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

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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.

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Acknowledgements

This work was (partially) supported by the Faculty of Veterinary Medicine, Kasetsart University, and the Kasetsart University Research and Development Institute (grant number 32.60), Thailand.

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Authors and Affiliations

  1. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, 1 Malaiman Road, Kamphaeng Saen, Nakhon Pathom, 73140, Thailand

    Preeda Lertwatcharasarakul, Chaleow Salakij, Pimsuda Suwanasaeng & Raveewan Ploypan

  2. Prasu-Arthorn Animal Hospital, Faculty of Veterinary Science, Mahidol University, 999 Phutthamonthon 4 Road, Salaya, Nakhon Pathom, 73170, Thailand

    Panjaporn Prasopsom

  3. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, 50 Phahonyothin Road, Bangkhen, Bangkok, 10900, Thailand

    Chaiyan Kasorndorkbua

  4. Kasetsart University Raptor Rehabilitation Unit, Faculty of Veterinary Medicine, Kasetsart University, 1 Malaiman Road, Kamphaeng Saen, Nakhon Pathom, 73140, Thailand

    Chaiyan Kasorndorkbua, Pattarapong Jakthong & Malisa Santavakul

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  1. Preeda Lertwatcharasarakul
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Correspondence to Chaleow Salakij.

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Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

Approval was granted by the Institutional Laboratory Animal Care and Use Committee of Kasetsart University, Thailand under protocol number ACKU 01560 (Date 25 September, 2014) and ACKU59-VET-027 (Date 13 September, 2016). The national guideline for using raptors was approved every 5-year since 2012 to 2019 by the Department of National Parks, Wildlife and Plant Conservation, Ministry of Natural Resources and Environment, Thailand.

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11686_2021_403_MOESM1_ESM.jpg

Supplementary Fig. 1 General diagram for measurement of a the length (A-top) and the width (A-bottom) of macrogametocytes; b the gametocytes and host cell which are used for the description of species of L. toddi; width and length of nucleus of host cell (d and D); width and length of gametocyte (l and L); width and length of cytoplasmic process (r and R) (Valkiünas 2005). c Diagrams of roundish (1) and fusiform (2) morphs of the Leucocytozoidae to demonstrate the morphological parameters used and the method of measurement. (PMxD = maximum diameter of the parasite; PMiD = minimum diameter of the parasite; PNMxD = maximum diameter of the parasite nucleus; NMiD = minimum diameter of the parasite nucleus, HPCD = maximum diameter of host-parasite complex; HPCL = maximum length of host-parasite complex (fusiform morph); (Bennett et al. 1991) (JPG 692 KB)

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Lertwatcharasarakul, P., Salakij, C., Prasopsom, P. et al. Molecular and Morphological Analyses of Leucocytozoon Parasites (Haemosporida: Leucocytozoidae) in Raptors From Thailand. Acta Parasit. 66, 1406–1416 (2021). https://doi.org/10.1007/s11686-021-00403-6

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