Novel phylogenetic clade of avian <i>Haemoproteus</i> parasites (Haemosporida, Haemoproteidae) from Accipitridae raptors, with description of a new <i>Haemoproteus</i> species

Harl, Josef; Fauchois, Anaïs; Puech, Marie-Pierre; Gey, Delphine; Ariey, Frédéric; Izac, Brigitte; Weissenböck, Herbert; Chakarov, Nayden; Iezhova, Tatjana; Valkiūnas, Gediminas; Duval, Linda

doi:10.1051/parasite/2023066

Cited by 3 publications

(1 citation statement)

References 62 publications

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“…The identification is supported by the MalAvi database ( Bensch et al., 2009 ), which contains data of over 5,000 unique avian haemosporidian lineages. The widely used detection method, a nested PCR targeting the barcoding region ( Hellgren et al., 2004 ), is well-suited for simple screening due to its high sensitivity, although limitations have already been discovered in the detection of Haemosporida from Accipitridae ( Harl et al., 2022 , 2024 ). Additionally, there exist two further limitations: a short target fragment (479 bp), which limits its suitability for phylogenetic work, and insufficient detection of mixed infections with parasites of different genera ( Bernotiene et al., 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites

Musa,

Hemberle,

Bensch

et al. 2024

Front. Cell. Infect. Microbiol.

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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.

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Section: Introductionmentioning

confidence: 99%

Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites

Musa,

Hemberle,

Bensch

et al. 2024

Front. Cell. Infect. Microbiol.

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A morphological survey of avian Haemosporida in Colombian wild birds

Arroyave Pérez,

Cifuentes-Rincón,

Roldán Carvajal

et al. 2025

Rev. colomb. cienc. pecu.

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Introduction: Infection by protozoan haemoparasites causing malaria in birds is chronic and can lead to physical, reproductive, and behavioral alterations, resulting in a decline in the populations of affected wild birds. Objective: To identify parasitic agents causing avian malaria in a wild bird population in Jardin (Antioquia, Colombia) through blood smears and to explore bird-level factors associated with positivity. Methods: A descriptive cross-sectional study was conducted with non-probabilistic convenience sampling. Blood samples were obtained from wild birds captured with mist nets. Each bird was characterized according to its genus, species, sex, and age group. Three blood smears per bird were prepared and examined for parasitic structures from Haemoproteus, Leucocytozoon, and Plasmodium genera. Descriptive statistics and the association between study variables and the dichotomous outcome of blood smear analysis (positive or negative) using Fisher's exact test. Results: A total of 46 wild birds from 20 different species of the orders Passeriformes and quasi-Passeriformes were captured at six different locations in the study municipality (between 1,665 and 2,053 m.a.s.l.) in July 2022. Structures compatible with Haemosporida causing avian malaria were found, with a general infection frequency of 34.8% (16/46). Among the birds, 32.6% (15/46) were positive for Plasmodium spp., 6.5% (3/46) for Haemoproteus spp., and 4.3% (2/46) for Leucocytozoon spp. In addition, 6.5% (3/46) of the birds were coinfected with 2 or 3 of the agents of interest. An association was found between Plasmodium spp. infection and age group when the bird was defined as an adult (p=0.050). Conclusion: This study contributes to the knowledge of haemoparasites in wild birds in Colombia, reporting the presence of avian malaria in the study area. Further research is required on the molecular identification of protozoan haemoparasites, pathogenicity, the health status of these birds, and the attributable impact on their populations.

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Mitochondrial and apicoplast genome copy abundances of haemosporidian parasites are explained by host species and parasitic lineage

Brandler,

Chakarov

2024

Front. Ecol. Evol.

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Endosymbiotic organelles, such as mitochondria and plastids, contain own remnant genomes (nucleoids), whose variable abundance in cells may be adaptive to the physiological necessities and functions of the cells. Unicellular apicomplexan parasites contain one mitochondrium and one apicoplast with variable genome copy numbers. We measured the abundance of mitochondrial, apicoplast and nuclear genome copies in a set of avian blood samples infected with haemosporidian blood parasites, belonging to the three main genera Plasmodium, Haemoproteus and Leucocytozoon. We designed general primers suitable for qPCR, amplifying fragments of the mitogenome, plastome and nuclear genomes of avian malaria and related haemosporidian parasites. We measured the amplification of these fragments in 153 samples of 23 avian host species and infected with 33 parasitic lineages. We estimate on average several hundred mitochondrial genome copies and several tens of apicoplast copies per haploid gametocyte cell with substantial variation among samples. Host species appeared to differ in their mitogenome abundance while parasitic lineages differed in plastome abundance per cell (per nuclear copy signal). We did not find consistent differences between parasite genera or higher avian taxa. Parasite lineages and host bird species did not differ consistently in infection intensity, estimated from parasite to host nuclear signals, which may indicate that samples were taken at different stages of infection. However, this and similar results remain to be cross-validated with in-situ imaging techniques. The novel molecular tools introduced here offer avenues for the characterization of nucleoid abundance of haemosporidian parasites over environmental conditions and parasitic developmental stages. Such measures will improve our understanding of parasite physiology, ecology, the coadaptation and coevolution with hosts and suggest possible augmentations to standard methods in the research field.

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Novel phylogenetic clade of avian Haemoproteus parasites (Haemosporida, Haemoproteidae) from Accipitridae raptors, with description of a new Haemoproteus species

Cited by 3 publications

References 62 publications

Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites

Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites

A morphological survey of avian Haemosporida in Colombian wild birds

Mitochondrial and apicoplast genome copy abundances of haemosporidian parasites are explained by host species and parasitic lineage

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