Morphological, histological and gene-expression analyses on stolonization in the Japanese Green Syllid, Megasyllis nipponica (Annelida, Syllidae).

Benthic annelids belonging to the family Syllidae (Annelida, Errantia, Phyllodocida) exhibit a unique reproduction mode called "schizogamy" or "stolonization", in which the posterior body part filled with gametes detaches from the original body, as a reproductive unit (stolon) that autonomously swims and spawns. In this study, morphological and histological observations on the developmental processes during stolonization were carried out in Megasyllis nipponica. Results suggest that the stolon formation started with maturation of gonads, followed by the formation of a head ganglion in the anteriormost segment of the developing stolon. Then, the detailed stolon-specific structures such as stolon eyes and notochaetae were formed. Furthermore, expression profiles of genes involved in the anterior-posterior identity (Hox genes), head determination, germ-line, and hormone regulation were compared between anterior and posterior body parts during the stolonization process. The results reveal that, in the posterior body part, genes for gonadal development were up-regulated, followed by hormone-related genes and head-determination genes. Unexpectedly, Hox genes known to identify body parts along the anterior-posterior axis showed no significant temporal expression changes. These findings suggest that during stolonization, gonad development induces the head formation of a stolon, without up-regulation of anterior Hox genes.

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.

Fusceulimoides kohtsukai gen. et sp. nov., a Minute Eulimid Gastropod Parasitic on the Little Brittle Star Ophiactis savignyi in Central Japan.

Gastropods of over a dozen genera in the family Eulimidae have been identified as parasites of brittle stars, and many more remain to be discovered and described for a comprehensive understanding of the evolutionary history of their host-parasite relationships. In this study, we describe Fusceulimoides kohtsukai gen. et sp. nov., parasitic on the little brittle star, Ophiactis savignyi (Ophiactidae), in Kanagawa, central Japan. The new genus is distinguished from other eulimid genera by the combination of following seven conchological characters: (1) a very small size of up to 1.7 mm high, (2) a colorless translucent appearance, (3) a conical pupiform shape with a paucispiral protoconch, (4) slightly convex teleoconch whorls, (5) a remarkably large body whorl occupying 65-70% of the total shell height, (6) a broad, somewhat squarish and laterally expanded aperture with a strongly curved outer lip, and (7) a developed parietal callus without an indentation or depression in the umbilical area. A multi-locus molecular phylogeny revealed its distant relationship to Hemiliostraca + Pyramidelloides, a previously known clade of ophiuroid parasites, thereby suggesting multiple origins of this host-parasite association in Eulimidae.

Growth in two deep-sea associates: the octocoral Pseudogorgia bellona and the euryalid snake star Asteroschema ajax.

The deep-sea octocoral Pseudochrysogorgia bellona was recently described from specimens sampled on the Chesterfield Plateau, off New Caledonia. It is morphologically and genetically similar to the con-familial Metallogorgia melanotrichos, which is known to closely associate with a species of brittle star, Ophiocreas oedipus. These latter two species have never been observed separately and are thought to grow synchronously. The morphological similarity between M. melanotrichos and P. bellona makes the latter another possible host for ophiuroids. However, no brittle star was associated with P. bellona specimens from the type collection. In 2017, 130 P. bellona colonies were sampled near the type locality, and 98% were associated with Asteroschema ajax, a species closely related to O. oedipus. Mitochondrial DNA analysis confirmed the morphological identifications of both P. bellona and A. ajax. Uni- and multivariate statistical analyses were used to characterize the morphological space of both species to test if larger ophiuroids are associated with larger corals. Two variables were measured to estimate the size of the coral (total height and diameter of the skeletal axis at its base) and 9 variables were used to characterize the brittle star (disc and arm morphology). Morphological variables representing the size for both species were significantly correlated (Spearman rank correlation coefficient: 50%, p < 0.001), suggesting that larger ophiuroids indeed associate with larger corals. This is one of the rare studies that allowed comparison of growth in associated deep-sea invertebrates.

Parasitism-Induced Intersexuality in a Sexually Dimorphic Varunid Crab, Ptychognathus ishii (Decapoda: Varunidae).

Although many animals that perform sexual reproduction exhibit sexual dimorphism, individuals with intersex traits between the traits of males and females appear in some species, depending on environmental factors. Ptychognathus ishii, a varunid crab, exhibits distinctive sexual dimorphism in the morphology of its abdomen, chelipeds and setal tufts on the chelipeds. In this study, however, we report for the first time that intersex individuals with intermediate characters between those of males and females were occasionally found in wild populations. Morphological features of intersex individuals are described. Their taxonomic positions are identified based on DNA sequences of part of the mitochondrial cytochrome c oxidase I (COI) gene. It was shown that the intersexuality was induced by entoniscid parasites, because all intersex individuals were parasitized by entoniscid isopods, identified as Entionella sp. The apparent correlation between parasitism and morphological anomalies suggests that the parasitic isopods affect physiological conditions, leading to the feminization of male hosts.

Description of a New Brooding Species of Ophiodelos (Echinodermata: Ophiuroidea) from Japan.

A new species of brittle star, Ophiodelos okayoshitakai, is described from two specimens collected in Sagami Bay, central-eastern Japan. Photographic examination of the holotype specimen of the sole other congener, Ophiodelos insignis Koehler, 1930, indicates that Ophiodelos okayoshitakai sp. nov. is distinguished from O. insignis by i) the disc stumps covering on the dorsal side of the disc, ii) the dorsal and ventral arm plates being separated from each other on the proximal arm regions, iii) the dorsal arm plate being smooth, iv) the arm spines at proximal portion of the arm being six in number and smooth in shape, and v) the number and shape of the tentacle scales at proximal portion of the arm being up to two and spine-shaped adradially and oval abradially. Detailed morphological observations of this new species suggest the inclusion of Ophiodelos, whose familial affiliation remains unclear, in the suborder Ophiacanthina. More than 10 juveniles of various sizes were found in the disc of Ophiodelos okayoshitakai sp. nov., indicating a brooding reproduction. This is the first report of the genus Ophiodelos from Japanese waters. We also provided a nucleotide sequence for part of the cytochrome c oxidase subunit I (COI) gene in O. okayoshitakai sp. nov. for future studies of DNA barcoding and phylogeny.

A New Species of the Genus Ophiomonas Djakonov (Echinodermata: Ophiuroidea: Amphilepididae) from the Deep-Sea of Japan.

A new species, Ophiomonas shinseimaruae, is described based on five specimens collected from deep water settings, southeast of Cape Erimo, Hokkaido, Japan. Ophiomonas shinseimaruae sp. n. is distinguished from other congeners based on the following characters: elongate semi-circular and separated radial shields; triangular oral shields; flat and broad tentacle scales on the second tentacle pore; octagonal dorsal arm plates, approximately three times wider than long on proximal portion of the arm; and three arm spines present proximally on the arm. This is the first record of the genus Ophiomonas from Japanese waters. The COI nucleotide sequence for Ophiomonas shinseimaruae sp. n. is provided.

Morphogenesis and histogenesis during the arm regeneration in a basket star Astrocladus dofleini (Euryalida, Ophiuroidea, Echinodermata).

Basket stars, that is, Ophiuroidea in Echinodermata, exhibit distinctive morphological characteristics with their complicatedly branched arms that can regenerate immediately after mutilation. Although, in brittle stars, that is, ophiuroids with nonbranched arms, the arm regeneration process following accidental trauma or autotomy have been morphologically and histologically observed in several species, few studies have so far been carried out on the regeneration of branched arms in basket stars. In this study, the developmental and morphological features of arm regeneration in Astrocladus dofleini (Gorgonocephalidae, Euryalida, Euryophiurida), one of the most common basket star species in Japanese waters, was anatomically and histologically investigated. Results clearly showed the following phases during the arm regeneration: (a) repair phase, (b) early regenerative phase, (c) intermediate regenerative phase, (d) advanced regenerative Phase I, and (e) advanced regenerative Phase II. The morphogenetic process during the arm regeneration in the basket star showed similar patterns to those of nonbranched arms observed in other ophiuroids. However, differences were also seen between the two ophiuroid types, that is, there were some developmental features specific to the basket star. In the early regenerative phase, branching of coelomic cavities was observed prior to the formation of other tissues, probably inducing the later morphogenesis of branched arms. In addition, hard skeletal ossicles form rapidly at the advanced regenerative Phase II. These developmental features may have led the evolution of bizarre morphologies seen in basket stars, probably contributing to the adaptation to shallow waters from deep-sea habitats.

A taxonomic review of the genus Astrocladus (Echinodermata, Ophiuroidea, Euryalida, Gorgonocephalidae) from Japanese coastal waters.

Japanese species of the genus Astrocladus (Echinodermata, Ophiuroidea, Euryalida, Gorgonocephalidae) are reviewed. Astrocladus coniferus recently has two junior synonyms, A. dofleini Döderlein and A. pardalis Döderlein, however, status of these species has long been questioned. These species concepts have not been reviewed in recent years and no molecular phylogenetic analyses have been performed. Observations of the lectotype of A. coniferus, as well as the lectotype and four paralectotypes of A. dofleini and the holotype of A. pardalis have revealed that A. coniferus and A. pardalis are conspecific and morphologically distinguishable from A. dofleini. Astrocladus coniferus and A. dofleini are supported as distinct species by our molecular data. Additionally, we re-describe A. exiguus and A. annulatus, based on recently collected specimens and the holotype. We conclude that four species, A. annulatus, A. coniferus, A. dofleini, and A. exiguus are present in Japanese waters.

Species-specific nitrogen resorption proficiency in legumes and nonlegumes.

Nitrogen (N) resorption from senescing leaves enables plants to reuse N, thereby making them less dependent on current N uptake from the environment. Therefore, N resorption is important for survival and fitness, particularly for plants growing under low N supply. We studied N resorption from senescing leaves of 25 legumes and 25 nonlegumes in a temperate region of Japan to test the hypothesis that high N resorption has not evolved in legumes that fix atmospheric N2. The extent of N resorption was quantified by N resorption proficiency that is measured as the level to which leaf N concentration was reduced during senescence, i.e., the lower the senesced leaf N concentration, the lower the N loss through leaf fall and higher the N resorption proficiency. In support of the hypothesis, senesced leaf N concentration was higher in legumes than in nonlegumes, but there was considerable overlap between the groups. The higher senesced leaf N concentration of legumes was associated with a lower proportion of leaf N resorbed during senescence, particularly in species with higher leaf N concentrations. According to a hierarchical partitioning analysis, there was a large contribution of species to the total variance in the senesced leaf N concentration as opposed to a minor contribution of functional group (legume/nonlegume). This study reveals that legumes are not proficient at resorbing N from senescing leaves but that N2-fixation might not be the single most important determinant of N resorption.

The widely occurring brittlestar Ophiactis savignyi (Amphilepidida: Ophiactidae) as an epibiont on loggerhead sea turtle, Caretta caretta.

We describe the first occurrence of the widely occurring brittlestar Ophiactis savignyi (Müller Troschel, 1842) as epibionts on Caretta caretta (Linnaeus, 1758), the loggerhead sea turtles. On the sea turtle epibionts, the coronulid barnacles were well studied as listed in Hayashi (2013), and recently some crustaceans were collected from loggerhead sea turtles and described as new species (Tanabe et al. 2017; Tanaka and Hayashi 2019). In contrast, echinoderm epibionts listed from sea turtles are poorly understood (Table 1) and this study represents the novel discovery of an ophiuroid, which was not previously known to occur on the surface of this species.

A comprehensive taxonomic list of brittle stars (Echinodermata: Ophiuroidea) from submarine caves of the Ryukyu Islands, southwestern Japan, with a description of a rare species, Dougaloplus echinatus (Amphiuridae).

The ophiuroid species listed were collected by benthic faunal surveys from 2016 to 2018 in submarine caves of the Ryukyu Islands, southwestern Japan. The material includes 20 species from 8 families, Amphiuridae, Ophiocomidae, Ophiodermatidae, Ophiolepididae, Ophioleucidae, Ophiomyxidae, Ophionereididae and Ophiotrichidae. This is the first comprehensive report of submarine cave-dwelling ophiuroids from Japanese waters. Also provided is a redescription of a rare species, Dougaloplus echinatus (Ljungman, 1867), with a detailed description of its ossicle morphology. The species has previously been reported from Brazil, Australia, Eastern Indian Ocean to Southern China, in 11-118 m depth, and this is the first record of D. echinatus from Japanese waters. A brief discussion of the scarcity of submarine cave systems, and the need for their conservation, is provided.

First finding of anchialine and submarine cave dwelling brittle stars from the Pacific Ocean, with descriptions of new species of Ophiolepis and Ophiozonella (Echinodermata: Ophiuroidea: Amphilepidida).

Two new species, Ophiolepis cavitata n. sp. and Ophiozonella cavernalis n. sp., are described from the specimens collected in shallow water submarine caves in the Ryukyu Islands, southwestern Japan. Ophiolepis cavitata n. sp. is distinguished from the other congeners in having prominent tubercle-shaped knobs on dorsal arm plates; two sizes of disc scales on the aboral disc, the larger surrounded by 3-8 times smaller disc scales; larger scales concave; radial shields wider than long; a trio of distinct scales present distal to each pair of radial shields; oral shields partly concave; 3 arm spines on proximal portion of the arms. Ophiozonella cavernalis n. sp. is distinguished from the other congeners in having polygonal, uniform size disc scales; oral shields almost as long as wide, shallow concave proximal sides that end in a fairly sharp point medially; arms about 9 times longer than disc diameter; 3 cylindrical arm spines, of which the oral most one is slightly larger than other 2 spines. Ophiozonella cavernalis n. sp. is the second finding of an anchialine-endemic ophiuroid and the first finding from the Pacific Ocean.

A taxonomic review of the genus Astrodendrum (Echinodermata, Ophiuroidea, Euryalida, Gorgonocephalidae) with description of a new species from Japan.

A revision of the genus Astrodendrum Döderlein (Ophiuroidea, Euryalida, Gorgonocephalidae) is based on 14 specimens, including the holotypes of Astrodendrum galapagense A. H. Clark and Astroconus capense Mortensen and a syntype of Gorgonocephalus sagaminus Döderlein. A new species Astrodendrum spinulosum is described from the Pacific coast of Japan. It is distinguished from its congeners in having conical external ossicles on aboral side of the disc; bulges on lateral edges of proximal arms; 1 terminal projection on each arm spine of proximal portion of the arm; 2 or 3 secondary teeth on each hook-shaped arm spine of distal portion of the arm. A taxonomic key to all six species of the genus Astrodendrum is provided.

Euryalid brittle stars from the International Indian Ocean Expedition 1963-64 (Echinodermata: Ophiuroidea: Euryalida).

Fourteen species of euryalids are reported from collections made in the eastern and western Indian Ocean during the International Indian Ocean Expedition. Three new species are described: Astrocladus goodingi from the Comoro Islands, Astrocladus socotrana from the northwestern Indian Ocean near the island of Socotra, and Astrocyclus somaliensis from off northeastern Somalia. Range extensions are documented for Asteronyx luzonicus, Astroglymma sculptum, Astrocladus ludwigi, Asteromorpha capensis and Squamophis amamiensis.

Description of a New Subfamily, Astrocloninae (Ophiuroidea: Euryalida: Gorgonocephalidae), Based on Molecular Phylogeny and Morphological Observations.

Previous phylogenetic studies of the family Gorgonocephalidae (brittle stars and basket stars) have identified three subfamilies, Astrotominae, Astrothamninae, and Gorgonocephalinae. The genus Astroclon was tentatively assigned to the subfamily Astrothamninae in previous studies, but its morphology is enigmatic and molecular data of the genus was insufficient. Therefore, the systematic position of Astroclon required confirmation to reconstruct the accurate systematics of the Euryalida. In the present study, we sought to clarify the subfamilial classification in the family Gorgonocephalidae (Echinodermata: Ophiuroidea: Euryalida). We revisited molecular phylogenetics of the Gorgonocephalidae including Astroclon and the major taxa in the family. The molecular analysis supported monophyly of the two species of Astroclon and its clear separation from Astrothamninae. The two Astroclon species were also distinguished from the other two subfamilies, Astrotominae and Gorgonocephalinae. Astroclon and three other subfamilies were divided in the basal position of the Gorgonocephalidae, and were assigned to subfamilies. A new subfamily, Astrocloninae was monotypically established for Astroclon in addition to the previous three subfamilies. Morphology of the first ventral arm plates and tentacle pores also supported distinctiveness of the new subfamily.

Non-destructive morphological observations of the fleshy brittle star, Asteronyx loveni using micro-computed tomography (Echinodermata, Ophiuroidea, Euryalida).

The first morphological observation of a euryalid brittle star, Asteronyx loveni, using non-destructive X-ray micro-computed tomography (µCT) was performed. The body of euryalids is covered by thick skin, and it is very difficult to observe the ossicles without dissolving the skin. Computed tomography with micrometer resolution (approximately 4.5-15.4 µm) was used to construct 3D images of skeletal ossicles and soft tissues in the ophiuroid's body. Shape and positional arrangement of taxonomically important ossicles were clearly observed without any damage to the body. Detailed pathways inside the vertebral ossicles, lateral arm plates, and arm spines for passage of nerves and water vascular structures were observed. Inter-vertebral muscles were also observed. Forms and 3D arrangements of many important taxonomical characters of the euryalids were scrutinized by µCT in high enough resolution for taxonomic description of ophiuroids.

A taxonomic review of the genus Astrohelix Döderlein, 1930 including the synonymy of the subgenus Asteroporpa (Astromoana) Baker, 1980 to Astrohelix.

The holotypes of Astrohelix bellator bellator and Astrohelix bellator paucidens (Echinodermata: Ophiuroidea: Euryalida: Gorgonocephalidae) are redescribed. Based on these observations Asteroporpa (Astromoana) with Astrohelix is proposed. With the elevation of two subspecies of Asteroporpa (Astrohelix) bellator bellator and A. (Astrohelix) bellator paucidens to species level, A. (Astrohelix) comprises six species, A. (Astrohelix) bellator, A. (Astrohelix) indicus, A. (Astrohelix) koyoae, A. (Astrohelix) muricatopatella, A. (Astrohelix) paucidens and A. (Astrohelix) reticulata. These species can be distinguished by the shape of the ossicles on the aboral disc, on the oral plates, and length of terminal projection of cone-shaped epidermal ossicles around the lateral interradial disc. A taxonomic key to all six species of the subgenus A. (Astrohelix) is provided.

Molecular phylogeny based on increased number of species and genes revealed more robust family-level systematics of the order Euryalida (Echinodermata: Ophiuroidea).

Previous molecular analysis of the order Euryalida (Echinodermata: Ophiuroidea), has identified three monophyletic families, the Euryalidae, Asteronychidae and Gorgonocephalidae. However, family-level relationships have remained unresolved due to inadequate taxon sampling and insufficient molecular markers. Here, we present a family-level revision of the Euryalida based on sequences from mitochondrial genes (16S rRNA and COI) and a nuclear gene (18S rRNA) from 83 euryalid ophiuroids. The monophyly of the three families, Euryalidae, Asteronychidae and Gorgonocephalidae is confirmed. The Euryalidae and Asteronychidae+Gorgonocephalidae are assigned to superfamilies, the Euryalidea and the Gorgonocephalidea, respectively. Three subclades within the family Gorgonocephalidae are identified and assigned to three subfamilies; Astrotominae includes Astrocrius, Astrohamma and Astrotoma, Astrothamninae (subfamily nov.) includes Astrothamnus and Astrothrombus with Gorgonocephalinae including the remaining genera. Morphological characters are consistent with the newly recognised superfamilies and subfamilies.

A new genus Squamophis of Asteroschematidae (Echinodermata, Ophiuroidea, Euryalida) from Australia.

Squamophis, a new genus of brittle star is described. Two species are included in the genus: Squamophis amamiensis (Okanishi & Fujita, 2009) from south-western Japan and Squamophis albozosteressp. n. from north-western Australia. Squamophisgen. n. is distinguished from the other genera of the family Asteroschematidae by the following characters: each radial shield is single-layered and is completely covered by plate-shaped epidermal ossicles, and the relative length of the longest arm spine throughout the arms is as long as the length of the corresponding arm segment. Squamophis albozosteressp. n. is distinguished from Squamophis amamiensis in having white, slightly domed, plate-shaped epidermal ossicles on the aboral side of the body, the ossicles on aboral and lateral portion of the arms form transverse rows, and the other part of aboral side of disc and basal to middle portion of arms are brown but tip of the arms are light purple.