J Vet Sci.  2018 Mar;19(2):280-289. 10.4142/jvs.2018.19.2.280.

Haemoproteus in barn and collared scops owls from Thailand

• Salakij C
• Pornpanom
• Lertwatcharasarakul P
• Kasorndorkbua C
• Salakij J

• KMID: 2407627
• DOI: http://doi.org/10.4142/jvs.2018.19.2.280

Abstract

The barn owl (BO) and the collared scops owl (CSO) are common nocturnal raptors throughout Thailand. Blood samples from 23 adult BOs and 14 CSOs were collected and processed for complete blood cell counts and parasite morphological examinations. Two Haemoproteus-positive samples were processed for ultrastructural observation. Polymerase chain reaction (PCR) analysis for a partial cytochrome b gene (cytb) from Haemoproteus was performed in all samples. Haemoproteus presence detected by light microscopy was lower than that detected by PCR (30.4% and 34.8%, respectively, in BO; and 50.0% and 78.6%, respectively, in CSO). Comparative hematology revealed that Haemoproteus-positive BOs had higher mean cell hemoglobin concentration, total leukocyte, absolute heterophil, basophil, and monocyte counts than Haemoproteus-negative BOs, but no significant differences between Haemoproteus-negative and -positive CSOs. Monocyte ultrastructure analysis revealed a role in the elimination of gametocytes. Morphologically, the Haemoproteus in 3 BOs and 6 CSOs were identified as H. noctuae, while that in 1 CSO was identified as H. syrnii. Phylogenetic analysis indicated the Haemoproteus spp. in 8 BOs and 7 CSOs were not closely related to H. noctuae or H. syrnii, and the cytb of 2 CSOs was that of H. syrnii. These results should be useful for study of Haemoproteus.

Keyword

Otus lettia; Tyto alba; avian malaria; cytochrome b; raptor

MeSH Terms

Adult
Animals
Basophils
Blood Cell Count
Cytochromes b
Erythrocyte Indices
Hematology
Humans
Leukocytes
Malaria, Avian
Microscopy
Monocytes
Parasites
Polymerase Chain Reaction
Raptors
Strigiformes*
Thailand*
Cytochromes b

Figure

• 

  Fig. 1 Light micrographs of Haemoproteus noctuae gametocytes from 2 barn owls (A: KU120 and B: KU132) and 2 collared scops owls (C and D: KU181 and E: KU222) and Haemoproteus syrnii gametocytes from KU127 collared scops owl (F). (A and C) Halteridial-shaped microgametocytes. (B) Banana-shaped microgametocyte. (D) Circumnuclear macrogametocyte. (E) Extra-erythrocyte round (arrow) macrogametocyte. (F) Banana-shaped macrogametocyte. Wright's stain. Scale bars = 10 µm (A–F).

• 

  Fig. 2 Transmission electron micrographs of Haemoproteus noctuae gametocytes in KU71 barn owl (A and B) and KU222 collared scops owl (C and D). (A) Circumnuclear gametocyte in red blood cell (RBC) showing parasitophorous vacuole membrane, homogeneous dense or osmiophilic bodies, and pigment granules (arrows). (B) Halteridial gametocyte in RBC showing nucleus and one pigment granule (arrow). Parasitophorous vacuole membrane of growing gametocytes did not touch the nuclei or cell membrane of infected RBC. (C) Free macro- (lower left) and micro- (upper right) gametocytes showing the nucleus and pigment granules (arrows) in microgametocyte. (D) Monocyte containing a phagocytized gametocyte with pigment granules (arrows). The monocyte contained many mitochondria near the phagocytized parasite. Uranyl acetate and lead citrate stains used for all specimens. Scale bars = 1 µm (A, C, and D), 0.5 µm (B). *Nucleus. †Monocyte. ‡Mitochondria.

• 

  Fig. 3 Phylogenetic tree based on partial cytochrome b gene (346 bp) sequences of Haemoproteus from 8 barn owls (●) and 11 collared scops owls (○) from Thailand. The sequences in bold were obtained in the present study. Numbers on the branches indicate the percent of replicates that reproduced the topology for each clade.

• 

  Fig. 4 Comparison of partial amino acid sequences of the cytochrome b (cytb) region inferred from previous reports and our nucleotide sequences obtained from Haemoproteus-infected owls. These amino acids were located on 1259–1373 residues of cytb protein compared with the complete genome of cytb gene of Haemoproteus columbae (NC 012448). Haemoproteus spp. in our study showed specific amino acids (isoleucine at the 1264 and 1281 positions, threonine at the 1282 position, alanine at the 1288 position [only in collared scops owl KU234], and methionine at the 1288 position).

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