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Figure 3 in Re-evaluation of the systematic position of the order Prostomatida (Protista: Ciliophora), with the establishment of two new genera and two new species

Figure 3. Platina marina gen. nov., sp. nov. from life (A–L), after protargol staining (M, O–R) and after 4ʹ,6-diamidino-2-phenylindole (DAPI) staining (N). A, ventral view of a representative individual, showing general appearance of body. B, the caudal cilium (arrow). C, H, I, lateral view, showing shape variants and transverse bulge located on dorsal side (arrowheads). D–G, ventral view, showing shape variants. J, K, compressed individuals, showing the silverline system of the ventral side (arrows) and dorsal side (arrowheads), and perioral cilia (double arrowheads). L, silverline system of the dorsal side. M, N, showing the macronucleus and micronucleus (arrow). O, ciliary pattern of the anterior portion of the cell, showing the circumoral kinety (arrowhead) and perioral kineties (arrow). P, the dividing line between the ventral side and the dorsal side, showing the only transverse row of kinetids on the dorsal side (arrowhead). Q, R, side view of the holotype specimen, showing ciliary pattern of the cell, circumoral kinety (arrowheads) and the only transverse row of kinetids on the dorsal side (arrow) except for perioral kineties. Scale bars: 25 μm in A–I; 10 μm in Q, R.

Figure 6 in Re-evaluation of the systematic position of the order Prostomatida (Protista: Ciliophora), with the establishment of two new genera and two new species

Figure 6. Maximum likelihood (ML) tree inferred from small subunit ribosomal RNA (SSU rRNA) gene sequences, showing the phylogenetic positions of the three newly sequenced species (bold font). Numbers near the nodes represent the ML bootstrap values and Bayesian inference (BI) posterior probabilities, respectively. Fully supported (100/1.00) branches are marked with filled circles. Asterisks (*) indicate disagreements between the ML and BI trees. The scale bar corresponds to five substitutions per 100 nucleotide positions.

Figure 2 in Re-evaluation of the systematic position of the order Prostomatida (Protista: Ciliophora), with the establishment of two new genera and two new species

Figure 2. Platina marina gen. nov., sp. nov. from life (A, F, G) and after protargol staining (B–E, H). A, ventral view of a representative individual, showing general appearance of body. B, C, dorsal (C) and ventral (B) side view of the holotype specimen, showing ciliary pattern of the cell, circumoral kinety (arrows) and the gap (arrowhead) in circumoral kinety. D, ciliary pattern of the anterior portion of the cell, showing the circumoral kinety (CK), perioral kineties (PK) and the only transverse row of kinetids on the dorsal side (arrowhead). E, showing the dividing line (arrow) between ventral side and dorsal side. F, lateral view (a) and ventral side view (b) of the same specimen, showing dorsoventrally flattened body and transverse bulge located on the dorsal side (arrowhead). G, silverline system of dorsal side. H, silverline system of ventral side. I, a schematic diagram of an apical view of the cell, showing the circumoral kinety (double arrowhead), perioral kineties (arrowheads) and the only transverse row of kinetids on the dorsal side (arrow). Scale bars: 15 μm.

Figure 1 in Re-evaluation of the systematic position of the order Prostomatida (Protista: Ciliophora), with the establishment of two new genera and two new species

Figure 1. Maps showing the location and photographs showing the appearance of the sampling sites. A, location of Qingdao and sampling sites. B, photograph of intertidal area near Zhanqiao Pier where Parametacystis pulchra gen. nov., sp. nov. and Platina marina gen. nov., sp. nov. were collected. C, photograph of a small pond in Taipingjiao Park where Apsiktrata gracilis was collected. S, salinity; T, temperature.

Figure 4 in Re-evaluation of the systematic position of the order Prostomatida (Protista: Ciliophora), with the establishment of two new genera and two new species

Figure 4. Drawing (A–E) and photomicrographs (F–L) of Parametacystis pulchra gen. nov., sp. nov. from life (A, D, F–H) and after protargol staining (B, C, E, I–L). A, lateral view of a representative individual, showing general appearance of body and protruding caudal vacuole (arrow). B, C, side view of the holotype specimen, showing ciliary pattern of the cell and circumoral kinety (arrowheads). D, showing the pattern of the silverline system and cortical granules. E, ciliary pattern of the anterior portion of the cell, showing the circumoral kinety (CK) and the perioral kineties (PK). F, showing the protruding caudal vacuole (arrow). G, lateral view of a slightly contracted individual, showing the furrows in ectoplasm (arrowheads). H, compressed individual, showing the pattern of the silverline system and cortical granules (arrows). I, lateral view of holotype specimen, showing ciliary pattern and macronucleus (Ma). J, showing the macronucleus and micronucleus (arrow). K, L, showing the circumoral kinety (arrowheads) and perioral kineties (arrows). Scale bars: 20 μm.

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FIGURE 3 in Hidden life cycle stages of the trypanosomatid Blastocrithidia triatomae Cerisola et al. 1971 (Protista: Excavata: Kinetoplastea) unveiled by a new cultivation method

FIGURE 3. Cladograms inferred from morphological (A) and 18S-rDNA (B) data to show the unambiguous separation of Blastocrithidia from Trypanosoma. Numbers associated to nodes are group contradiction frequency differences (GC) obtained from 1.000 replicates (values> 0 are considered as reliable). GenBank/NCBI accession codes are displayed right of strain names in B.

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FIGURE 1 in Hidden life cycle stages of the trypanosomatid Blastocrithidia triatomae Cerisola et al. 1971 (Protista: Excavata: Kinetoplastea) unveiled by a new cultivation method

FIGURE 1. Ontogeny of main stages found in B. triatomae in MBT medium. (A, B, C) Epimastigotes. (D, E, F) Epimastigotes originating scleromastigotes. G. Scleromastigotes dividing. (H, I) Scleromastigotes differentiation into epimastigotes. (J, K) Epimastigotes differentiation into sinemastigotes. (L, M) Sinemastigotes. Legends: f—flagellum, k—kinetoplast, n—nucleus. Bar = 10µm.

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FIGURE 2 in Hidden life cycle stages of the trypanosomatid Blastocrithidia triatomae Cerisola et al. 1971 (Protista: Excavata: Kinetoplastea) unveiled by a new cultivation method

FIGURE 2. Scanning electron microscopy of B. triatomae culture stages. A. epimastigote with curled flagellum (arrow). (B, C, D) Straphangers. Arrow indicates a sinemastigote (B, C); arrowhead shows a scleromastigote (B, D); asterisk shows scleromastigote bacillus-like stages (B, D). Note the disposition of these stages on the epimastigote flagellum, suggesting its ontogenesis (arrowhead). (E, F, G) Microepimastigotogenesis. Bacillus-like emitting an axial projection (E) and posterior flagellum (F). G. process in a more advanced stage. H. Binary division originating epimastigotes-like. I. Scleromastigotes in division. Note the cytoplasmic projections connecting the cells (arrowhead). Bars: (A, C, D, G, H) = 5µm; B = 2µm; (E, F, I) 1µm.

FIGURE 5. F in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 5. F. angusta angusta from live material (a–c, g) and after silver staining (d–f). Ventral-lateral view (a–c). Infraciliatures of the ventral (d), dorsal (e) and buccal (f) areas. Part of a pellicle, to show extrusomes (g). Abbreviations: AL: argentophilic line; C: Cilia; CV: contractile vacuole; D: diatom; E: extrusome; EP: excretory pore; Ma: macronucleus; Mi: micronucleus; OA: oral apparatus; P1–P3: peniculi 1, 2, and 3; PK: postoral kineties (arrowheads in f); PM: paroral membrane; PoS: postoral suture; PrS: preoral suture; Cyp: cytopyge; VK1–3: vestibular kineties 1, 2, and 3. Arrowheads mark postoral kineties. Scale bar = 70 µm.

FIGURE 4. F in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 4. F. acuminata from live material (a–c, g) and after silver staining (d–f). Ventral view of a representative specimen (a–e). Prominent characteristic of the anterior spot view of a representative specimen (a) and cytoplasm views of typical individuals (a–d). Ventral and dorsal views of the somatic ciliature and views of the oral ciliature; silver nitrate staining (e, f, i), silver carbonate staining (g, h, j). View of carrot-shaped macronucleus (k). View of an argyrome part (l). Abbreviations: AS: anterior spot; CV: contractile vacuole; D: diatom; E: extrusome; EP: excretory pores; FA: filamentous algae; Ma: macro nucleus; Mi: micronucleus; OA: oral apparatus; P: peniculi; VK: vestibular kineties. Scale bar = 20 µm.

FIGURE 9 in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 9. BI and ML analyses based on SSU rDNA gene sequences. The sequences of Frontonia populations in study are indicated in bold. Numbers at the nodes indicate the BI posterior probability and the ML bootstrap values (BI/ML). Nodes that were fully supported (100% ML, 1.00 BI) are represented by solid circles. A dash indicates a value less than 0.50 (BI) or 50% (ML). Bar, 8 substitutions per 100 nucleotide positions.

FIGURE 2. F in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 2. F. leucas from life (a–g) and after silver staining (h–k). Cytoplasm and contractile vacuole views in typical F. leucas (a–g). Ventral (h) and dorsal (i) views of the infraciliature and nuclear apparatus. Argyrome (j). Detailed structures of buccal region (k). Abbreviations: CC: collecting canal; CV: contractile vacuole; D: diatom; EP: excretory pore; FA: filamentous algae; FV: food vacuole; Ma: macronucleus; OA: oral apparatus; P1-P3: peniculi 1, 2, and 3; PM: paroral membrane; PoS: postoral suture; PrS: preoral suture; VK1-3: vestibular kineties 1, 2, and 3. Scale bar = 120 µm.

FIGURE 1. F in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 1. F. leucas from life (a–c, g) and after silver staining (d–f). Ventral-lateral view (a–c). Infraciliatures of the ventral (d), dorsal (e) and buccal (f) areas. Part of pellicle, to show extrusomes (g). Abbreviations: AL: argentophilic line; C: Cilia; CC: collecting canal; CV: contractile vacuole; D: diatom; E: extrusome; EP: excretory pore; FA: filamentous algae; FV: food vacuole; Ma: macronucleus; Mi: micronucleus; OA: oral apparatus; P1-P3: peniculi 1, 2, and 3; PK: postoral kineties (arrowheads in f); PM: paroral membrane; PoS: postoral suture; PrS: preoral suture; VK1-3: vestibular kineties 1, 2, and 3. Scale bar = 150 µm.

FIGURE 8. F in Morphology and molecular phylogeny of four Frontonia species from Turkey (Protista, Ciliophora)

FIGURE 8. F. anatolica from live material (a–f) and after silver staining (g–k). Ventral views of typical individuals (a–e). Cytoplasm with diatom and ingested algae (c–e). Part of a pellicle, to show extrusomes (f). Ventral, lateral, and dorsal views (g, h, i) of the somatic ciliature and oral ciliature views (j). View of an argyrome part (k). Abbreviations: CV: contractile vacuole; D: diatom; E: extrusome; EP: excretory pore; FV: food vacuole; Ma: macronucleus; Mi: micronucleus; OA: oral apparatus; P1–P3: peniculi 1, 2, and 3; PM: paroral membrane; PoS: postoral suture; PrS: preoral suture; VK1–3: vestibular kineties 1, 2, and 3. Scale bar = 40 µm.

FIGURE 5 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 5. Photomicrographs of Dysteria monostyla (A–E) and D. crassipes (F–I) from live (A, B, F, and G) and after protargol impregnation (C–E and H, I). A. Left lateral view of a representative individual, arrowheads show contractile vacuoles (CV). B. Cell surface, arrow refers to the transverse stripe, arrowhead shows bar-shaped ectosymbiotic bacteria. C. Left lateral view of ciliary pattern. D. Anterior part of cell. E. Ventral view of mid-body. F. Left lateral view of a representative individual, arrows point to contractile vacuoles. G. Rod-shaped ectosymbiotic bacteria attached on cell surface (arrowheads). H, I. Left lateral views of ciliature. Abbreviations: Co—circumoral kineties; EF—equatorial fragment; FvK—frontoventral kineties; Lfleft frontal kineties; LK—left kineties; Ma—macronucleus; Pr—preoral kinety; RK—right kineties; TF—terminal fragment. Scale bars: 30 μm (A, C, F and H).

FIGURE 4 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 4. Photomicrographs of Chlamydodon bourlandi from life (A, D, and E in bright field, B, C, F, and G by differential interference contrast), after fluorescent staining (H), and after protargol staining (I–L). A, G. Ventral views of different individuals. B. Nematodesmal rods from a compressed cell, arrowheads indicate C-shaped clasps. C. Ventral view of part of a compressed cell showing macronucleus, arrow denotes the cross-striated band. D. Cortical granules on dorsal side, arrowhead marks the pigment spot. E. Cortical granules (arrowhead) on ventral side. F. Ventral view of cell, arrowheads indicate the pigment spot. H. Macronucleus. I. Ventral view of ciliature and macronucleus. J, K. Ventral view of the anterior part of ciliature showing oral structure. L. Dorsal view of the anterior body, arrowheads indicate the terminal fragments. Abbreviations: Cocircumoral kineties; Cs—cytostome; Ma—macronucleus; NR—nematodesmal rods; Pr—preoral kinety. Scale bars: 100 μm.

FIGURE 3 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 3. Photomicrographs of Chlamydodon pararoseus sp. n. from life (A, D, and E by differential interference contrast, B, C, and F in bright field), after fluorescent staining (G), and after protargol staining (H–L). A, B. Ventral views of slightly compressed individuals, arrow in (A) marks the cytostome, and arrowheads indicate contractile vacuoles, and arrows in (B) refer to the sub-equatorial fossa. C, E. Dorsal views, arrow marks the cytostome, and arrowheads mark the pigment spot. D, F. Ventral views of individuals showing variations in body shape. G. Macronucleus. H. Ventral ciliature. I. Oral structure. J. Barren kinetosomes in oral area. K. Terminal fragment. L. Sub-equatorial fossa after protargol staining. Abbreviations: Co—circumoral kineties; Ks—barren kinetosomes; Ma—macronucleus; Pr—preoral kinety; TF—terminal fragment. Scale bars: 40 μm.

FIGURE 1 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 1. Locations of the sample sites. A. Overview of locations. B. A mangrove wetland in Zhanjiang. C. A small puddle of the Dongpo Lake, Haikou. D, E. Intertidal zone in Qingdao, arrow indicates the sampling pool. E. The same tidal pool as shown by arrow in D.

FIGURE 6 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 6. Phylogenetic tree of Maximum-likelihood (ML) based on small SSU rRNA gene sequences with Bayesian inference (BI) tapped on it. The four new sequences in the present paper are highlighted in bold. Numbers at nodes show ML bootstrap values and the BI posterior probabilities (ML/BI). Values with full support (ML/BI, 100/1.00) are marked with solid circle. The scale bar corresponds to 5 substitutions per 100 nucleotide positions. * Guangdong population of Chlamydodon bourlandi, ** misidentification, should be Chlamydodon bourlandi.

FIGURE 2 in Morphology and SSU rDNA sequences of four cyrtophorian ciliates from China, with description of a new species (Protista, Ciliophora, Phyllopharyngea)

FIGURE 2. Drawings of Chlamydodon pararoseus sp. n. from life (A–D) and after protargol staining (E–G). A. Ventral side of a representative individual. Dorsal (B) and ventral (C) views of a compressed individual, arrows denote the cross-striated band (CSB) and the arrowheads mark pigment spots. D. Cyrtos. E, F. Ventral (E) and dorsal (F) views of the same holotype specimen, arrow indicates the sub-equatorial fossa. G. Ventral view of anterior cell, arrow marks the suture, arrowheads denote barren kinetosomes. Abbreviations: Co—circumoral kineties; CSB—cross-striated band; LK—left kineties; Ma—macronucleus; NR—nematodesmal rods; PoK—postoral kineties; Pr—preoral kinety; RK—right kineties; TF—terminal fragments. Scale bars: 40 μm.