Postembryonic development and lifestyle shift in the commensal ribbon worm.

BACKGROUND: Various morphological adaptations are associated with symbiotic relationships between organisms. One such adaptation is seen in the nemertean genus Malacobdella. All species in the genus are commensals of molluscan hosts, attaching to the surface of host mantles with a terminal sucker. Malacobdella possesses several unique characteristics within the order Monostilifera, exhibiting the terminal sucker and the absence of eyes and apical/cerebral organs, which are related to their adaptation to a commensal lifestyle. Nevertheless, the developmental processes that give rise to these morphological characteristics during their transition from free-living larvae to commensal adults remain uncertain. RESULTS: In the present study, therefore, we visualized the developmental processes of the internal morphologies during postembryonic larval stages using fluorescent molecular markers. We demonstrated the developmental processes, including the formation of the sucker primordium and the functional sucker. Furthermore, our data revealed that sensory organs, including apical/cerebral organs, formed in embryonic and early postembryonic stages but degenerated in the late postembryonic stage prior to settlement within their host using a terminal sucker. CONCLUSIONS: This study reveals the formation of the terminal sucker through tissue invagination, shedding light on its adhesion mechanism. Sucker muscle development likely originates from body wall muscles. Notably, M. japonica exhibits negative phototaxis despite lacking larval ocelli. This observation suggests a potential role for other sensory mechanisms, such as the apical and cerebral organs identified in the larvae, in facilitating settlement and adhesive behaviors. The loss of sensory organs during larval development might reflect a transition from planktonic feeding to a stable, host-associated lifestyle. This study also emphasizes the need for further studies to explore the phylogenetic relationships within the infraorder Amphiporiina and investigate the postembryonic development of neuromuscular systems in closely related taxa to gain a more comprehensive understanding of ecological adaptations in Nemertea.

A New Species of Branchellion (Hirudinea: Piscicolidae) Parasitizing the Gills of Short-Tail Stingrays (Batoidea: Dasyatidae) from the West Pacific.

A new fish leech, Branchellion brevicaudatae sp. n., is described based on specimens parasitizing the gills of the short-tail stingray, Bathytoshia brevicaudata (Hutton, 1875), collected from Japanese waters. The new species can be distinguished from other congeners by having: i) pulsating vesicles emerging from posterior base of branchiae, one pair per somite; ii) dorsal white spots, not arranged in longitudinal row; and iii) blackish body. A phylogenetic tree based on partial sequences of the mitochondrial cytochrome c oxidase subunit I gene from the new species and other piscicolid worms showed that the new species is sister to Branchellion torpedinis Savigny, 1822. This is the first record of Branchellion Savigny, 1822 from Japanese waters.

Three new species in Tetrastemma Ehrenberg, 1828 (Nemertea, Monostilifera) from sublittoral to upper bathyal zones of the northwestern Pacific.

Monostiliferous nemerteans in the genus Tetrastemma Ehrenberg, 1828 are generally characterized as having four eyes, and they occur worldwide, from the intertidal zone to the deep-sea bottom. Recent extensive sampling of Tetrastemma has explored the high species diversity, including many undescribed forms, but phylogenic analysis has revealed non-monophyly of the genus. We herein describe three new species of the genus (T.albumsp. nov., T.personasp. nov., and T.shohoensesp. nov.) from northwestern Pacific waters based on specimens collected by dredging or by use of a remotely operated vehicle at depths of 116-455 m. Since anatomical and histological characters traditionally used in systematics of the genus are sometimes interspecifically uniform, a histology-free approach is applied for the species descriptions in this study. To confirm the generic affiliation of the new species, a molecular phylogenetic analysis based on partial sequences of cytochrome c oxidase subunit I, 16S rRNA, 18S rRNA, 28S rRNA, and histone H3 genes was performed. Our result shows that all three new species are nested in a subclade formed by species from the North Pacific and American Atlantic, inferring that geographic distribution does not reflect the cladogenesis of Tetrastemma. Furthermore, two Tetrastemma species with a cylindrical stylet basis, T.freyaeChernyshev et al., 2020 from off the coast of India and Hawaii and T.shohoensesp. nov. from Shoho Seamount, Japan, constitute a clade in the resulting tree.

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.

First eumonostiliferous nemertean from the Nishi-Shichito Ridge, Genrokunemertes obesa gen. et sp. nov. (Eumonostilifera, Nemertea).

Nemerteans are mostly marine, benthic invertebrates, inhabiting intertidal to hadal zones. Recently, they have been recognized from deep sea with environmental DNA (eDNA) metabarcoding of sediment samples whereas any records from the locations and/or the water depth have not been documented in nemertean taxonomic publications to date. It suggests that there are major gaps between deep-sea nemertean fauna observed with eDNA and taxonomic knowledge. During a research expedition in 2021, we obtained a single specimen of eumonostiliferous nemertean from the southern part of Genroku Seamount Chain, the Nishi-Shichito Ridge, where any nemertean species have never been reported. Subsequent morphological and molecular examination reveal that the species is placed in a new genus and herein described as Genrokunemertes obesa gen. et sp. nov. It resembles shallow-water-dwelling Kurilonemertes and Typhloerstedia, but differs from the former in lacking eyes and the latter in possessing well-developed cephalic glands and lacking accessory nerves of lateral nerve cords. In terms of genetic distances based on partial sequences of the cytochrome c oxidase subunit I gene, G. obesa gen. et sp. nov. is closest to Monostilifera sp. Owase collected from Japan, which is characterized by large four eyes; the COI distance is higher than commonly observed interspecific divergences in eumonostiliferans.

First record of the decapod-egg predator Ovicides paralithodis (Nemertea, Carcinonemertidae) from the snow crab Chionoecetes opilio (Decapoda, Brachyura).

The carcinonemertid monostiliferan Ovicides paralithodis Kajihara and Kuris, 2013 was originally described as an egg predator of the red-king crab Paralithodes camtschaticus (Tilesius, 1815) in the Sea of Okhotsk, the Bering Sea, and the Gulf of Alaska. In the present study, several carcinonemertid specimens were obtained from the egg mass of the snow crab Chionoecetes opilio (O. Fabricius, 1788) in the Sea of Japan. Partial sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI) determined from two specimens of the carcinonemertid were identical with a barcode sequence from the holotype of O. paralithodis, indicating that the host range of the species covers at least the two decapod species, P. camtschaticus and C. opilio.

Description of Tubulanus misakiensis sp. nov. (Nemertea: Palaeonemertea) from Sagami Bay, Japan.

The palaeonemertean genus Tubulanus Renier, 1804 currently harbors 35 species inhabiting the intertidal to the hadal zones. During a faunal survey in Sagami Bay, Japan, a single specimen of undescribed tubulanid having uniformly pale-colored body with a single dark-colored stripe on the dorsal surface was collected by dredging at depths of 116-211 m. Based on the body coloration and some anatomical features, it can be differentiated from the 35 congeners and is thus described herein as Tubulanus misakiensis sp. nov. In a molecular phylogenetic tree constructed with sequences available in public databases representing five markers (16S rRNA, cytochrome c oxidase subunit I [COI], 18S rRNA, 28S rRNA [not obtained for T. misakiensis], and histone H3), T. misakiensis was sister to Tubulanus izuensis Hookabe, Asai, Nakano, Kimura and Kajihara, 2020 and nested within a clade along with seven unidentified specimens from the abyssal and hadal zones (3428-9577 m); T. misakiensis and T. izuensis differed by 11.7% uncorrected p-distance in terms of 657-bp COI sequences.

The Phylogenetic Position of Branchamphinome (Annelida, Amphinomidae) with a Description of a New Species from the North Pacific Ocean.

A new species of amphinomid polychaete, Branchamphinome kohtsukai sp. nov., is described from Japanese waters, 29-211 m in depth. The species is distinguishable from other congeners by the following features: i) branchiae with four-six filaments in midbody segments; ii) two pairs of eyes not coalescent; iii) the ventral side of the first four chaetigers broadly pigmented. This is the first record of Branchamphinome from the North Pacific Ocean. We provide a phylogenetic tree based on the sequences of four genes (COI, 16S, 18S, 28S) and discuss amphinomids' relationships.

Lacydoniidae (Annelida) Off the Coast of North-eastern Japan: A Description of Lacydonia japonica sp. nov.

Lacydonia japonica sp. nov. (Annelida, Lacydoniidae) is described based on material found in sediments collected off the Pacific coast of northern Honshu, Japan, at depths of 262 m and 407 m. The sediments were obtained by a remotely operated vehicle equipped with a suction sampler during a Tohoku Ecosystem-Associated Marine Sciences (TEAMS) project in 2019. Lacydonia japonica sp. nov. belongs to the eyeless group of lacydoniids and is discriminated from the morphologically most similar congener, Lacydonia papillata Uschakov, 1958 by its reddish pigments on both the dorsal and ventral parapodial cirri and four pigment spots on the pygidium. To assess the phylogenetic position of the new species among other lacydoniids for which sequence data are available in public databases, analyses were performed using the mitochondrial cytochrome c oxidase subunit I (COI) and 16S rRNA, as well as the nuclear 18S rRNA and 28S rRNA genes. We additionally obtained some lacydoniids by sledging off western Japan, but these were severely fragmented and broken during collection. Using the paucity of morphological data, they were left unidentified as Lacydonia sp. but included in the molecular analyses. Genetic distances between Lacydonia eliasoni Hartmann-Schröder, 1996, Lacydonia japonica, and Lacydonia sp. off western Japan were 10.4-17.1% uncorrected p-distance (11.3-18.6% K2P) in terms of 658-bp COI sequences.

Taxonomic Reappraisal of Lineus longifissus Auct. (Nemertea: Pilidiophora) from Japan for the First Time in 122 Years.

Having been reported in 1898 for the first time from Japanese waters, the lineid heteronemertean Lineus longifissus auct. is known to inhabit tidal flats under the influence of the warm Kuroshio Current along the coasts of Honshu and southwestward, characteristically with a uniformly raisin-colored to black body lacking a caudal cirrus. The taxonomic identity of the Japanese L. longifissus auct. has been questioned by specialists because of some obvious morphological differences between Lineus longifissus (Hubrecht, 1887) s.str. (now in Heteronemertes Chernyshev, 1995), originally described from the subantarctic region. In the present study, we describe the Japanese L. longifissus auct. as Corsoua takakurai sp. nov. Before the present study, the genus Corsoua Corrêa, 1963 had been monotypic with the type species Corsoua kristenseni Corrêa, 1963 from the Caribbean. We infer the phylogenetic position of Corsoua takakurai within Lineidae based on the mitochondrial 16S rRNA and cytochrome c oxidase subunit I, and the nuclear 18S rRNA, 28S rRNA, and histone H3 genes.

Molecular systematics of the heteronemertean genus Dushia (Nemertea, Pilidiophora), with descriptions of D. wijnhoffae sp. nov. and D. nigra species complex comb. nov.

The heteronemertean genus Dushia Corrêa, 1963 was established for what was identified as D. atra (Girard, 1851) (originally Meckelia atra) based on material from littoral, shallow waters in Curaçao, while the nominal species Meckelia atra was originally described from deep water off Florida Cape. In this paper, we conclude that the type species for Dushia has been misidentified. Based on specimens from the Caribbean, we establish D. wijnhoffae Schwartz Norenburg sp. nov. to represent the true identity of the genus, according to Article 70.3.2 of the International Code of Zoological Nomenclature; Meckelia atra should be regarded as a nomen dubium. While the genus has remained monotypic since its establishment, our molecular analysis discovered a second member-or rather a group of members-from the West Pacific. This 'group of members', herein termed Dushia nigra (Stimpson, 1855) species complex comb. nov., involves i) at least two genetically separated biological entities, 0.136-0.148 (p-distance) and 0.152-0.168 (K2P) apart in terms of 513-bp COI sequences, which we interpret as likely to represent cryptic species, ii) three color forms, orange, brown, and black, with the last one occurring most frequently, and iii) four nominal species, Meckelia nigra Stimpson, 1855 (now Cerebratulus niger), Meckelia rubella Stimpson, 1855 (now Cerebratulus rubellus), Micrura formosana Yamaoka, 1939, and Micrura japonica Iwata, 1952. At present, however, we have no objective ground as to which of the four potentially available names (i.e., formosana, japonica, nigra, and rubella) should be allotted to the two cryptic species discovered in the analysis, because i) a single locality can harbor two cryptic species, ii) a single cryptic species may contain three different color morphs (i.e., orange, brown, black), and iii) no data from the type localities for these four nominal species are available at the moment. Our multi-locus analysis of heteronemerteans-for which 16S rRNA, COI, 18S rRNA, 28S rRNA, histone H3 genes are available in public databases-shows that Dushia wijnhoffae sp. nov. and Dushia nigra species complex comb. nov. form a clade, which is closely related to Gorgonorhynchus albocinctus Kajihara, 2015 and an undetermined heteronemertean that has been misidentified as Cerebratulus leucopsis (Coe, 1901). Members of Dushia thus show a vicariant Caribbean-West-Pacific distribution, indicating that the lineage predates the rise of the Isthmus of Panama.

A Population Without Females: Males of Baseodiscus delineatus (Nemertea: Heteronemertea) Reproduce Asexually by Fragmentation.

The valenciniid heteronemertean Baseodiscus delineatus (Delle Chiaje, 1825) was originally described from Naples, Italy, and shows a circumglobal distribution from tropical to temperate seas in both hemispheres. To investigate its reproductive biology, we performed intermittent year-round sampling from 2014 to 2018 in Misaki on the Pacific coast of Honshu, Japan. Of the 40 specimens obtained, 11 were males, while 29 were immature. No female specimen was confirmed during the sampling period. We additionally observed that B. delineatus is fissiparous. A male individual in captivity reproduced asexually by spontaneous posterior fragmentation, an ability that is described here for the first time among Valenciniidae. Autotomy occurred every 2-10 days, with each of the tail fragments having undergone anterior regeneration, which took about 24-36 days before completion of internal organs, such as ocelli, cerebral organs, brain, alimentary canal, rhynchocoel, and proboscis. A review of the literature suggests that the species was absent in the Misaki region 120 years ago. We assume a recent settlement of a male founder, which has putatively reproduced asexually to yield a clonal, unisexual population in Misaki.