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

A new sexannulate species of the genus Orobdella Oka, 1895, Orobdella mononokesp. n., is described on the basis of five specimens collected from Yakushima Island, Japan. Orobdella mononokesp. 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, tRNA(Cys), tRNA(Met), 12S rDNA, tRNA(Val) and 16S rDNA markers show that Orobdella mononokesp. 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 new quadrannulate species of Orobdella (Hirudinida,Arhynchobdellida, Orobdellidae) from western Honshu,Japan

A new quadrannulate species of Orobdella Oka, 1895, Orobdella naraharaetmagarum sp. n., from the mountainous region of western Honshu, Japan is described. Orobdella naraharaetmagarum is a small species with a body length of less than 5 cm. Phylogenetic analyses using nuclear 18S rRNA and histone H3, as well as mitochondrial cytochrome c oxidase subunit I, tRNA^Cys, tRNA^Met, 12S rRNA, tRNA^Val, 16S rRNA, tRNA^Leu and NADH dehydrogenase subunit 1 markers indicated that the present new species is the sister species of the quadrannulate Orobdella esulcata Nakano, 2010. Furthermore, mitochondrial DNA genealogy within Orobdella naraharaetmagarum demonstrated that this new species is divided into eastern and western lineages.     

A new quadrannulate species of Orobdella (Hirudinida,Arhynchobdellida, Orobdellidae) from central Honshu,Japan

A new quadrannulate species of Orobdella, Orobdella masaakikuroiwai sp. n., from the mountainous region of central Honshu, Japan is described. This is only the second small species known within this genus, with a body length of less than 4 cm for mature individuals. Phylogenetic analyses using nuclear 18S rDNA and histone H3 as well as mitochondrial COI, tRNA^Cys, tRNA^Met, 12S, tRNA^Val, 16S, and ND1 markers showed that Orobdella masaakikuroiwai sp. n. is the sister species of the quadrannulate Orobdella whitmani Oka, 1895. Phylogenetic relationships within Orobdella masaakikuroiwai sp. n. conducted using mitochondrial markers reveled a distinction between eastern and western phylogroups.     

Holotype redescription of Mimobdella japonica (Hirudinida, Arhynchobdellida, Erpobdelliformes) and taxonomic status of the genus Mimobdella

Mimobdella japonica Blanchard, 1897, the type species of the genus Mimobdella Blanchard, 1897, is redescribed based on the holotype. This species is characterized by the following characteristics: mid-body somites novem-annulate, two post-anal annuli, male gonopore in XI/XII, female gonopore in XII/XIII, 9 annuli (one full somite) between gonopores, strepsilaematous pharynx and three myognaths with stylets, possessing post-crop caeca in pairs, ovisacs reaching to XXI a2. The genus Mimobdella is placed under the family Salifidae, not Gastrostomobdellidae or Erpobdellidae, according to its possessing three myognaths bearing pharyngeal stylets.     

Chromosome identification and karyotype analysis of Podophyllum hexandrum Roxb. ex Kunth using FISH

Podophyllum hexandrum is an important high altitude medicinal plant from Himalaya. Somatic chromosomes of this species were studied to delineate and physical mapping of repetitive rDNA sites to provide landmarks in chromosome identification. The karyotype formula of this species was found to be 6m + 2sm + 2st + 2t with secondary constriction in the chromosome 1 and 7. The FISH analysis of rDNA sites showed 4 sites for 18S rDNA and 2 sites for 5S rDNA. The chromosome number 1, 2, 5 and 6 can be identified based on 18S rDNA sites in their short arm and chromosome 1 and 2 can be identified by 5S rDNA site in the centromere region. The estimated genome size of this plant is 16.07 pg (1C).     

A new species of the genus Orobdella (Hirudinida: Arhynchobdellida: Gastrostomobdellidae) from Kumamoto, Japan, and a redescription of O. whitmani with the designation of the lectotype

Orobdella whitmani has three syntype localities, Mt. Kinkazan, Gifu Pref., Mt. Kodaijiyama, Kyoto Pref. and Mt. Kimposan, Kumamoto Pref. The specimens from Mt. Kimposan can be distinguished from those from Mt. Kinkazan and Mt. Kodaijiyama by an annulation of somite XXVI, and possession of a tubular gastroporal duct. Specimens from Mt. Kimposan are described as representing a new species, Orobdella esulcata. Orobdella whitmani is redescribed with designation of a specimen from Mt. Kinkazan deposited at The University Museum, The University of Tokyo, as the lectotype.     

Two new ballistoconidium-forming yeast species, Bullera melastomae and Bullera formosana, found in Taiwan

Two yeast strains, the cells of which contained xylose and Q-10 as the major ubiquinone, were isolated from a plant leaf collected in Taiwan. These yeasts were found to represent two new species of the genus Bullera in the Hymenomycetes. Identification was based on the sequence analysis of the 18S rDNA, the internal transcribed spacer (ITS) regions and the D1/D2 domain of 26S rDNA. The yeasts are named Bullera melastomae sp. nov. and Bullera formosana sp. nov. In the phylogenetic trees based on 18S rDNA and D1/D2 domain of 26S rDNA sequences, these two species constitute a cluster connected with Dioszegia cluster in the Cryptococcus luteolus lineage.     

The karyotype of Nothoscordum arenarium Herter (Gilliesioideae, Alliaceae): A populational and cytomolecular analysis

The genus Nothoscordum Kunth comprises approximately 20 species native to South America. Karyologically, the genus is remarkable for its large chromosomes and Robertsonian translocations. Variation in chromosome number has been recorded in a few polyploid species and it is unknown among diploids. This study presents the chromosome number and morphology of 53 individuals of seven populations of N. arenarium Herter (2n = 10). In addition, karyotype analyses after C-banding, staining with CMA and DAPI, and in situ hybridization with 5S and 45S rDNA probes were performed in six individuals from one population. All individuals exhibited 2n = 10 (6M + 4A), except for one tetraploid (2n = 20, 12M + 8A) and one triploid (2n = 15, 9M + 6A) plant. C-banding revealed the presence of CMA(+) /DAPI (-) heterochromatin in the short arm and in the proximal region of the long arm of all acrocentric chromosomes. The 45S rDNA sites co-localized with the CMA (+) regions of the acrocentrics short arms, while the 5S rDNA probe only hybridized with the subterminal region of a pair of metacentric chromosomes. A change in the pattern of CMA bands and rDNA sites was observed in only one individual bearing a reciprocal translocation involving the long arm of a metacentric and the long arm of an acrocentric chromosome. These data suggest that, despite isolated cases of polyploidy and translocation, the karyotype of N. arenarium is very stable and the karyotypic instability described for other species may be associated with their polyploid condition.     

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island,Korea

There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.     

Identification of parasite DNA in common bile duct stones by PCR and DNA sequencing

We attempted to identify parasite DNA in the biliary stones of humans via PCR and DNA sequencing. Genomic DNA was isolated from each of 15 common bile duct (CBD) stones and 5 gallbladder (GB) stones. The patients who had the CBD stones suffered from cholangitis, and the patients with GB stones showed acute cholecystitis, respectively. The 28S and 18S rDNA genes were amplified successfully from 3 and/or 1 common bile duct stone samples, and then cloned and sequenced. The 28S and 18S rDNA sequences were highly conserved among isolates. Identity of the obtained 28S D1 rDNA with that of Clonorchis sinensis was higher than 97.6%, and identity of the 18S rDNA with that of other Ascarididae was 97.9%. Almost no intra-specific variations were detected in the 28S and 18S rDNA with the exception of a few nucleotide variations, i.e., substitution and deletion. These findings suggest that C. sinensis and Ascaris lumbricoides may be related with the biliary stone formation and development.     

Expanded description of Neoechinorhynchus (Hebesoma) manubrianus (Acanthocephala: Neoechinorhynchidae) from marine fish in Halong Bay, Vietnam

Neoechinorhynchus manubrianus Amin, Ha & Ha, 2011 (Acanthocephala: Neoechinorhynchidae) (formerly Neoechinorhynchus manubriensis Amin, Ha & Ha, 2011), was recently described based on optical microscopy of four males and two females (none was gravid) from caroun croaker, Johnius carouna (Cuvier), flower croaker, Nibea albiflora (Richardson), and silver croaker, Pennabia argentata (Houttuyen) (Sciaenidae) in Halong Bay, Vietnam. Subsequently, many more specimens became available from N. albiflora that were studied using SEM. SEM studies showed many additional features that were not possible to discern with optical microscopy. These included the prominent angulation of the anterior trunk, the presence of (1) anterio-dorsal and (2) undulating mid-lateral fin-like protrusions of the body wall, uniquely shaped eggs as well as details of trunk micropores, proboscis, bursa, and female gonopore. Microscopical examination of eggs from the new collection demonstrated the presence of polar prolongation of fertilization membrane which places N. manubriensis in the subgenus Hebesoma. The features of trunk angulation, trunk fins, and egg morphology further distinguish N. manubriensis from all other species of Neoechinorhynchus Stiles and Hassall, 1905 from Vietnam or from any where else in the world.     

First report of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from marine fish of the eastern seaboard of Vietnam, with the description of six new species

The occurrence of acanthocephalans of the genus Neoechinorhynchus Stiles and Hassall, 1905 in Vietnamese waters is reported for the first time. Six new species are described from seven species of marine fish of the families Belonidae, Clupeidae, Megalopidae, Mugilidae, and Sciaenidae, collected in Halong Bay of the eastern seaboard of Vietnam in 2008 and 2009. These are Neoechinorhynchus (Neoechinorhynchus) plaquensis n. sp. characterized by dermal plaques covering the entire trunk; Neoechinorhynchus manubriensis n. sp. with very long anterior proboscis hooks having roots with prominent anterior manubria and very small and equal middle and posterior hooks, two pseudo-retractors in the receptacle, simple vagina, and terminal gonopore; Neoechinorhynchus pennahia n. sp. with equal anterior and middle proboscis and somewhat smaller posterior hooks, and terminal female gonopore; Neoechinorhynchus ampullata with many giant nuclei in the body wall and lemnisci and a parareceptacle structure complex which includes pumping ampullas reported for the first time; Neoechinorhynchus (Neoechinorhynchus) longinucleatus n. sp. with very long giant nuclei in the Lemnisci, anteriorly twisted vagina, and subterminal female gonopore. Neoechinorhynchus (Neoechinorhynchus) ascus n. sp. is the second species of Neoechinorhynchus found with the parareceptacle structure/ampulla complex. Neoechinorhynchus (Neoechinorhynchus) johnii Yamaguti, 1929 of Bilqees, 1972 is not N. johnii because of proboscis armature and other discrepancies with the Yamaguti material. Notes on host distribution and feeding habits are also included.     

First report of Xiphinema brevicolle Lordello et Costa, 1961 (Nematoda, Longidoridae) in Japan

Mixed populations of Xiphinema americanum-group species were detected from a root zone soil sample of Japanese holly, Ilex crenata, during a survey for plant-parasitic nematodes of commercial ornamental plant nurseries in Chiba Prefecture, Japan. From the result of the morphological study, the species were identified as Xiphinema brevicolle and Xiphinema sp. This is the first record of Xiphinema brevicolle in Japan. Morphometrics of Xiphinema brevicolle generally agree with those of the type specimens and the topotype specimens. Xiphinema sp. morphometrically resembles Xiphinema paramonovi except for tail length. The mitochondrial COI region, the nuclear 18S rDNA and the nuclear large subunit rDNA D2/D3 region of the species were sequenced and compared in the molecular study. For the COI region, PCR primers were newly designed to obtain longer sequences, ca. 900 bp, than previously used. Sequence identities of COI, 18S and D2/D3 regions between these two populations were 84.0-84.1%, 99.9% and 98.1-98.2%, respectively. Phylogenetic analyses of maximum likelihood trees were carried out to compare genetic relationships among the group and some suggestions were made on the Xiphinema brevicolle-subgroup.     

New species, new records and new morphological characters of the genus Tillicera Spinola from China (Coleoptera, Cleridae, Clerinae)

Two new species of the genus Tillicera Spinola, 1841 from China are described and illustrated: Tillicera sensibilissp. n. from Yunnan (also from Myanmar, Thailand and Laos) and Tillicera weniisp. n. from Taiwan. Tillicera bibalteata Gorham, 1892, Tillicera hirsuta (Pic, 1926) and Tillicera michaeli Gerstmeier & Bernhard, 2010 are newly recorded from China. Tillicera auratofasciata (Pic, 1927) is newly recorded in some provinces of China. A key to species of the genus from China is provided. Relationships between species are discussed with emphasis on characters of male phallus, female internal reproductive organs and pit-like sensilla in male terminal antennomere, which is discovered in Tillicera for the first time. The present generic definition of Tillicera is discussed as well. Photos of terminalia of the previously known species are also provided for comparison.     

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 28S D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenic tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.     

Review of the Parasa undulata (Cai, 1983) species group with the first conifer-feeding larva for Limacodidae and descriptions of two new species from China and Taiwan (Lepidoptera,Limacodidae)

Although the caterpillars are well-known for the stings and magnificent coloration, the systematics of Limacodidae is historically neglected and chaotic due to the difficulty in matching the larval with adult stages as well as the very conservative and convergent adult morphology. One of the biggest taxonomic problems surrounds a collective group from Southeastern Asia, termed the “green limacodid moths”, which harbours at least 90 species placed in the genus Parasa Walker, 1859 and 14 “subunits”. The P. undulata group was previously composed of 3 species from China and Taiwan, and characterized only by wing pattern. This species group is extensively studied herein with two new species described, i.e. P. viridiflamma sp. n. (Taiwan) and P. minwangi sp. n. (S. China), and discovery of female genitalia of three species, presenting new phylogenetic insights in this potentially paraphyletic genus. In addition, one limacodid larva was found to be feeding exclusively on Picea (Pinaceae) in Taiwan. Its identity, Parasa pygmy Solovyev, 2010 in P. undulata group, is confirmed through matching its COI sequence to the adult. This discovery is also biologically significant because the previous known host breadth of Parasa was of polyphagy on various angiosperm plant families. This case, therefore, represents the first record of conifer-feeding behavior in this family as well as the first of specialized herbivory in the genus. Meanwhile, the background match between Picea leaves and larval coloration is shared with other Picea-feeding insects. This phenomenon is worth of further investigation in the aspect of convergent evolution of crypsis associated with a particular plant.     

Phylogenetic Relationships of Globodera millefolii,G. artemisiae,and Cactodera salina Based on ITS Region of Ribosomal DNA

Globodera millefolii and G. artemisiae are interesting because their type localities (Estonia and Russia, respectively) are geographically distant from those of the potato cyst nematodes and other Globodera species that seem to have originated in the Western world, and because the type host for each is a member of Compositae rather than Solanaceae. Sequence data for ITS1, ITS2, and 5.8S ribosomal DNA (ITS rDNA) for G. millefolii and G. artemisiae were nearly identical to sequence data for Cactodera salina from the rhizosphere of the estuary plant Salicornia bigelovii in Sonora, Mexico. The ITS rDNA sequences of these three species were all about 94% similar to those of two other Cactodera species for which ITS rDNA data were obtained. Phylogenetic analysis indicated that, based on the ITS rDNA data, G. millefolii and G. artemisiae are more closely related phylogenetically to the Cactodera species than to other nominal Globodera species. The molecular data further suggest that the genus Cactodera may comprise two or more morphologically similar but separate groups.     

Perlesta ephelida, a new Nearctic stonefly species (Plecoptera, Perlidae)

A new Nearctic species of Perlidae (Insecta, Plecoptera), Perlesta ephelidasp. n., is described from the male, female, and egg stages. This species has been previously reported as, or confused with, Perlesta shubuta Stark from several central and eastern U.S. states. Perlesta ephelida is distinctive from Perlesta shubuta and other regional Nearctic congeners mainly according to male genitalic and egg characteristics. Perlesta ephelida is a widely-distributed eastern Nearctic species, whereas Perlesta shubuta appears to be restricted to a narrow latitudinal belt in the Gulf Coast region from Louisiana east conservatively to the Florida panhandle. The egg of Perlesta shubuta is depicted with scanning electron microscopy for the first time.     

A New Species of Pristionchus (Rhabditida: Diplogastridae) and Its Bacterial Symbiont from Yixing,China

A new nematode species, Pristionchus entomophilus n. sp., was collected during a soil sample survey in Yixing of Jiangsu province, eastern China. P. entomophilus n. sp. is distinguished by its unique characteristics. This new species is mainly hermaphroditic, with males seldom found. The new nematode has a similar body length but has much narrower body width compared with P. pacificus. Its body is covered with longitudinal ridges: 12 ridges on head, 13 or 14 ridges in the middle, 11 and 7 ridges in front and rear of the anus, respectively. The eurystomatous form mouth includes a triangular dorsal tooth, a large claw-like right subventral tooth, and a row of five ventral denticles placed opposite the dorsal tooth. Only eight pairs of genital papillae and a pair of phasmids are present in the tail of the male as the sixth pair of papillae having seemingly been degenerated and lost. Molecular phylogenetic trees based on 18S rDNA confirmed that the new species belongs to the genus Pristionchus and is most closely related to P. pacificus. Moreover, the new species was found to be occasionally associated with the entomopathogenic bacterial strain 09FLYB1 of Serratia nematodophila and be able to stably transfer the bacterial strain for several generations.