Massive Infection of Lungs with Exo-Erythrocytic Meronts in European Robin Erithacus rubecula during Natural Haemoproteus attenuatus Haemoproteosis

SIMPLE SUMMARY: Haemoproteus parasites are cosmopolitan bird pathogens belonging to the order Haemosporida (Apicomplexa). A majority of the described species are transmitted by Culicoides biting midges, which inject infective stages (sporozoites) in birds during blood meals. The sporozoites initiate tissue merogony, resulting in numerous merozoites, part of which penetrate red blood cells and produce blood stages (gametocytes), which are infective for vectors. The blood stages of Haemoproteus parasites have been relatively well-investigated, although tissue stages and patterns of their development remain unidentified in the majority of Haemoproteus species. Nevertheless, they often damage various organs which makes them important for bird health. This study contributes new knowledge about tissue merogony of Haemoproteus attenuatus, which parasitize birds of the Muscicapidae. Naturally infected European robins Erithacus rubecula were caught in Lithuania during autumnal migration. Parasites were identified using morphological features of gametocytes and DNA sequence analysis. Organs of infected birds were examined using histological methods. Tissue stages (meronts) were present only in the lungs, where they were numerous and markedly varied in shape, size and maturation stage. Description of meronts was provided and molecular phylogenetic analysis identified closely related lineages that could present similar exo-erythrocytic development in lungs. Lung damage caused by meronts of H. attenuatus and closely related lineages is worth attention due to their possible implications on a bird’s health. ABSTRACT: Haemoproteus species are widespread avian blood parasites belonging to Haemoproteidae (Haemosporida). Blood stages of these pathogens have been relatively well-investigated, though exo-erythrocytic (tissue) stages remain unidentified for the majority of species. However, recent histopathological studies show that haemoproteins markedly affect bird organs during tissue merogony. This study investigated the exo-erythrocytic development of Haemoproteus (Parahaemoproteus) attenuatus (lineage hROBIN1), the common parasite of flycatchers (Muscicapidae). Naturally infected European robins Erithacus rubecula were examined. Parasite species and lineage were identified using microscopic examination of blood stages and DNA sequence analysis. Parasitaemia intensity varied between 0.8 and 26.5% in seven host individuals. Organs of infected birds were collected and processed for histological examination. Tissues stages (meronts) were seen in six birds and were present only in the lungs. The parasites were usually located in groups and were at different stages of maturation, indicating asynchronous exo-erythrocytic development. In most parasitized individuals, 100 meronts were observed in 1 cm(2) section of lungs. The largest meronts reached 108 µm in length. Mature meronts contained numerous roundish merozoites of approximately 0.8 µm in diameter. Megalomeronts were not observed. Massive merogony and resulting damage of lungs is a characteristic feature during H. attenuatus infections and might occur in related parasite lineages, causing haemoproteosis.