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.

Theama japonica sp. nov., an Interstitial Polyclad Flatworm Showing a Wide Distribution along Japanese Coasts.

We establish a new interstitial polyclad species, Theama japonica sp. nov., based on specimens collected from coarse-sandy habitats in three Japanese main islands (Hokkaido, Honshu, and Shikoku) along the coasts of the Pacific Ocean and the Sea of Japan. Theama japonica is characterized by i) two pairs of cerebral eyespots and four to six precerebral eyespots; ii) eosinophilic secretion glands distributed in the distal half of the inner ventral part of the prostatic vesicle; iii) a conical penis papilla, bent up dorsally, with a sclerotized inner wall; iv) the prostatic sheath with an inner angular fold on the dorso-distal side; and v) the external cilia longer dorsally than ventrally. Partial sequences of the cytochrome c oxidase subunit I (COI) gene from 20 specimens collected at eight localities along Japanese coasts represented 19 haplotypes. The uncorrected p-distances among these COI haplotypes fell within intraspecific variations observed in other polyclads. A network analysis based on these COI haplotypes suggested a geographically non-cohesive genetic structure of the species, possibly indicating the species' high dispersibility. Molecular phylogenetic analyses based on a concatenated dataset of 18S and 28S rDNA sequences showed T. japonica formed a clade with other Theama species. The resulting tree also indicates that our new species is more closely related to Theama sp. from Colombia than species from Panama and Croatia.

Investigating the diversity of bioluminescent marine worm Polycirrus (Annelida), with description of three new species from the Western Pacific.

Bioluminescence, a phenomenon observed widely in organisms ranging from bacteria to metazoans, has a significant impact on the behaviour and ecology of organisms. Among bioluminescent organisms, Polycirrus, which has unique emission wavelengths, has received attention, and advanced studies such as RNA-Seq have been conducted, but they are limited to a few cases. In addition, accurate species identification is difficult due to lack of taxonomic organization. In this study, we conducted comprehensive taxonomic survey of Japanese Polycirrus based on multiple specimens from different locations and described as three new species: Polycirrus onibi sp. nov., P. ikeguchii sp. nov. and P. aoandon sp. nov. The three species can be distinguished from the known species based on the following characters: (i) arrangement of mid-ventral groove, (ii) arrangement of notochaetigerous segments, (iii) type of neurochaetae uncini, and (iv) arrangement of nephridial papillae. By linking the bioluminescence phenomenon with taxonomic knowledge, we established a foundation for future bioluminescent research development. We also provide a brief phylogenetic tree based on cytochrome c oxidase subunit I (COI) sequences to discuss the evolution of bioluminescence and the direction of future research.

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.

Descriptions of two new species of Armatoplana (Polycladida: Stylochoplanidae) from the coasts of Japan, with their phylogenetic positions in Leptoplanoidea.

We describe two new species of Armatoplana Faubel, 1983, namely, A. albomaculata sp. nov. and A. kaburakii sp. nov., from Japan. This is the first record of the genus from the West Pacific. Armatoplana albomaculata sp. nov. has the following characteristics: i) no nuchal tentacles; ii) white spots on the dorsal surface of the body; iii) an elongated oval prostatic vesicle directing posteriorly but curving dorsally in the distal part; iv) a long, curved penis stylet; and v) a small, oval Langs vesicle without accessory vesicles. Armatoplana kaburakii sp. nov. is distinguished from other congeners by having i) no nuchal tentacles; ii) a large, elongated Langs vesicle without accessory vesicles; and iii) gonopores opening closely to each other. We propose to re-circumscribe Armatoplana so that it would not become a junior synonym of Candimboides Prudhoe, 1982 and Phylloplana Laidlaw, 1903. We provide partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene as DNA barcodes for the two new species. Our phylogenetic analyses based on concatenated sequences of the 16S, 18S, and 28S ribosomal DNA and COI revealed that A. albomaculata sp. nov. and A. kaburakii sp. nov. were sister taxa; however, they did not form a monophyletic clade with Armatoplana divae (Marcus, 1947) and Armatoplana leptalea (Marcus, 1947).

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 clade that was composed of all the other Prosthiostomum species included in the analysis.

Description of Pericelis flavomarginata sp. nov. (Polycladida: Cotylea) and its predatory behavior on a scaleworm.

We describe a new species of polyclad flatworm, Pericelis flavomarginata sp. nov., from the intertidal and subtidal zones along localities on the Pacific coast of Japan. Pericelis flavomarginata sp. nov. is characterized by i) the dorsal surface of the body fringed by a lemon-yellow line except for the tip of tentacles, with a narrow brown midline running from the anterior edge of the body to the posterior end of the pharynx, ii) the pair of marginal tentacles with the tips extending and tapering, and iii) the presence of a common gonopore. Our molecular phylogenetic analysis revealed that selected Pericelis species were divided into two clades, each of which may be agreed with a characteristic dorsal color pattern. Additionally, we report an observation on the feeding behavior of P. flavomarginata sp. nov. on the polychaete Iphione muricata (Savigny in Lamarck, 1818).

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.