The association between haemosporidian infection and non-breeding moult location in great reed warblers revisited by combining feather stable isotope profiles and geolocator data.

Stable isotope analysis provides valuable insights into the ecology of long-distance migratory birds during periods spent away from a specific study site. In a previous study, Swedish great reed warblers (Acrocephalus arundinaceus) infected with haemosporidian parasites differed in feather isotope ratios compared to non-infected birds, suggesting that infected and non-infected birds spent the non-breeding season in different locations or habitats. Here, we use a novel dataset comprising geolocator data, isotopes, and haemosporidian infection status of 92 individuals from four Eurasian populations to investigate whether parasite transmission varies with geography or habitats. We found that the probability of harbouring Plasmodium and Leucocytozoon parasites was higher in birds moulting in the eastern region of the non-breeding grounds. However, no geographic pattern occurred for Haemoproteus infections or overall infection status. In contrast to the previous study, we did not find any relationship between feather isotope ratios and overall haemosporidian infection for the entire current dataset. Plasmodium-infected birds had lower feather δ15N values indicating that they occupied more mesic habitats. Leucocytozoon-infected birds had higher feather δ34S values suggesting more coastal sites or wetlands with anoxic sulphate reduction. As the composition and prevalence of haemosporidian parasites differed between the old and the current dataset, we suggest that the differences might be a consequence of temporal dynamics of haemosporidian parasites. Our results emphasize the importance of replicating studies conducted on a single population over a restricted time period, as the patterns can become more complex for data from wider geographical areas and different time periods.

First evidence for development of Plasmodium relictum (Grassi and Feletti, 1891) sporozoites in the salivary glands of Culex modestus Ficalbi, 1889.

The competence of insect vectors to transmit diseases plays a key role in host-parasite interactions and in the dynamics of avian malaria and other haemosporidian infections (Apicomplexa, Haemosporida). However, the presence of parasite DNA in the body of blood-sucking insects does not always constitute evidence for their competence as vectors. In this study, we investigate the susceptibility of wild-caught mosquitoes (Culex spp.) to complete sporogony of Plasmodium relictum (cyt b lineage SGS1) isolated from great tits (Parus major L., 1758). Adult female mosquitoes were collected with a CO2 bait trap overnight. A set of 50 mosquitoes was allowed to feed for 3 h at night on a single great tit infected with P. relictum. This trial was repeated on 6 different birds. The bloodfed mosquitoes that survived (n = 68) were dissected within 1-2 days (for ookinetes, n = 10) and 10-33 days post infection (for oocysts and sporozoites, n = 58) in order to confirm the respective parasite stages in their organs. The experiment confirmed the successful development of P. relictum (cyt b lineage SGS1) to the stage of sporozoites in Culex pipiens L., 1758 (n = 27) and in Culex modestus (n = 2). Our study provides the first evidence that C. modestus is a competent vector of P. relictum isolated from great tits, suggesting that this mosquito species could also play a role in the natural transmission of avian malaria.

Low intensity blood parasite infections do not reduce the aerobic performance of migratory birds.

Blood parasites (Haemosporidia) are thought to impair the flight performance of infected animals, and therefore, infected birds are expected to differ from their non-infected counterparts in migratory capacity. Since haemosporidians invade host erythrocytes, it is commonly assumed that infected individuals will have compromised aerobic capacity, but this has not been examined in free-living birds. We tested if haemosporidian infections affect aerobic performance by examining metabolic rates and exercise endurance in migratory great reed warblers (Acrocephalus arundinaceus) experimentally treated with Plasmodium relictum pGRW04 and in naturally infected wild birds over consecutive life-history stages. We found no effect of acute or chronic infections on resting metabolic rate, maximum metabolic rate or exercise endurance in either experimentally treated or free-living birds. Oxygen consumption rates during rest and while undergoing maximum exercise as well as exercise endurance increased from breeding to migration stages in both infected and non-infected birds. Importantly, phenotypic changes associated with preparation for migration were similarly unaffected by parasitaemia. Consequently, migratory birds experiencing parasitaemia levels typical of chronic infection do not differ in migratory capacity from their uninfected counterparts. Thus, if infected hosts differ from uninfected conspecifics in migration phenology, other mechanisms besides aerobic capacity should be considered.

Migratory birds along the Mediterranean - Black Sea Flyway as carriers of zoonotic pathogens.

At the crossroad between Europe, Asia, and Africa, Bulgaria is part of the Mediterranean - Black Sea Flyway (MBSF) used by millions of migratory birds. In this study, bird species migrating through Bulgaria were investigated as carriers of zoonotic pathogens. In total, 706 birds belonging to 46 species were checked for the presence of various bacterial pathogens (Campylobacter, Yersinia, Salmonella, Listeria, Escherichia coli, Staphylococcus aureus, Francisella tularensis, Coxiella burnetii, Borrelia burgdorferi, and Brucella spp.). From 673 birds we investigated fecal samples, from the remaining 33, blood samples. We detected Campylobacter 16S rDNA gene in 1.3% of birds, but none were of pathogenic Campylobacter jejuni and Campylobacter coli species. Escherichia coli 16S rDNA gene was found in 8.8% of the birds. Out of 34 birds that transported Yersinia enterocolitica strains (5.05%), only 1 carried a pathogenic isolate. Three birds (0.4%) were carriers of nonpathogenic Salmonella strains. Four avian samples (0.6%) were positive for Listeria monocytogenes and 1 (0.15%) was positive for Brucella spp. None of the birds tested carried the tick-borne pathogens C. burnetii or B. burgdorferi sensu lato. Antibiotic-resistant strains were detected, suggesting that migratory birds could be reservoirs and spreaders of bacterial pathogens as well as antibiotic resistance genes.

Detecting local transmission of avian malaria and related haemosporidian parasites (Apicomlexa, Haemosporida) at a Special Protection Area of Natura 2000 network.

Avian haemosporidian parasites (Apicomplexa, Haemosporida) are widespread pathogens that cause malaria (Plasmodium spp.) and other haemosporidioses (Haemoproteus spp. and Leucocytozoon spp.) in birds. The Special Protection Area Durankulak Lake (SPA DL) is a coastal lake in northeast Bulgaria, part of the Natura 2000 network that was declared as important area for wintering, breeding and migratory birds. Despite a number of conservation efforts outlined for the SPAs of Natura 2000 network, the potential threats and influences of haemosporidians and other parasites on occurring birds were not considered. In the present study, we aim to investigate the richness of haemosporidian parasites in birds captured in the protected area and to report the parasite species/DNA lineages that undergo local transmission in the region. We used both microscopic examination and PCR-based methods to diagnose haemosporidian infections in juvenile (captured in the year of hatching) and adult birds. The overall prevalence of haemosporidian parasites was significantly higher in the adult birds compared to juveniles. We identified five out of 21 recorded cytochrome b (cyt b) parasite lineages that are locally transmitted in the SPA DL (one of the genus Haemoproteus Kruse, 1890 and four of genus Plasmodium Marchiafava and Celli, 1885): cyt b lineages hRW2 of Haemoproteus belopolskyi, pSGS1 of Plasmodium relictum, pCOLL1, pYWT4 and pPADOM01 of Plasmodium (Haemamoeba) spp. It is likely that the majority of the parasites with local transmission are widespread host generalists and that host exchange is rather frequent among the birds inhabiting SPA DL.

Host migration strategy and blood parasite infections of three sparrow species sympatrically breeding in Southeast Europe.

Mobile hosts like birds occupy a wide array of habitats in which they encounter various vector and parasite faunas. If the infection probability for vector-borne parasites varies among seasons and biomes, a migratory life can critically influence the infections of a host. The growing body of literature on avian blood parasites suggests that host migrations do not only influence prevalence of infection but can also evoke higher infection intensities and increased parasite diversity in migrant compared to resident host species. We investigated the prevalence, intensity and diversity of Plasmodium and Haemoproteus infections in three closely-related and sympatrically breeding sparrow species with different migration strategies ranging from residential house sparrow and partially migratory tree sparrow to the obligate migratory Spanish sparrow. With a prevalence of 49%, the migratory Spanish sparrows were significantly less frequently infected than the resident house sparrows (82%). The partially migratory tree sparrow showed an intermediate prevalence of 60%. The parasitaemias were similar in all three host species and indicated mostly chronic but also few acute infections. While we found Plasmodium parasites in all three sparrow species, only Spanish sparrows were infected with Haemoproteus parasites in our study. With nine clearly identified parasite lineages in our study and the highest number of lineages per infected individuals (i.e. relative diversity), Spanish sparrows harboured the most diverse parasite fauna. Our results suggest that migration strategies can affect Plasmodium and Haemoproteus infections of sparrows resulting in a lower parasite prevalence and higher parasite diversity in migratory hosts-at least during our host's breeding period. A general scope for all annual cycle periods and across various bird taxa remains to be elucidated.

Genomic evidence of demographic fluctuations and lack of genetic structure across flyways in a long distance migrant, the European turtle dove.

BACKGROUND: Understanding how past climatic oscillations have affected organismic evolution will help predict the impact that current climate change has on living organisms. The European turtle dove, Streptopelia turtur, is a warm-temperature adapted species and a long distance migrant that uses multiple flyways to move between Europe and Africa. Despite being abundant, it is categorized as vulnerable because of a long-term demographic decline. We studied the demographic history and population genetic structure of the European turtle dove using genomic data and mitochondrial DNA sequences from individuals sampled across Europe, and performing paleoclimatic niche modelling simulations. RESULTS: Overall our data suggest that this species is panmictic across Europe, and is not genetically structured across flyways. We found the genetic signatures of demographic fluctuations, inferring an effective population size (Ne) expansion that occurred between the late Pleistocene and early Holocene, followed by a decrease in the Ne that started between the mid Holocene and the present. Our niche modelling analyses suggest that the variations in the Ne are coincident with recent changes in the availability of suitable habitat. CONCLUSIONS: We argue that the European turtle dove is prone to undergo demographic fluctuations, a trait that makes it sensitive to anthropogenic impacts, especially when its numbers are decreasing. Also, considering the lack of genetic structure, we suggest all populations across Europe are equally relevant for conservation.

Molecular characterisation of three avian haemoproteids (Haemosporida, Haemoproteidae), with the description of Haemoproteus (Parahaemoproteus) palloris n. sp.

DNA barcoding (molecular characterisation) is a useful tool for describing the taxonomy and systematics of organisms. Over 250 species of avian haemosporidian parasites have been described using morphological characters, yet molecular techniques based on polymerase chain reaction (PCR) suggest this diversity is underestimated. Moreover, molecular techniques are particularly useful for the detection of chronic infections and tissue stages of these parasites. Species delimitation is problematic among haemosporidians, and many questions about the mechanisms and patterns of speciation, host specificity and pathogenicity are still unresolved. Accumulation of additional genetic and morphological information is needed to approach these questions. Here, we combine microscopic examination with PCR-based methods to develop molecular characterisation of Haemoproteus (Parahaemoproteus) manwelli Bennett, 1978 and Haemoproteus (Parahaemoproteus) gavrilovi Valkiunas & Iezhova, 1990, both of which parasitise the bee-eater Merops apiaster L. We also describe a new species, Haemoproteus (Parahaemoproteus) palloris n. sp., from the blood of the willow warbler Phylloscopus trochilus (L.). We performed phylogenetic analyses with a set of mitochondrial cytochrome b (cyt b) gene lineages, which have been linked to parasite morphospecies and are available in the MalAvi database. Our findings show that morphological characters, which have been traditionally used in the description of haemosporidians, exhibit phylogenetic congruence. This study contributes to a better understanding of avian haemosporidian diversity and provides new molecular markers (cyt b and apicoplast gene sequences) for the diagnostics of inadequately investigated haemosporidian infections.

Plasmodium spp.: an experimental study on vertebrate host susceptibility to avian malaria.

The interest in experimental studies on avian malaria caused by Plasmodium species has increased recently due to the need of direct information about host-parasite interactions. Numerous important issues (host susceptibility, development of infection, the resistance and tolerance to avian malaria) can be answered using experimental infections. However, specificity of genetically different lineages of malaria parasites and their isolates is largely unknown. This study reviews recent experimental studies and offers additional data about susceptibility of birds to several widespread cytochrome b (cyt b) lineages of Plasmodium species belonging to four subgenera. We exposed two domesticated avian hosts (canaries Serinus canaria and ducklings Anas platyrhynchos) and also 16 species of common wild European birds to malaria infections by intramuscular injection of infected blood and then tested them by microscopic examination and PCR-based methods. Our study confirms former field and experimental observations about low specificity and wide host-range of Plasmodium relictum (lineages SGS1 and GRW11) and P. circumflexum (lineage TURDUS1) belonging to the subgenera Haemamoeba and Giovannolaia, respectively. However, the specificity of different lineages and isolates of the same parasite lineage differed between species of exposed hosts. Several tested Novyella lineages were species specific, with a few cases of successful development in experimentally exposed birds. The majority of reported cases of mortality and high parasitaemia were observed during parasite co-infections. Canaries were susceptible mainly for the species of Haemamoeba and Giovannolaia, but were refractory to the majority of Novyella isolates. Ducklings were susceptible to three malaria infections (SGS1, TURDUS1 and COLL4), but parasitaemia was light (<0.01%) and transient in all exposed birds. This study provides novel information about susceptibility of avian hosts to a wide array of malaria parasite lineages, outlining directions for future experimental research on various aspects of biology and epidemiology of avian malaria.

Degree of associations among vectors of the genus Culicoides (Diptera: Ceratopogonidae) and host bird species with respect to haemosporidian parasites in NE Bulgaria.

The occurrence of haemosporidians in biting midges of the genus Culicoides is examined in North-East Bulgaria in order to reveal their potential role for parasite transmission. A PCR-based technique amplifying part of the mitochondrial cytochrome b gene of the parasite is applied on naturally infected biting midges. Totally, 640 parous individuals of four species and 95 blood-fed individuals of six species of Culicoides are examined for the presence of DNA of haemosporidians. Haemosporidian genetic lineages are identified in individuals of three insect species: Culicoides alazanicus (12 lineages, nine lineages of Haemoproteus and three lineages of Plasmodium), Culicoides festivipennis and Culicoides circumscriptus (with two and one lineages of Haemoproteus, respectively). Two genetic lineages of Haemoproteus are recorded in more than one vector species. These results demonstrate variations in the specificity of Haemoproteus genetic lineages to their potential vectors, since some lineages are recorded in a single vector species and others occur in two or more vector species.

Molecular characterization of five widespread avian haemosporidian parasites (Haemosporida), with perspectives on the PCR-based detection of haemosporidians in wildlife.

Haemosporidians (Haemosporida) are cosmopolitan in birds. Over 250 species of these blood parasites have been described and named; however, molecular markers remain unidentified for the great majority of them. This is unfortunate because linkage between DNA sequences and identifications based on morphological species can provide important information about patterns of transmission, virulence, and evolutionary biology of these organisms. There is an urgent need to remedy this because few experts possess the knowledge to identify haemosporidian species and few laboratories are involved in training these taxonomic skills. Here, we describe new mitochondrial cytochrome b markers for the polymerase chain reaction (PCR)-based detection of four widespread species of avian Haemoproteus (Haemoproteus hirundinis, Haemoproteus parabelopolskyi, Haemoproteus pastoris, Haemoproteus syrnii) and 1 species of Plasmodium (Plasmodium circumflexum). Illustrations of blood stages of the reported species are given, and morphological and phylogenetic analyses identify the DNA lineages that are associated with these parasites. This study indicates that morphological characters, which have been traditionally used in taxonomy of avian haemosporidian parasites, have a phylogenetic value. Perspectives on haemosporidian diagnostics using microscopic and PCR-based methods are discussed, particularly the difficulties in detection of light parasitemia, coinfections, and abortive parasite development. We emphasize that sensitive PCR amplifies more infections than can be transmitted; it should be used carefully in epidemiology studies, particularly in wildlife parasitology research. Because molecular studies are describing remarkably more parasite diversity than previously expected, the need for traditional taxonomy and traditional biological knowledge is becoming all the more crucial. The linkage of molecular and morphological approaches is worth more of the attention of researchers because this approach provides new knowledge for better understanding insufficiently investigated lethal diseases caused by haemosporidian infections, particularly on the exoerythrocytic (tissue) and vector stages. That requires close collaboration between researchers from different fields with a common interest.

Two new species of Haemoproteus Kruse, 1890 (Haemosporida, Haemoproteidae) from European birds, with emphasis on DNA barcoding for detection of haemosporidians in wildlife.

Two new species of Haemoproteus Kruse, 1890 (Haemosporida, Haemoproteidae) are described: Haemoproteus (Parahaemoproteus) homovelans n. sp. from Grey-faced Woodpecker, Picus canus Gmelin, and Haemoproteus (Parahaemoproteus) concavocentralis n. sp. recorded in Hawfinch, Coccothraustes coccothraustes (Linnaeus), both sampled in Bulgaria. The morphology of the gametocytes and their host-cells are described and mitochondrial cytochrome b (cyt b) gene sequences are generated. Haemoproteus homovelans possesses circumnuclear gametocytes lacking volutin granules. This parasite is particularly similar to Haemoproteus velans Coatney & Roudabush, 1937 also possessing circumnuclear gametocytes that are, however, overfilled with volutin. Haemoproteus concavocentralis can be readily distinguished from all described avian haemoproteids due to the presence of an unfilled concave space between the central part of advanced gametocytes and erythrocyte nucleus. Bayesian phylogenetic analyses of 40 haemosporidian cyt b lineages showed close relationships of H. concavocentralis (hHAWF2) with a group of Haemoproteus spp. possessing gametocytes that are pale-stained with Giemsa. The lineage hPICAN02 of H. homovelans clustered with parasites infecting non-passerine birds. Phylogenetic analyses support the current subgeneric classification of the avian haemoproteids and suggest that cyt b lineage hPIPUB01 (GenBank EU254552) has been incorrectly assigned to Haemoproteus picae Coatney & Roudabush, 1937, a common parasite of corvid birds (Passeriformes). This study emphasises the importance of combining molecular techniques and light microscopy in the identification and field studies of avian haemosporidian parasites. Future development of barcodes for molecular identification of haemoproteids will allow better diagnostics of these infections, particularly in veterinary studies addressing insufficiently investigated tissue pathology caused by these parasites.

A new approach to evaluate multimodal orientation behaviour of migratory passerine birds recorded in circular orientation cages.

Circular orientation cages have been used for several decades to record the migratory orientation of passerine migrants, and have been central to the investigation of the functional characteristics of the biological compasses used for orientation. The use of these cages offers unique possibilities to study the migratory behaviour of songbirds, but suffers from statistical limitations in evaluating the directions of the activity recorded in the cages. The migratory activity has been reported to vary, including complex multimodal orientation of migratory passerines tested in orientation cages irrespective of species studied. The currently applied circular statistical methods fail to describe orientation responses differing from unimodal and axial distributions. We propose for the first time a modelling procedure enabling the analysis of multimodal distributions at either an individual or a group level. In this paper we compare the results of conventional methods and the recommended modelling approach. Migratory routes may be more complex than a simple migratory direction, and multimodal behaviour in migratory species at the individual and population levels can be advantageous. Individuals may select the expected migratory direction, but may also return to safer sites en route, i.e. sites already known, which provide food and/or shelter in reverse directions. In individual birds, several directions may be expressed in the same test hour. At the species level, multimodal orientation may give an opportunity to expand the range or may refer to differential migration route preferences in different populations of birds. A conflicting experimental situation may also result in a different preferential orientation. In this paper we suggest a statistical solution to deal with these types of variations in orientation preference.

First data on the genetic diversity of avian haemosporidians in China: cytochrome b lineages of the genera Plasmodium and Haemoproteus (Haemosporida) from Gansu Province.

A total of 76 birds belonging to 23 species and 14 families was examined for the presence of Plasmodium spp. and Haemoproteus spp. Birds were trapped at four localities in Gansu Province, China, in June-July 2011. DNA was isolated from blood samples and parasite detection, and identification was based on PCR assays and sequences of 479 bp of cyt b gene. The total prevalence of haemosporidians was 21.0%. Haemoproteus spp. were detected in 14 birds (prevalence 18.4%). The lineage CYAPIC1 from Cyanopica cyanus, Parus major, Passer montanus and Pyrrhocorax pyrrhocorax was new; it is genetically distinct and probably represents a new species of the genus Haemoproteus. Three lineages represented known species: RBS4 (from Lanius tephronotus), a lineage of Haemoproteus lanii; COLL2 (from Turdus mupinensis), a lineage of Haemoproteus pallidus and TURDUS2 (from Turdus rubrocanus), a lineage of Haemoproteus minutus. The lineage RBS5 (from Lanius cristatus and L. tephronotus) differs by 1.4% from RBS4 and probably represents an intraspecific entity of H. lanii. The lineages TUCHR1 (recorded from T. mupinensis), WW1 (recorded from Upupa epops) and YWT2 (recorded from Motacilla flava) have not been linked to any known species for the moment. Only one bird was positive for Plasmodium (prevalence 1.4%), i.e. P. major infected with the lineage GRW4 of Plasmodium relictum. The latter lineage has been considered by previous studies as typical for migratory birds and having transmission in tropical areas only; its record in a sedentary bird in China suggests its transmission in temperate latitudes.

Molecular characterization of haemosporidian parasites (Haemosporida) in yellow wagtail (Motacillaflava), with description of in vitro ookinetes of Haemoproteus motacillae.

In studies of haemosporidian systematics and taxonomy, a combination of microscopic examination and molecular identification has been recommended. The yellow wagtail (Motacillaflava) is a frequently used species in studies of host-parasite interactions; blood parasites of this bird have been well studied using microscopic examination of blood films, but data on molecular characterization of the parasites are scattered. Here we present the first study linking several haemosporidian cyt b sequences with their morphospecies parasitizing the yellow wagtail, combining light microscopy and nested PCR methods. The lineage hYWT1 of Haemoproteus motacillae was the most prevalent parasite; it was recorded in 38% of the birds. The latter lineage and the lineages hYWT2, hYWT3, hYWT5 and hYWT6 form a well-supported clade on the phylogenetic tree and likely represent intraspecific genetic variation of H. motacillae, with genetic divergence of 0.3 - 1.5 % among these lineages. Microscopic examination of smears prepared during an in vitro experiment, which was designed for observation of developing ookinetes, showed that H. motacillae ookinetes were present 5 hours after exposure of blood to air at 18-20° C. Ookinetes of this parasite belong to a group of large Haemoproteus spp. ookinetes (19.9 µm in length on average). Illustrations of the parasites are given. Two of reported Plasmodium lineages, pCOLLI and pYWT4 are phylogenetically closely related to the widespread host generalist parasites Plasmodium relictun (cyt b lineages pSGS 1, pGRW4 and pGRW11), but it was not possible to establish this morphologically. This study contributes to the molecular identification of avian haemosporidian parasites and provides information on morphology of H. motacillae ookinetes, which is additional information assisting the microscopic identification of this species.

Primary peak and chronic malaria infection levels are correlated in experimentally infected great reed warblers.

Malaria parasites often manage to maintain an infection for several months or years in their vertebrate hosts. In humans, rodents and birds, most of the fitness costs associated with malaria infections are in the short initial primary (high parasitaemia) phase of the infection, whereas the chronic phase (low parasitaemia) is more benign to the host. In wild birds, malaria parasites have mainly been studied during the chronic phase of the infection. This is because the initial primary phase of infection is short in duration and infected birds with severe disease symptoms tend to hide in sheltered places and are thus rarely caught and sampled. We therefore wanted to investigate the relationship between the parasitaemia during the primary and chronic phases of the infection using an experimental infection approach. We found a significant positive correlation between parasitaemia in the primary peak and the subsequent chronic phase of infection when we experimentally infected great reed warblers (Acrocephalus arundinaceus) with Plasmodium ashfordi. The reason for this association remains to be understood, but might arise from individual variation in exoerythrocytic parasite reservoirs in hosts, parasite antigenic diversity and/or host genetics. Our results suggest that the chronic phase parasitaemia can be used to qualitatively infer the parasitaemia of the preceding and more severe primary phase, which is a very important finding for studies of avian malaria in wild populations.

Effects of blood parasite infections on spatiotemporal migration patterns and activity budgets in a long-distance migratory passerine.

How blood parasite infections influence the migration of hosts remains a lively debated issue as past studies found negative, positive, or no response to infections. This particularly applies to small birds, for which monitoring of detailed migration behavior over a whole annual cycle has been technically unachievable so far. Here, we investigate how bird migration is influenced by parasite infections. To this end, we tracked great reed warblers (Acrocephalus arundinaceus) with multisensor loggers, characterized general migration patterns as well as detailed flight bout durations, resting times and flight heights, and related these to the genus and intensity of their avian haemosporidian infections. We found migration distances to be shorter and the onset of autumn migration to be delayed with increasing intensity of blood parasite infection, in particular for birds with Plasmodium and mixed-genus infections. Additionally, the durations of migratory flight bout were prolonged for infected compared to uninfected birds. But since severely infected birds and particularly birds with mixed-genus infections had shorter resting times, initial delays seemed to be compensated for and the timing in other periods of the annual cycle was not compromised by infection. Overall, our multisensor logger approach revealed that avian blood parasites have mostly subtle effects on migratory performance and that effects can occur in specific periods of the year only.

Seasonal dynamics of haemosporidian (Apicomplexa, Haemosporida) parasites in house sparrows Passer domesticus at four European sites: comparison between lineages and the importance of screening methods.

Infectious diseases often vary seasonally in a predictable manner, and seasonality may be responsible for geographical differences in prevalence. In temperate regions, vector-borne parasites such as malaria are expected to evolve lower virulence and a time-varying strategy to invest more in transmission when vectors are available. A previous model of seasonal variation of avian malaria described a double peak in prevalence of Plasmodium parasites in multiple hosts resulting from spring relapses and transmission to susceptible individuals in summer. However, this model was rejected by a study describing different patterns of seasonal variation of two Plasmodium spp. at the same site, with the double peak only apparent when these species were combined. Here, we assessed the seasonal variation in prevalence of haemosporidian parasites (Plasmodium, Haemoproteus and Leucocytozoon) in house sparrows (Passer domesticus) sampled across 1 year at four temperate European sites spanning a latitudinal range of 17°. We showed that parasite prevalence and diversity decreased with increasing latitude, but the parasite communities differed between sites, with only one Plasmodium lineage (P_SGS1) occurring at all sites. Moreover, the nested PCR method commonly used to detect and identify haemosporidian parasites strongly underestimated co-infections of Haemoproteus and Plasmodium, significantly biasing the pattern of seasonal variation, so additional molecular methods were used. Finally, we showed that: (i) seasonal variation in prevalence of haemosporidian parasites varied between study sites and parasite lineages/species/genera, describing further cases where the double peak model is not met; (ii) the seasonal dynamics of single lineages (P_SGS1) varied between sites; and (iii) unexpectedly, seasonality was greatest at the southernmost site, a pattern that was mostly driven by lineage H_PADOM05. Limitations of the genotyping methods and consequences of pooling (parasite lineages, sites and years) in studies of haemosporidian parasites are discussed and recommendations proposed, since these actions may obscure the patterns of prevalence and limit ecological inferences.

Avian haemosporidians from rain forests in Madagascar: Molecular and morphological data of the genera Plasmodium, Haemoproteus and Leucocytozoon.

The diversity of the haemosporidian genera Plasmodium, Haemoproteus and Leucocytozoon in birds from rain forests in Madagascar is characterized combining techniques of PCR and microscopy and based on the examination of 72 host individuals of 23 species in 15 families. High total prevalence of haemosporidians (68%) is detected, with Leucocytozoon infections being predominant (59.7%) and lower comparable prevalence of Plasmodium (18.0%) and Haemoproteus (23.6%) infections. Using mitochondrial cytochrome b (cytb) marker, 23 genetically distinct lineages are identified: 9 of Plasmodium spp., 6 of Haemoproteus spp. and 8 of Leucocytozoon spp. Fifteen of all lineages have not been reported by previous studies. This study provides the first data on haemosporidian morphological and molecular diversity found in the endemic families Vangidae and Bernieriidae. Two haemoproteid species, Haemoproteus fuscae Mello and Fonseca, 1937 and H. killangoi Bennett and Peirce, 1981, are redescribed based on the present samples and linked to the cytb lineages hCELEC01 and hZOSMAD01, respectively. Phylogenetic analysis is performed to test the relationship of the discovered new lineages with parasites from closely related avian hosts suggesting that multiple colonisation of hosts by haemosporidian parasites has occurred on the island.

Miniaturized multi-sensor loggers provide new insight into year-round flight behaviour of small trans-Sahara avian migrants.

BACKGROUND: Over the past decade, the miniaturisation of animal borne tags such as geolocators and GPS-transmitters has revolutionized our knowledge of the whereabouts of migratory species. Novel light-weight multi-sensor loggers (1.4 g), which harbour sensors for measuring ambient light intensity, atmospheric pressure, temperature and acceleration, were fixed to two long-distance migrant bird species - eurasian hoopoe (Upupa epops) and great reed warbler (Acrocephalus arundinaceus). Using acceleration and atmospheric pressure data recorded every 5 and 30 min, respectively, we aimed at reconstructing individual diurnal and seasonal patterns of flight activity and flight altitude and thereby, at describing basic, yet hitherto unknown characteristics of migratory flight behaviour. Furthermore, we wanted to characterise the variability in these migration characteristics between individuals, species and migration periods. RESULTS: The flight duration from breeding to sub-Saharan African non-breeding sites and back was more variable within than between the species. Great reed warblers were airborne for a total of 252 flight hours and thus, only slightly longer than eurasian hoopoes with 232 h. With a few exceptions, both species migrated predominantly nocturnally - departure around dusk and landing before dawn. Mean flight altitudes were higher during pre- than during post-breeding migration (median 1100 to 1600 m a.s.l.) and flight above 3000 m occurred regularly with a few great reed warblers exceeding 6000 m a.s.l. (max. 6458 m a.s.l.). Individuals changed flight altitudes repeatedly during a flight bout, indicating a continuous search for (more) favourable flight conditions. CONCLUSIONS: We found high variation between individuals in the flight behaviour parameters measured - a variation that surprisingly even exceeded the variation between the species. More importantly, our results have shown that multi-sensor loggers have the potential to provide detailed insights into many fundamental aspects of individual behaviour in small aerial migrants. Combining the data recorded on the multiple sensors with, e.g., remote sensing data like weather and habitat quality on the spatial and temporal scale will be a great step forward to explore individual decisions during migration and their consequences.