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The role of plumage ornamentation as a signal of parasite infection is a key issue in the evolution of animal behaviour. Despite the undeniable role of host–parasite interactions in sexual selection, it is still unclear how parasites influence the variance in the expression of females displaying multiple ornaments. Here, by taking advantage of a long-term and individual-based, monitored population of pied flycatchers, Ficedula hypoleuca , in a Mediterranean area, we investigated the variation in the expression of wing patch (size) and forehead patch (occurrence and size) in relation to infections with haemosporidian parasites in breeding adult females. Haemosporidian infection was related to wing patch size, and this association varied in direction and magnitude depending on the origin of birds (locally born versus immigrant) and the type of breeding habitat (oakwood versus pine plantation). Specifically, differences in wing patch size were most evident among immigrant females breeding in the pine plantation, with noninfected females expressing larger wing patches than infected ones. Regarding the forehead patch, its occurrence was also modulated by haemosporidian infection through two-way interactions with bird origin (lower occurrence in locally born females) and breeding habitat type (lower occurrence among infected than uninfected females in the oakwood, whereas the opposite was true in the pine plantation). Only bird age predicted forehead patch size. Therefore, in addition to haemosporidian infection, the expression of these ornaments was related to extrinsic (breeding habitat) and intrinsic (age, origin) factors, which suggests the existence of trade-offs between physiological responses to infection and the expression of secondary sexual traits. Overall, our results suggest that plumage ornaments provide independent and reliable information on the haemosporidian infection status of female pied flycatchers. • Whether plumage ornaments are a sign of parasitic infections in females is unclear. • We assessed the links between two female ornaments and haemosporidian infection. • The expression of both ornaments was related to infection, origin, habitat and age. • However, the correlations between these factors differed depending on the ornament. • This suggests different signals for ornamentation – infection response trade-offs. [ABSTRACT FROM AUTHOR]

Introduction: Leucocytozoon is an intracellular blood parasite that affects various bird species globally and is transmitted by blackfly vectors. This parasite is responsible for leucocytozoonosis, a disease that results in significant economic losses due to reduced meat and egg production. There is limited knowledge about the epidemiological pattern of leucocytozoonosis and its causative species in Egypt, particularly in pigeons. Methods: The current study involved the collection of 203 blood samples from domestic pigeons from various household breeders and local markets across Qena Province, Upper Egypt. Samples were initially examined for potential Leucocytozoon infection using blood smears, followed by an evaluation of associated risk factors. Molecular identification of the parasite in selected samples (n = 11), which had initially tested positive via blood smears, was further refined through nested PCR and sequence analysis of the mitochondrial cytochrome b gene to ascertain the Leucocytozoon species present. Additionally, histopathological examination of the liver, spleen, and pancreas was conducted on animals that tested positive by blood smears. Results: Interestingly, 26 out of 203 samples (12.08%) had confirmed Leucocytozoon infections based on microscopic analysis. Additionally, all 11 samples that initially tested positive via blood smears were confirmed positive through nested PCR analysis, and their sequencing revealed the presence of Leucocytozoon sabrazesi, marking the first report of this parasite in Egypt. The study into potential risk factors unveiled the prevalence of Leucocytozoon spp. seems host gender-dependent, with males exhibiting a significantly higher infection rate (33.33%). Additionally, adult birds demonstrated a significantly higher infection prevalence than squabs, suggesting an age-dependent trend in prevalence. Seasonality played a significant role, with the highest occurrence observed during summer (37.25%). Histopathological examination revealed the presence of numerous megaloschizonts accompanied by lymphocytic infiltration and multiple focal areas of ischemic necrosis. Conclusion: To our knowledge, this is the first study to shed light on the epidemiological characteristics and molecular characterization of leucocytozoonosis in pigeons in Egypt. Further research endeavors are warranted to curb the resurgence of Leucocytozoon parasites in other avian species across Egypt, thereby refining the epidemiological understanding of the disease for more effective control and prevention measures. [ABSTRACT FROM AUTHOR]

Abstract Background Studies on haemosporidian diversity, including origin of human malaria parasites, malaria's zoonotic dynamic, and regional biodiversity patterns, have used target gene approaches. However, current methods have a trade-off between scalability and data quality. Here, a long-read Next-Generation Sequencing protocol using PacBio HiFi is presented. The data processing is supported by a pipeline that uses machine-learning for analysing the reads. Methods A set of primers was designed to target approximately 6 kb, almost the entire length of the haemosporidian mitochondrial genome. Amplicons from different samples were multiplexed in an SMRTbell® library preparation. A pipeline (HmtG-PacBio Pipeline) to process the reads is also provided; it integrates multiple sequence alignments, a machine-learning algorithm that uses modified variational autoencoders, and a clustering method to identify the mitochondrial haplotypes/species in a sample. Although 192 specimens could be studied simultaneously, a pilot experiment with 15 specimens is presented, including in silico experiments where multiple data combinations were tested. Results The primers amplified various haemosporidian parasite genomes and yielded high-quality mt genome sequences. This new protocol allowed the detection and characterization of mixed infections and co-infections in the samples. The machine-learning approach converged into reproducible haplotypes with a low error rate, averaging 0.2% per read (minimum of 0.03% and maximum of 0.46%). The minimum recommended coverage per haplotype is 30X based on the detected error rates. The pipeline facilitates inspecting the data, including a local blast against a file of provided mitochondrial sequences that the researcher can customize. Conclusions This is not a diagnostic approach but a high-throughput method to study haemosporidian sequence assemblages and perform genotyping by targeting the mitochondrial genome. Accordingly, the methodology allowed for examining specimens with multiple infections and co-infections of different haemosporidian parasites. The pipeline enables data quality assessment and comparison of the haplotypes obtained to those from previous studies. Although a single locus approach, whole mitochondrial data provide high-quality information to characterize species pools of haemosporidian parasites.

Simple Summary: Feathers are essential for a bird's flight, insulation, communication, and camouflage. They degrade over time, so birds must moult regularly. This study examined how avian haemosporidian infection and the size of the uropygial gland affect feather quality and growth rate in two migratory bird species in southwestern Spain—the house martin (Delichon urbicum) and the sand martin (Riparia riparia). We found that house martins had the highest haemosporidian infection rates, likely due to their large colony size. Infection only decreased feather quality in house martins and did not affect the feather growth rate in any of the two hirundinids. Additionally, feather growth rate was positively linked to feather quality, but only in house martins. Finally, we found no connection between the uropygial gland size and feather quality or feather growth rate. These results show, for the first time, that avian haemosporidian parasites can negatively impact the feather quality of migratory birds, thus potentially affecting their flight and survival. Further research is needed to fully understand these relationships. Bird feathers have several functions, including flight, insulation, communication, and camouflage. Since feathers degrade over time, birds need to moult regularly to maintain these functions. However, environmental factors like food scarcity, stress, and parasite infections can affect feather quality and moult speed. This study examined the impact of avian haemosporidian infection and uropygial gland volume, as well as feather quality and feather growth rate in two migratory hirundine species captured in southwestern Spain—the house martin (Delichon urbicum) and sand martin (Riparia riparia). Our findings showed that the prevalence of infection varied among species, with house martins having the highest rates, possibly due to their larger colony size. Moreover, haemosporidian infection had a different impact on each species; infected house martins exhibited lower feather quality than healthy individuals, although this outcome was not observed in sand martins. Furthermore, no effect of infection on feather growth rate was observed in both hirundinids. Additionally, feather growth rate only correlated positively with feather quality in house martins. Finally, no link was observed between uropygial gland volume and feather quality or feather growth rate in any of the species in this study. These findings highlight the effect of haemosporidian infections on the plumage of migratory birds, marking, for the first time, how avian haemosporidian infection is shown to adversely impact feather quality. Even so, further research is needed to explore these relationships more deeply. [ABSTRACT FROM AUTHOR]

Avian haemosporidian parasites are useful model organisms to study the ecology and evolution of parasite-host interactions due to their global distribution and extensive biodiversity. Detection of these parasites has evolved from microscopic examination to PCR-based methods, with the mitochondrial cytochrome b gene serving as barcoding region. However, standard PCR protocols used for screening and identification purposes have limitations in detecting mixed infections and generating phylogenetically informative data due to short amplicon lengths. To address these issues, we developed a novel genusspecific nested PCR protocol targeting avian haemosporidian parasites. The protocol underwent rigorous testing utilizing a large dataset comprising blood samples from Malagasy birds of three distinct Passeriformes families. Furthermore, validation was done by examining smaller datasets in two other laboratories employing divergent master mixes and different bird species. Comparative analyses were conducted between the outcomes of the novel PCR protocol and those obtained through the widely used standard nested PCR method. The novel protocol enables specific identification of Plasmodium, Haemoproteus (Parahaemoproteus), and Leucocytozoon parasites. The analyses demonstrated comparable sensitivity to the standard nested PCR with notable improvements in detecting mixed infections. In addition, phylogenetic resolution is improved by amplification of longer fragments, leading to a better understanding of the haemosporidian biodiversity and evolution. Overall, the novel protocol represents a valuable addition to avian haemosporidian detection methodologies, facilitating comprehensive studies on parasite ecology, epidemiology, and evolution. [ABSTRACT FROM AUTHOR]

Background: Studies on haemosporidian diversity, including origin of human malaria parasites, malaria's zoonotic dynamic, and regional biodiversity patterns, have used target gene approaches. However, current methods have a trade-off between scalability and data quality. Here, a long-read Next-Generation Sequencing protocol using PacBio HiFi is presented. The data processing is supported by a pipeline that uses machine-learning for analysing the reads. Methods: A set of primers was designed to target approximately 6 kb, almost the entire length of the haemosporidian mitochondrial genome. Amplicons from different samples were multiplexed in an SMRTbell® library preparation. A pipeline (HmtG-PacBio Pipeline) to process the reads is also provided; it integrates multiple sequence alignments, a machine-learning algorithm that uses modified variational autoencoders, and a clustering method to identify the mitochondrial haplotypes/species in a sample. Although 192 specimens could be studied simultaneously, a pilot experiment with 15 specimens is presented, including in silico experiments where multiple data combinations were tested. Results: The primers amplified various haemosporidian parasite genomes and yielded high-quality mt genome sequences. This new protocol allowed the detection and characterization of mixed infections and co-infections in the samples. The machine-learning approach converged into reproducible haplotypes with a low error rate, averaging 0.2% per read (minimum of 0.03% and maximum of 0.46%). The minimum recommended coverage per haplotype is 30X based on the detected error rates. The pipeline facilitates inspecting the data, including a local blast against a file of provided mitochondrial sequences that the researcher can customize. Conclusions: This is not a diagnostic approach but a high-throughput method to study haemosporidian sequence assemblages and perform genotyping by targeting the mitochondrial genome. Accordingly, the methodology allowed for examining specimens with multiple infections and co-infections of different haemosporidian parasites. The pipeline enables data quality assessment and comparison of the haplotypes obtained to those from previous studies. Although a single locus approach, whole mitochondrial data provide high-quality information to characterize species pools of haemosporidian parasites. [ABSTRACT FROM AUTHOR]

IntroductionLeucocytozoon is an intracellular blood parasite that affects various bird species globally and is transmitted by blackfly vectors. This parasite is responsible for leucocytozoonosis, a disease that results in significant economic losses due to reduced meat and egg production. There is limited knowledge about the epidemiological pattern of leucocytozoonosis and its causative species in Egypt, particularly in pigeons.MethodsThe current study involved the collection of 203 blood samples from domestic pigeons from various household breeders and local markets across Qena Province, Upper Egypt. Samples were initially examined for potential Leucocytozoon infection using blood smears, followed by an evaluation of associated risk factors. Molecular identification of the parasite in selected samples (n = 11), which had initially tested positive via blood smears, was further refined through nested PCR and sequence analysis of the mitochondrial cytochrome b gene to ascertain the Leucocytozoon species present. Additionally, histopathological examination of the liver, spleen, and pancreas was conducted on animals that tested positive by blood smears.ResultsInterestingly, 26 out of 203 samples (12.08%) had confirmed Leucocytozoon infections based on microscopic analysis. Additionally, all 11 samples that initially tested positive via blood smears were confirmed positive through nested PCR analysis, and their sequencing revealed the presence of Leucocytozoon sabrazesi, marking the first report of this parasite in Egypt. The study into potential risk factors unveiled the prevalence of Leucocytozoon spp. seems host gender-dependent, with males exhibiting a significantly higher infection rate (33.33%). Additionally, adult birds demonstrated a significantly higher infection prevalence than squabs, suggesting an age-dependent trend in prevalence. Seasonality played a significant role, with the highest occurrence observed during summer (37.25%). Histopathological examination revealed the presence of numerous megaloschizonts accompanied by lymphocytic infiltration and multiple focal areas of ischemic necrosis.ConclusionTo our knowledge, this is the first study to shed light on the epidemiological characteristics and molecular characterization of leucocytozoonosis in pigeons in Egypt. Further research endeavors are warranted to curb the resurgence of Leucocytozoon parasites in other avian species across Egypt, thereby refining the epidemiological understanding of the disease for more effective control and prevention measures.

Simple Summary: Over thirty percent of bird species are undergoing population declines and are threatened with extinction in Portugal. Several reasons have been proposed to explain this decrease, such as the impact of human activities on natural environments and pathogens affecting the health of wildlife, domestic animals, and humans. Wildlife rescue and rehabilitation centres play an essential role in the conservation of endangered species. Despite wildlife rehabilitation centres providing valuable information on disease prevalence and transmission, the information on haemosporidian infection is still very scarce for birds admitted in these centres. In this study, we discovered new malaria parasites in birds admitted to wildlife rehabilitation centres in Portugal. We also revealed infection in bird species that were previously unknown to be infected with malaria parasites. Birds admitted to rehabilitation due to debilitating disease were more frequently infected with malaria. Furthermore, we demonstrate that the malaria infection extends the required period for medical treatment in these birds, which imposes additional economic costs for the rehabilitation and reduces the survival probabilities of the bird. These findings stress the importance of the study of malaria parasites in wildlife rehabilitation centres, also helping to design protocols and interventions to preserve endangered species. In the last decade, over 40% of bird species in Europe have experienced poor and bad conservation status, with more than 30% of bird species in mainland Portugal threatened with extinction. Along with anthropogenic factors, parasites and pathogens such as avian haemosporidians have been suggested to be responsible for these avian population declines. Wildlife rehabilitation centres play an essential role in species conservation and preservation. Moreover, animals admitted for rehabilitation can provide valuable information regarding transmission and pathogenicity of many diseases that affect wild birds that are rarely sampled in nature. However, reports of haemosporidians in captive birds are still limited. Here, we explored the prevalence and genetic diversity of avian haemosporidians in 89 birds from 29 species admitted to rehabilitation centres in Portugal, showing an overall infection prevalence of 30.3%. The prevalence of infection was higher in Strigiformes and in birds admitted to rehabilitation centres due to debilitating diseases. Remarkably, 30% of the infected bird species have not been found to harbour malaria parasites in preceding studies. We detected 15 different haemosporidian lineages infecting a third of bird species sampled. Notably, 2 out of these 15 detected haemosporidian lineages have not been obtained previously in other studies. Furthermore, we also identified nine new host–parasite interactions representing new host records for these haemosporidian parasites. Finally, our results revealed that birds infected with haemosporidians require longer rehabilitation treatments, which increase the economic costs for rehabilitation and may impair their survival prospects. These findings emphasise the importance of integrating haemosporidian infection considerations into rehabilitation protocols, highlighting the challenges posed by these infections in avian conservation and rehabilitation, including economic and logistical demands. [ABSTRACT FROM AUTHOR]

Avian haemosporidian parasites are widespread globally and infect numerous wild bird species. However, they have primarily been studied in passerine birds. Accordingly, the prevalence and diversity of these parasites in birds of prey remain understudied. This lack of research is primarily due to the international protection status of many of these species, their sparse distribution across ecosystems and difficulty to capture in the wild. The aim of this study was to examine the prevalence and diversity of haemosporidian parasites in two species of birds of prey, namely white‐tailed eagle and lesser spotted eagle. To achieve this, a non‐invasive approach was employed, involving the extraction of DNA from blood spots present in moulted feathers. Freshly moulted feathers were collected from the ground under nests or within the nests of these birds during the breeding season. A visible blood spot located at the junction of the calamus and rachis was removed and fixed in SET buffer for molecular analysis. The identification of haemosporidian parasites (Plasmodium, Haemoproteus and Leucocytozoon) was conducted using PCR‐based methods. Overall, parasite DNA was successfully detected in shed feathers that were kept in their original form at least decade prior to analysis. Among the studied individuals, resident white‐tailed eagles showed significantly lower infection rates of haemosporidian parasites compared to migratory lesser spotted eagles. A total of nine genetic lineages of haemosporidian parasites were detected, with seven of them being new to science based on partial sequences of the cytb gene. Additionally, the phylogenetic relationships among these new lineages and previously described ones were established. These findings highlight the suitability of non‐invasive sampling for investigating the prevalence and diversity of haemosporidian parasites in wild birds of prey populations. Moreover, this approach holds promise for studying other challenging‐to‐reach and protected bird species. According to our research, there is a greater chance of finding haemosporidian parasites in freshly gathered feathers. [ABSTRACT FROM AUTHOR]

Haemosporidians are a group of vector-borne parasites belonging to the order Haemosporida. These parasites infect avian hosts and require blood-sucking insects (Diptera) for transmission. The occurrence and diversity of haemosporidian parasites are shaped primarily by the specificity of the parasite and the susceptibility of the host/vector. In this study, the presence and distribution of haemosporidians in blood samples from birds in urbanized and natural habitats were estimated using microscopic and molecular approaches. Birds in urbanized habitats were infected with four different species of Plasmodium, P. relictum, P. vaughani, P. matutinum, and P. circumflexum, and one species of Haemoproteus, H. parabelopolskyi, and Leucocytozoon sp. The species H. attenuatus, H. concavocentralis, H. minutus, H. pallidus, H. noctuae, and H. tartakovskyi were additionally identified in birds in natural habitats. Typically, juvenile birds are essential markers of parasite species transmitted in the study area. The juveniles in the urbanized habitats carried P. relictum, P. vaughani, P. circumflexum, H. parabelopolskyi, and Leucocytozoon species. The most abundant parasite was H. parabelopolskyi, which was found in both habitat types. The prevalence of Haemoproteus/Plasmodium species determined by nested PCR in birds in natural habitats (43.80%; 53/121) was significantly greater than that in birds in urbanized habitats (21.94%; 43/196) (p < 0.05). There was no significant difference in the infection rate of Leucocytozoon sp. between the habitat types (p > 0.05; 10/121 vs. 19/196). [ABSTRACT FROM AUTHOR]

Haemosporidians are a diverse group of vector-borne parasitic protozoa that have a global distribution and some of them live in the avian body as a benign parasite, with no symptoms. Avian Haemosporidians, include three main genera, Plasmodium, Haemoproteus, and Leucocytozoon. These blood parasites are transmitted across different regions through migratory birds. We studied the prevalence of avian haemosporidian parasites in both resident and migratory birds in the central and southern regions of Iraq. Our study represents pioneering research on Haemosporidian parasites in a wide range of avian species, including 248 samples from 47 species of both resident and migratory birds, collected from five cities in central and southern Iraq. We identified and determined the prevalence of three distinct haemosporidian genera, Haemoproteus spp., Leucocytozoon spp., and Plasmodium spp. Overall parasite infection was higher in resident hosts than in migrants. The prevalence of Plasmodium spp. was found to be highest among resident birds, with a rate of 20.1%, while the highest rate of Leucocytozoon spp. was observed in migratory birds, with a rate of 8.2%. Our study holds promising opportunities for understanding the impact of migratory and resident hosts on the dynamics of infectious diseases in wildlife.

The nectarivorous common sunbird asity (Neodrepanis coruscans) is phylogenetically closely related to the frugivorous velvet asity (Philepitta castanea), yet it shares similar habitat and foraging behaviour as the Malagasy sunbirds (Cinnyris spp.). As ecological factors have been shown to influence blood parasite prevalence, it should be tested whether parasite abundance, prevalence and diversity of N. coruscans are more similar to the sunbirds than to its relative. Therefore, blood samples (n = 156) and smears (n = 60) were tested for different blood parasites (Haemosporida, trypanosomes, filarioid nematodes) using molecular and microscopic methods. High prevalence of haemosporidian parasites was observed in all bird taxa, with rates ranging from 23% in N. coruscans to 84.6% in C. notatus. The Malagasy Cinnyris spp. exhibited a high occurrence of mixed haemosporidian infections (>76%) with various specialized lineages. Within the Philepittidae family, no Haemoproteus infection was detected and just a few cases of mixed infections. Nectariniidae species predominantly had specialized haemosporidian lineages, while Philepittidae had infections mainly caused by generalist lineages. These findings emphasize the diverse range of blood parasites in Nectariniidae, while additionally highlighting the high diversity of trypanosomes and filarioid nematodes in Philepittidae. Additionally, several newly discovered haemosporidian lineages, Trypanosoma isolates and filarioid nematode isolates were identified. Notably, Philepittidae exhibited a lower prevalence of avian haemosporidian parasites compared to Nectariniidae, possibly due to potential resistance mechanisms. Despite N. coruscans sharing similar habitat and behavioural ecology with both Cinnyris spp., it closely resembles its relative, P. castanea, in all aspects of haemosporidian parasitism.

Leucocytozoon parasites (Haemosporida, Leucocytozoidae) are haemosporidians whose diversity, exo-erythrocytic development and potential vectors are the least studied. The knowledge about their exo-erythrocytic development and pathogenicity is fragmentary, resulting in an incomplete comprehension of the impact of these parasites on avian hosts. For a long time, Leucocytozoon infections were considered benign to wild birds, even though they were virulent in poultry and responsible for some wild bird population declines. This study aimed to investigate the presence of Leucocytozoon species exo-erythrocytic stages in song thrushes Turdus philomelos using conventional histological techniques (sections stained by H&E) and chromogenic in situ hybridization (CISH). Tissues from ten birds (seven naturally infected and three opportunistic samplings) were examined using both methods. Parasite lineages were identified from blood samples using PCR-based techniques. Leucocytozoon species meronts were found in five individuals (in four birds using H&E staining protocol, and in three in CISH-treated histological sections). Meronts were found mainly in the kidneys, but some meronts were also present in the lungs. It was possible to observe different maturation stages of meronts in the same bird individual, indicating an asynchronous development. Cytomeres were readily visible in developing meronts. One megalomeront-like structure was present close to a blood vessel in the heart. It was covered with a prominent capsular-like wall. No inflammatory reaction or necrosis was seen in the tissues surrounding the meronts or the megalomeront-like structure. We could confirm the transmission of three Leucocytozoon lineages (lTUPHI14, lSTUR1 and lTUPHI13) in Europe, and add evidence of the transmission of two Plasmodium lineages, including Plasmodium circumflexum (pTURDUS1), and Haemoproteus asymmetricus (hTUPHI01). We call for further research to better understand Leucocytozoon parasite exo-erythrocytic development.

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. [ABSTRACT FROM AUTHOR]

In addition to the presence of a suitable host and vector, the prevalence of haemosporidians is influenced by several important factors, including the environmental conditions of the habitat, which depend on broader geographic characteristics. The aim of this study is to perform a preliminarily assessment of the distribution of blood parasites in birds from the mountainous area of Zhongar Alatau NP and to find potential new sites for research on their ecology in Kazakhstan. The results of this research constitute the first report on the occurrence of blood parasites from this area. A total of 58 birds, from the order Passeriformes and one individual from the order Caprimulgiformes, were examined during the study. The overall prevalence of infections caused by haemosporidian parasites (Haemoproteus, Leucocytozoon) was 18.6%. Neither the genus Plasmodium nor the presence of trypanosomes and microfilariae was detected in the birds examined. Three birds (5.1% prevalence) were infected with parasites of the genus Haemoproteus, in all eleven positive birds the analyses showed the presence of parasites of the genus Leucocytozoon (18.6% prevalence). The presence of parasites genus Haemoproteus was detected only in birds that were also infected with Leucocytozoon parasites. More infections with parasites of the genus Leucocytozoon are predicted due to the higher altitude and ecological factors at the capture sites, which are more favourable for the development of vectors of this genus. The species Haemoproteus majoris was detected in the host Emberiza cioides and species Haemoproteus minutus in host Turdus merula. Other species of this genus in the hosts Cyanistes cyanus and Turdus atrogularis were not determined. The species Leucocytozoon fringilinarum was detected in the hosts Cyanistes cyanus and Parus major, Leucocytozoon dubreuili was detected in Turdus atrogularis and Turdus merula. In the other host species Aegithalos caudatus, Emberiza cioides and Periparus aterus, it was not possible to dermine the species of the genus Leucocytozoon. [ABSTRACT FROM AUTHOR]

In a 2-yr study on prevalence of Haemosporidia in an avian community in Ithaca, New York, USA, we tested the hypothesis that apparent seasonal variation in prevalence is influenced by the detection protocol. We confirmed a higher detection of Haemosporidia using a molecular diagnosis technique (PCR) than by microscopy; this further increased when the PCR test was triplicated. Microscopic examination and PCR techniques have different specificity and sensitivity and therefore different probabilities of detecting hemoparasites. Birds with chronic infections or sampled during winter often have very low parasitemia, and such infections may be missed by microscopy but detected by PCR. Haemosporidian prevalence was higher during the breeding season than during the nonbreeding season regardless of the method used. Detection of Leucocytozoon spp. infection from blood smears using microscopy was challenging. [ABSTRACT FROM AUTHOR]

[Display omitted] • Geographic range is more important than bird migratory behaviour for parasite diversity. • Patterns differed between Plasmodium , Haemoproteus and Leucocytozoon parasites. • Species resident in both Africa and Eurasia (RB) had higher diversity than all other groups. • RB and migrants had higher Haemoproteus diversity than single-continent residents. Avian blood parasites, from the genera Plasmodium, Haemoproteus and Leucocytozoon , are predicted to alter their range and prevalence as global temperatures change, and host and vector ranges shift. Understanding large-scale patterns in the prevalence and diversity of avian malaria and malaria-like parasites is important due to an incomplete understanding of their effects in the wild, where studies suggest even light parasitaemia can potentially cause rapid mortality, especially in naïve populations. We conducted phylogenetically controlled analyses to test for differences in prevalence and lineage diversity of haemoparasite infection (for Plasmodium , Haemoproteus and Leucocytozoon) in and between resident and migratory species along the African-Eurasian flyway. To test whether migratory strategy or range size drives differences in parasite prevalence and diversity between resident and migrant species, we included three categories of resident species: Eurasian only (n = 36 species), African only (n = 41), and species resident on both continents (n = 17), alongside intercontinental migrants (n = 64), using a subset of data from the MalAvi database comprising 27,861 individual birds. We found that species resident on both continents had a higher overall parasite diversity than all other categories. Eurasian residents had lower Plasmodium diversity than all other groups, and both migrants and species resident on both continents had higher Haemoproteus diversity than both African and Eurasian residents. Leucocytozoon diversity did not differ between groups. Prevalence patterns were less clear, with marked differences between genera. Both Plasmodium and Leucocytozoon prevalence was higher in species resident on both continents and African residents than in migrants and Eurasian residents. Haemoproteus prevalence was lower in Eurasian residents than species resident on both continents. Our findings contrast with previous findings in the North-South American flyway, where long-distance migrants had higher parasite diversity than residents and short-distance migrants, although we found contrasting patterns for parasite diversity to those seen for parasite prevalence. Crucially, our results suggest that geographic range may be more important than migratory strategy in driving parasite diversity within species along the African-Palaearctic flyway. Our findings differ between the three parasite genera included in our analysis, suggesting that vector ecology may be important in determining these large-scale patterns. Our results add to our understanding of global patterns in parasite diversity and abundance, and highlight the need to better understand the influence of vector ecology to understand the drivers of infection risk and predict responses to environmental change. [ABSTRACT FROM AUTHOR]

Haemosporidian parasites that infect birds (Apicomplexa: Haemosporida) are blood parasites that require an invertebrate host (vector) and a vertebrate host for their lifecycle and cause malaria-like diseases. This group of parasites has provided valuable insights into host specificity, virulence, and parasite dispersal. Additionally, they have played a significant role in reshaping our understanding of the evolutionary history of apicomplexans. In order to accurately identify species and to address phylogenetic questions such as the timing of the haemosporidian radiation, the use of a sufficiently large genetic data set is crucial. However, acquiring this genetic data poses significant challenges. In this research, a sensitive nested PCR assay was developed. This assay allows for the easy amplification of complete mitochondrial genomes of haemosporidian parasites in birds, even during the chronic stage of infection. The effectiveness of this new nested PCR assay was evaluated using blood and tissue samples of birds with verified single parasite infections from previous studies. The approach involves amplifying four overlapping fragments of the mitochondrial genome and requires DNA extracts from single-infected samples. This method successfully amplified the complete mitochondrial genomes of 24 distinct haemosporidian parasite lineages found in various bird species. This data is invaluable for conducting phylogenetic analyses and accurately defining species. Furthermore, this study proposes the existence of at least 15 new haemosporidian parasite species based on the genetic information obtained. Data regarding pGRW04, previously categorized as Plasmodium relictum like pSGS1 and pGRW11, indicates that the pGRW04 lineage is actually a separate, hidden Plasmodium species. [ABSTRACT FROM AUTHOR]

Leucocytozoon parasites (Haemosporida, Leucocytozoidae) are haemosporidians whose diversity, exo-erythrocytic development and potential vectors are the least studied. The knowledge about their exo-erythrocytic development and pathogenicity is fragmentary, resulting in an incomplete comprehension of the impact of these parasites on avian hosts. For a long time, Leucocytozoon infections were considered benign to wild birds, even though they were virulent in poultry and responsible for some wild bird population declines. This study aimed to investigate the presence of Leucocytozoon species exo-erythrocytic stages in song thrushes Turdus philomelos using conventional histological techniques (sections stained by H&E) and chromogenic in situ hybridization (CISH). Tissues from ten birds (seven naturally infected and three opportunistic samplings) were examined using both methods. Parasite lineages were identified from blood samples using PCR-based techniques. Leucocytozoon species meronts were found in five individuals (in four birds using H&E staining protocol, and in three in CISH-treated histological sections). Meronts were found mainly in the kidneys, but some meronts were also present in the lungs. It was possible to observe different maturation stages of meronts in the same bird individual, indicating an asynchronous development. Cytomeres were readily visible in developing meronts. One megalomeront-like structure was present close to a blood vessel in the heart. It was covered with a prominent capsular-like wall. No inflammatory reaction or necrosis was seen in the tissues surrounding the meronts or the megalomeront-like structure. We could confirm the transmission of three Leucocytozoon lineages (lTUPHI14, lSTUR1 and lTUPHI13) in Europe, and add evidence of the transmission of two Plasmodium l ineages, including Plasmodium circumflexum (pTURDUS1), and Haemoproteus asymmetricus (hTUPHI01). We call for further research to better understand Leucocytozoon parasite exo-erythrocytic development. [Display omitted] • First report of Leucocytozoon exo-erythrocytic stages in song thrushes. • Meronts of Leucocytozoon infecting song thrushes are common in the kidneys. • A megalomeront-like structure of unusual morphology was found in one bird. • Chromogenic in situ hybridization is essential to confirm parasite identity. • Confirmed the transmission of three Leucocytozoon lineages (lTUPHI14, lSTUR1 and lTUPHI13) in Europe. [ABSTRACT FROM AUTHOR]

Avian haemosporidian parasites are useful model organisms to study the ecology and evolution of parasite-host interactions due to their global distribution and extensive biodiversity. Detection of these parasites has evolved from microscopic examination to PCR-based methods, with the mitochondrial cytochrome b gene serving as barcoding region. However, standard PCR protocols used for screening and identification purposes have limitations in detecting mixed infections and generating phylogenetically informative data due to short amplicon lengths. To address these issues, we developed a novel genus-specific nested PCR protocol targeting avian haemosporidian parasites. The protocol underwent rigorous testing utilizing a large dataset comprising blood samples from Malagasy birds of three distinct Passeriformes families. Furthermore, validation was done by examining smaller datasets in two other laboratories employing divergent master mixes and different bird species. Comparative analyses were conducted between the outcomes of the novel PCR protocol and those obtained through the widely used standard nested PCR method. The novel protocol enables specific identification of Plasmodium, Haemoproteus (Parahaemoproteus), and Leucocytozoon parasites. The analyses demonstrated comparable sensitivity to the standard nested PCR with notable improvements in detecting mixed infections. In addition, phylogenetic resolution is improved by amplification of longer fragments, leading to a better understanding of the haemosporidian biodiversity and evolution. Overall, the novel protocol represents a valuable addition to avian haemosporidian detection methodologies, facilitating comprehensive studies on parasite ecology, epidemiology, and evolution.

Abstract Avian haemosporidian parasites are widespread globally and infect numerous wild bird species. However, they have primarily been studied in passerine birds. Accordingly, the prevalence and diversity of these parasites in birds of prey remain understudied. This lack of research is primarily due to the international protection status of many of these species, their sparse distribution across ecosystems and difficulty to capture in the wild. The aim of this study was to examine the prevalence and diversity of haemosporidian parasites in two species of birds of prey, namely white‐tailed eagle and lesser spotted eagle. To achieve this, a non‐invasive approach was employed, involving the extraction of DNA from blood spots present in moulted feathers. Freshly moulted feathers were collected from the ground under nests or within the nests of these birds during the breeding season. A visible blood spot located at the junction of the calamus and rachis was removed and fixed in SET buffer for molecular analysis. The identification of haemosporidian parasites (Plasmodium, Haemoproteus and Leucocytozoon) was conducted using PCR‐based methods. Overall, parasite DNA was successfully detected in shed feathers that were kept in their original form at least decade prior to analysis. Among the studied individuals, resident white‐tailed eagles showed significantly lower infection rates of haemosporidian parasites compared to migratory lesser spotted eagles. A total of nine genetic lineages of haemosporidian parasites were detected, with seven of them being new to science based on partial sequences of the cytb gene. Additionally, the phylogenetic relationships among these new lineages and previously described ones were established. These findings highlight the suitability of non‐invasive sampling for investigating the prevalence and diversity of haemosporidian parasites in wild birds of prey populations. Moreover, this approach holds promise for studying other challenging‐to‐reach and protected bird species. According to our research, there is a greater chance of finding haemosporidian parasites in freshly gathered feathers.

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.

Bird feathers have several functions, including flight, insulation, communication, and camouflage. Since feathers degrade over time, birds need to moult regularly to maintain these functions. However, environmental factors like food scarcity, stress, and parasite infections can affect feather quality and moult speed. This study examined the impact of avian haemosporidian infection and uropygial gland volume, as well as feather quality and feather growth rate in two migratory hirundine species captured in southwestern Spain—the house martin (Delichon urbicum) and sand martin (Riparia riparia). Our findings showed that the prevalence of infection varied among species, with house martins having the highest rates, possibly due to their larger colony size. Moreover, haemosporidian infection had a different impact on each species; infected house martins exhibited lower feather quality than healthy individuals, although this outcome was not observed in sand martins. Furthermore, no effect of infection on feather growth rate was observed in both hirundinids. Additionally, feather growth rate only correlated positively with feather quality in house martins. Finally, no link was observed between uropygial gland volume and feather quality or feather growth rate in any of the species in this study. These findings highlight the effect of haemosporidian infections on the plumage of migratory birds, marking, for the first time, how avian haemosporidian infection is shown to adversely impact feather quality. Even so, further research is needed to explore these relationships more deeply.

Hemosporidian parasite communities are broadly similar in Boulder County, Colorado, between two common songbirds--the Black-capped Chickadee (Poecile atricapillus) and Mountain Chickadee (Poecile gambeli). However, Mountain Chickadees appear more likely to be infected with Plasmodium and potentially experience higher infection burdens with Leucocytozoon in contrast to Black-capped Chickadees. We found that elevation change (and associated ecology) drives the distributions of these parasite genera. For Boulder County chickadees, environmental factors play a more important role in structuring hemosporidian communities than host evolutionary differences. However, evolutionary differences are likely key to shaping the probability of infection, infection burden, and whether an infection remains detectable over time. We found that for recaptured birds, their infection status (i.e. presence or absence of detectable parasite infection) tends to remain consistent across capture periods. We sampled 235 chickadees between 2017 and 2021 across a ~1,500-m elevation gradient from low elevation (i.e. the city of Boulder) to comparatively high elevation (i.e. the CU Boulder Mountain Research Station). It is unknown whether long-term hemosporidian abundance trends have changed over time in our sampling region. However, we ask whether potentially disparate patterns of Plasmodium susceptibility and Leucocytozoon infection burden could be playing a role in the negative population trends of Mountain Chickadees. [ABSTRACT FROM AUTHOR]

Simple Summary: Chickens can be infected by several avian blood parasites (Plasmodium, Haemoproteus, Trypanosoma and microfilaria) that can cause a big impact on poultry production. However, some of them are known to cause a high impact on poultry production (Plasmodium and Leucocytozoon), while others still require further investigation. Raising backyard chickens is a common practice in Thailand, and the low-biosecurity system in which they are kept favors the transmission of vector-borne diseases, which include several blood parasites. The spread of such infections can compromise production, resulting in economic impact. This study aimed to report the molecular prevalence, lineage diversity and morphology of blood parasites infecting backyard chickens in three different provinces in Thailand. We found a high prevalence of Plasmodium sp. and Leucocytozoon sp. infections, while Trypanosoma and microfilaria had a lower prevalence. Plasmodium gallinaceum and Leucocytozoon macleani were present in the studied individuals as well as Trypanosoma, which resembles T. calmettei. The buffy-coat method and molecular analysis were shown to be valuable diagnostic tools for blood parasites in chickens. These results can be used to promote awareness of parasite infections in the study area. Avian malaria and leucocytozoonosis can cause fatal diseases, whereas avian trypanosomiasis is reported to be harmless in chickens. Backyard chickens can be infected by several pathogens, including blood parasites, that may shed to industrial poultry production, with a consequently higher economic impact. This study aimed to investigate the presence of several blood parasites (Plasmodium, Leucocytozoon and Trypanosoma) in backyard chickens raised in Southern Thailand, using PCR-based detection and microscopic methods. From June 2021 to June 2022, 57 backyard chickens were sampled. Fresh thin blood smears were prepared from 11 individuals, and buffy coat smears were prepared from 55 of them. Both thin blood smears and buffy coat smears were used for microscopic analysis. Two nested PCR protocols that amplify a fragment of cytochrome b (cytb) and small subunit rRNA (SSU rRNA) genes were used to identify Haemosporida and Trypanosoma parasites, respectively. The number of positive samples was higher with the application of nested PCR than when buffy coat smears were used. Three new Plasmodium lineages (GALLUS47-49) and thirteen Leucocytozoon lineages (GALLUS50-62) were found. Trophozoites, meronts and gametocytes of Plasmodium gallinaceum (GALLUS01) were present in one thin blood smear. All thin blood smears revealed Leucocytozoon infections, but only three samples were a single infection. These three samples revealed the presence of fusiform host cell–parasite complexes, of which the morphological features resembled those of Leucocytozoon macleani (possible synonym is Leucocytozoon sabrazesi), while the cytb showed that this parasite is closely related to the lineage GALLUS06-07, described as Leucocytozoon schouteni. The Trypanosoma prevalence was 33.33%; it was present in only one of the thin blood smears, and it resembles Trypanosoma calmettei. This study showed the prevalence of a high diversity of Plasmodium (64.91%) and Leucocytozoon (89.47%) in Thai chickens. Both nested-PCR and buffy coat smear can be used as the diagnostic tool for the testing of Plasmodium, Leucocytozoon and Trypanosoma for parasitic control in backyard chickens and poultry farms. The information on the parasite species that can be found in chickens raised in Southern Thailand was also considered as the baseline information for further study. [ABSTRACT FROM AUTHOR]

Animals are frequently coinfected with multiple parasites concurrently, and advances in our sampling of these complex intra-host parasite communities have revealed important ecological impacts on their hosts. However, the spatial distributions and environmental determinants of parasite coinfection remain infrequently studied. Here, we investigated the drivers of haemosporidian blood parasite coinfection in the Bicknell's thrush (Catharus bicknelli) and grey-cheeked thrush (Catharus minimus), parapatric sister species that occur across a broad latitudinal range in northern North America. Using 298 samples from across the distributions of these species, we found high overall infection (86%) and coinfection (41%) rates within host populations. Coinfection rates within populations were highly variable across sampling sites, ranging from 7 to 75%. Latitude was a more important predictor of coinfection frequency than host species identity, with coinfections becoming more abundant at higher latitudes. The 2 host species exhibited similar parasite faunas, and an analysis of the co-occurrence patterns among haemosporidians showed that host species identity was largely not a factor in structuring which parasites were found within coinfections. To our knowledge, this is the first study to illustrate a reverse latitudinal gradient in coinfection frequency in a eukaryotic parasite system. Further work is necessary to determine whether vector ecology or some other factor is the primary proximate driver of this pattern.

Abstract The Brown-capped Rosy-Finch is an endangered high alpine specialist that breeds on cliffs in the Rocky Mountains of North America. We know little about the parasites that infect these birds but filling this knowledge gap is important for understanding their population decline. The aim of this study was to survey haemosporidian parasite infections in Brown-capped Rosy-Finches. We sampled 104 Brown-capped Rosy-Finches during their breeding season at six sampling sites spanning the Colorado Rocky Mountains where they are nearly endemic. We used nested PCR methods to screen birds for parasite infections, and Sanger sequencing data were used to identify parasite lineages. Four of the sampling sites had birds with infections. Females were more often infected than males (prevalence = 15.6% and 9.7%, respectively). We observed twice as many infected birds sampled in July compared to birds sampled in June (prevalence = 12.5% and 6.25%, respectively). The percent of infected birds by sampling site ranged from 0 to over 20%. In total we identified eight different genetic lineages of haemosporidian parasites infecting Brown-capped Rosy-Finches—seven were in the genus Leucocytozoon, and one was in the genus Haemoproteus. Network analysis clearly separates Haemoproteus from Leucocytozoon, with Leucocytozoon lineages comprising two major clusters. Based on reports made to the MalAvi database, all lineages in Cluster 1 have the same reported insect vector, Simulium silvestre. We report that Brown-capped Rosy-Finches experience infections with haemosporidian parasites and that the percentage of infected birds differed markedly between sampling sites. We hypothesize that vector ecology and associated variation in climate at sampling sites explain our observations.

In the last decade, over 40% of bird species in Europe have experienced poor and bad conservation status, with more than 30% of bird species in mainland Portugal threatened with extinction. Along with anthropogenic factors, parasites and pathogens such as avian haemosporidians have been suggested to be responsible for these avian population declines. Wildlife rehabilitation centres play an essential role in species conservation and preservation. Moreover, animals admitted for rehabilitation can provide valuable information regarding transmission and pathogenicity of many diseases that affect wild birds that are rarely sampled in nature. However, reports of haemosporidians in captive birds are still limited. Here, we explored the prevalence and genetic diversity of avian haemosporidians in 89 birds from 29 species admitted to rehabilitation centres in Portugal, showing an overall infection prevalence of 30.3%. The prevalence of infection was higher in Strigiformes and in birds admitted to rehabilitation centres due to debilitating diseases. Remarkably, 30% of the infected bird species have not been found to harbour malaria parasites in preceding studies. We detected 15 different haemosporidian lineages infecting a third of bird species sampled. Notably, 2 out of these 15 detected haemosporidian lineages have not been obtained previously in other studies. Furthermore, we also identified nine new host–parasite interactions representing new host records for these haemosporidian parasites. Finally, our results revealed that birds infected with haemosporidians require longer rehabilitation treatments, which increase the economic costs for rehabilitation and may impair their survival prospects. These findings emphasise the importance of integrating haemosporidian infection considerations into rehabilitation protocols, highlighting the challenges posed by these infections in avian conservation and rehabilitation, including economic and logistical demands.

Haemosporidians are a group of vector-borne parasites belonging to the order Haemosporida. These parasites infect avian hosts and require blood-sucking insects (Diptera) for transmission. The occurrence and diversity of haemosporidian parasites are shaped primarily by the specificity of the parasite and the susceptibility of the host/vector. In this study, the presence and distribution of haemosporidians in blood samples from birds in urbanized and natural habitats were estimated using microscopic and molecular approaches. Birds in urbanized habitats were infected with four different species of Plasmodium, P. relictum, P. vaughani, P. matutinum, and P. circumflexum, and one species of Haemoproteus, H. parabelopolskyi, and Leucocytozoon sp. The species H. attenuatus, H. concavocentralis, H. minutus, H. pallidus, H. noctuae, and H. tartakovskyi were additionally identified in birds in natural habitats. Typically, juvenile birds are essential markers of parasite species transmitted in the study area. The juveniles in the urbanized habitats carried P. relictum, P. vaughani, P. circumflexum, H. parabelopolskyi, and Leucocytozoon species. The most abundant parasite was H. parabelopolskyi, which was found in both habitat types. The prevalence of Haemoproteus/Plasmodium species determined by nested PCR in birds in natural habitats (43.80%; 53/121) was significantly greater than that in birds in urbanized habitats (21.94%; 43/196) (p < 0.05). There was no significant difference in the infection rate of Leucocytozoon sp. between the habitat types (p > 0.05; 10/121 vs. 19/196).

Haemosporidians are a diverse group of vector-borne parasitic protozoa that have a global distribution and some of them live in the avian body as a benign parasite, with no symptoms. Avian Haemosporidians, include three main genera, Plasmodium, Haemoproteus, and Leucocytozoon. These blood parasites are transmitted across different regions through migratory birds. We studied the prevalence of avian haemosporidian parasites in both resident and migratory birds in the central and southern regions of Iraq. Our study represents pioneering research on Haemosporidian parasites in a wide range of avian species, including 248 samples from 47 species of both resident and migratory birds, collected from five cities in central and southern Iraq. We identified and determined the prevalence of three distinct haemosporidian genera, Haemoproteus spp., Leucocytozoon spp., and Plasmodium spp. Overall parasite infection was higher in resident hosts than in migrants. The prevalence of Plasmodium spp. was found to be highest among resident birds, with a rate of 20.1%, while the highest rate of Leucocytozoon spp. was observed in migratory birds, with a rate of 8.2%. Our study holds promising opportunities for understanding the impact of migratory and resident hosts on the dynamics of infectious diseases in wildlife. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Host–parasite interactions include effects on both proximate and ultimate levels: parasite infections affect individual's fitness and play a significant role in shaping the life history of host species. Global environmental changes as well as significant shifts in abiotic factors might impact the dynamics of parasite–host interactions, especially in Arctic regions, where the climate is changing at an alarming rate. With global warming, parasites and their vectors are predicted to spread to polar latitudes, and it is crucial to follow the changes occurring in the ecosystems in the era of global changes. We studied blood parasites (Haemosporidae) of passerine birds (Passeriformes: Aves) in southwest Yamal (north-western Siberia) using genetic and morphological methods. We found an overall parasite prevalence of 76.3%, with highest values for Leucocytozoon (72.0%) and lower values for Parahaemoproteus and Plasmodium (8.9 and 8.2%, respectively). We determined 26 genetic lineages in total, five of them were novel. The most common parasite lineages were TRPIP2 (18%), BT1 (14%), novel ACAFLA06 (13%), BT2 (7%), novel ACAFLA07 (6%), BT4 (5%) for Leucocytozoon; SISKIN1 (4%) for Parahaemoproteus; TURDUS1 (4%) for Plasmodium. For redpoll (Acanthis flammea), brambling (Fringilla montifringilla), bluethroat (Luscinia svecica) and little bunting (Emberiza pusilla) overall prevalence exceeded 90%. We also found significant differences in prevalence and lineage composition between sample sites, species and sexes, and a positive correlation between number of infections and host body mass. Our study provides knowledge about haemosporidian infections in the region, which had been barely studied for blood parasites. Gathered information is essential for the future monitoring and assessing potential shifts due to global change. [ABSTRACT FROM AUTHOR]

Blood parasites of the genus Leucocytozoon (Leucocytozoidae) only inhabit birds and represent a readily distinct evolutionary branch of the haemosporidians (Haemosporida, Apicomplexa). Some species cause pathology and even severe leucocytozoonosis in avian hosts, including poultry. The diversity of Leucocytozoon pathogens is remarkable, with over 1400 genetic lineages detected, most of which, however, have not been identified to the species level. At most, approximately 45 morphologically distinct species of Leucocytozoon have been described, but only a few have associated molecular data. This is unfortunate because basic information about named and morphologically recognized Leucocytozoon species is essential for a better understanding of phylogenetically closely related leucocytozoids that are known only by DNA sequence. Despite much research on haemosporidian parasites during the past 30 years, there has not been much progress in taxonomy, vectors, patterns of transmission, pathogenicity, and other aspects of the biology of these cosmopolitan bird pathogens. This study reviewed the available basic information on avian Leucocytozoon species, with particular attention to some obstacles that prevent progress to better understanding the biology of leucocytozoids. Major gaps in current Leucocytozoon species research are discussed, and possible approaches are suggested to resolve some issues that have limited practical parasitological studies of these pathogens. [ABSTRACT FROM AUTHOR]

Haemosporidian parasites are significant global drivers of avian disease and infections are affected by complex ecological and biological interactions. We quantified variation in haemosporidian infection prevalence of Plasmodium, Haemoproteus, and Leucocytozoon in 311 Song Sparrows (Melospiza melodia) from 2 breeding populations across years, seasons, age classes, and sexes, finding significant differences in infection prevalence across years, seasons, and sites, as well as sex differences in Leucocytozoon infection prevalence. We also found that birds were more likely to be infected with Plasmodium than Leucocytozoon and coinfections were more likely than expected. Many avian haemosporidian studies use data collected over a short period of time at one location, and few studies investigate multi-year infection dynamics across populations. However, such long-term studies are important because ecological factors can affect the distribution and prevalence of vector-borne pathogens like malaria. Because biological factors can also affect the likelihood of exposure to vector-borne pathogens, it is also important to determine how host traits influence infection dynamics. Our results demonstrate that the prevalence and composition of avian haemosporidian infections can be highly variable, making sampling regimes a key consideration when drawing conclusions from study results. To better understand the fitness consequences of haemosporidian infection in avian hosts, future work should take ecological and biological variation into account when evaluating the effects of infection on aspects of condition and quality, reproductive success, and survival. [ABSTRACT FROM AUTHOR]

The commonly occurring haemoparasites in pigeons include Plasmodium, Leucocytozoon and Haemoproteus which cause decrease in production and performance of the birds. Due to the dearth of knowledge on prevalence of these parasites in pigeons in Pakistan, current survey-based study was planned to assess the prevalence of above-mentioned parasites in pigeons in selected rural areas of District Faisalabad, Punjab, Pakistan. The morphological identification of parasites was done under microscope by comparing it with standard keys. Out of 384 samples, 56.51% were found positive for haemoparasites. According to town-wise distribution, Jinnah town, Iqbal town, Madina town and Lyallpur town prevalence was 61.24, 48.86, 55.56, and 59.26%, respectively. According to genus-wise distribution, 48.14, 48.21, and 64.92% prevalence of mixed infection, genera Plasmodium/Haemoproteus and Leucocytozoon were observed, respectively. The prevalence was found high in female birds (65.9%) as compared to male birds (51.18%). The prevalence was higher in Cheena (69.02%) and Golden breeds (76.57%) as compared to Sialkoti (32.89%) and Dub wali (37.17%). The prevalence was found higher in squabs (74.15%) as compared to adults (49.6%) and growers (52.56%), in scavengers feeding pattern (77.31%) as compared to feeders (35.78%), and in open coop type (76.68%) as compared to closed coop type (36.12%). The values of Hb, PCV, and RBC's count were lower, while WBC's counts were higher in infected pigeons. The values of uric acid, creatinine, albumin and bilirubin were lower while gamma-glutamyl transferase (GGT), alanine transaminase (ALT), aspartate transaminase (AST) and glucose were higher in infested birds as compared to non- infected birds. The study indicated the impact of haemoparasites on the hematological parameters of the birds and the data generated could help in formulating effective control strategies against haemoparasites particularly Plasmodium, Leucocytozoon and Haemoproteus. [ABSTRACT FROM AUTHOR]

Host–parasite distribution and interactions depend on geography, evolutionary associations, species composition, and their response to environmental variables. In this sense, host specificity as a key parasite's life history trait may determine parasite community composition but can be difficult to identify in nature. The haemosporidian–avian host communities of tropical mountain systems offer an opportunity to better understand how network interactions influence parasite–host specialization. By using frequency (i.e., prevalence) and interaction intensity (i.e., number of parasites per infected host) data, we calculated two quantitative indices to evaluate haemosporidian host specialization at both community (H2′) and haemosporidian lineages and bird species levels (d′). Additionally, we evaluated alpha (SI) and beta diversity (βWN) of haemosporidian–bird interactions along four distinct elevations (representing different vegetation types) in Central Veracruz, Mexico. We recorded 607 birds from 88 species in two sampling years, registering 78 haemosporidian cytochrome b lineages by PCR in 125 positive samples from 38 bird species. Haemosporidian–bird interactions showed high specialization and modular network structure for each vegetation type. The degree of specialization and modularity was stronger when considering interaction intensity (i.e., parasitemia). Species‐level specialization was higher for birds than for haemosporidians at the two vegetation types with highest interaction richness (i.e., pine–oak forest and tropical deciduous forest). There were high interaction beta diversity values (~1) among vegetation types, reflecting turnover in both avian and parasite communities. Our findings suggest that vegetation type (proxy for the local environmental conditions) constrains the distribution of hosts and parasites, and that intraspecies infection intensity plays an important role in the transmission dynamics and interactions of the haemosporidian community. [ABSTRACT FROM AUTHOR]

The Brown-capped Rosy-Finch is an endangered high alpine specialist that breeds on cliffs in the Rocky Mountains of North America. We know little about the parasites that infect these birds but filling this knowledge gap is important for understanding their population decline. The aim of this study was to survey haemosporidian parasite infections in Brown-capped Rosy-Finches. We sampled 104 Brown-capped Rosy-Finches during their breeding season at six sampling sites spanning the Colorado Rocky Mountains where they are nearly endemic. We used nested PCR methods to screen birds for parasite infections, and Sanger sequencing data were used to identify parasite lineages. Four of the sampling sites had birds with infections. Females were more often infected than males (prevalence = 15.6% and 9.7%, respectively). We observed twice as many infected birds sampled in July compared to birds sampled in June (prevalence = 12.5% and 6.25%, respectively). The percent of infected birds by sampling site ranged from 0 to over 20%. In total we identified eight different genetic lineages of haemosporidian parasites infecting Brown-capped Rosy-Finches—seven were in the genus Leucocytozoon, and one was in the genus Haemoproteus. Network analysis clearly separates Haemoproteus from Leucocytozoon, with Leucocytozoon lineages comprising two major clusters. Based on reports made to the MalAvi database, all lineages in Cluster 1 have the same reported insect vector, Simulium silvestre. We report that Brown-capped Rosy-Finches experience infections with haemosporidian parasites and that the percentage of infected birds differed markedly between sampling sites. We hypothesize that vector ecology and associated variation in climate at sampling sites explain our observations. [ABSTRACT FROM AUTHOR]

Abstract Host–parasite distribution and interactions depend on geography, evolutionary associations, species composition, and their response to environmental variables. In this sense, host specificity as a key parasite's life history trait may determine parasite community composition but can be difficult to identify in nature. The haemosporidian–avian host communities of tropical mountain systems offer an opportunity to better understand how network interactions influence parasite–host specialization. By using frequency (i.e., prevalence) and interaction intensity (i.e., number of parasites per infected host) data, we calculated two quantitative indices to evaluate haemosporidian host specialization at both community (H2′) and haemosporidian lineages and bird species levels (d′). Additionally, we evaluated alpha (SI) and beta diversity (βWN) of haemosporidian–bird interactions along four distinct elevations (representing different vegetation types) in Central Veracruz, Mexico. We recorded 607 birds from 88 species in two sampling years, registering 78 haemosporidian cytochrome b lineages by PCR in 125 positive samples from 38 bird species. Haemosporidian–bird interactions showed high specialization and modular network structure for each vegetation type. The degree of specialization and modularity was stronger when considering interaction intensity (i.e., parasitemia). Species‐level specialization was higher for birds than for haemosporidians at the two vegetation types with highest interaction richness (i.e., pine–oak forest and tropical deciduous forest). There were high interaction beta diversity values (~1) among vegetation types, reflecting turnover in both avian and parasite communities. Our findings suggest that vegetation type (proxy for the local environmental conditions) constrains the distribution of hosts and parasites, and that intraspecies infection intensity plays an important role in the transmission dynamics and interactions of the haemosporidian community.

The Neotropics are highly diverse in avian species. Neotropical countries contribute a large part of the estimated diversity of haemosporidian parasites reported for the planet's tropical zones. However, sampling is limited and biased, illustrated by only 30% of the genetic records (barcodes) from non-passerines, most of them not linked to a nominal species. This paper aimed to perform the molecular and morphological characterization of the haemosporidians that infect non-passerine birds from Colombia deposited in the biological collection named "Grupo de Estudio Relación Parásito Hospedero (GERPH)". We analyzed 1239 samples from twelve biomes and two animal care facilities. Phylogenetic relationships using barcodes and mitochondrial genomes were estimated. In addition, the reports of haemosporidian infections in non-passerine birds from the Neotropics recorded after 1978 were summarized. We reported the presence of thirteen morphological haemosporidian species, four potential new species deposited in GERPH, a host range expansion for two Plasmodium species, and a barcode sequence for Haemoproteus caprimulgi. We confirmed the species associated with 56 molecular lineages reported in other neotropical countries at the genus level. Thus, biological collections and curated databases such as MalAvi are essential to support integrative approaches demanded in modern taxonomy. [ABSTRACT FROM AUTHOR]

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Simple Summary: Parasitism is one of the most common life strategies on Earth, where the host and the parasite establish a successful relationship and continually adapt to each other. Most of the studies on wild birds show that those with hemoparasites are usually asymptomatic due to this host–parasite coevolution, so blood parasites are often detected as incidental laboratory findings. Most of these studies have been performed mainly in passerines and migratory species, but nocturnal raptors seem to be more exposed to blood parasite vectors than other avian species due to their behavior and distribution. Blood samples were collected from 134 individuals during a four-year period to assess the occurrence of blood parasites and parasitemia in different species of nocturnal raptors and their effect on hematological parameters. Thirty-five percent (95% CI: 27.5–43.5%) of individuals included in the study tested positive for at least one hemoparasite genus, and 11.2% showed coinfection. Leucocytozoon was the genus most frequently detected (32.1%), followed by Haemoproteus (11.2%), Trypanosoma and Plasmodium (2.2% each). The Eurasian eagle-owl (Bubo bubo) was the species with the highest prevalence (94.7%). Moderate anemia and an increase in leukocyte counts were detected in the positive birds. Moreover, the positive animals showed a poor body condition score. Most hemoparasites hosted by wild birds appear to be harmless, but most of the blood parasite studies in avian wildlife are mainly focused on passerines or migratory species. This study aimed to assess the occurrence of blood parasites in nocturnal raptors (Strigiformes order) and their effect on hematological parameters. A total of 134 blood samples were collected during a four-year period for hematological analysis and hemoparasite detection and quantification by microscopical examination of the samples. Overall, the occurrence of hemoparasites was 35.1%, with Leucocytozoon being the most frequently detected (32.1%), followed by Haemoproteus (11.2%), Trypanosoma and Plasmodium (2.2% each). Among the different bird species, the Eurasian eagle-owl (Bubo bubo) showed the highest blood parasite positivity (94.7%). In barn owls, the positive birds displayed a lower hematocrit measurement and body condition score than the non-parasitized ones (p = 0.007 and p = 0.005, respectively), especially those parasitized by Leucocytozoon. Moreover, the analysis of the magnitude of this association revealed that the presence of hemoparasites is five times more frequent in barn owls with a 2/5 body condition score. Despite the host–parasite coevolution in Strigiformes, our results show a correlation between the presence of hemoparasites and some health parameters, including blood parameters. [ABSTRACT FROM AUTHOR]

Simple Summary: A total of 122 chickens (66 chickens during the breeding season and 56 chickens during the non-breeding season) from tropical Hainan Island, China were sampled to test for haemosporidian infections. We showed that chickens have a higher incidence of haemosporidian infection and a greater diversity of haemosporidian parasite lineages during the breeding season relative to the non-breeding season. Reproduction is believed to contribute to the frequently observed seasonal cycles in parasite loads in many organisms, as an investment in reproduction by the host could result in a higher susceptibility to parasites. In this study, we examined the impact of breeding season on haemosporidian infection in free-range chickens (Gallus gallus domesticus). We sampled a total of 122 chickens (66 chickens during the breeding season of April 2017 and 56 chickens during the non-breeding season of January 2017) to test for haemosporidian infections. The result showed that 56 out of 66 chickens examined during the breeding season tested positive for parasites (84.8% parasite prevalence), whereas 39 out of 56 chickens tested positive for parasites during the non-breeding season (69.6% parasite prevalence). Moreover, among the 11 Leucocytozoon lineages and 2 Plasmodium lineages identified, the parasite lineages that infected chickens during the breeding season were more diversified than those that affected chickens during the non-breeding season. This study indicated that chickens have a higher incidence of haemosporidian infection and a greater diversity of haemosporidian parasite lineages during the breeding season relative to the non-breeding season. [ABSTRACT FROM AUTHOR]

Avian haemosporidian parasites have received considerable attention in ecology and evolution as a result of their wide distribution and ease of detection. However, conventional PCR-based detection methods may sometimes underestimate haemosporidian mixed infections, which are frequent in natural populations. This underestimation is due to differences in PCR sensitivity for detection of lineages within the mixed infections. Therefore, we designed new primers to amplify sequences that were not detected by the conventional primers and examined if our primers were useful for accurate detection of mixed infections. Blood samples were collected from 32 wild birds captured in Hokkaido, and 16 of these were positive for Leucocytozoon using the conventional primers, while 15 were positive using our primers. All positively amplified samples were sequenced, and we found that the conventional primers detected 16% (5/32) of multiple infections and none of them was a novel lineage, whereas our primers detected 44% (14/32) of multiple infections and ten of them were novel lineages. A phylogenetic analysis showed that the new primers can detect a wide range of Leucocytozoon lineages compared with that detected by the conventional primers. The results indicate that our primers are particularly suitable for revealing unique strains from multiple infections. Highly variable multiple infections in the same population of birds at the same location were found for the first time. We revealed a higher diversity of Leucocytozoon lineages in nature than expected, which would provide more information to better understand parasite diversity and host-vector interactions in wildlife. [ABSTRACT FROM AUTHOR]

Haemosporida, vector-transmitted blood parasites, can have various effects and may also exert selection pressures on their hosts. In this study we analyse the presence of Haemosporida in a previously unstudied migratory seabird species, the red-throated diver Gavia stellata. Red-throated divers were sampled during winter and spring in the eastern German Bight (North Sea). We used molecular methods and data from a related tracking study to reveal (i) if red-throated divers are infected with Haemosporida of the genera Leucocytozoon, Plasmodium and Haemoproteus, and (ii) how infection and prevalence are linked with the breeding regions of infected individuals. Divers in this study were assigned to western Palearctic breeding grounds, namely Greenland, Svalbard, Norway and Arctic Russia. We found a prevalence of Leucocytozoon of 11.0% in all birds sampled (n = 45), of 33.0% in birds breeding in Norway (n = 3) and of 8.3% in birds breeding in Arctic Russia (n = 25). For two birds that were infected no breeding regions could be assigned. We identified two previously unknown lineages, one each of Plasmodium and Leucocytozoon. Haemosporida have not been detected in birds from Greenland (n = 2) and Svalbard (n = 2). In summary, this study presents the first record of Haemosporida in red-throated divers and reports a new lineage of each, Plasmodium and Leucocytozoon GAVSTE01 and GAVSTE02, respectively. [ABSTRACT FROM AUTHOR]

Abstract Background Hepatozoonosis is a common tick-borne illness reported from all over the world. The infection has been well documented in dogs and cats, and has also been identified in wild canids and felids. India is home to many canid species; however, the incidence of Hepatozoonosis in wild canids is rarely reported. A wide variety of protocols have been discussed for the clinical management of the infection in companion animals; however, the suitability of treatment protocols in wild canids is understudied. The current case report highlights the clinical management of Hepatozoonosis in an Indian jackal and molecular investigation to provide vital insights into the epidemiology of the disease. Case Presentation A paraplegic Indian jackal was rescued from Melghat Tiger Reserve, Maharashtra, India. The animal had extensive decubital ulcers on the left pin bone and could not walk; however, the animal was active and dragged the hindlimb during locomotion. The vital parameters, blood and serum investigations were normal. Post physiotherapy, massage and infrared therapy, the animal could walk but started knuckling, resulting in injuries. Eight weeks into rehabilitation, the animal had a steep fall in haemoglobin concentration, platelet count, weight loss and was diagnosed with Hepatozoonosis. Considering the altered vital parameters, the jackal was rationally treated with Doxycyclin @ 20 mg/Kg O.D. (Once Daily) for 45 days along with supportive therapy. The jackal recovered after the treatment and led a normal life. Conclusion Mono-drug regime using Doxycycline was effective in the alleviation of H. canis infection in jackal. The drug was effective in alleviating the clinical presentation without alteration of vital parameters. The molecular investigation provided qualitative inputs in understanding the epidemiology of Hepatozoon in wild canids.

Avian malaria and leucocytozoonosis can cause fatal diseases, whereas avian trypanosomiasis is reported to be harmless in chickens. Backyard chickens can be infected by several pathogens, including blood parasites, that may shed to industrial poultry production, with a consequently higher economic impact. This study aimed to investigate the presence of several blood parasites (Plasmodium, Leucocytozoon and Trypanosoma) in backyard chickens raised in Southern Thailand, using PCR-based detection and microscopic methods. From June 2021 to June 2022, 57 backyard chickens were sampled. Fresh thin blood smears were prepared from 11 individuals, and buffy coat smears were prepared from 55 of them. Both thin blood smears and buffy coat smears were used for microscopic analysis. Two nested PCR protocols that amplify a fragment of cytochrome b (cytb) and small subunit rRNA (SSU rRNA) genes were used to identify Haemosporida and Trypanosoma parasites, respectively. The number of positive samples was higher with the application of nested PCR than when buffy coat smears were used. Three new Plasmodium lineages (GALLUS47-49) and thirteen Leucocytozoon lineages (GALLUS50-62) were found. Trophozoites, meronts and gametocytes of Plasmodium gallinaceum (GALLUS01) were present in one thin blood smear. All thin blood smears revealed Leucocytozoon infections, but only three samples were a single infection. These three samples revealed the presence of fusiform host cell–parasite complexes, of which the morphological features resembled those of Leucocytozoon macleani (possible synonym is Leucocytozoon sabrazesi), while the cytb showed that this parasite is closely related to the lineage GALLUS06-07, described as Leucocytozoon schouteni. The Trypanosoma prevalence was 33.33%; it was present in only one of the thin blood smears, and it resembles Trypanosoma calmettei. This study showed the prevalence of a high diversity of Plasmodium (64.91%) and Leucocytozoon (89.47%) in Thai chickens. Both nested-PCR and buffy coat smear can be used as the diagnostic tool for the testing of Plasmodium, Leucocytozoon and Trypanosoma for parasitic control in backyard chickens and poultry farms. The information on the parasite species that can be found in chickens raised in Southern Thailand was also considered as the baseline information for further study.