Pre-migratory flights in migrant songbirds: the ecological and evolutionary importance of understudied exploratory movements.

Across the animal kingdom, from honeybees to cranes to beavers, exploratory movements to exploit resources, scout prospective territories, or otherwise gain valuable experiences and information that promote fitness have been documented. For example, exploratory movements to investigate potential dispersal targets have been observed in roe deer, Northern cardinals, and tigers alike. However, despite how widespread these movements are, a cohesive definition of exploratory movements has been lacking. We first provide a clear definition of exploratory movements, and use one particular group-migratory songbirds-to catalogue exploratory movements across the annual cycle. The exceptional mobility of migratory songbirds results in exploratory movements not only at a local scale, but also on a regional scale, both in and out of the breeding season. We review the extent to which these movements are made within this group, paying particular attention to how such movements confer fitness benefits, as by securing high-quality territories, prospecting for extra-pair paternity, or even exploiting ephemeral resources. We then zoom in one step further to a particular exploratory movement that has been, to date, almost completely overlooked within this group: that of pre-migratory flights. These flights, which occur during the transitional period between the stationary breeding period and the onset of migration, occur at night and may not be made by all individuals in a population-reasons why these flights have been heretofore critically understudied. We provide the first definition for this behaviour, summarise the current knowledge of this cryptic movement, and hypothesise what evolutionary/ecological advantages conducting it may confer to the individuals that undertake it. As these flights provide experience to the individuals that undertake them, we expect that birds that make pre-migratory flights are better equipped to survive migration (direct fitness benefits) and, due to orientation/navigation abilities, may also reach preferred territories on breeding and wintering grounds faster (indirect fitness benefits). We hope to encourage ecologists to consider such hidden movements in their research concepts and to enhance the framework of movement ecology by this behaviour due to its presumed high biological importance to the annual cycle of birds.

Automated component analysis in DOSY NMR using information criteria.

This study introduces a model selection technique based on Bayesian information criteria for estimating the number of components in a mixture during Diffusion-Ordered Spectroscopy (DOSY) Nuclear Magnetic Resonance (NMR) data analysis. As the accuracy of this technique is dependent on the efficiency of parameter estimators, we further investigate the performance of the Weighted Least Squares (WLS) and Maximum a Posteriori (MAP) estimators. The WLS method, enhanced with meticulously tuned L2-regularization, effectively detects components when the difference in self-diffusion coefficients is more than two-fold, especially when the component with the smaller coefficient has a larger weight ratio. The MAP method, strengthened by a substantial database of prior information, exhibits outstanding precision, decreasing this threshold to 1.5 times. Both estimators provide weight ratio estimates with standard deviations of approximately around 1 percentage point, although the MAP method tends to overestimate the component with a larger self-diffusion coefficient. Deviations from the expected values can exceed 10 percentage points, often due to inaccuracies in component detection. The error estimates are determined using data resampling techniques derived from a large-scale 1000-point experiment and an additional five measurements from a single-component mixture. This approach allowed us to thoroughly examine data distribution characteristics, thereby laying a robust groundwork for future refinement efforts.

Experimental study of newly described avian malaria parasite Plasmodium (Novyella) collidatum n. sp., genetic lineage pFANTAIL01 obtained from South Asian migrant bird.

BACKGROUND: Avian malaria parasites are microorganisms parasitizing erythrocytes and various tissues of the birds; they are common and distributed worldwide. These parasites are known to infect birds of different taxa and be the cause of the deaths of birds in the wild and in captivity. The species of parasites with the ability to colonize new territories and infect local non-migratory birds are of particular interest. This scenario is likely in temperate zones of Europe, because of climate change and its contribution in spreading vectors of southern origin, which can be involved in the transmission of malaria parasites. In the present study, a tropical Plasmodium parasite from a naturally infected long-distance migrant bird was isolated and tested for its ability to develop in common species of mosquitoes and European short-distance migrant birds. METHODS: Plasmodium sp. (pFANTAIL01) was isolated on the Curonian spit of the Baltic sea coast from the naturally infected Common rosefinch, Carpodacus erythrinus in June 2019. The parasite was described based on the morphological features of its blood stages, the partial mitochondrial cytochrome b gene and development after experimental infection of birds and mosquitoes. The parasite was inoculated into Eurasian siskins, Carduelis spinus. Parasitaemia, haematocrit and weight of birds were monitored. At the end of the survey, internal organs were collected to study exoerythrocytic stages of this parasite. Experimental infection of mosquitoes Culex pipiens form molestus and Culex quinquefasciatus was applied to study sporogonic development of the parasite. RESULTS: Based on morphological features, the parasite was described as a new species, Plasmodium collidatum n. sp., and attributed to subgenus Novyella. It was revealed that the obtained pFANTAIL01 lineage is a generalist parasite infecting a wide range of avian hosts and most likely is transmitted in South and Southeast (SE) Asia and Oceania. In Europe, this strain was recorded only in adult migratory birds wintering in South Asia. This parasite developed high parasitaemia in experimentally infected siskins and caused 25 % mortality. Exoerythrocytic stages of pFANTAIL01 were found in the lungs, liver, spleen and kidney of the deceased birds. Sporogonic development did not occur in Cx. pipiens form molestus and Cx. quinquefasciatus mosquitoes. CONCLUSIONS: Plasmodium collidatum is a highly virulent for Eurasian siskin and completes its development in these birds, which can be considered as a potential vertebrate host if the transmission of the infection starts occurring in Europe and temperate zones.

Culicoides biting midges involved in transmission of haemoproteids.

BACKGROUND: Culicoides biting midges (Diptera, Ceratopogonidae) are known vectors of avian Haemoproteus parasites. These parasites cause diseases, pathology and even mortality in birds. The diversity of biting midges in Europe is great, but only four Culicoides species are known to be vectors of avian Haemoproteus parasites. In general, our knowledge about the role of the particular Culicoides species in the transmission of Haemoproteus parasites remains insufficient. Information gaps hinder a better understanding of parasite biology and the epizootiology of parasite-caused diseases. The aim of this study was to determine new Culicoides species involved in the transmission of Haemoproteus parasites. METHODS: Biting midges were collected using a UV trap as well as sticky traps installed in bird nest boxes. Individual parous females were diagnosed for the presence of haemoproteids using both PCR-based and microscopic methods. RESULTS: We collected and dissected 232 parous Culicoides females from 9 species using a UV trap and 293 females from 11 species from bird nest boxes. Culicoides obsoletus was the dominant species collected using a UV trap, and Culicoides kibunensis dominated among midges collected in nest boxes. PCR-based screening showed that 5.2% of parous biting midges collected using a UV trap and 4.4% of midges collected from nest boxes were infected with avian haemosporidian parasites. Haemoproteid DNA was detected in C. kibunensis, Culicoides pictipennis, Culicoides punctatus, Culicoides segnis and Culicoides impunctatus females. The sporozoites of Haemoproteus minutus (genetic lineages hTURDUS2 and hTUPHI01) were detected in the salivary glands of two C. kibunensis females using microscopy, and this finding was confirmed by PCR. CONCLUSIONS: Culicoides kibunensis was detected as a new natural vector of Haemoproteus minutus (hTURDUS2 and hTUPHI01). Haemoproteid DNA was detected in females from five Culicoides species. This study contributes to the epizootiology of avian Haemoproteus infections by specifying Culicoides species as vectors and species that are likely to be responsible for the transmission of haemoproteids in Europe.

Dynamics of blood stage and sporozoite-induced malarial infections in experimentally infected passerines.

Complex experimental studies of vertebrate host, vector, and parasite interactions are essential in understanding virulence, but are difficult or impossible to conduct if vector species are unknown. Subinoculation of erythrocytic meronts of avian malarial parasites into susceptible hosts can avoid this problem, but this approach omits early exoerythrocytic stages, e.g. cryptozoites and metacryptozoites, that normally develop from sporozoites. A fundamental question that has remained unanswered is whether blood stage and sporozoite-induced malarial infections lead to differences in the dynamics of parasitemia in acute infections, patterns of parasite development, and host mortality. Here we demonstrate in a Carduelis spinus - Plasmodium relictum (genetic lineage pSGS1) system that experimental infections using inoculation of infected blood and using mosquito bite show similar peaks of parasitemias, but some measures of parasite development in the vertebrate host differ. Infected birds from all groups show decreased activity during the peak of parasitemia. There is no doubt that experimental infections using vectors provide the most precise information about the development of a parasite and its virulence in the host, but experimental infections using blood stages demonstrate similar parasitemias and effects on the host. These results are important for further experimental research of malarial parasites, especially studying avian Plasmodium parasites with unknown vectors.

Interrupted breeding in a songbird migrant triggers development of nocturnal locomotor activity.

Long-distance avian migrants, e.g. Eurasian reed warblers (Acrocephalus scirpaceus), can precisely schedule events of their annual cycle. However, the proximate mechanisms controlling annual cycle and their interplay with environmental factors are poorly understood. We artificially interrupted breeding in reed warblers by bringing them into captivity and recording birds' locomotor activity for 5-7 days. Over this time, most of the captive birds gradually developed nocturnal locomotor activity not observed in breeding birds. When the birds were later released and radio-tracked, the individuals with highly developed caged activity performed nocturnal flights. We also found that reed warblers kept indoors without access to local cues developed a higher level of nocturnal activity compared to the birds kept outdoors with an access to the familiar environment. Also, birds translocated from a distant site (21 km) had a higher motivation to fly at night-time after release compared to the birds captured within 1 km of a study site. Our study suggests that an interrupted breeding triggers development of nocturnal locomotor activity in cages, and the level of activity is correlated with motivation to perform nocturnal flights in the wild, which can be restrained by familiar environment.

The widespread biting midge Culicoides impunctatus (Ceratopogonidae) is susceptible to infection with numerous Haemoproteus (Haemoproteidae) species.

BACKGROUND: Haemoproteus parasites are widespread, and some species cause disease in wild and domestic birds. However, the insect vectors remain unknown for the majority of species and genetic lineages of avian Haemoproteus. This information is crucial for better understanding the biology of haemoproteids, the epidemiology of haemoproteosis, and the development of morphological characters of sporogonic stages in wildlife haemosporidian parasites. It remains unclear whether the specificity of Haemoproteus parasites for vectors is broad or the transmission of a given parasite can be restricted to a single or few species of vectors. The aim of this study was to examine the sporogonic development of four species of common European avian haemoproteids in the common biting midge Culicoides impunctatus. METHODS: Wild-caught females of C. impunctatus were infected experimentally by allowing them to take blood meals on naturally infected Muscicapa striata, Cyanistes caeruleus, Ficedula hypoleuca and Motacilla flava harbouring mature gametocytes of Haemoproteus balmorali (genetic lineage hSFC9), H. majoris (hPARUS1), H. motacillae (hYWT1) and H. pallidus (hPFC1), respectively. Infected insects were collected, maintained under laboratory conditions and dissected daily in order to detect the development of ookinetes, oocysts and sporozoites. Microscopic examination and polymerase chain reaction based methods were used to detect the parasites. Bayesian analysis was applied to identify phylogenetic relationships among Haemoproteus lineages. RESULTS: All investigated parasites completed sporogony in C. impunctatus, indicating broad susceptibility of this biting midge for numerous Haemoproteus parasites. Ookinetes, oocysts and sporozoites were reported, described and compared morphologically. The investigated parasite species can be distinguished at the sporogony stage, particularly with regards to the morphology and rate of development of mature ookinetes. Analysis of data from the literature, and this study, shows that 12 genetically distantly related Haemoproteus parasites complete sporogony in C. impunctatus. CONCLUSIONS: Susceptibility of C. impunctatus is broad for Haemoproteus parasites, indicating that this biting midge is an important natural vector of numerous species of avian haemoproteids in Europe. Some Haemoproteus species can be readily distinguished using morphological characters of ookinetes and sporozoites, as well as the rate of ookinete development. These characters can be used for the identification of Haemoproteus species during sporogony in vectors, and are worth more attention in these parasite taxonomy studies at the species levels.

Parallel telomere shortening in multiple body tissues owing to malaria infection.

Several studies have shown associations between shorter telomere length in blood and weakened immune function, susceptibility to infections, and increased risk of morbidity and mortality. Recently, we have shown that malaria accelerates telomere attrition in blood cells and shortens lifespan in birds. However, the impact of infections on telomere attrition in different body tissues within an individual is unknown. Here, we tested whether malarial infection leads to parallel telomere shortening in blood and tissue samples from different organs. We experimentally infected siskins (Spinus spinus) with the avian malaria parasite Plasmodium ashfordi, and used real-time quantitative polymerase chain reaction (PCR) to measure telomere length in control and experimentally infected siskins. We found that experimentally infected birds showed faster telomere attrition in blood over the course of infection compared with control individuals (repeatedly measured over 105 days post-infection (DPI)). Shorter telomeres were also found in the tissue of all six major organs investigated (liver, lungs, spleen, heart, kidney, and brain) in infected birds compared with controls at 105 DPI. To the best of our knowledge, this is the first study showing that an infectious disease results in synchronous telomere shortening in the blood and tissue cells of internal organs within individuals, implying that the infection induces systemic stress. Our results have far-reaching implications for understanding how the short-term effects of an infection can translate into long-term costs, such as organ dysfunction, degenerative diseases, and ageing.

The Strategy to Survive Primary Malaria Infection: An Experimental Study on Behavioural Changes in Parasitized Birds.

Avian malaria parasites (Haemosporida, Plasmodium) are of cosmopolitan distribution, and they have a significant impact on vertebrate host fitness. Experimental studies show that high parasitemia often develops during primary malaria infections. However, field studies only occasionally reveal high parasitemia in free-living birds sampled using the traditional methods of mist-netting or trapping, and light chronic infections predominate. The reason for this discrepancy between field observation and experimental data remains insufficiently understood. Since mist-netting is a passive capture method, two main parameters determine its success in sampling infected birds in wildlife, i. e. the presence of parasitized birds at a study site and their mobility. In other words, the trapping probability depends on the survival rate of birds and their locomotor activity during infection. Here we test (1) the mortality rate of wild birds infected with Plasmodium relictum (the lineage pSGS1), (2) the changes in their behaviour during presence of an aerial predator, and (3) the changes in their locomotor activity at the stage of high primary parasitemia.We show that some behavioural features which might affect a bird's survival during a predator attack (time of reaction, speed of flush flight and take off angle) did not change significantly during primary infection. However, the locomotor activity of infected birds was almost halved compared to control (non-infected) birds during the peak of parasitemia. We report (1) the markedly reduced mobility and (2) the 20% mortality rate caused by P. relictum and conclude that these factors are responsible for the underrepresentation of birds in mist nets and traps during the stage of high primary parasitemia in wildlife. This study indicates that the widespread parasite, P. relictum (pSGS1) influences the behaviour of birds during primary parasitemia. Experimental studies combined with field observations are needed to better understand the mechanisms of pathogenicity of avian malaria parasites and their influence on bird populations.

Under cover of darkness: nocturnal life of diurnal birds.

Songbirds are generally considered diurnal, although many species show periodic nocturnal activity during migration seasons. From a breeding-range perspective, such migratory species appear to be diurnal because they are observed to nest and feed their young during the day. But are they really exclusively diurnal? The authors tested how a passerine long-distance migrant, the Eurasian reed warbler, schedules movements during the breeding period by tracking birds in 2 experimental situations: 1) Birds experienced simulated nest loss and were monitored during their search for alternative locations, and 2) birds were translocated to reed beds at distances from 2 to 21 km and tracked during homing. The simulated unpredictable events disrupted normal breeding, forced birds to move over relatively long distances, and triggered rapid change in diel activity. In all but 1 case, birds resorted to nocturnality to find their way home and to search for new places to breed. Nocturnality during the breeding season indicates that songbird schedules are far more flexible than previously assumed. The reasons for nocturnal movements are poorly understood. Among the presumed advantages, the reduced predation pressure at night stands out because it is advantageous for movements on local as well as global scales. Predation may be particularly relevant for inhabitants of fragmented habitats, which encounter unfavorable conditions when crossing gaps in their preferred habitat. Therefore, similar selection pressures around the year may have favored the evolution of a general circadian mechanism for switches to nocturnality. Furthermore, the novel finding of homing and dispersal at night may give leads toward understanding the still enigmatic navigational abilities of songbirds.

Nocturnal life of young songbirds well before migration.

In songbirds, nocturnal activity is believed to be a characteristic feature of migration. However, unlike experimental conditions where the onset of nocturnal restlessness is defined as a shift of activity leading up to the dark period, this behaviour has, until now, not been observed in natural conditions. Here we studied the nocturnal behaviour of radio-tagged juvenile Eurasian reed warblers (Acrocephalus scirpaceus) during the pre-migratory period. The birds started nocturnal flights at the age of 38 days, whereas migration did not commence until they were at least 50 days old. The birds left their natal site by nocturnal flights and repeatedly returned to it. Such shuttle movements suggest the existence of a previously unknown period of nocturnal activity. Motivation to perform such night flights gradually increases with age. We relate the function of these nocturnal pre-migratory flights to the development of a stellar compass, necessary for detecting the compass direction towards winter quarters and for the formation of a navigational target, which will be used during return (spring) migration.