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FIGURE 4 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 4. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype). (A–E) Transverse sections through post-cerebral, pre-oral region, showing the morphology of various blood vessels; black arrowhead on (B) pointing to anterolateral diverticula from sub-rhynchocoelic vessel. (F) Magnification of (B), showing the interwoven longitudinal and circular muscle fibres in the rhynchocoel wall under vascular plug; asterisks (*) indicate lumen of anterolateral diverticula from sub-rhynchocoelic vessel. (G) Transverse section through the foregut region showing the vascular plug. (H) Magnification of (G), showing (1) rhynchocoel inner longitudinal muscle, (2) rhynchocoel outer circular muscle, (3) longitudinal muscle plate, and (4) transverse muscle above foregut.

FIGURE 8 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 8. An unrooted maximum-likelihood tree based on T92 + G model with 496-bp partial sequences of the 16S rRNA gene from Cerebratulus orochi sp. nov. and 10 other DDBJ/EMBL/GenBank entries that showed highest max/total scores as a result of a MegaBLAST search (Morgulis et al. 2008) at the NCBI website (https://blast.ncbi.nlm.nih.gov). Numbers near nodes show bootstrap values with 1000 replicates. Red horizontal bars indicate conspecifics according to species delimitation analyses by employing ABGD (Puillandre et al. 2012) and PTP (Zhang et al. 2013), which resulted in the same outcome.

FIGURE 3 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 3. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype). (A) Transverse section through intestinal region showing ovary containing mature oocytes. (B) Transverse section through intestinal region showing dorsoventral muscles (indicated by black arrowheads) running both proximal and lateral to lateral nerve cord; white arrowhead showing lateral edge of body. (C) Transverse section through proboscis showing heterotype musculature. (D) Transverse section through proboscis showing muscle cross (indicated by white arrowheads). (E) Pseudocnidae.

FIGURE 2 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 2. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype). (A) Transverse section through foregut region showing body-wall musculature; asterisk (*) indicating rhynchocoel outer circular muscle; black arrowhead pointing to rhynchocoel inner longitudinal muscle. (B) Transverse section through foregut region showing cutis structure; arrowheads indicating diagonal muscles. (C) Tangential section through foregut region showing subepidermal musculature. (D) Tangential section through intestinal region showing subepidermal musculature. (E) Transverse section through precerebral region.

FIGURE 1 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 1. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype), photographs taken in life under anaesthetized state by Yuki Oya. (A) Entire body. (B) Head, ventral view. (C) Magnification of rhynchodaeal opening. (D) Tail.

FIGURE 7. A in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 7. A maximum-likelihood (ML) tree showing the phylogenetic position of Cerebratulus orochi sp. nov. among the "Cerebratulus clade" (Chernyshev & Polyakova 2019) based on concatenated 16S rRNA, COI, 18S rRNA, 28S rRNA, and histone H3 genes. Nodal support is represented by ML bootstrap value with 1000 replicates and Bayesian posterior probability; solid, black circles indicate full support.

FIGURE 6 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 6. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype). (A) Transverse section through brain. (B) Transverse section through foregut region showing lateral nerve cord; white arrowheads indicating muscle fibres; black arrowheads showing radial muscle fibres. (C) Transverse section through foregut region showing excretory collecting tubules; black arrowheads showing somatic muscles.

FIGURE 5 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 5. Cerebratulus orochi sp. nov., ICHUM 6078 (holotype). (A) Transverse section through posterior region of brain. (B) Magnification of (A) showing different types of neuronal cells. (C) Magnification of (A) showing neurochord cell. (D) Transverse section through anterior tip of head showing three frontal organs.

FIGURE 9 in Redescription of Cerebratulus marginatus auct. (Nemertea: Pilidiophora) from Hokkaido, Japan, as a new species

FIGURE 9. An unrooted maximum-likelihood tree based on HKY + I model with 583-bp partial sequences of the COI gene from Cerebratulus orochi sp. nov. and 10 other DDBJ/EMBL/GenBank entries that showed highest max/total scores as a result of a MegaBLAST search (Morgulis et al. 2008) at the NCBI website (https://blast.ncbi.nlm.nih.gov). Numbers near nodes show bootstrap values with 1000 replicates. Red horizontal bars indicate conspecifics according to species delimitation analyses by employing ABGD (Puillandre et al. 2012) and PTP (Zhang et al. 2013), which resulted in the same outcome.

Fig. 4 in Sperm morphology and some aspects of acrosomal complex development in four species of Heteronemertea (Pilidiophora, Nemertea)

Fig. 4. Ultrastructural aspects of the acrosomal complex development in Kulikovia alborostrata (A) Primary spermatocyte (sc) (B) Fragment of primary spermatocyte with Golgi body (Gb) located near a pair of centrioles (c). Arrows indicate small proacrosomal vesicles (C) Dividing spermatocyte (D) Higher magnification of proacrosomal vesicles (arrows) (E) Spermatid with single large rounded acrosomal vesicle (av) (F) Spermatids with transformed acrosomal vesicles in posterior and anterior positions. Abbreviations: chr, chromatin; m, mitochondrion; n, nucleus; pnf, posterior nuclear fossa. Scale bars: A, F = 2 μm, B, C, E = 1 μm, D = 0.5 μm.

Fig. 1 in Sperm morphology and some aspects of acrosomal complex development in four species of Heteronemertea (Pilidiophora, Nemertea)

Fig. 1. Fine organization of spermatozoon in Micrura bella (A–C) SEM (D–J) TEM (A) General view of spermatozoon (B) Sperm head with visible acrosomal region (a), slightly curveted nucleus (n) and midpiece (mp) with mitochondria rounded or slightly elongated along anterior/posterior axis. Flagellum (f) is posteriorly oriented (C) Longitudinal section along sperm head that consists of acrosomal complex (ac), nucleus, midpiece containing mitochondria, and centriolar complex (cc) (D) Longitudinal section along acrosomal complex containing large electron-dense vesicle in apical position (dv), thin layer of electron-dense material surrounding the electron-dense vesicle (arrows), small vesicles with moderate electron density (mv) located beneath plasma membrane and lower than the region of large vesicle; flocculent subacrosomal material of moderate electron density (asterisk) fills the central region immediately under the electron-dense vesicle (E) Higher magnification of large acrosomal vesicle in longitudinal projection. Its basal part forms invagination (F) Transverse section across acrosomal complex at the level of small vesicles. The vesicles are located peripherally forming a ring, often open. Central part is filled by subacrosomal material (asterisk) (G) Transverse section across acrosomal complex at the level of electron-dense vesicle. The vesicle is surrounded by electron-dense ring (arrow) (H) Longitudinal section along midpiece with centriolar complex represented by proximal (pc) and distal centrioles (dc) and mitochondria. The basal part of the nucleus forms posterior nuclear fossa (pnf) and invaginations hosting mitochondria (arrow) at sites of their contact with the nucleus (I) Transverse section across midpiece containing five mitochondria (J) Pericentriolar complex of distal centriole formed by satellite fibers (sf) (K) Transverse section through flagellum. Scale bars: A = 5 μm, B, C =2 μm, D-G =0.2 μm, H-J = 0.5 μm, K = 0.1 μm.

Figure 1 in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 1. Maximum-likelihood (ML) tree showing the phylogenetic relationships among 41 newly sequenced specimens of pilidiophorans (indicated with solid blue circles). Numbers near nodes are support values, ML bootstrap/Bayesian inference (BI) posterior probability. Nodes with yellow triangles are fully supported, with 100% ML bootstrap and 1.00 BI posterior probability. New species names are indicated in bold. Some nodes were polytomous in the BI tree (indicated by hyphens in place of posterior probability values).

Figure 7. A in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 7. A, Baseodiscus narusei sp. nov., holotype, ICHUM 6310, entire body, showing characteristic lateral edges; B, Baseodiscus paracelensis sp. nov., holotype, MIMB 33132, entire body; C, Baseodiscus aff. marmoratus (Bürger, 1890), anterior end of body, ventral view, head to the left; D, E, Baseodiscus ohtsukai sp. nov., holotype, ICHUM, 6327, drawings of anterior end of body viewed dorsally (D) and ventrally (E); F, G, Baseodiscus urgorrii sp. nov., holotype, MCZ IZ-135319, anterior end of body viewed dorsally (F) and ventrolaterally (G). Photos by T. Naruse (A), A. V. Chernyshev (B, C), and G. Giribet (F, G).

Figure 5. Cephalomastax brevis Iwata, 1957, ICHUM 6267. A in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 5. Cephalomastax brevis Iwata, 1957, ICHUM 6267. A, anterior fragment in contracted state, lateral view; B, anterior end of body, ventral view; C, transverse section showing proboscis; D, magnification of anterior proboscis musculature (arrowhead, radial muscle fibre); E, transverse section showing rhynchocoel musculature. Photos by H. Kajihara.

Figure 3. A in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 3. A, Baseodiscus aff. marmoratus (Bürger, 1890) (voucher DNA, ICHUM 6322); B, Baseodiscus aff. maculosus (Bürger, 1895a) (voucher DNA, ICHUM 6324); C–E, Baseodiscus delineatus (Delle Chiaje, 1822-1829), ICHUM 6326; F, Baseodiscus cf. curtus (Hubrecht, 1879), ICHUM 6328; G, H, Baseodiscus urgorrii sp. nov., holotype, MCZ IZ-135319, entire body (G) and protruding proboscis (indicated by an arrowhead, H); I, Baseodiscus punnetti (Coe, 1904), specimen different from any voucher specimens in this study; J, K, Baseodiscus kakuii sp. nov., holotype, ICHUM 6334, anterior body (J) and

Figure 4. A–C in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 4. A–C, Eopilidion misakiense gen. et sp. nov., holotype, ICHUM 6303, anterior end of body, ventral view, head to the right (A), frontal view (B), squeezed-slide preparation (C); D, Valencinura jambio sp. nov., holotype, ICHUM 6305, anterior end of body, ventral view. Photos by H. Kajihara.

Figure 6. A–E in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 6. A–E, Baseodiscus takakurai Gibson, 1995, ICHUM 6308, A, anterior end of body, ventral view; B, middle body region, showing appearance of body markings in differently contracted states; C, juvenile, showing dorsal mottling; D, juvenile, with head viewed ventrolaterally, showing two ocelli; E, transverse section of proboscis; F, Baseodiscus komatsui sp. nov., holotype, NSNM NMNS-Ne 1, entire body. Photos by H. Kajihara (A–E) and H. Komatsu (F).

Figure 2. A in Exploring the basal topology of the heteronemertean tree of life: establishment of a new family, along with turbotaxonomy of Valenciniidae (Nemertea: Pilidiophora: Heteronemertea)

Figure 2. A, Eopilidion misakiense gen. et sp. nov., holotype, ICHUM 6303, lateral view, head to the right; B, Oxypolella hiebertae sp. nov., holotype, MIMB 42256; C, Cephalomastax brevis Iwata, 1957, ICHUM 6304; D, E, Valencinura jambio sp. nov., holotype, ICHUM 6305, anterior body fragment (D) and enlargement of intestinal region (E) showing yellowish proboscis (indicated by arrowheads); F, Baseodiscus takakurai Gibson, 1995, ICHUM 6306; G, Baseodiscus profundus sp. nov., holotype, MIMB 42257; H, I, Baseodiscus narusei sp. nov., holotype, ICHUM 6310, anterior end of body, dorsal (H) and ventral (I) views; J, Baseodiscus paracelensis sp. nov., holotype, MIMB 33132; K, Baseodiscus komatsui sp. nov., holotype, NMNS-Ne 1; L, Baseodiscus unicolor Stiasny-Wijnhoff, 1925; M, Baseodiscus giribeti sp. nov., holotype, MCZ IZ-135324; N, Baseodiscus cf. amboinensis (Staub, 1900); O, Baseodiscus hemprichii (Ehrenberg, 1828-1831), specimen from Okinawa, different from any voucher specimens in this study; P, Baseodiscus zebra sp. nov., holotype, RUMF-ZN-00001; Q, Baseodiscus quinquelineatus (Quoy & Gaimard, 1833), ICHUM 6319. Photographs by H. Kajihara (A, C, D, E, F, O), A. V. Chernyshev (B, G, J), T. Naruse (H, I), H. Komatsu (K), G. Giribet (L, M), D. Uyeno (P), and R. Yoshida (Q).