Molecular and Morphological Analyses of Leucocytozoon Parasites (Haemosporida: Leucocytozoidae) in Raptors From Thailand

Abstract

Introduction: Leucocytozoon spp. causes a vector-borne disease that is nonpathogenic in domestic and wild birds. To date, there was no report of leucocytozoonosis in raptors from Thailand. Methods: This study was carried out to perform morphological and molecular analyses of Leucocytozoon in 400 raptors at a rehabilitation center at Kasetsart University, Thailand during a 7-year period. The nested PCR was used to amplify the cytochrome b gene of Leucocytozoon with primers HaemNF1 and HaemNR3 as the primary reaction. Results: The light microscopic examination revealed Leucocytozoon gametocytes in five raptors; three diurnal raptors [two Crested Goshawks (CGs, Accipiter trivirgatus) and one Eastern Imperial Eagle (EIE, Aquila heliaca)], and two nocturnal raptors (one Oriental Scops-Owl (OSO, Otus sunia,) and one Short-eared Owl, Asio flammeus) and two species were identified: Leucocytozoon danilewskyi in both owl species and L. californicus in two CGs. The PCR method revealed more infection rate (2.0%, 8/400) than the light microscopic method including one Barred Eagle-Owl (BEO, Bubo sumatranus), one Brown Hawk Owl (BHO, Ninox scutulata) and one OSO. A phylogeny revealed that sequences from one SEO and one OSO were clustered with L. danilewskyi and the three Leucocytozoon sequences from diurnal raptors were clustered with L. californicus. The other three sequences from a BHO, a BEO and an OSO were ambiguous. Conclusion: This study combined morphological, morphometric and molecular phylogenetic analyses to identify L. danilewskyi in two species of owls, L. californicus in three diurnal raptors, and unknown species in three other owls, representing the first records of leucocytozoon infection in raptors from Thailand.