Haemoproteus velans identi

Groff, Tierra C., Lorenz, Teresa J., Crespo, Rocio, Iezhova, Tatjana, Valkiūnas, Gediminas & Sehgal, Ravinder N. M., 2019, Haemoproteosis lethality in a woodpecker, with molecular and morphological characterization of Haemoproteus velans (Haemosporida, Haemoproteidae), International Journal for Parasitology: Parasites and Wildlife 10, pp. 93-100 : 96-97

3.2. Northern Flicker Haemoproteus velans identi fi cation

Sampling from 139 live birds in 2016 and 2017 revealed three adult Northern Flickers with Haemoproteus infections. All three samples had an identical genetic sequences and infections were confirmed with microscopy. The genetic sequence was two base pairs different or 0.417% divergent from the sequence of the White-headed woodpecker. When morphologically comparing gametocytes in blood smears from individuals from this study and from the Downy woodpecker (EU24552), all samples were identified as H. velans . There were no blood smears available from the White-headed woodpecker. Given the genetic similarity Haemoproteus parasites in the Northern Flicker, White-headed woodpecker, and Downy woodpecker samples and also the morphological similarity of gametocytes from the Northern Flicker and Downy woodpecker, it is probable that all records of haemoproteids in these birds were H. velans .

Only one morphotype was present in all three of the positive Northern Flicker slides examined ( Fig. 3 ). The original description of H. velans is fragmentary, and morphometric data are incomplete ( Coatney and Roudabush, 1937; Greiner et al., 1977; Valki ū nas, 2005). Here, we provide additional information on the morphology of this parasite ( Fig. 3 ) and morphometrics of its fully-grown gametocytes and their host cells ( Table 2) from the type vertebrate host, the Northern Flicker. It should be noted that blood stages were identical in the main morphological features to those described by Coatney and Roudabush (1937) and Valki ū nas (2005), their description is not repeated here. However, several additional features in gametocyte morphology were reported. First, there were more amoeboid gametocyte cells in some individual avian hosts than in previous descriptions (see Fig. 3g, k ). Second, volutin was readily visible, but present markedly unequally in individual birds. Third, the number of pigment granules is greater in our material than in the original description. Due to marked volutinization of parasite cytoplasm, it is often difficult to calculate the number of pigment granules in gametocytes of this parasite (Valki ū nas, 2005). Gametocytes in the type material of H. velans are overfilled with prominent volutin, and pigment granules are poorly visible due to marked fading of hemozoin in these old preparations ( Dimitrov et al., 2014). We calculated pigment granules in mature gametocytes with a relatively small amount of volutin, and these data show that mature gametocytes contain on average approximately 35 and 28 pigment granules in macro-and microgametocytes, respectively ( Table 2). This is greater than was reported in the original description, in which 18 and 21 pigment granules were reported on average in macro- and microgametocytes, respectively ( Coatney and Roudabush, 1937; Valki ū nas, 2005).

Maximum-likelihood predictions placed H. velans into a clade with Haemoproteus species parasitizing non-passeriform birds belonging to Strigiformes, Ciconiiformes, Columbiformes, and European members of Piciformes with 96% bootstrap support ( Fig. 4 ). However, the relationships between these species did not have significant bootstrap support. The NOFL1 (Northern Flicker) and WHWO1 (White-headed woodpecker) lineages of H. velans formed a clade with 100% bootstrap support with the likely misidentified sequence from the Downy woodpecker (EU254552, H. picae ) and the unidentified Haemoproteus sequence from the Red-headed woodpecker (AF465590).

Figures

Fig. 3. Haemoproteus velans (lineage NOFL1) from the blood of the Northern Flicker (Colaptes auratus): a, b - young gametocytes; c-h - macrogametocytes; i-l - microgametocytes. Long simple arrows – nuclei of parasites. Simple arrowhead – pigment granules. Triangle arrowheads – volutin granules. Short simple wide arrow – unfilled space in poles of erythrocytes. Giemsa-stained thin blood films. Scale bar = 10 μm.

Fig. 4. Consensus tree displaying Haemoproteus velans phylogenetic relationships as predicted by Maximum-likelihood inference, using GTR + I+Γ substitution model in PAUP* v.4.0a.b161. Maximum-likelihood bootstrap values> 70 are indicated. Lineages detected in the current study are indicated by red boxes. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)