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Fig. 8 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 8 Phylogenetic tree based on the 18S, 5.8S, and 28S rRNA genes, showing the systematic positions of Metaradiophrya speculorum sp. n. and Maupasella mucronata. The subclass Hymenostomatia was used to a posteriori root the tree. Bootstrap values for maximum likelihood conducted in IQTrees as well as posterior probabilities for Bayesian inferences conducted in Phycas and MrBayes were mapped onto the

Fig. 7 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 7 Phylogenetic tree based on the 18S rRNA gene and the ITS region, showing the systematic positions of Metaradiophrya speculorum sp. n. and Maupasella mucronata within the subclass Astomatia. The tree was a posteriori rooted according to Fig. 6. Bootstrap values for maximum likelihood conducted in IQTrees as well as posterior probabilities for Bayesian inferences conducted in Phycas and MrBayes were mapped

Fig. 5 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 5 Phylogenetic tree based on the 18S rRNA gene, showing the systematic positions of Metaradiophrya speculorum sp. n. and Maupasella mucronata within the class Oligohymenophorea. The subclass Peniculia was used to a posteriori root the tree. Bootstrap values for maximum likelihood conducted in IQTrees as well as posterior probabilities for Bayesian inferences conducted in Phycas and

Fig. 2 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 2 Metaradiophrya speculorum sp. n., holotype specimen in vivo. a Overview, showing the general body organization. Arrow marks the arched skeletal ridge; arrowheads denote the contractile vacuoles. b, d–f Details showing the ciliary pattern, the skeletal system, the nuclear apparatus, and the multiple contractile vacuoles (arrowheads). The skeletal system consists of a fibrillar hook and numerous fibers, which

Fig. 1 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 1 Metaradiophrya speculorum sp. n., holotype specimen in vivo. a, b Semi-schematic diagram of the ventral and the dorsal side, showing the ciliary pattern, the nuclear apparatus, as well as the skeletal system. c Ventral view, showing the general body organization. Arrow marks the arched skeletal ridge; arrowheads denote the contractile vacuoles. d Detail of the skeletal system, which consists of a fibrillar hook and

Fig. 6 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 6 Phylogenetic tree based on the 18S rRNA gene, showing the systematic positions of Metaradiophrya speculorum sp. n. and Maupasella mucronata isolated from endogeic lumbricid earthworms. The phylogenetic tree suggests that the evolution of endosymbiotic astome ciliates has proceeded through specialization to ecological groups of their host earthworms. Haptophrya planariarum and Dexiotricha spp. were used to a posteriori root the tree. Bootstrap

Fig. 4 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 4 Maupasella mucronata, Slovak specimens in vivo. a, b, g Overviews, showing the ciliary pattern, the skeletal system, and the nuclear apparatus. c Detail of the thorn and its supporting fibers. d The supporting fibers of the attachment apparatus of the opisthe are formed at the anterior end of the broken somatic ciliary rows. e Somatic kineties are narrowly arranged and composed of very densely spaced basal bodies. f

Fig. 3 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 3 Maupasella mucronata, Slovak specimens in vivo. a, d Semischematic diagrams of the ventral side, showing the ciliary pattern and the nuclear apparatus of representative specimens. b, c Ventral view, showing the ciliary pattern and the nuclear apparatus of a mid-divider. e, f Semi-schematic diagrams, showing the general body organization. Arrows mark the thorn, which consists of two skeletal fibers arranged in

Fig. 9 in Multi-gene phylogeny of the subclass Astomatia (Protista: Ciliophora) refreshed with two rare astome ciliates from the digestive tube of endogeic earthworms

Fig. 9 Putative secondary structure of ITS2 molecules of Metaradiophrya speculorum sp. n. and Maupasella mucronata as well as comparison of stems of helix III between Metaradiophrya speculorum and Metaradiophrya lumbrici

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Figure 1 in Morphology and molecular phylogeny of a Chinese population of Rubrioxytricha guamensis Kumar et al., 2018 (Ciliophora: Hypotrichia)

Figure 1. (A–R) Morphology of Chinese population of Rubrioxytricha guamensis from life (A, D–L) and after protargol staining (B, C, M–R). (A) Ventral view of a representative individual. (B, C) Ventral (B) and dorsal (C) view of the same specimen showing the ciliary pattern and nuclear apparatus, arrowhead shows buccal cirrus. (D) Distribution of cortical granules (arrowheads). (E) Shows flexibility of the body; arrowheads represent the spherical yellow cortical granules at the body margin. (F) Ventral view of anterior cell part. (G) Posterior cell part, arrow shows the slightly enlarged transverse cirri. (H) Various cytoplasmic inclusions, ie crystals, lipid droplets. (I) Mid-portion of a slightly squeezed cell, arrows show two macronuclear nodules. (J) Arrow marks the contractile vacuole. (K, L) Ventral views of different cells, showing body shape. (M, N) Ventral (M) and dorsal (N) view of the same specimen, showing ciliature and nuclear apparatus. (O) Ventral view of the anterior end of body. (P) Ventral view of the mid-portion of body, showing left and right marginal rows and postoral ventral cirri. (Q) Ventral view of the posterior portion of cell. (R) Dorsal view of the posterior end of cell, showing caudal cirrus. AZM, adoral zone of membranelles; CC, caudal cirrus; E, endoral membrane; FC, frontal cirri; FVC, frontoventral cirri; LMR, left marginal row; Ma, macronuclear nodules; P, paroral membrane; PTVC, pretransverse ventral cirri; PVC, postoral ventral cirri; RMR, right marginal row; TC, transverse cirri; 1–4, dorsal kineties 1–4. Scale bars = 30 µm.

Figure 6 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 6. Maximum likelihood (ML) tree inferred from the SSU rDNA sequences showing the systematic position of Sterkiella paratricirrata n. sp. (in red). The sequences of other Sterkiella species are indicated in bold. Numbers near nodes are bootstrap values for maximum-likelihood and posterior probability values for Bayesian inference (BI). '-' at nodes indicate disagreement between the two methods. The scale bar corresponds to 0.01 expected substitutions per site.

Figure 5 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 5. Photomicrographs of Sterkiella paratricirrata n. sp. in morphogenesis after protargol impregnation. (a, b) Ventral views of early dividers, to show the oral primordium in each divider (arrowheads). (c) Ventral view of an early divider, the arrowhead points to the disaggregating buccal cirrus, and the arrow shows cirrus V/4 dedifferentiates. (d) Arrowheads mark the frontoventraltransverse anlagen. Note that anlagen V and VI formed de novo in proter. (e) The arrow marks cirri III/2 and IV/3 disaggregate to involve in the formation of primordial streaks. (f) Arrowheads indicate the right marginal anlagen originate earlier than the left ones. (g, h) Ventral (g) and dorsal (h) view of a same divider, showing the anlagen for the left and right marginal cirral rows (arrowheads in g), parental undulating membranes dedifferentiate to form the undulating membranes anlage (arrow in g), cirrus V/3 is not involved in primordia formation (circle in g), and dorsal kineties anlagen develop intrakinetally (arrowheads in h). (i–k) Ventral (i, k) and dorsal (j) views of middle dividers, showing the frontoventral-transverse anlagen fragmentise to cirri. Arrowheads in (i) mark the leftmost frontal cirrus separated from the undulating membranes anlage, the arrows in (i) point the newly formed dorsomarginal kinety anlage, and arrows in (j) show the third dorsal kinety anlage splitting to form dorsal kineties 3 and 4. Note the macronuclear nodules fusing into a single mass. (l–o) Ventral (l, n) and dorsal (m, o) views of late dividers (n and o show the proter of a same divider). Note that the new ciliary structures move towards their final positions, arrowheads in (m) indicate the caudal cirri. 1–6, dorsal kineties 1–6. Scale bars = 50 µm.

Figure 4 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 4. Middle and late stages of morphogenesis of Sterkiella paratricirrata n. sp. after protargol staining. (a–d) Ventral (a, c) and dorsal (b, d) views of two middle dividers, showing frontoventraltransverse anlagen differentiate into cirri and the leftmost frontal cirrus is separated from the undulating membranes anlage. Arrowheads in (a) mark the anlagen for dorsomarginal kineties. Arrowheads in (d) show the third dorsal kinety anlage splits to form dorsal kineties 3 and 4. Note the macronuclear nodules fusing into a single mass. (e–h) Ventral (e, g) and dorsal (f, h) views of two late dividers to show 16 frontoventral-transverse cirri migrating towards their final positions. Dotted lines connect cirri that develop from the same cirral streak. Note caudal cirri formed, and macronuclear nodules divide. LMA, left marginal anlagen; LMR, left marginal row; RMA, right marginal anlagen; RMR, right marginal row. Scale bars = 50 µm.

Figure 2 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 2. Photomicrographs of Sterkiella paratricirrata n. sp. from life (a–h) and after staining with protargol (i–l). (a–d) Ventral views, showing the different body shapes, arrow in (a) marks the contractile vacuole. (e) Ventral view of the posterior portion of a cell, showing transverse cirri. (f) Showing nuclear apparatus. (g, h) Resting cyst, showing the cyst wall (opposed arrowheads in g). Squeezed cyst (h) with cytoplasm released, showing the polygon wrinkles on the surface. (i) Ventral view of the buccal field. (j) Dorsal view shows three caudal cirri (arrow). (k, l) Ventral (k) and dorsal (l) view of the holotype specimen, showing the general cirral pattern, arrowheads in (k) mark the dorsal kineties. Ma, macronuclear nodules; P, paroral; PTVC, pretransverse ventral cirri; PVC, postoral ventral cirri. Scale bars = 15 µm (g, h) and 50 µm (a–d, k, l).

Figure 1 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 1. Morphology of Sterkiella paratricirrata n. sp. from life (a, b) and after protargol staining (c, d). (a) Ventral view of a representative individual. (b) Resting cyst. (c, d) Ventral (c) and dorsal (d) view of the holotype specimen, showing the infraciliature and nuclear apparatus. AZM, adoral zone of membranelles; CC, caudal cirri; CV, contractile vacuole; LMR, left marginal row; Ma, macronuclear nodules; Mi, micronuclei; RMR, right marginal row; I/1, II/3, III/3, frontal cirri; II/2, buccal cirrus; III/2, IV/ 3, VI/3, VI/4, frontoventral cirri; IV/2, V/4, V/3, postoral ventral cirri; V/2, VI/2, pretransverse ventral cirri; IV/1, V/1, VI/1, transverse cirri; 1–6, dorsal kineties 1–6. Scale bars = 10 µm (b) and 50 µm (a, c, d).

Figure 3 in Morphology, morphogenesis and molecular phylogeny of the new soil ciliate Sterkiella paratricirrata n. sp. (Ciliophora, Hypotrichia, Oxytrichidae)

Figure 3. Early stages of morphogenesis of Sterkiella paratricirrata n. sp. after protargol staining. (a, b) Ventral view of very early dividers to show oral primordium in each opisthe (arrowheads), note that the cirrus V/4 dedifferentiates to form anlagen V and VI (arrow in b), and the buccal cirrus begins to be disorganised. (c) Ventral view of an early divider, the arrowhead shows cirri III/3 and IV/3 are involved to form anlagen III and IV, while anlagen V and VI develop de novo in proter. (d, e) Ventral (d) and dorsal (e) view of an early divider, showing the cirri II/2, III/2 and IV/3 are involved in the formation of six primordial streaks (arrowheads in d), and the dorsal kineties anlagen start to develop intrakinetally within dorsal kineties 1–3 in both proter and opisthe (arrowheads in e). (f–h) Ventral (f, g) and dorsal (h) views of early dividers, showing five cirral streaks formed (arrows in f, g), the right marginal anlagen originate, and the dorsal kineties anlagen elongate (arrowheads in h). (i) Ventral view to show the marginal anlagen develop intrakinetally. LMA, left marginal anlagen; RMA, right marginal anlagen. Scale bars = 50 µm.

Figure 4 in Morphology and morphogenesis of a saline soil ciliate, Cladotricha niesseniae sp. nov. (Ciliophora, Hypotrichia)

Figure 4. Morphogenesis in Cladotricha niesseniae sp. nov. after protargol staining. (a,b) Ventral and dorsal view of a late divider, to show the formation of the ciliature. Note the formation of the caudal cirri (arrowheads). LMA, left marginal anlagen; Ma, macronuclear nodules; Mi, micronuclei; RMA, right marginal anlagen. Scale bar: 30 μm.

Figure 3 in Morphology and morphogenesis of a saline soil ciliate, Cladotricha niesseniae sp. nov. (Ciliophora, Hypotrichia)

Figure 3. Morphogenesis in Cladotricha niesseniae sp. nov. after protargol staining. (a) Ventral view of an early divider, showing the oral primordium in the opisthe. (b) Ventral view of an early divider; arrows denote frontoventral-transverse cirral anlagen and arrowhead marks the right marginal anlage. (c,d) Ventral and dorsal view of a slightly later divider, to show the frontoventral-transverse cirral anlagen (arrows) and marginal anlagen (arrowheads). Note that dorsal kineties anlagen developed intrakinetally. (e,f) Ventral and dorsal view of a middle divider; arrows mark frontoventral-transverse cirral anlagen (e) and dorsal kineties anlagen (f). (g,h) Ventral and dorsal view of a late divider, to show formation of the caudal cirri (arrowheads). DKA, dorsal kineties anlagen; LMA, left marginal anlagen; Ma, macronuclear nodules; Mi, micronuclei; OP, oral primordium; RMA, right marginal anlagen. Scale bars: 30 μm.

Figure 5 in Morphology and morphogenesis of a saline soil ciliate, Cladotricha niesseniae sp. nov. (Ciliophora, Hypotrichia)

Figure 5. Photomicrographs of Cladotricha niesseniae sp. nov. during morphogenesis, after protargol staining. (a) Ventral view of an early divider, showing oral primordium of opisthe. (b,c) Ventral views of an early divider. Arrows denote the frontoventral-transverse cirral anlagen; arrowhead marks the differentiation of opisthe's oral primordium. (d) Dorsal view to show nuclear apparatus. (e) Dorsal view; arrows depict intrakinetal development of dorsal kineties anlagen. (f) Ventral view, to denote the frontoventral-transverse cirral anlagen (arrow) and the undulating membranes anlage (arrowhead). (g,j) Dorsal views, to mark the fusion of macronuclear nodules and the dorsal kineties anlagen (arrows). (h,i) Ventral views, to show the formation of the buccal cirri (arrow in h) and marginal anlagen (arrows in i). (k, n) Dorsal views, to show macronuclear nodules, the dorsal kineties anlagen (arrows) and the caudal cirri (arrowheads). (l,m) Ventral views of a late divider, to show the formation of the ciliature. Arrow marks the newly formed adoral zone of membranelles for the opisthe. LMA, left marginal anlagen; Ma, macronuclear nodules; Mi, micronuclei; OP, oral primordium; RMA, right marginal anlagen. Scale bars: 20 μm.

Figure 1 in Morphology and morphogenesis of a saline soil ciliate, Cladotricha niesseniae sp. nov. (Ciliophora, Hypotrichia)

Figure 1. Morphology of Cladotricha niesseniae sp. nov. from life (a,b) and after protargol staining (c–e). (a) Ventral view of a representative specimen. (b) Various specimens showing flexibility and variability in body shape. (c) Ventral view of holotype specimen, to show frontoventral ciliature. (d,e) Ventral (d) and dorsal (e) view of holotype specimen, showing ciliature and nuclear apparatus. (f,g) Sample site (f) and surrounding area (f). AZM, adoral zone of membranelles; BC, buccal cirri; CC, caudal cirri; E, endoral; FC, frontal cirri; LMR, left marginal row; Ma, macronuclear nodule; Mi, micronucleus; P, paroral; RMR, right marginal row; III–V, frontoventral cirral rows III–V; 1–3, dorsal kineties 1–3. Scale bars: a,d,e = 50 μm; b,c = 30 μm.