Delimiting the polymorphic congeners of the genus Oerstedia Quatrefages, 1864 (Nemertea, Hoplonemertea), and descriptions of three new species from the Northwest Pacific

Three new species of the monostiliferous hoplonemertean genus Oerstedia Quatrefages, 1864, are herein described using morphological and molecular data—Oerstedia pseudoculata sp. nov., from Akkeshi Bay and Oshoro Bay, Hokkaido, Japan, and from Aniwa Bay, Sakhalin, Russia; Oerstedia rugosa sp. nov. from Sagami Bay, Misaki, Kanagawa, Japan, and Van Phong Bay, Vietnam; and Oerstedia viridifusca sp. nov. from Manazuru, Kanagawa, Japan. As to the external morphology, O. pseudoculata sp. nov. can be differentiated from O. oculata only by its bright-orange ocelli visible on both sides of the head, and a proboscis pore opening at the ventral tip of the head. These two sister species repeat each other’s color patterns, a phenomenon that can be explained by Vavilov’s law of homologous series. Oerstedia rugosa sp. nov. can be identified by its carmine or deep-red to brownish-red body with several longitudinal, intertwined white lines or wrinkles running from the head to the posterior body, and by 17–23 vaguely bordered white bands composed of variedly sized dots encircling the body, arranged at irregular intervals. Oerstedia viridifusca sp. nov. can be distinguished from other Oerstedia by (i) the entire body flecked with minute greenish-brown dots, especially densely on the anterior portion of the dorsal surface, but sparsely on the posterior half of the ventral surface; (ii) a collar-like portion encircling the body along the posterior cephalic furrow where the greenish-brown dots are absent; (iii) the anterolateral edges of the head lacking the greenish-brown dots; and (iv) the ocelli being brownish-orange in color. Oerstedia phoresiae (Kulikova, 1987) is reported for the first time from Japan, in addition to its previous distribution record in Russia and in South Korea. Phylogenetic analyses based on the 16S, 18S, 28S ribosomal RNA, cytochrome c oxidase subunit I, and histone H3 genes show that the new species are true congeners of the genus Oerstedia with O. pseudoculata sp. nov. and O. viridifusca sp. nov. nested within the clade Paroerstediella whereas O. rugosa sp. nov. in the clade Oerstedia. This taxonomic work emphasizes the importance of DNA barcode sequence in the taxonomy and systematics of the polymorphic congeners of the genus Oerstedia

Flatworm cocoons in the abyss: same plan under pressure

While knowledge of early ontogeny in abyssal animals is highly limited in general, it was completely lacking for abyssal, free-living platyhelminths. We discovered flatworm egg capsules (or ‘cocoons') on rocks collected at depths of 6176–6200 m on the abyssal slope of the Kuril–Kamchatka Trench, northwestern Pacific. The egg capsules were black and spherical, around 3 mm in diameter, and contained three to seven individuals (n = 4) at the same developmental stage, either the spherical (putative early embryo) or vermiform (putative late embryo) stages. A molecular phylogenetic analysis based on 18S and 28S rRNA sequences revealed that the flatworms belong in suborder Maricola in Tricladida and suggested that they may have colonized from shallow to deep waters. This study provides the deepest record for free-living flatworms and the first information on their early life stages in the abyssal zone, which were very similar to those in shallow-water forms. This similarity in development between the relatively benign shallow-water and the extreme abyssal environments suggests that triclads adapting to the latter faced primarily physiological and/or ecological adaptive challenges rather than developmental ones

A giant new species of Enchiridium (Polycladida, Prosthiostomidae) from southwestern Japan

We describe a new species of polyclad flatworm, Enchiridium daidai sp. nov., from the rocky subtidal zone in the East China Sea along the coasts of the Kyushu and Okinawa Islands, Japan. Enchiridium daidai sp. nov. is characterized by i) the entire periphery of the dorsal surface narrowly fringed with orange, ii) a marginal-eyespot band extending to the position of the mouth (about anterior one-eighth of body), and iii) two prostatic vesicles covered by a common muscle sheath, which is penetrated by the ejaculatory duct. We performed a molecular phylogenetic analysis based on 945-bp 28S rDNA sequences of 16 species of Prosthiostomidae currently available in public databases in addition to those of E. daidai sp. nov. and Prosthiostomum torquatum Tsuyuki et al., 2019. In the resulting tree, our new species was nested in a clade composed of Enchiridium species. The tree topology was in favor of a taxonomic view that Enchiridium should be defined by having i) a common muscle sheath that encloses two prostatic vesicles and ii) marginal eyespots that may or may not surround the periphery of the dorsal surface

File S3. Concatenated 18S+28S sequences used for the maximum-likelihood analysis, reduced to 2440 positions (1–1458 for 18S; 1459–2440 for 28S) by removing alignment-ambiguous sites with Gblocks under “relaxed” parameters. from Flatworm cocoons in the abyss: same plan under pressure

While knowledge of early ontogeny in abyssal animals is highly limited in general, it was completely lacking for abyssal, free-living platyhelminths. We discovered flatworm egg capsules (or ‘cocoons') on rocks collected at depths of 6176–6200 m on the abyssal slope of the Kuril-Kamchatka Trench, northwestern Pacific. The egg capsules were black and spherical, around 3 mm in diameter, and contained three to seven individuals (n = 4) at the same developmental stage, either the spherical (putative early embryo) or vermiform (putative late embryo) stages. A molecular phylogenetic analysis based on 18S and 28S rRNA sequences revealed that the flatworms belong in suborder Maricola in Tricladida and suggested that they may have colonized from shallow to deep waters. This study provides the deepest record for free-living flatworms and the first information on their early life stages in the abyssal zone, which were very similar to those in shallow-water forms. This similarity in development between the relatively benign shallow-water and the extreme abyssal environments suggests that triclads adapting to the latter faced primarily physiological and/or ecological adaptive challenges rather than developmental ones

First record of Stylostomum ellipse (Dalyell, 1853) (Platyhelminthes, Polycladida) from the Pacific Ocean

The polyclad flatworm Stylostomum ellipse (Dalyell, 1853) has hitherto been recorded from the Antarctic region, Mediterranean Sea, Patagonian region, Scandinavia, South Africa, and South Georgia Island. In this study, we report S. ellipse for the first time from the Pacific Ocean based on specimens collected in Hokkaido, northern Japan. Our specimens are morphologically identifiable as S. ellipse, but may represent a biologically different species from a population of the Mediterranean Sea. This is because, based on the previous genetic data of other cotylean species, the observed uncorrected p-distance 0.02160 between the two distinct populations in terms of a partial 972 bp region of the 28S rDNA sequence may be great enough to separate the species biologically

Reversible shifts between interstitial and epibenthic habitats in evolutionary history : Molecular phylogeny of the marine flatworm family Boniniidae (Platyhelminthes: Polycladida: Cotylea) with descriptions of two new species

Tiny animals in various metazoan phyla inhabit the interstices between sand and/or gravel grains, and adaptive traits in their body plan, such as simplification and size reduction, have attracted research attention. Several possible explanations of how such animals colonized interstitial habitats have been proposed, but their adaptation to this environment has generally been regarded as irreversible. However, the actual evolutionary transitions are not well understood in almost all taxa. In the present study, we show reversible evolutionary shifts from interstitial to epibenthic habitats in the lineage of the polyclad flatworm genus Boninia. In addition, we establish two new species of this genus found from different microhabitats on a single beach in Okinawa Island, Japan: (i) the interstitial species Boninia uru sp. nov. from gravelly sediments and (ii) the epibenthic species Boninia yambarensis sp. nov. from rock undersurfaces. Our observations suggest that rigid microhabitat segregation exists between these two species. Molecular phylogenetic analyses based on the partial 18S and 28S rDNA sequences of the new Boninia species and four other congeners, for which molecular sequences were available in public databases [Boninia antillara (epibenthic), Boninia divae (epibenthic), Boninia neotethydis (interstitial), and an unidentified Boninia sp. (habitat indeterminate)], revealed that the two interstitial species (B. neotethydis and B. uru sp. nov.) were not monophyletic among the three epibenthic species. According to ancestral state reconstruction analysis, the last common ancestor of the analyzed Boninia species inhabited interstitial realms, and a shift to the epibenthic environment occurred at least once. Such an interstitial to noninterstitial evolutionary route seems to be rare among Animalia; to date, it has been reported only in acochlidian slugs in the clade Hedylopsacea. Our phylogenetic tree also showed that the sympatric B. uru sp. nov. and B. yambarensis sp. nov. were not in a sister relationship, indicating that they colonized the same beach independently rather than descended in situ from a common ancestor that migrated and settled at the beach

A new species of slender flatworm in the genus Eucestoplana and a record of E. cf. cuneata (Platyhelminthes, Polycladida) from the Okinawa Islands, Japan, with an inference of their phylogenetic positions within Cestoplanidae

In this study, we describe a new species of elongated marine flatworm, Eucestoplana ittanmomen sp. nov., collected from the intertidal zone of the Okinawa Islands, Japan. Eucestoplana ittanmomen sp. nov. is distinguished from other congeners based on the following characteristics: i) its translucent body lacking coloration, ii) its dome-shaped penis sheath, iii) the absence of cilia on the inner wall of the male atrium except outside the penis sheath, and iv) the presence of an adhesive organ at the posterior end of the body. Additionally, we report the occurrence of E. cf. cuneata (Sopott-Ehlers & Schmidt, 1975) in Japan; E. cuneata has previously been documented in the Galapagos and Fiji Islands. We conducted phylogenetic analyses to infer the positions of the two Eucestoplana species within Cestoplanidae using a concatenated dataset comprising partial 18S and 28S rDNA sequences from E. cf. cuneata and E. ittanmomen sp. nov. from Japan, as well as four known Cestoplana species with sequences available in public databases. Our phylogenetic analyses revealed that Cestoplana and Eucestoplana were reciprocally monophyletic. Furthermore, the genetic distance of the 16S rDNA sequences supported the genetic independence of the two sister species, E. cf. cuneata and E. ittanmomen sp. nov

Description of a New Species of the Marine Flatworm Prosthiostomum (Platyhelminthes: Polycladida) and its Three Known Congeners from Misaki, Japan, with Inference of Their Phylogenetic Positions within Prosthiostomidae

The present study provides morphological descriptions of four species of Prosthiostomum (Polycladida, Prosthiostomidae)-P auratum Kato, 1937; P. hibana sp. n.; P. cf. ostreae Kato, 1937; and P. vulgare Kato, 1938-based on specimens collected among branching coralline algae and kelp holdfasts in Misaki, Japan. The new species P. hibana sp. n. is characterized by i) the dorsal surface of the body covered with numerous orange maculae, some of which coalesce together to form larger ones; ii) a pair of linear cerebral-eyespot clusters, each consisting of relatively few (7-9) cerebral eyespots; iii) 3-4 pairs of ventral eyespots embedded in parenchyma: iv) the inner wall of the male atrium deeply ruffled; v) the lumen of the seminal vesicle being narrow and elongated in shape; and vi) a large sucker situated in the center of the body. We remark on some morphological characters that were not mentioned in the original description of P. auratum. We infer the phylogenetic positions of these four species within Prosthiostomidae using the maximum-likelihood analysis based on partial 28S rRNA and COI gene sequences determined de novo, in addition to those that are currently available in public databases. In the resulting tree, the four species-P. auratum, P. hibana sp. n., P. cf. ostreae, and P. vulgare-were nested in a Glade that was composed of all the other Prosthiostomum species included in the analysis

File S2. Aligned 28S sequences used for the maximum-likelihood analysis, trimmed in MEGA7 to the shortest length among the sequences. from Flatworm cocoons in the abyss: same plan under pressure

While knowledge of early ontogeny in abyssal animals is highly limited in general, it was completely lacking for abyssal, free-living platyhelminths. We discovered flatworm egg capsules (or ‘cocoons') on rocks collected at depths of 6176–6200 m on the abyssal slope of the Kuril-Kamchatka Trench, northwestern Pacific. The egg capsules were black and spherical, around 3 mm in diameter, and contained three to seven individuals (n = 4) at the same developmental stage, either the spherical (putative early embryo) or vermiform (putative late embryo) stages. A molecular phylogenetic analysis based on 18S and 28S rRNA sequences revealed that the flatworms belong in suborder Maricola in Tricladida and suggested that they may have colonized from shallow to deep waters. This study provides the deepest record for free-living flatworms and the first information on their early life stages in the abyssal zone, which were very similar to those in shallow-water forms. This similarity in development between the relatively benign shallow-water and the extreme abyssal environments suggests that triclads adapting to the latter faced primarily physiological and/or ecological adaptive challenges rather than developmental ones