a result of a re-colonization process after the last glaciation. Thus we predicted greater levels of genetic differentiation among the SW/SE populations. However, a lack of genetic differentiation was found between SW and SE populations, suggesting that interbreeding likely occurs among Blackcaps with different migratory orientations across a large area; therefore the SW/SE migratory divide can be seen as diffuse, broad band and is, at best, a weak isolating barrier. Conversely, weak, albeit significant genetic differentiation was evident between NW and SW migrants breeding sympatrically in southern Germany, suggesting a stronger isolating mechanism may be acting in this population. Populations located within/near the SW/SE contact zone were the least genetically divergent from NW migrants, confirming NW migrants likely originated from within the contact zone. Significant isolation-by-distance was found among eastern Blackcap populations (i.e. SE migrants), but not among western populations (i.e. NW and SW migrants), revealing different patterns of genetic divergence among Blackcap populations in Europe. We discuss possible explanations for the genetic structure of European Blackcaps and how gene flow influences the persistence of divergent migratory behaviors.