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).

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 genusBoninia. 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 speciesBoninia urusp. nov. from gravelly sediments and (ii) the epibenthic speciesBoninia yambarensissp. 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 newBoniniaspecies and four other congeners, for which molecular sequences were available in public databases [Boninia antillara(epibenthic),Boninia divae(epibenthic),Boninia neotethydis(interstitial), and an unidentifiedBoniniasp. (habitat indeterminate)], revealed that the two interstitial species (B.neotethydisandB.urusp. nov.) were not monophyletic among the three epibenthic species. According to ancestral state reconstruction analysis, the last common ancestor of the analyzedBoniniaspecies 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 sympatricB.urusp. nov. andB.yambarensissp. nov. were not in a sister relationship, indicating that they colonized the same beach independently rather than descendedin situfrom a common ancestor that migrated and settled at the beach.

Among ~1300 species of world nemerteans, seven species in five genera of lineid heteronemerteans have been known to possess a branched proboscis. In this paper, we describe the eighth branched-proboscis species: Gorgonorhynchus citrinus sp. nov. from Okinawa, Japan. We also report Gorgonorhynchus cf. repens Dakin & Fordham, 1931 with uniformly orange body, as a new member for the Japanese nemertean fauna. We infer the phylogenetic relationships between these forms and other members of Lineidae McIntosh, 1874 for which partial sequences of the mitochondrial 16S rRNA and cytochrome c oxidase subunit I, and the nuclear 18S rRNA, 28S rRNA, and histone H3 genes are available in public databases, along with newly sequenced data of another branched-proboscis heteronemertean, Polydendrorhynchus zhanjiangensis (Yin & Zheng, 1984) from China. In the resulting tree, Gorgonorhychus Dakin & Fordham, 1931 was sister group to non-branched-proboscis Dushia Corrêa, 1963, whereas P. zhanjiangensis was sister group to likewise non-branched-proboscis Cerebratulus lacteus (Leidy, 1851). http://zoobank.org/urn:lsid:zoobank.org:pub:685992C5-F595-4C28-9178-256D945E595A

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.

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.

We describe a new species of polyclad flatworm, Prosthiostomum torquatum sp. nov., from the rocky intertidal zone in Shirahama, on the Pacific coast of middle Honshu, Japan. Prosthiostomum torquatum is characterised by a unique dorsal colouration, which is comprised of i) numerous orange maculae and blue dots dispersed all over the dorsal surface, ii) a transverse dark-brown line in the anterior part of the body running slightly anterior to another transverse white line, both of which are slightly bent backwards at mid-point, and iii) an incomplete, mesh-like, median band made by dark-brown pigments, fading away posteriorly. By this dorsal colour pattern, the new species can be distinguished from similar congeners such as P. trilineatum Yeri and Kaburaki, 1920 and P. komaii Kato, 1944. We performed a molecular phylogenetic analysis based on 462-bp partial mitochondrial cytochrome c oxidase subunit I (COI) sequences of four species of Prosthiostomidae currently available in public databases in addition to that of P. torquatum. In the resulting tree, P. torquatum was sister to Lurymare clavocapitata Marquina, Aguado, and Noreña, 2015 originally described from Lizard Island, Australia. While these two share a similar dorsal colouration, P. torquatum can be distinguished from L. clavocapitata by i) the absence of a common muscle bulb/sheath enclosing the whole male copulatory apparatus, ii) the median mesh-like band comprised of dark-brown pigments, and iii) COI uncorrected p-distance being 0.094. As a result, our phylogenetic tree indicates the possibility that Prosthiostomum and Lurymare as currently diagnosed may not be monophyletic, and that the common muscle bulb enclosing the whole male copulatory apparatus may not be appropriate as a diagnostic character for Lurymare.