A new sexannulate species of Orobdella (Hirudinida, Arhynchobdellida, Orobdellidae) from Yakushima Island, Japan

Abstract

A new sexannulate species of the genus Orobdella Oka, 1895, Orobdella mononoke sp. n., is described on the basis of five specimens collected from Yakushima Island, Japan. Orobdella mononoke sp. n. differs from other sexannulate Orobdella species in its possessing the following combination of characters: dorsal surface bicolor in life, I–XIII, XXVII and caudal sucker grayish purple, XIV–XXVI amber, male gonopore at XI c11/c12, female gonopore at XIII b2, 8 + 1/2 between gonopores, tubular but bulbous at junction with crop gastroporal duct, epididymides in XV–XIX, and atrial cornua ovate. Phylogenetic analyses using nuclear 18S rDNA and histone H3, and mitochondrial COI, tRNACys, tRNAMet, 12S rDNA, tRNAVal and 16S rDNA markers show that Orobdella mononoke sp. n. is closely related to Orobdella esulcata Nakano, 2010 from Kyushu, Japan, and two species, Orobdella dolichopharynx Nakano, 2011 and Orobdella shimadae Nakano, 2011, from the Ryukyu Archipelago, Japan.

A molecular phylogeny of Pseudocrangonyx from Japan, including a new subterranean species (Crustacea, Amphipoda, Pseudocrangonyctidae)

“…Those for the other sequences were performed using the modified method outlined by Nakano (2012) and ; reactions were performed using a T-100 Thermal Cycler (Bio-Rad). When using a PC-320 Thermal Cycler, the PCR mixtures were heated to 94°C for 7 min, followed by 35 cycles at 94°C (45 s), 42°C (1 min) and 72°C (1 min), and a final extension at 72°C (7 min).…”

Four new species of the genus Orobdella from Shikoku and Awajishima island, Japan (Hirudinida, Arhynchobdellida, Orobdellidae)

“…The mid-body somite annulation of the known sexannulate Orobdella species was described as being composed of b1, b2, a2, b5, c11 and c12 (Nakano 2011b(Nakano , 2012a. However, the annulation of somites VII [a1 = a2 < b5 (ventrally c9 = c10) > b6] and VIII [b1 = b2 = a2 < b5 (c9 = c10) > b6] of one specimen, KUZ Z1359, clearly suggests that b5 is divided into c9 and c10, and b6 remains undivided in O. yamaneae.…”

“…Primer sets for the PCR and cycle sequencing (CS) reactions used in this study were as follows: for 18S rRNA, A and L (PCR and CS), C and Y (PCR and CS), as well as O and B (PCR and CS) (Apakupakul et al 1999); for histone H3 (H3), H3aF and H3bR (PCR and CS) (Colgan et al 1998); for cytochrome c oxidase subunit I (COI), LCO1490 (PCR and CS) and HCO2198 (CS) (Folmer et al 1994), and LCO-in (CS) and HCO-out (PCR and CS) (Nakano 2012b) or HCO-outout (PCR and CS) (Nakano 2012a , and 12SA-in and 12SB-out (PCR and CS) (Nakano 2012b); for tRNA Leu and NADH dehydrogenase subunit 1 (ND1) (tRNA Leu -ND1), LND3000 and HND1932 (PCR and CS) (Light and Siddall 1999). The PCR reactions and DNA sequencing were performed using the modified method mentioned in Nakano (2012a). The PCR reactions were performed using a GeneAmp PCR System 2700 and 9700 (Applied Biosystems) as well as a T100 Thermal Cycler (Bio-Rad).…”

“…According to taxonomic studies on sexannulate Orobdella species (Nakano 2011b(Nakano , 2012a, Orobdella okanoi differs from the four known sexannulate species, O. dolichopharynx Nakano, 2011b, O. ijimai Oka, 1895, O. mononoke Nakano, 2012aand O. shimadae Nakano, 2011b, as well as the new sexannulate species described below in having the following characteristics (Table 4): dorsal surface reddish, somite VII quinquannulate, somite VIII sexannulate, 8 + 1/2 annuli between gonopores, pharynx reaching to XIV, gastroporal duct bulbous, epididymides in XV to XVII, pre-atrial loop absent and atrial cornua ellipsoid. Orobdella okanoi is clearly distinguished from quadrannulate and octannulate species in having sexannulate mid-body somites.…”

First Record of <i>Mimobdella japonica</i> (Hirudinida: Arhynchobdellida: Salifidae) from Hachijojima Island, Izu Islands, Japan, with a Comment on the Genetic Diversity of the Species

“…Primer sets for the PCR and cycle sequencing (CS) reactions used in this study were as follows: for cytochrome c oxidase subunit I (COI), LCO 1490 and HCO 2198 (PCR and CS) (Folmer et al 1994), and LCO-in and HCO-out (PCR and CS) (Nakano 2012a); for tRNA Cys , tRNA Met , 12S ribosomal RNA, tRNA Val , and 16S ribosomal RNA (tRNA Cys -16S), 12SA-out and 12SB-in (PCR and CS), and 12SA-in and 12SB-out (PCR and CS) (Nakano 2012a); for tRNA Leu and NADH dehydrogenase subunit 1 (tRNA Leu -ND1), LND3000 and HND1932 (PCR and CS) (Light and Siddall 1999). The PCR reactions and DNA sequencing were performed using the modified method outlined by Nakano (2012b). The PCR reactions were performed using a GeneAmp PCR System 2700 and a GeneAmp PCR System 9700 (Applied Biosystems, Waltham, USA) as well as a T100 Thermal Cycler (Bio-Rad, Hercules, USA).…”