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Introduction. In contact with insects, people present a wide range of reactions, local or systemic, whether caused by insect bites or a venom injections, sucking blood or exposure to insect?s body and its secretions. Simulium erythrocephalum is an aggressive anthropophilic species, and following its bite humans may develop purpuric macules, edema and erythema surrounding the bite site. Our case series indicates frequent occurrence of Simulium erythrocephalum bites in Serbia. Case Series. We present a series of 30 cases reviewed between April and July 2006, who developed a local reaction at the site of this insect?s bite. In all patients, the skin lesions were located on the extremities, mainly the lower legs. The number of bites varied from two to more than ten. In all patients the bites occurred either during outdoor activities on the riverside or their residence was near the Danube. The therapy with antihistamines, local and/or systemic corticosteroids, as well as symptomatic therapy, with compression and limb elevation was recommended. The skin lesions regressed in one week in most patients, but in few they lasted for several weeks. Conclusion. In addition to its impact on human health, Simulium erythrocephalum also has a role in veterinary medicine. Insect bites in animals can cause significant livestock losses, occasionally resulting in animal mortality. Apart from this, Simulium erythrocephalum may also be a vector in the transmission of the parasite from the genus of Leucocytozoon that infests birds.

© 2022, The Author(s), under exclusive licence to Springer Nature B.V.Infections of avian haemosporidian parasites are regularly identified by molecular methods including multiplex PCR, which allows researchers to distinguish mixed infections of parasites from multiple genera. Here we extend the utility of a previously designed multiplex PCR by designing a primer set specific to parasites of the subgenus Haemoproteus (genus: Haemoproteus). The updated one-step multiplex PCR protocol we describe here allows for the detection of the genera Plasmodium and Leucocytozoon and the two subgenera (Haemoproteus and Parahaemoproteus) of the genus Haemoproteus. A sensitivity analysis showed that the multiplex PCR could amplify DNA of parasites in the subgenus Haemoproteus at very low levels of infection. We used this multiplex PCR to identify haemosporidian infections in 250 adult domestic pigeons (Columba livia) in Turkey. All samples were also screened by microscopy and a widely used nested PCR to compare with the results of multiplex PCR, to detect low levels of parasitemia, and to identify possible abortive infections. In total, 71 pigeons (28.4%) were found to be infected by all three methods. The multiplex PCR protocol successfully detected and discriminated both subgenera Haemoproteus and Parahaemoproteus infections. We compared our results with previous host species records to assess the host specificity of the parasite lineages we found. Our findings provide novel data on the prevalence of avian haemosporidians in domestic pigeons and demonstrate the utility of the new one-step multiplex PCR protocol for the determination of mixed avian haemosporidian infections. We expect that this protocol will contribute to a better understanding of the distribution, epizootiology, and ecology of avian haemosporidians.

In this study, we explore blood parasite prevalence, infection intensity, and co-infection levels in an urban population of feral pigeonsColumba liviain Cape Town. We analyze the effect of blood parasites on host body condition and the association between melanin expression in the host’s plumage and parasite infection intensity and co-infection levels. Relating to the haemosporidian parasite itself, we study their genetic diversity by means of DNA barcoding (cytochrome b) and show the geographic and host distribution of related parasite lineages in pigeons worldwide. Blood from 195C. liviaindividuals was collected from April to June 2018. Morphometric measurements and plumage melanism were recorded from every captured bird. Haemosporidian prevalence and infection intensity were determined by screening blood smears and parasite lineages by DNA sequencing. Prevalence ofHaemoproteusspp. was high at 96.9%. The body condition of the hosts was negatively associated with infection intensity. However, infection intensity was unrelated to plumage melanism. Thecytochrome bsequences revealed the presence of fourHaemoproteuslineages in our population of pigeons, which show high levels of co-occurrence within individual birds. Three lineages (HAECOL1, COLIV03, COQUI05) belong toHaemoproteus columbaeand differ only by 0.1% to 0.8% in thecytochrome bgene. Another lineage (COLIV06) differs by 8.3% from the latter ones and is not linked to a morphospecies, yet. No parasites of the generaLeucocytozoonandPlasmodiumwere detected.

Metatranscriptomics is a powerful method for studying the composition and function of complex microbial communities. The application of metatranscriptomics to multi-species parasite infections is of particular interest, as research on parasite evolution and diversification has been hampered by technical challenges to genome-scale DNA sequencing. In particular, blood parasites of vertebrates are abundant and diverse though they often occur at low infection intensities and exist as multi-species infections, rendering the isolation of genomic sequence data challenging. Here, we use birds and their diverse haemosporidian parasites to illustrate the potential for metatranscriptome sequencing to generate large quantities of genome-wide sequence data from multiple blood parasite species simultaneously. We used RNA-Seq on 24 blood samples from songbirds in North America to show that metatranscriptomes can yield large proportions of haemosporidian protein-coding gene repertoires even when infections are low-intensity (

One captive musophagid bird at a zoological garden in Japan showed clinical symptoms and was found to be infected with avian haemosporidia. We subsequently collected blood from all musophagid birds kept in the garden and examined for avian haemosporidia using both microscopic and molecular examination. Only Haemoproteus gametocytes were observed in the blood of two Guinea turaco (Tauraco persa). Three genetic lineages of Haemoproteus were identified from three Guinea turacos and one genetic lineage of Leucocytozoon was identified from a grey plantain-eater (Crinifer piscator). Detected Haemoproteus lineages were all identical and completely different from those previously reported in Japan, suggesting that these birds were infected in their original habitat. This is the first record of Haemoproteus infection in Guinea turacos.

Background Small poultry flock ownership has become a popular hobby in Europe and North America in recent years but there is a general lack of information regarding bird health and welfare. This retrospective analysis of routine post-mortem cases of non-commercial anseriform poultry aimed at providing information on causes of mortality mostly in relation to mortality events. For this purpose, birds that were submitted for routine post-mortem diagnostics to the National Veterinary Institute (SVA) in Sweden in 2011–2020 were retrospectively reviewed to determine main causes of mortality. Results Records from 79 necropsy submissions involving 120 birds (domestic ducks n = 41, Muscovy ducks n = 45, hybrid ducks n = 2 and domestic geese n = 32) were retrieved and analysed. Most submissions (72.2%) represented flock disease events and unexpected mortality was the most common cause of submission (70.9% of submissions). Twenty-two submissions (27.8%) were referred by veterinarians. There was a wide range of diagnoses of infectious and noninfectious aetiologies. Infectious causes of mortality included parasitic (19.2%), bacterial (13.3%), fungal (10.0%) and viral infections (3.3%) (at bird level of all 120 birds). Some of these infections such as duck virus enteritis (DVE), highly pathogenic influenza (HPAI H5N8) in Muscovy ducks and leucocytozoonosis (Leucocytozoon sp.) in all three species were most likely acquired from contact with wild free-living waterfowl. Generalised yeast infection (Muscovy duck disease) was diagnosed in Muscovy ducks and in a Muscovy duck/domestic duck hybrid. Other diseases were related to generalised noninfectious causes (27.5% of all birds) including diseases such as kidney disease, amyloidosis, cardiac dilatation, reproductive diseases and idiopathic inflammatory conditions. Nutritional or management-related diseases were diagnosed in 14.2% of all birds including rickets and gastrointestinal impaction/obstruction. Congenital/developmental, neoplastic, toxic and traumatic causes of mortality were rare. Conclusions The information obtained in this study can be used to identify and evaluate risks and help owners and veterinarians to prevent disease and provide adequate veterinary care for non-commercial anseriform poultry.

Avian haemosporidian parasites can cause malaria-like symptoms in their hosts and have been implicated in the demise of some bird species. The newly described Matryoshka RNA viruses (MaRNAV1 and MaRNAV2) infect haemosporidian parasites that in turn infect their vertebrate hosts. MaRNAV2 was the first RNA virus discovered associated with parasites of the genus Leucocytozoon. By analyzing metatranscriptomes from the NCBI SRA database with local sequence alignment tools, we detected two novel RNA viruses; we describe them as MaRNAV3 associated with Leucocytozoon and MaRNAV4 associated with Parahaemoproteus. MaRNAV3 had ~59% amino acid identity to the RNA-dependent RNA-polymerase (RdRp) of MaRNAV1 and ~63% amino acid identity to MaRNAV2. MaRNAV4 had ~44% amino acid identity to MaRNAV1 and ~47% amino acid identity to MaRNAV2. These findings lead us to hypothesize that MaRNAV_like viruses are widespread and tightly associated with the order Haemosporida since they have been described in human Plasmodium vivax, and now two genera of avian haemosporidians.

Haemosporidian parasites may impact avian health and are subject to shifts in distribution and abundance with changing ecologic conditions. Therefore, understanding variation in parasite prevalence is important for evaluating biologically meaningful changes in infection patterns and associated population level impacts. Previous research in western Alaska, US, indicated a possible increase in Leucocytozoon spp. infection between Emperor Geese (Anser canagicus) sampled in 1996 (

Aim: Macroecological analyses provide valuable insights into factors that influence how parasites are distributed across space and among hosts. Amid large uncertainties that arise when generalizing from local and regional findings, hierarchical approaches applied to global datasets are required to determine whether drivers of parasite infection patterns vary across scales. We assessed global patterns of haemosporidian infections across a broad diversity of avian host clades and zoogeographical realms to depict hotspots of prevalence and to identify possible underlying drivers. Location: Global. Time period: 1994–2019. Major taxa studied: Avian haemosporidian parasites (genera Plasmodium, Haemoproteus, Leucocytozoon and Parahaemoproteus). Methods: We amalgamated infection data from 53,669 individual birds representing 2,445 species world-wide. Spatio-phylogenetic hierarchical Bayesian models were built to disentangle potential landscape, climatic and biotic drivers of infection probability while accounting for spatial context and avian host phylogenetic relationships. Results: Idiosyncratic responses of the three most common haemosporidian genera to climate, habitat, host relatedness and host ecological traits indicated marked variation in host infection rates from local to global scales. Notably, host ecological drivers, such as migration distance for Plasmodium and Parahaemoproteus, exhibited predominantly varying or even opposite effects on infection rates across regions, whereas climatic effects on infection rates were more consistent across realms. Moreover, infections in some low-prevalence realms were disproportionately concentrated in a few local hotspots, suggesting that regional-scale variation in habitat and microclimate might influence transmission, in addition to global drivers. Main conclusions: Our hierarchical global analysis supports regional-scale findings showing the synergistic effects of landscape, climate and host ecological traits on parasite transmission for a cosmopolitan and diverse group of avian parasites. Our results underscore the need to account for such interactions, in addition to possible variation in drivers across regions, to produce the robust inference required to predict changes in infection risk under future scenarios.

Avian Haemosporidian parasites - Plasmodium, Haemoproteus, Leucocytozoon and Fallisia - have a wide distribution except for Antarctica. Leucocytozoon sp. has been poorly described in Brazil, and few studies have indicated infections in birds from the Atlantic Forest, Pantanal, Pampa and Amazon biomes. This study describes, for the first time, the occurrence of Leucocytozoon infection in red-legged seriemas (Cariama cristata) in the Brazilian savanna (Cerrado biome) using molecular diagnosis. Leucocytozoon spp. lineage CARCRI01 was detected in three C. cristata, a non-migratory bird, confirming transmission in mid-elevation areas in central Brazil. Further studies are needed to certify whether infections in red-legged seriemas were not abortive and to elucidate Leucocytozoon infection at low altitudes in the Brazilian lands.

Urbanization is an important human-driven process that leads to biodiversity loss and alters the interactions between organisms, including disease transmission. Although urbanization affects both host and vector communities, the effects on vector-borne pathogens are still poorly understood. Here, we monitored variation in prevalence and richness of three common blood parasites in birds (Plasmodium, Haemoproteus and Leucocytozoon) from localities with different land uses (urban, rural and natural) during two consecutive years (2013 and 2014). Overall, 1400 juvenile house sparrows (Passer domesticus) from 15 localities in southern Spain were included in this study. Haemoproteus and Leucocytozoon prevalence was higher in 2013 than in 2014, particularly in urban and natural habitats for the case of Leucocytozoon. Prevalence was correlated between years for Haemoproteus and Leucocytozoon, independently of the habitat. Additionally, rural habitats harboured significantly higher Haemoproteus lineage richness compared to urban and natural habitats during 2014. Leucocytozoon lineage richness was negatively correlated between years in rural habitats but positively correlated in urban and natural habitats in comparison. Parasite lineages found in birds were homogeneously distributed along habitats and years and the common lineages prevalence were not influenced by them. Our results highlight different patterns of infection depending on the parasite genera probably related to the composition and density of vector communities. The specific reproductive environmental requirements of the different groups of vectors involved in the transmission may be affected by climatic conditions and landscape features.

Emperor Geese (Anser canagicus) are iconic waterfowl endemic to Alaska and adjacent areas of northeastern Russia that are considered to be near threatened by the International Union for Conservation. This species has been identified as harboring diverse viruses and parasites which have, at times, been associated with disease in other avian taxa. To better assess if disease represents a vulnerability for Emperor Geese breeding on the Yukon-Kuskokwim Delta, Alaska, we evaluated if haemosporidian parasites were associated with decreased mass or survival among adult female nesting birds captured during 2006–2016. Through molecular analyses, we detected genetically diverse Leucocytozoon, Haemoproteus, and Plasmodium parasites in 28%, 1%, and 1% of 607 blood samples screened in triplicate, respectively. Using regression analysis, we found evidence for a small effect of Leucocytozoon infection on the mass of incubating adult female Emperor Geese. The estimated mass of infected individuals was approximately 43 g (95% CI: 20–67 g), or approximately 2%, less than uninfected birds when captured during the second half of incubation (days 11–25). We did not, however, find support for an effect of Leucocytozoon infection on survival of adult female nesting Emperor Geese using a multi-state hidden Markov framework to analyze mark-resight and recapture data. Using parasite mitochondrial DNA cytochrome b sequences, we identified 23 haplotypes among infected Emperor Geese. Leucocytozoon haplotypes clustered into three phylogenetically supported clades designated as ‘L. simondi clade A’, ‘L. simondi clade B’, and ‘other Leucocytozoon’. We did not find evidence that parasites assigned to any of these clades were associated with differential mass measures among nesting adult female Emperor Geese. Collectively, our results provide negligible evidence for Leucocytozoon parasites as causing detrimental effects to adult female Emperor Geese breeding on the Yukon-Kuskokwim Delta., Graphical abstract Image 1, Highlights • Emperor Geese are consistently infected with Leucocytozoon parasites. • We found evidence for a small effect of infection on mass of incubating females. • We did not find support for an effect of Leucocytozoon infection on survival.

Bio-invasions are a major threat to biodiversity and ecosystems globally and may contribute to the proliferation of emerging infectious diseases. We examined the prevalence and phylogenetic diversity of avian haemosporidian parasites infecting the non-native house sparrows (Passer domesticus) and the native southern grey-headed sparrows (Passer diffusus). Blood samples from 104 sparrows (74 house sparrows and 30 southern grey-headed sparrows) mist-netted inside and around the Kruger National Park were used. Genomic DNA was extracted from each blood sample and subjected to nested PCR analyses, Sanger sequencing and phylogenetic analyses. Overall, 35.57% (37/104) of the birds sampled were infected with at least one haemosporidian parasites. Southern grey-headed sparrows had a higher parasite prevalence (60%) than house sparrows (24.3%). A total of 16 parasite lineages were identified, of which eight were novel lineages. Whereas Haemoproteus spp. showed the highest lineage diversity, Leucocytozoon spp. were the most prevalent parasites, albeit with significant differences between sparrow species. A single Plasmodium sp. infection was recorded in a southern grey-headed sparrow. In support of the enemy release hypothesis, we found that prevalence on non-native house sparrows was lower than prevalence recorded in their region of origin and also that they were infected only by indigenous parasites lineages.

Within animal hybrid zones, parasites may determine competitive outcomes between host species and thus affect hybridization dynamics. We addressed this hypothesis by evaluating haemosporidian prevalence and community composition in a rapidly moving hybrid zone between Black-capped Chickadees (Poecile atricapillus) and Carolina Chickadees (P. carolinensis). Using molecular methods, we screened for haemosporidians in multiple chickadee populations across the hybrid zone and investigated whether parasite prevalence varied as a function of admixture among these birds. We identified 36 parasite lineages from 3 haemosporidian genera (Plasmodium, Haemoproteus, and Leucocytozoon) but found no genera or lineages more likely to infect any particular chickadee taxon. Instead, haemosporidian prevalence varied across sites and seasons: Leucocytozoon was more prevalent during chickadees' breeding season, whereas Haemoproteus prevalence peaked during nonbreeding periods. Leucocytozoon infected proportionally fewer birds at the leading edge of the hybrid zone than near its center. However, haemosporidian communities were similar among chickadee populations, and evidence for parasite exchanges between chickadee taxa was lacking. These results underscore the complexity of bird–parasite relationships and suggest that haemosporidians are unlikely to play a major role in the ongoing movement of this hybrid zone.LAY SUMMARYWe tested whether patterns of parasite diversity and prevalence support the hypothesis that parasite communities may determine competitive outcomes within an iconic avian hybrid zone.We used molecular methods to detect and identify haemosporidian lineages infecting Black-capped, Carolina, and hybrid chickadees across their hybrid zone in southeastern Pennsylvania.Analyses revealed 36 unique haemosporidian lineages, the prevalence and richness of which varied little across chickadee taxa.For some parasites, locality and season predicted haemosporidian prevalence.Haemosporidians are likely not key drivers underlying movement of this chickadee hybrid zone.

Black flies (Diptera: Simuliidae) are among the most bothersome blood-sucking dipterans causing severe irritation and distress to poultry, wild birds, animals, and humans globally. These insects are vectors of viruses, bacteria, parasitic protozoans, and nematodes of humans and animals. Parasitic protozoa belonging to Haemosporida (Apicomplexa) are distributed worldwide and black flies are the principal vectors of avian haemosporidian parasites of the genus Leucocytozoon, a common parasite of birds. Based on the detection of parasite DNA in insects, 13 black fly species were reported to be potential vectors of Leucocytozoon in Europe. Information about which species of Simulium can play a role in the transmission of Leucocytozoon parasites is insufficient and needs to be developed. The aim of our study was to determine which black fly species are involved in the transmission of Leucocytozoon parasites in the Eastern Europe. The black fly females were collected in Lithuania using entomological net. They were morphologically identified, dissected to prepare salivary glands preparations, and then screened for the presence of Leucocytozoon parasites using microscopy and PCR-based methods. In all, we collected 437 black fly females belonging to eight species. The DNA of Leucocytozoon (genetic lineage lCOCO18) was detected in one of analysed females identified as Simulium maculatum. All salivary gland preparations were negative for the presence of Leucocytozoon sporozoites. Our results included S. maculatum as a potential vector of Leucocytozoon parasites. Increasing the knowledge on vector ecology, behaviour and improving collection methods may be the key to understand the evolution and diversity of these parasites.

The persistence of a parasite species in an ecological community is determined both by its environmental tolerance and host breadth. The relative contribution of these niche characteristics to parasite community structure is challenging to parse because host persistence is also a consequence of extrinsic environmental factors. We investigated haemosporidian parasites (genera Plasmodium, Leucocytozoon, and Haemoproteus) in a clade of avian hosts in eastern North America. Species in this clade of Catharus thrushes occupy specific elevational bands in a non‐phylogenetically determined manner. This allowed us to tease apart the effects of host relatedness and elevation on parasite community structure, diversity, and infection prevalence. We screened blood and tissue samples from 414 adult birds from four mountain ranges in the Appalachian Highlands for blood parasites using a cytochrome‐b‐nested PCR protocol and identified parasite lineages by sequencing. We found an overall infection prevalence of 88.4% and identified a total of 38 parasite lineages including six novel lineages. Parasite community patterns varied by genus. Host relatedness rather than elevational zone predicted the beta turnover and phylobeta turnover of Leucocytozoon parasites, indicating that closely related rather than geographically proximate host species had more similar parasites. This pattern was not seen in Plasmodium parasites because the diversity of this parasite genus varied considerably in the high elevational zones among mountain ranges, that is, a sky‐island effect. Haemoproteus parasites were rare in this study system. Our study suggests that the mechanisms that underlie community structuring vary between haemosporidian genera due to differences in the degree of host sharing among lineages.

Understanding factors that influence the spatial and temporal distributions of blood parasites is important to help predict how host species and their parasites may respond to global change. Factors that may influence parasite distributions are land cover and host dispersal patterns, which may result in exposure of a host to novel parasites, or escape from parasites of their origin. We screened golden-winged warblers from across the United States and Canada for blood parasites, and investigated whether land-use patterns or host dispersal affected the prevalence and composition of haemosporidian assemblages. Parasite prevalence varied strongly with study area, and areas with high agricultural cover had a significantly higher prevalence of Leucocytozoon and Parahaemoproteus parasites. Lineages of Parahaemoproteus and Leucocytozoon were genetically differentiated among study areas, and prevalence and composition of parasite assemblages indicated an increase in parasite prevalence and accumulation of unique parasite lineages from the southeast to the northwest. This matches the historical range expansion and natal dispersal patterns of golden-winged warblers, and suggests that golden-winged warblers may have been sensitive to novel parasites as they dispersed. The high prevalence and diversity of parasite lineages in the north-west extent of their breeding range (Manitoba) indicates that this population may face unique pressures.

Background Plasmodium species feature only four to eight nuclear ribosomal units on different chromosomes, which are assumed to evolve independently according to a birth-and-death model, in which new variants originate by duplication and others are deleted throughout time. Moreover, distinct ribosomal units were shown to be expressed during different developmental stages in the vertebrate and mosquito hosts. Here, the 18S rDNA sequences of 32 species of avian haemosporidian parasites are reported and compared to those of simian and rodent Plasmodium species. Methods Almost the entire 18S rDNAs of avian haemosporidians belonging to the genera Plasmodium (7), Haemoproteus (9), and Leucocytozoon (16) were obtained by PCR, molecular cloning, and sequencing ten clones each. Phylogenetic trees were calculated and sequence patterns were analysed and compared to those of simian and rodent malaria species. A section of the mitochondrial CytB was also sequenced. Results Sequence patterns in most avian Plasmodium species were similar to those in the mammalian parasites with most species featuring two distinct 18S rDNA sequence clusters. Distinct 18S variants were also found in Haemoproteus tartakovskyi and the three Leucocytozoon species, whereas the other species featured sets of similar haplotypes. The 18S rDNA GC-contents of the Leucocytozoon toddi complex and the subgenus Parahaemoproteus were extremely high with 49.3% and 44.9%, respectively. The 18S sequences of several species from all three genera showed chimeric features, thus indicating recombination. Conclusion Gene duplication events leading to two diverged main sequence clusters happened independently in at least six out of seven avian Plasmodium species, thus supporting evolution according to a birth-and-death model like proposed for the ribosomal units of simian and rodent Plasmodium species. Patterns were similar in the 18S rDNAs of the Leucocytozoon toddi complex and Haemoproteus tartakovskyi. However, the 18S rDNAs of the other species seem to evolve in concerted fashion like in most eukaryotes, but the presence of chimeric variants indicates that the ribosomal units rather evolve in a semi-concerted manner. The new data may provide a basis for studies testing whether differential expression of distinct 18S rDNA also occurs in avian Plasmodium species and related haemosporidian parasites.

Avian communities from South America harbor an extraordinary diversity of Leucocytozoon species (Haemosporida, Leucocytozoidae). Here, of 890 birds sampled, 10 (1.2%) were infected with Leucocytozoon parasites. Among them, two new species were discovered and described. Leucocytozoon grallariae sp. nov. and Leucocytozoon neotropicalis sp. nov. were found in non-migratory highland passeriforms belonging to the Grallaridae and Cotingidae, respectively. They both possess gametocytes in fusiform host cells. However, due to combining microscopic examination and molecular detection, it was revealed that these parasites were present in co-infections with other Leucocytozoon species, which gametocytes develop in roundish host cells, therefore exhibiting two highly distant parasite lineages isolated from the same samples. Remarkably, the lineages obtained by cloning the mtDNA genomes were not captured by the classic nested PCR, which amplifies a short fragment of cytochrome b gene. Phylogenetic analyses revealed that the lineages obtained by the classic nested PCR clustered with parasites possessing gametocytes in roundish host cells, while the lineages obtained by the mtDNA genome PCR protocol were closely related to Leucocytozoon parasites possessing gametocytes in fusiform host cells. These findings suggest problems with the sensitivity of the molecular protocols commonly used to detect Leucocytozoon species. A detailed analysis of the primers used in the classic nested PCR revealed a match with DNA sequences from those parasites that possess gametocytes in roundish host cells (i.e., Leucocytozoon fringillinarum), while they differ with the orthologous regions in the mtDNA genomes isolated from the samples containing the two new species. Since these are mixed infections, none of the lineages detected in this study can be assigned accurately to the new Leucocytozoon morphospecies that develops in fusiform host cells. However, phylogenetic analyses allowed us to hypothesize their most probable associations. This study highlights the need for developing detection methods to assess the diversity of Leucocytozoon parasites accurately., Graphical abstract Image 1, Highlights • Molecular diversity of Leucocytozoon is underestimated. • We described two new Leucocytozoon species infecting passerines endemic of Neotropics. • Commonly PCR protocols failed to detect Leucocytozoon lineages from Neotropical region.

Avian haemosporidian infections are widespread and can result in the decline of wild bird populations or in some cases contribute to extinction of species. We determined the prevalence and genetic diversity of avian haemosporidia in 93 samples from 22 landbird species from South Africa (N = 76) and West Africa (N = 17), of which six are intra-African migrants and one is a Palearctic migrant. The samples were analysed for the presence of avian haemosporidian DNA using real-time quantitative PCR (qPCR) and nested PCR assays targeting specific mitochondrial genes of these parasites. The cytochrome b (cytb) gene was sequenced for all samples that tested positive and phylogenetic analysis was conducted in order to determine the relationship of the new sequences with previously published sequences from the MalAvi database. The overall prevalence of avian haemosporidiosis was 68.82% (95% CI: 56.4%–78.87%) and 82.80% (95% CI: 65.68%–86.11%) as determined by qPCR and nested PCR respectively. Eighteen (19.36%; 95% CI; 10.78%–29.97%) samples had mixed infections. Infection prevalence of all haemosporidian spp. were significantly higher (p, Graphical abstract Image 1, Highlights • Avian haemosporidian infections in free ranging birds are common and diverse. • The prevalence was 68.82% and 82.80% by qPCR and nested PCR assays respectively. • Eighteen (19.36%) samples had mixed infections. • Seventeen cytochrome b lineages were identified from 14 avian species.

Background Avian haemosporidia are obligate blood parasites with an ample range of hosts worldwide. To understand how host communities may influence the diversity of parasites of the neotropics, the spatial genetic variation of avian Plasmodium, Haemoproteus, and Leucocytozoon was examined between areas of host endemism and along the elevational gradient in the tropical Andes. Methods A total of 1686 accessions of the cytochrome b gene of avian haemosporidia were selected from 43 publications, that further provides additional information on 14.2% of bird species in the Neotropics. Haplotype groups were identified using a similarity-based clustering of sequences using a cut-off level ≥ 99.3% of sequence identity. Phylogenetic-based analyses were implemented to examine the spatial genetic structure of avian haemosporidia among areas of host endemism and the elevation gradient in the tropical Andes. Results The areas of avian endemism, including the tropical Andes, can explain the differential distribution of the haemosporidia cytochrome b gene variation. In the tropical Andes region, the total number of avian haemosporidia haplotypes follows a unimodal pattern that peaks at mid-elevation between 2000 and 2500 m above sea level. Furthermore, the haplotype assemblages of obligate blood parasites tend to overlap towards mid-elevation, where avian host diversity tends to be maximized. Conclusions Spatial analyses revealed that richness and turnover in haemosporidia suggest an association with montane host diversity, according to elevation in the tropical Andes. In addition, the spatial distribution of haemosporidia diversity is closely associated with patterns of host assemblages over large geographical scale in the tropical Andes and areas of avian endemism nearby. Electronic supplementary material The online version of this article (10.1186/s12936-019-2699-9) contains supplementary material, which is available to authorized users.

Prevalence responses to anthropic factors differ across hosts and parasite species. We here analyzed the spatiotemporal variation of avian haemosporidian prevalence in bird assemblages of the Mooswald forest (i.e., urban greenspace; Freiburg, Germany), in response to local environmental features (e.g., water sources, human presence (visited)/absence (unvisited)) and bird-level traits (e.g., body condition, age, sex) in 2 years. We used a nested PCR protocol (mitochondrial (mt)DNA cytochrome b (cyt b) gene) and microscopy to determine haemosporidian infections. Prevalence was analyzed using a general linear multi-model (glmulti) approach with Akaike information criterion corrected for small samples (AICc), with subsequent model inferences using a GLMM on the best selected model, considering bird species as a random factor. Analyses were conducted for the main understory bird species (Blackcap – Sylvia atricapilla, Chaffinch – Coereba flaveola, Great Tit – Parus major, Blue Tit – Cyanistes caeruleus, European Robin – Erithacus rubecula, Blackbird – Turdus merula, Song Thrush – Turdus philomelos). We further conducted spatial autocorrelation analyses for all haemosporidian infections, and classification and regression trees (CARTs) for focal species. We analyzed a total of 544 samples of seven bird species. In 2011 prevalence for Haemoproteus/Plasmodium was 25.8% and 11.7% for Leucocytozoon. In 2013 prevalence for Haemoproteus/Plasmodium was 26.5% and 35.5% for Leucocytozoon. Haemosporidian prevalence was significantly different between some focal species. There was a negative association between distance to the nearest water source and prevalence in the year 2011, and the opposite pattern for the year 2013. However, when analyzed for the six focal species separately, such a relationship could change from a negative to a positive one, or there could be no relationship at all. For Leucocytozoon there was higher prevalence in the section of the forest visited by humans. We did not find spatial autocorrelation for prevalence across the study site, but there were statistically significant local spatial clusters in the visited section. Although there were similar responses of prevalence to some factors, infection patterns were generally bird species-specific. Thus, prevalence is a labile epidemiological parameter, varying spatiotemporally in an idiosyncratic way.

Haemosporidian parasites causing malaria-like diseases in birds are globally distributed and have been associated with reduced host fitness and mortality in susceptible bird species. This group of parasites has not only enabled a greater understanding of host specificity, virulence, and parasite dispersal, but has also been crucial in restructuring the evolutionary history of apicomplexans. Despite their importance, genomic resources of avian haemosporidians have proved difficult to obtain, and they have, as a result, been lagging behind the congeneric Plasmodium species infecting mammals. In this review, I discuss recent genomic advances in the field of avian malaria research, and outline outstanding questions that will become possible to investigate with the continued successful efforts to generate avian haemosporidian genomic data.