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Fig. 4 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 4. Comparison of fully grown gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of Curruca communis (A, B), Haemoproteus sittae (unknown lineage) from the blood of Sittae europaea (C, D) and Haemoproteus dolniki (unknown lineage) from the blood of Fringilla coelebs (E, F). Note: the elongate and markedly attenuated nuclei of infected erythrocytes containing advanced gametocytes of the new species (A, B), which is not the case in other Haemoproteus parasites (C–F). All images are from type specimens of these species. Symbols are the same as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.

Fig. 7 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 7. High gametocytaemia of Haemoproteus palloris (lineage hWW1) from the blood of a willow warbler Phylloscopus trochilus. Note that young gametocytes are absent, and the parasitaemia consists exclusively of mature fully grown macro- and microgametocytes, indicating a synchronous parasite development and probable absence of recent maturation of tissue meronts, which are the only source of merozoites for young gametocyte development during Haemoproteus infections. Triangle arrowhead – macrogametocytes; triangle wide arrowheads – microgametocytes. Long arrows – parasite nuclei. Giemsa-stained preparations. Scale bar = 10 µm.

Fig. 2 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 2. Gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of its type host, the common whitethroat Curruca communis: A-H – macrogametocytes, I-L – microgametocytes. Note: the markedly attenuated width of infected erythrocytes (D–G) containing advanced gametocytes, compared to uninfected erythrocytes, and the presence of predominantly oval or elongate pigment granules in fully grown gametocytes (F–H, J-L). All images are from the hapantotype. Long arrows – parasite nuclei. Short arrows – vacuoles. Arrowheads – pigment granules. Short simple wide arrow – nucleolus. Long triangle wide arrows – unfilled spaces between erythrocyte nuclei and gametocytes. Giemsa-stained thin blood films. Scale bar = 10 µm.

Fig. 6 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 6. Bayesian Inference tree (A) based on partial (478 bp) cytb sequences of Haemoproteus angustus n. sp. (lineage hCWT7) and the 20 closest related Haemoproteus lineages. Bayesian posterior probabilities and Maximum Likelihood bootstrap values were indicated above and below nodes, respectively. For each lineage, representative GenBank accession numbers and MalAvi lineage codes (if available) are indicated as well as the most common bird host. The scale bar indicates the expected mean number of substitutions per site according to the model of sequence evolution applied. Images B and C show the Median-Joining DNA haplotype network of partial (478 bp) cytb sequences of H. angustus hCWT7 and the 20 closest related Haemoproteus lineages. The upper image (B) shows the host distribution, and the lower image (C) depicts the geographic distribution according to the United Nations geoscheme. Each circle represents a unique haplotype/lineage. The frequency of each lineage is indicated for all haplotypes with more than one record and roughly corresponds to the size of circles. Bars on branches indicate the number of substitutions between two haplotypes. Small white circles represent median vectors, which are hypothetical (often ancestral or unsampled) sequences required to connect existing haplotypes with maximum parsimony.

Fig. 3 in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 3. Comparison of fully grown gametocytes of Haemoproteus angustus n. sp. (lineage hCWT7) from the blood of Curruca communis (A–D), H. belopolskyi (lineage hHIICT1) from the blood of Hippolais icterina (E–H) as well as H. parabelopolskyi (hSYAT2) (I–L), H. pallidulus (hSYAT3) (M–P), H. homogeneae (hSYAT16) (Q–T) and H. majoris (hWW2) (U–X) from the blood of Sylvia atricapilla. Note: the markedly attenuated gametocytes of the new species (A–D), which are not present in other Haemoproteus parasites (E–X); the fully grown gametocytes of H. belopolskyi (F, H) and H. parabelopolskyi (J, L) are bigger than those of H. angustus n. sp.; the pigment granules are predominantly roundish and small in H. pallidulus (M–P); the fully grown gametocytes are small and do not reach poles of infected erythrocytes in H. homogeneae (R, T); the erythrocyte nuclei are displaced laterally by H. majoris (V, X) – all these feature are not characteristic of H. angustus sp. nov. Symbols are the same as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.

Fig. 1. Bayesian phylogenetic tree constructed using partial cytochrome b in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 1. Bayesian phylogenetic tree constructed using partial cytochrome b sequences of 61 lineages of Haemoproteus, 4 lineages of Plasmodium, and Leucocytozoon sp. lSISKIN2 as outgroup. Posterior probabilities higher than 0.8 are indicated close to the respective nodes. Red font indicates the parasite lineage described in this publication. Vertical bars (A–D) show groups of closely related lineages, which complete development and produce gametocytes only in non-passerines (A, D), both non-passerines and passerines (B), and only passerines (C). Blue font indicates Haemoproteus species, which develop in non-passerine avian hosts, which are indicated by symbols (● – Psittaciformes; ∎ - Coraciiformes; ▴ - Strigiformes; ◆ - Anseriformes; ★ - Charadriiformes; ♥ - Pelecaniformes; ⋄ - Piciformes; ⊠ - Sphenisciformes; Ω - Musophagiformes; § - Trochiliformes; Ψ – Falconiformes; Σ – Columbiformes; Φ - Galliformes). Lineage names were provided (according to MalAvi database), followed by parasite species names and sequence GenBank accession numbers.

Fig. 5. Haemoproteus angustus n in Unexpected absence of exo-erythrocytic merogony during high gametocytaemia in two species of Haemoproteus (Haemosporida: Haemoproteidae), including description of Haemoproteus angustus n. sp. (lineage hCWT7) and a report of previously unknown residual bodies during in vitro gametogenesis

Fig. 5. Haemoproteus angustus n. sp. (lineage hCWT7) gametogenesis (A–H) and ookinete development (I–L) in vitro: A, B – initial stages of rounding up of gametocytes after the exposure of infected blood to air; C, E – rounded up macrogametocyte (C) and microgametocyte (E); F – the exflagellation; G – microgamete; H – fertilization; I – initial stage of ookinete development; J – medium differentiated ookinete; K – nearly mature ookinete with a residual body; L – mature ookinete without residual body. Note: the presence of several small residual bodies in erythrocytes containing gametocytes, which were preparing to escape from infected erythrocytes (A-C, E) – a unique character of this species during the initial stage of gametogenesis. Short triangle wide arrows – residual bodies; triangle arrowhead – macrogamete; triangle wide arrowheads – microgametes; short barbed arrows – finger-like outgrowth; long barbed arrows – apical end of developing ookinete; simple wide arrowhead – residual body of ookinete. Other symbols as in Fig. 2. Giemsa-stained thin blood films. Scale bar = 10 µm.

Fig. 2. Macroscopic examination. A in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study

Fig. 2. Macroscopic examination. A pale enlarged liver (Li); B hyperplastic nodules on the surface of the large spleen (S), inflated kidneys (K), and hypostatic congestion of left lung (Lu).

Fig. 5 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study

Fig. 5. Maximum-likelihood tree analysis based on partial cytb gene sequences (433 bp) of Haemoproteus from the studied sequence (linage BUTBUT15) and other sequences obtained from MalAvi database. Numbers on the branches indicate the percent of replicates that reproduced the topology for each clade. The percentages of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. Black circle indicates sequence obtained from the study.

Fig. 1 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study

Fig. 1. Gametocytes of haemosporidians (arrows) from the blood of the common buzzards. A-C mature macrogametocytes of L. buteonis in fusiform host cells; A host cell nucleus is not distorted but displaced laterally; B&C host cell nucleus is lateral and flattened; D young gametocyte of Haemoproteus. Giemsa stained thin blood films. Scale bar = 10 μm.

Fig. 4 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study

Fig. 4. Megalomeront of Haemoproteus sp. (linage BUTBUT15) from the kidney of a common buzzard. A&B same megalomeront of different magnifications (arrows). The structure is covered with a semi-thick capsule-like wall and contained highly eosinophilic irregularly-shaped cytomeres which have merozoites. Note that the host cell nucleus is not visible inside or close to megalomeront. Scale bars = 200 μm (A); 80 μm (B).

Fig. 3 in Exo-erythrocytic stages of Haemoproteus sp. in common buzzard (Buteo buteo): A histopathological and molecular study

Fig. 3. Meronts of Haemoproteus sp. (linage BUTBUT15) from the lung of a common buzzard. A-D different shapes of pulmonary meronts (arrows). Each meront is surrounded by a thin wall and contained numerous basophilic round to oval merozoites. Note that the host cell nucleus is not visible in meronts. Scale bar = 40 μm.