The Usutu Virus (USUV) is a mosquito-borne flavivirus originated in Africa. The virus circulates in Germany since 2010. It is primarily transmitted and maintained in the natural cycle by Culex mosquitoes and primarily affects birds, particularly Eurasian blackbird (Turdus merula), leading to significant mortality. Several studies have reported a high co-infection rate of European birds with both USUV and haemosporidians. Haemosporidians are blood parasites which maintain an enzootic life cycle with birds via different arthropod vectors. This study conducted screenings of birds from Germany received through a citizen\'s science project for both, USUV and haemosporidians between 2016 and 2021. The prevalence of USUV reached its peak in 2018, when it was first detected throughout most parts of Germany rather than being limited to localised hotspots. Subsequently, USUV prevalence consistently declined. On the other hand, the prevalence of haemosporidians initially declined between 2016 and 2019, but experienced a subsequent increase in the following years, exhibiting a more or less inverse pattern compared to the prevalence of USUV. In 2020, a statistically significant positive association between both pathogens was found, which was also detected across all years combined, indicating if at all a weak relationship between these pathogens.

Malaria remains the most important arthropod-borne infectious disease globally. The causative agent, Plasmodium, is a unicellular eukaryote that develops inside red blood cells. Identifying new Plasmodium parasite species that infect mammalian hosts can shed light on the complex evolution and diversity of malaria parasites. Bats feature a high diversity of microorganisms including seven separate genera of malarial parasites. Three species of Plasmodium have been reported so far, for which scarce reports exist. Here we present data from an investigation of Plasmodium infections in bats in the western Guinean lowland forest in Sierra Leone. We discovered a new Plasmodium parasite in the horseshoe bat Rhinolophus landeri. Plasmodium cyclopsi infections in a member of leaf-nosed bats, Doryrhina cyclops, exhibited a high prevalence of 100%. Phylogenetic analysis of complete mitochondrial genomes and nine nuclear markers recovered a close relationship between P. cyclopsi and the new Plasmodium parasite with the rodent species Plasmodium berghei, a widely used in vivo model to study malaria in humans. The data suggests that the \"rodent/bat\" Plasmodium (Vinckeia) clade represents a diverse group of malarial parasites that would likely expand with a systematic sampling of small mammals in tropical Africa. Identifying the bat Plasmodium repertoire is central to our understanding of the evolution of Plasmodium parasites in mammals.

The Australian skink Egernia stokesii had been recognised as a host of two species of Plasmodium, Plasmodium mackerrasae and P. circularis; nevertheless, molecular data are available for only a single haemosporidian species of this host. Its sequences are labelled as \"Plasmodium sp.\" or \"Plasmodium mackerrasae\", but morphological characteristics of this isolate are unavailable. Phylogenetic analyses of these sequences placed them into the clade of the genus Haemocystidium. In this study, blood samples of six E. stokesii were analysed by both, molecular and microscopic methods to clarify the haemosporidia of this lizard. Application of these approaches offered discordant results. Whereas sequence analysis clustered our isolates with lizard species of Haemocystidium, morphology of blood stages is more akin to Plasmodium than Haemocystidium. However, limited sampling, indistinguishable nuclei/merozoites and risk of possible hidden presence of mixed infection prevent reliable species identification of detected parasites or their description as new species of Haemocystidium.

Avian malaria parasites or haemosporidia are found in bird species worldwide. This special issue focuses on 3 most commonly studied genera: Haemoproteus, Plasmodium and Leucocytozoon. Seven research articles and reviews are provided to illustrate the breadth of knowledge of the diversity of avian malaria parasites in different regional habitats and across bird species, and the use of avian haemosporidian systems to examine host–parasite eco-evolutionary questions.

Accipitriform raptors are significant indicators of biodiversity and environmental health. Currently, most of the studies on avian haemosporidian parasites are on passerine birds, and data on raptors is constricted, with similarities both around the world and in Turkey. This study aimed to investigate the presence of Haemoproteus and Plasmodium spp. in raptors by microscopy and nested PCR technique. The study material consisted of 47 accipitriform raptors (Buteo buteo: 14, Buteo rufinus: 7, Clanga pomarina: 8, Circaetus gallicus: 12, Milvus migrans: 6). The prevalence of haemosporidian infection was 12.8% (6/47, 1 from Buteo buteo, 4 from Clanga pomarina, 1 from Milvus migrans) microscopically and 14.9% (7/47) molecularly. One Circaetus gallicus, microscopically found to be negative, probably due to low parasitemia, was molecularly found to be positive. All PCR-positive amplicons were bidirectionally sequenced, and the identification of lineages of the isolates and phylogenetic analysis were performed using the MalAvi and GenBank databases. The study revealed H-MILANS02 lineage in Buteo buteo, H-MILANS02 and P-CIAE1 lineages in Clanga pomarina, P-GRW06 lineage in Circaetus gallicus, and P-RTSR1 lineage in Milvus migrans, respectively. While this study removes the uncertainty regarding the reporting of the H-MILANS02 lineage in Turkey, it is also the first report to reveal 3 different Plasmodium spp. CytB lineages in raptors. Moreover, the fact that the P-GRW06 lineage (Plasmodium elongatum) detected in passerine birds was detected in a raptor, Circaetus gallicus, draws attention to the need for further investigations on host-parasite interaction and gives clues about the host-shifting ability of this parasite.

Haemosporidians are a widespread group of blood parasites transmitted by vectors. Despite their relevance for bird conservation, few studies have been conducted in the Amazonia and even less in terrestrial wild birds. We analysed blood samples from 168 game birds, collected from 2008 to 2015 by subsistence hunters of an indigenous rural community in the Peruvian Amazonia. DNA was tested for Haemoproteus spp., Plasmodium spp. and Leucocytozoon spp. and positive amplicons were sequenced and curated for phylogenetic analysis. Haemosporidian prevalence was 72% overall, 66.7% for Haemoproteus spp. and 5.4% for Plasmodium spp. and respectively by bird species: Spix\'s Guan (Penelope jacquacu, n = 72) 87.5% and 0%, Razor-billed Curassow (Mitu tuberosum, n = 45) 77.8% and 6.7%, White-winged Trumpeter (Psophia leucoptera, n = 20) 6.3% and 12.5%, Blue-throated Piping-guan (Pipile cumanensis, n = 16) 73.3% and 6.7%, and Great Tinamou (Tinamus major, n = 15) 10% and 15%. Leucocytozoon spp. was not found. P. leucoptera and T. major were less likely to be infected with Haemoproteus spp. Fruit abundance had a negative association with Haemoproteus spp. prevalence and precipitation was negatively associated with Plasmodium spp. prevalence. The 106 sequences examined represented 29 lineages, 82.8% of them were new lineages (Plasmodium n = 3, Haemoproteus n = 21). Novel host-parasite associations and lineages were unveiled, including probably new species of Plasmodium spp. Our results highlight the scientific value of alternative sampling methods and the collaboration with local communities.

Investigating the impact of parasitism on host phenotype is key to understanding parasite transmission ecology, host behavioural ecology and host-parasite coevolution. Previous studies have provided evidence that avian odour is one such phenotypic trait, as mosquitoes that vector the haemosporidian blood parasite Plasmodium tend to prefer birds that are already infected. Preen oil is a major source of avian odour, yet studies to date have not identified differences in preen oil odour based on the presence or absence of haemosporidian infection. Because preen oil can vary with physiological dynamics, we predicted that the composition of preen oil odours might vary according to parasite load, rather than solely by the presence or absence of infection. We used gas chromatography-mass spectrometry to characterize the composition of volatile compounds in preen oil taken from female dark-eyed juncos, Junco hyemalis carolinensis, and asked whether their composition varied with relative haemosporidian parasite load, which we assessed using quantitative PCR. We identified a subset of volatile compounds (a \'blend\') and two specific compounds that varied with increasing parasite load. Importantly, the quantity of these compounds did not vary based on parasite presence or absence, suggesting that birds with low parasite loads might be phenotypically indistinguishable from uninfected birds. The volatile blend associated with parasite load also varied with sampling date, suggesting a possible seasonal relapse of chronic infections triggered by shifts in junco host reproductive state. Furthermore, we found a positive relationship between parasite load and a volatile blend shown in a previous study to predict reproductive success in juncos. This is the first study to demonstrate quantitative differences in avian host odour based on haemosporidian parasite load. Our findings highlight the importance of focusing on parasite load, rather than solely presence or absence, in investigating host-parasite interactions.

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.

Mammalian haemosporidian parasites are classified in ten genera, including Plasmodium, Hepatocystis and Nycteria. A high diversity of haemosporidian parasites has been described from bats, but our understanding of their prevalence, distribution and use of hosts remain fragmented. The haemosporidian parasites of bats in Cameroon have been largely understudied, but here, bats, sampled from different habitat types of the Central Region of Cameroon, were investigated for haemosporidian infections with a combination of microscopic and molecular phylogenetic analysis. An overall prevalence of 18.1% of haemosporidian infections was detected in a total of 155 investigated bats belonging to 14 bat species. For the first time Hepatocystis and Nycteria parasites were detected in bats from Cameroon and molecularly characterized. Hepatocystis infections were exclusively identified in the epauletted fruit bat host species Epomophorus pusillus with a high prevalence of 65.5%, whereas Nycteria infections could be detected in several hosts, namely: Doryrhina cyclops (60.0%), Rhinolophus landeri (20.0%) and one Nycteris grandis. This study unveils evidence that habitat types may play a role in transmission of Hepatocystis parasites on a local scale and it adds important information on the distribution and host specificity of the neglected haemosporidian genus Nycteria.

Endemic island species face unprecedented threats, with many populations in decline or at risk of extinction. One important threat is the introduction of novel and potentially devastating diseases, made more pressing due to accelerating global connectivity, urban development, and climatic changes. In the Galápagos archipelago two important wildlife diseases: avian pox (Avipoxvirus spp.) and avian malaria (Plasmodium spp. and related Haemosporidia) challenge endemic species. San Cristóbal island has seen a paucity of disease surveillance in avian populations, despite the island\'s connectedness to the continent and the wider archipelago. To survey prevalence and better understand the dynamics of these two diseases on San Cristóbal, we captured 1205 birds of 11 species on the island between 2016 and 2020. Study sites included urban and rural lowland localities as well as rural highland sites in 2019. Of 995 blood samples screened for avian haemosporidia, none tested positive for infection. In contrast, evidence of past and active pox infection was observed in 97 birds and identified as strains Gal1 and Gal2. Active pox prevalence differed significantly with contemporary climatic conditions, being highest during El Niño events (~11% in 2016 and in 2019 versus <1% in the La Niña year of 2018). Pox prevalence was also higher at urban sites than rural (11% to 4%, in 2019) and prevalence varied between host species, ranging from 12% in medium ground finches (Geospiza fortis) to 4% in Yellow Warblers (Setophaga petechial aureola). In the most common infected species (Small Ground Finch: Geospiza fuliginosa), birds recovered from pox had significantly longer wings, which may suggest a selective cost to infection. These results illustrate the threat future climate changes and urbanization may present in influencing disease dynamics in the Galápagos, while also highlighting unknowns regarding species-specific susceptibilities to avian pox and the transmission dynamics facilitating outbreaks within these iconic species.