Prevalence and composition of haemosporidians in an avian community from a World Heritage area: Associations with host foraging strata and forest regeneration.

Forest regeneration is becoming a powerful tool to combat land conversion which covers 30 % of the Neotropical territory. However, little is known about the effect of forest regeneration on vector-borne diseases. Here, we describe the haemosporidian lineage composition across a successional gradient within an Atlantic Forest bird community. We test whether forest successional stages, in addition to host life history traits affect haemosporidian infection probability. We sampled birds at 16 sampling units with different successional stages between 2017 and 2018 within a forest remnant located in Antonina, Paraná, Brazil. We captured bird individuals using mist-nets, identified them to the species level, and collected blood samples to detect and identify Plasmodium and Haemoproteus lineages based on molecular analysis. We used a Bayesian phylogenetic linear model with a Bernoulli distribution to test whether the haemosporidian infection probability is affected by nest type, foraging stratum, and forest successional stage. We captured 322 bird individuals belonging to 52 species and 21 families. We found 31 parasite lineages and an overall haemosporidian prevalence of 23.9 %, with most infections being caused by Plasmodium (21.7 % of prevalence). The Plasmodium probability of infection was associated with forest successional stage and bird foraging stratum. Birds from the secondary forest in an intermediate stage of succession are more likely to be infected by the parasites than birds from the primary forests (ß = 1.21, 95 % CI = 0.11 - 2.43), birds from upper strata exhibit a lower probability of infection than birds from lower foraging strata (ß = -1.81, 95 % CI = -3.80 - -0.08). Nest type did not affect the Plasmodium probability of infection. Our results highlight the relevance of forest succession on haemosporidian infection dynamics, which is particularly relevant in a world where natural regeneration is the main tool used in forest restoration.

Avian haemosporidians of the genera Plasmodium and Haemoproteus from resident and Neotropical migrant birds in Colombia.

Avian haemosporidians of the genera Plasmodium and Haemoproteus are a group of widely distributed blood parasites that can negatively affect the fitness of their hosts. Colombia contains the greatest diversity of birds on the planet, but knowledge about the associations between haemosporidian and its avifauna is scarce and fragmented. We collected blood samples from 255 birds (203 residents and 52 neotropical migrants) belonging to 27 families and 108 species. The study was conducted in six localities in the inter-Andean valleys of the Cauca and Magdalena rivers. Parasites of the genera Plasmodium and Haemoproteus were identified in the samples by morphological and molecular analysis of a fragment of the mitochondrial gene cyt b. Among the samples, 9.3% (n = 24) were positive for Plasmodium or Haemoproteus. Co-infection with Plasmodium and Haemoproteus was found in Red-eyed Vireo. Seventeen haemosporidian lineages were identified, five of which were reported for the first time in resident birds (Common Ground Dove, Checker-throated Stipplethroat, Tropical Kingbird, Pale-breasted Thrush, and Ruddy-breasted Seedeater) and one in the Summer Tanager (neotropical migrant). The research results confirm the wide diversity of haemosporidian present in tropical lowlands and the possible role of neotropical migratory birds in dissemination on haemosporidian along their migratory routes.

Higher body condition with infection by Haemoproteus parasites in Bananaquits (Coereba flaveola).

Parasite transmission is a heterogenous process in host-parasite interactions. This heterogeneity is particularly apparent in vector-borne parasite transmission where the vector adds an additional level of complexity. Haemosporidian parasites, a widespread protist, cause a malaria-like disease in birds globally, but we still have much to learn about the consequences of infection to hosts' health. In the Caribbean, where malarial parasites are endemic, studying host-parasites interactions may give us important insights about energetic trade-offs involved in malarial parasites infections in birds. In this study, we tested the consequences of Haemoproteus infection on the Bananaquit, a resident species of Puerto Rico. We also tested for potential sources of individual heterogeneity in the consequences of infection such as host age and sex. To quantify the consequences of infection to hosts' health we compared three complementary body condition indices between infected and uninfected individuals. Our results showed that Bananaquits infected by Haemoproteus had higher body condition than uninfected individuals. This result was consistent among the three body condition indices. Still, we found no clear evidence that this effect was mediated by host age or sex. We discuss a set of non-mutually exclusive hypotheses that may explain this pattern including metabolic syndrome, immunological responses leading to host tolerance or resistance to infection, and potential changes in consumption rates. Overall, our results suggest that other mechanisms, may drive the consequences of avian malarial infection.

Energy input, habitat heterogeneity and host specificity drive avian haemosporidian diversity at continental scales.

The correct identification of variables affecting parasite diversity and assemblage composition at different spatial scales is crucial for understanding how pathogen distribution responds to anthropogenic disturbance and climate change. Here, we used a database of avian haemosporidian parasites to test how the taxonomic and phylogenetic diversity and phylogenetic structure of the genera Plasmodium, Haemoproteus and Leucocytozoon from three zoogeographic regions are related to surrogate variables of Earth's energy input, habitat heterogeneity (climatic diversity, landscape heterogeneity, host richness and human disturbance) and ecological interactions (resource use), which was measured by a novel assemblage-level metric related to parasite niche overlap (degree of generalism). We found that different components of energy input explained variation in richness for each genus. We found that human disturbance influences the phylogenetic structure of Haemoproteus while the degree of generalism explained richness and phylogenetic structure of Plasmodium and Leucocytozoon genera. Furthermore, landscape attributes related to human disturbance (human footprint) can filter Haemoproteus assemblages by their phylogenetic relatedness. Finally, assembly processes related to resource use within parasite assemblages modify species richness and phylogenetic structure of Plasmodium and Leucocytozoon assemblages. Overall, our study highlighted the genus-specific patterns with the different components of Earth's energy budget, human disturbances and degree of generalism.

Molecular Survey of Haemosporidian Parasites in Procellariiformes Sampled in Southern Brazil, 2013-22.

The order Procellariiformes includes several species of seabirds that perform long-distance migrations crossing all the oceans. These movements may contribute to the dispersal and exchange of hemoparasites, such as haemosporidians. There is a lack of studies regarding the order Haemosporida in Procellariiformes, and, to date, only the genus Plasmodium has been reported. This survey investigated the occurrence of the three genera of haemosporidians, Plasmodium, Haemoproteus, and Leucocytozoon, in samples collected between 2013 and 2022 from 95 individuals of 14 species of Procellariiformes from southern Brazil, including live animals in rehabilitation centers, individuals caught as incidental bycatch, and carcasses found along the coast. A total of 171 samples of blood and fragments of liver and spleen were analyzed, with extracted DNA being subjected to a nested PCR followed by phylogeny analysis. All animals were negative for Plasmodium spp. and Leucocytozoon spp., but one Black-browed Albatross (Thalassarche melanophris) and one Manx Shearwater (Puffinus puffinus) specimen were positive for Haemoproteus spp. The sequences obtained from positive seabirds did not show 100% similarity with other known lineages available in the MalAvi database and thus were probably novel lineages. However, one sequence clustered together with Haemoproteus noctuae, a parasite from Strigiformes, while the other was grouped with Haemoproteus columbae, which is classically related to Columbiformes. These results suggest that both positive animals may have become infected when beached or in rehabilitation centers by a spillover of vectors from local birds. This highlights the importance of surveillance of the health of Procellariiformes regarding the possibility of dissemination of new pathogens in different bird populations.

Hemoparasite occurrence and hematological/serum chemistry variations in Podocnemis vogli turtles: A comparative analysis between wild-residing infected specimens and captive non-infected counterparts.

Reptiles show a high occurrence of hemoparasites in the wild; however, little is known about the impact of such infections on their hosts' physiology and health status. Podocnemis vogli is an ancient turtle distributed in South America, frequently infected by blood parasites. Specifically, we analyzed the hematological and serum chemistry parameters of 78 wild turtles. We compared these values with those obtained from non-infected turtles of the same species in ex-situ conditions, considering factors such as sex and coinfections. Two orders of hemoparasites were detected under microscopic analyses: Adelorina, represented by Haemogregarina sp. (98.72 ± 0.28%), and Haemosporida, represented by Haemocystidium sp. (30.77 ± 1.16%), the latter genus being always in coinfection with Haemogregarina. Significant differences were observed in 20 parameters between infected (free living) and uninfected (living in captivity) turtles. The ALP, PCV, Hb, and MCV were significantly different by sex; albumin, cholesterol, creatinine, percentage of heterophils, lymphocytes, and basophils differed in coinfection. This is the first report of reference intervals of P. vogli in the wild and the first study comparing hematological and blood biochemistry values between hemoparasite-infected and uninfected P. vogli turtles. However, the limited knowledge of these parameters in wild reptiles and the wide range of interval values makes it difficult to determine any significant impact on turtle health. Nevertheless, this study highlights the need for more in-depth research to explore the potential effects of blood parasite infections on turtles, including immune response and coevolution studies.

Leucocytozoon cariamae n. sp. and Haemoproteus pulcher coinfection in Cariama cristata (Aves: Cariamiformes): first mitochondrial genome analysis and morphological description of a leucocytozoid in Brazil.

The distribution of avian haemosporidians of the genus Leucocytozoon in the Neotropics remains poorly understood. Recent studies confirmed their presence in the region using molecular techniques alone, but evidence for gametocytes and data on putative competent hosts for Leucocytozoon are still lacking outside highland areas. We combined morphological and molecular data to characterize a new Leucocytozoon species infecting a non-migratory red-legged seriema (Cariama cristata), the first report of a competent host for Leucocytozoon in Brazil. Leucocytozoon cariamae n. sp. is distinguished from the Leucocytozoon fringillinarum group by its microgametocytes that are not strongly appressed to the host cell nucleus. The bird studied was coinfected with Haemoproteus pulcher, and we present a Bayesian phylogenetic analysis based on nearly complete mitochondrial genomes of these 2 parasites. Leucocytozoon cariamae n. sp. morphology is consistent with our phylogenetic analysis indicating that it does not share a recent common ancestor with the L. fringillinarum group. Haemoproteus pulcher and Haemoproteus catharti form a monophyletic group with Haemocystidium parasites of Reptilia, supporting the polyphyly of the genus Haemoproteus. We also discussed the hypothesis that H. pulcher and H. catharti may be avian Haemocystidium, highlighting the need to study non-passerine parasites to untangle the systematics of Haemosporida.

Co-infection with Leucocytozoon and Other Haemosporidian Parasites Increases with Latitude and Altitude in New World Bird Communities.

Establishing how environmental gradients and host ecology drive spatial variation in infection rates and diversity of pathogenic organisms is one of the central goals in disease ecology. Here, we identified the predictors of concomitant infection and lineage richness of blood parasites in New Word bird communities. Our multi-level Bayesian models revealed that higher latitudes and elevations played a determinant role in increasing the probability of a bird being co-infected with Leucocytozoon and other haemosporidian parasites. The heterogeneity in both single and co-infection rates was similarly driven by host attributes and temperature, with higher probabilities of infection in heavier migratory host species and at cooler localities. Latitude, elevation, host body mass, migratory behavior, and climate were also predictors of Leucocytozoon lineage richness across the New World avian communities, with decreasing parasite richness at higher elevations, rainy and warmer localities, and in heavier and resident host species. Increased parasite richness was found farther from the equator, confirming a reverse Latitudinal Diversity Gradient pattern for this parasite group. The increased rates of Leucocytozoon co-infection and lineage richness with increased latitude are in opposition with the pervasive assumption that pathogen infection rates and diversity are higher in tropical host communities.

Bird community effects on avian malaria infections.

In community assembly processes, interspecific interactions play an important role in shaping community diversity, especially at the local scale. Changes in species richness or abundance can modify local infectious disease dynamics, either reducing or increasing the risk of transmission within the community. This study evaluates the effects of bird community on avian haemosporidians infections in a Neotropical region. Bird samples were collected from areas surrounding three dams, and molecular analysis were performed to identify blood-parasitic haemosporidia infecting the birds. Generalized linear models were used to analyze the relationships between the bird community and the prevalence, number of infections, and richness of avian haemosporidian lineages. Non-significant effects of bird community dominance and richness on the prevalence of avian parasites and the number of infections of Haemoproteus were found. However, there was evidence of an amplification effect. Host dominance was associated with the total number of infections, the number Plasmodium infections and the expected richness of Plasmodium lineages, while the expected richness of Haemoproteus lineages was associated with the richness of bird species. These findings highlight the role of host community dominance and richness in the dynamics of parasite infections, potentially influenced by the availability of competent hosts. This study contributes significantly to our understanding of blood parasite diversity in tropical birds within a relatively understudied region of South America.

Prevalence and genetic diversity of avian haemosporidian parasites in islands within a mega hydroelectric dam in the Brazilian Amazon.

The Brazilian Amazon supports an extremely diverse avifauna and serves as the diversification center for avian malaria parasites in South America. Construction of hydroelectric dams can drive biodiversity loss by creating islands incapable of sustaining the bird communities found in intact forest sites. Besides anthropogenic actions, the presence of parasites can also influence the dynamics and structure of bird communities. Avian malaria (Plasmodium) and related haemosporidian parasites (Haemoproteus and Leucocytozoon) are a globally distributed group of protozoan parasites recovered from all major bird groups. However, no study to date has analyzed the presence of avian haemosporidian parasites in fragmented areas such as land bridge islands formed during artificial flooding following the construction of hydroelectric dams. The aim of this study is to assess the prevalence and molecular diversity of haemosporidians in bird communities inhabiting artificial islands in the area of the Balbina Hydroelectric Dam. The reservoir area covers 443,700 ha with 3546 islands on the left bank of the Uatumã River known to contain more than 400 bird species. We surveyed haemosporidian infections in blood samples collected from 445 understory birds, belonging to 53 species, 24 families, and 8 orders. Passeriformes represented 95.5% of all analyzed samples. We found a low overall Plasmodium prevalence (2.9%), with 13 positive samples (two Plasmodium elongatum and 11 Plasmodium sp.) belonging to eight lineages. Six of these lineages were previously recorded in the Amazon, whereas two of them are new. Hypocnemis cantator, the Guianan Warbling Antbird, represented 38.5% of all infected individuals, even though it represents only 5.6% of the sampled individuals. Since comparison with Plasmodium prevalence data prior to construction of Balbina is not possible, other studies in artificially flooded areas are imperative to test if anthropogenic flooding may disrupt vector-parasite relationships leading to low Plasmodium prevalence.

Host phylogeny and seasonality shapes avian haemosporidian prevalence in a Brazilian biodiverse and dry forest: the Caatinga.

The relationships between host phylogenetics, functional traits and parasites in wildlife remain poorly understood in the Neotropics, especially in habitats with marked seasonal variation. Here, we examined the effect of seasonality and host functional traits on the prevalence of avian haemosporidians (Plasmodium and Haemoproteus) in the Brazilian Caatinga, a seasonally dry tropical forest. 933 birds were evaluated for haemosporidian infections. We found a high parasitism prevalence (51.2%), which was correlated with phylogenetic relatedness among avian species. Prevalence varied drastically among the 20 well-sampled species, ranging from 0 to 70%. Seasonality was the main factor associated with infections, but how this abiotic condition influenced parasite prevalence varied according to the host-parasite system. Plasmodium prevalence increased during the rainy season and, after excluding the large sample size of Columbiformes (n = 462/933), Plasmodium infection rate was maintained high in the wet season and showed a negative association with host body mass. No association was found between non-Columbiform bird prevalence and seasonality or body mass when evaluating both Plasmodium and Haemoproteus or only Haemoproteus infections. Parasite community was composed of 32 lineages including 7 new lineages. We evidenced that even dry domains can harbour a high prevalence and diversity of vector-borne parasites and pointed out seasonality as a ruling factor.

Surveillance of avian malaria and related haemoparasites in common terns (Sterna hirundo) on the Atlantic coast of South America.

Haemosporidia (Apicomplexa, Haemosporida) are protozoa that infect vertebrate blood cells and are transmitted by vectors. Among vertebrates, birds possess the greatest diversity of haemosporidia, historically placed in 3 genera: Haemoproteus, Leucocytozoon and Plasmodium, the causative agent of avian malaria. In South America, existing data on haemosporidia are spatially and temporally dispersed, so increased surveillance is needed to improve the determination and diagnosis of these parasites. During the non-breeding season in 2020 and 2021, 60 common terns (Sterna hirundo) were captured and bled as part of ongoing research on the population health of migratory birds on the Argentinian Atlantic coast. Blood samples and blood smears were obtained. Fifty-eight samples were screened for Plasmodium, Haemoproteus and Leucocytozoon, as well as for Babesia parasites by nested polymerase chain reaction and by microscopic examination of smears. Two positive samples for Plasmodium were found. The cytochrome b lineages detected in the present study are found for the first time, and are close to Plasmodium lineages found in other bird orders. The low prevalence (3.6%) of haemoparasites found in this research was similar to those reported for previous studies on seabirds, including Charadriiformes. Our findings provide new information about the distribution and prevalence of haemosporidian parasites from charadriiforms in the southernmost part of South America, which remains understudied.

Sex-biased, but not plumage color-based, prevalence of haemosporidian parasites in free-range chickens.

Previous studies found a relationship between blood parasite infection and bird gender, with higher prevalence in males. Some studies also found a relationship between host plumage color and parasitic infection, while others did not. Here, we investigated the blood parasite prevalence in correlation with sex and plumage color in free-range chickens (Gallus gallus domesticus) in China. We analyzed a total of 297 blood samples, out of which 234 chickens tested positive for haemosporidian parasites, with 78.5% parasite prevalence. Out of 139 males, 118 tested positive with 84.8% parasite prevalence while 116 of 158 female samples tested positive (73.4%). Leucocytotozoon was the most frequent genus isolated (193 infected individuals /234 birds), followed by Plasmodium (41 infected individuals/234 birds), with no Haemoproteus parasites being detected. There were no significant differences in the body parameters and chicken color plumages with regards to the infection status. Our study indicated that blood parasite infection was significantly different between male and female chickens, with infection prevalent in males.

Morphometric and molecular characterization of an unpigmented haemosporidian parasite in the Neotropical turnip-tailed gecko (Thecadactylus rapicauda).

Morphological traits from blood stages have been the gold standard for determining haemosporidian parasite species. However, the status of some taxa and the value of such traits in parasites from reptiles remain contentious. The scarce sampling of these species worsens the situation, and several taxa lack molecular data. A survey was performed in the Magdalena Department in Colombia, where 16 species of reptiles were captured. A peculiar haemosporidian parasite was found in the Turnip-tailed gecko Thecadactylus rapicauda. This haemosporidian does not show malarial pigment in blood stages under light microscopy; thus, it fits the Garnia genus's characters belonging to the Garniidae. However, the phylogenetic analyses using a partial sequence of cytochrome b and the mitochondrial DNA placed it within the Plasmodium clade. Our findings suggest that many putative Garnia species belong to the genus Plasmodium, like the one reported here. This study either shows that visible malarial pigment in blood stages is not a diagnostic trait of the genus Plasmodium or malarial pigment might be present in an undetectable form under a light microscope. In any case, the current taxonomy of haemosporidian parasites in reptiles requires revision. This study highlights the importance of using molecular and morphological traits to address taxonomic questions at the species and genus levels in haemosporidian parasites from reptiles.

Blood parasites of passerines in the Brazilian Pampas and their implications for a potential population supplementation program for the endangered Yellow Cardinal (Gubernatrix cristata).

Espinilho savanna ("seasonal steppe savanna") is a unique vegetation formation of the Pampas biome that is found near the tri-border of Brazil, Uruguay, and Argentina. The Yellow Cardinal (Gubernatrix cristata) is a flagship species of this ecosystem, but it is classified as "critically endangered" in Brazil due to habitat loss and poaching for the illegal trade. Population supplementation through the release of individuals that were captive-bred or apprehended by authorities from the illegal trade has been considered as a conservation strategy for this species; however, the risk of pathogen introduction is a critical concern. We used microscopy and molecular methods to investigate the occurrence of blood parasites in wild passerines (n = 64, including three Yellow Cardinals) at Espinilho State Park, Rio Grande do Sul, Brazil, and in captive Yellow Cardinals (n = 30) at three facilities in Brazil. Haemosporidian parasites were detected in the blood smears of 10.9% of the wild passerines, comprising the morphospecies Haemoproteus erythrogravidus in Rufous-collared Sparrow (Zonotrichia capensis), H. quiscalus in Grayish Baywing (Agelaioides badius), and H. tyranni in Great Kiskadee (Pitangus sulphuratus); these are the southernmost records for these morphospecies and their first record for the Pampas biome. No haemosporidian parasites were detected in the blood smears of the Yellow Cardinals, wild or captive. Microfilariae were detected in the blood smears of 14.1% of the wild passerines, including all wild Yellow Cardinals, and in 43.3% of captive Yellow Cardinals. Trypanosoma sp. was detected in the blood smear of one captive Yellow Cardinal. Nested PCR and gene sequencing of the cyt-b gene of Haemoproteus/Plasmodium was used to test a subset of wild passerines and captive Yellow Cardinals, allowing for the molecular barcoding of H. quiscalus lineage AGEBAD04 and H. tyranni lineage PITSUL01; additionally, DNA identical to that of lineage PITSUL01 was detected in the blood of one captive Yellow Cardinal. This study provides valuable data to support the conservation management of the Yellow Cardinal and other threatened passerines from the Pampas and highlights the need for further studies on the epidemiology and pathology of filarioid worms and trypanosomes in passerines from this biome.

Beta diversity, prevalence, and specificity of avian haemosporidian parasites throughout the annual cycle of Chilean Elaenia (Elaenia chilensis), a Neotropical austral migrant.

Migratory birds are implicated in dispersing haemosporidian parasites over great geographic distances. However, their role in sharing these vector-transmitted blood parasites with resident avian host species along their migration flyway is not well understood. We studied avian haemosporidian parasites in 10 localities where Chilean Elaenia, a long-distance Neotropical austral migrant species, spends part of its annual cycle to determine local parasite transmission among resident sympatric host species in the elaenia's distributional range across South America. We sampled 371 Chilean Elaenias and 1,818 birds representing 243 additional sympatric species from Brazilian wintering grounds to Argentinian breeding grounds. The 23 haemosporidian lineages found in Chilean Elaenias exhibited considerable variation in distribution, specialization, and turnover across the 10 avian communities in South America. Parasite lineage dissimilarity increased with geographic distance, and infection probability by Parahaemoproteus decreased in localities harbouring a more diverse haemosporidian fauna. Furthermore, blood smears from migrating Chilean Elaenias and local resident avian host species did not contain infective stages of Leucocytozoon, suggesting that transmission did not take place in the Brazilian stopover site. Our analyses confirm that this Neotropical austral migrant connects avian host communities and transports haemosporidian parasites along its distributional range in South America. However, the lack of transmissive stages at stopover site and the infrequent parasite lineage sharing between migratory host populations and residents at breeding and wintering grounds suggest that Chilean Elaenias do not play a significant role in dispersing haemosporidian parasites, nor do they influence local transmission across South America.

Great-tailed Grackles (Quiscalus mexicanus) as a tolerant host of avian malaria parasites.

Great-tailed Grackles (Quiscalus mexicanus) are a social, polygamous bird species whose populations have rapidly expanded their geographic range across North America over the past century. Before 1865, Great-tailed Grackles were only documented in Central America, Mexico, and southern Texas in the USA. Given the rapid northern expansion of this species, it is relevant to study its role in the dynamics of avian blood parasites. Here, 87 Great-tailed grackles in Arizona (a population in the new center of the range) were screened for haemosporidian parasites using microscopy and PCR targeting the parasite mitochondrial cytochrome b gene. Individuals were caught in the wild from January 2018 until February 2020. Haemosporidian parasite prevalence was 62.1% (54/87). A high Plasmodium prevalence was found (60.9%, 53/87), and one grackle was infected with Haemoproteus (Parahaemoproteus) sp. (lineage SIAMEX01). Twenty-one grackles were infected with P. cathemerium, sixteen with P. homopolare, four with P. relictum (strain GRW04), and eleven with three different genetic lineages of Plasmodium spp. that have not been characterized to species level (MOLATE01, PHPAT01, and ZEMAC01). Gametocytes were observed in birds infected with three different Plasmodium lineages, revealing that grackles are competent hosts for some parasite species. This study also suggests that grackles are highly susceptible and develop chronic infections consistent with parasite tolerance, making them competent to transmit some generalist haemosporidian lineages. It can be hypothesized that, as the Great-tailed Grackle expands its geographic range, it may affect local bird communities by increasing the transmission of local parasites but not introducing new species into the parasite species pool.

Haemosporidians from a Neglected Group of Terrestrial Wild Birds in the Peruvian Amazonia.

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.

Molecular detection of Leucocytozoon in red-legged seriemas (Cariama cristata), a non-migratory bird species in the Brazilian Cerrado.

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.