Molecular Evidence for Cryptic Speciation in the Cyclophorus fulguratus (Pfeiffer, 1854) Species Complex (Caenogastropoda: Cyclophoridae) with Description of New Species

A high degree of intraspecific variation, both genetic and in shell morphology, of the operculate land snail Cyclophorus fulguratus (Pfeiffer, 1854) suggests that its classification as a single species warrants reconsideration. We sequenced two nuclear (18S and 28S) and two mitochondrial (16S and COI) genes of 46 C. fulguratus specimens and used them to estimate the phylogeny and to determine the validity of species boundaries. Molecular phylogenetic analyses revealed the presence of three lineages corresponding to three geographically disjunctive populations of C. fulguratus in Thailand. Likelihood tests of topologies significantly supported the non-monophyly of the C. fulguratus–complex and Bayesian species delimitation analysis significantly supported the potential representation as distinct species of these three lineages. Discriminant function analysis based on geometric-morphometrics of shell shape allowed for significant distinction of these three candidate species, although they revealed a considerable degree of overlap of shell shape reflecting their crypsis morphologically. The diagnostic characters are provided by color pattern, pattern of protoconch and pattern of jaw. In conclusion, the results support that the C. fulguratus s.l., as currently recognized, consists of three distinct species in Thailand: C. fulguratus s.s., C. rangunensis and C. abditus sp.nov., which are described herein.     

Taxonomic revision of the Elephant Pupinid snail genus Pollicaria Gould, 1856 (Prosobranchia,Pupinidae)

The status of species currently assigned to the Southeast Asian Elephant Pupinid snail genus Pollicaria Gould, 1856 is reassessed. Shell, radular and reproductive morphology are investigated and analysed with reference to karyotype patterns previously reported and to distribution patterns among the species. Six previously described species are recognised: Pollicaria gravida (Benson, 1856), Pollicaria myersii (Haines, 1855), Pollicaria mouhoti (Pfeiffer, 1862), Pollicaria elephas (Morgan, 1885), Pollicaria crossei (Dautzenberg & d’Hamonville, 1887) and Pollicaria rochebruni (Mabille, 1887). A new subspecies, Pollicaria mouhoti monochroma ssp. n.,is proposed and a dichotomous key to species is provided.     

Systematics of the freshwater leech genus Hirudinaria Whitman, 1886 (Arhynchobdellida,Hirudinidae) from northeastern Thailand

In total, 435 specimens of the Southeast Asian freshwater leech species within the Hirudinidae family were collected from 17 locations of various types of aquatic habitats in northeastern Thailand. They were all morphologically placed within the genus Hirudinaria Whitman, 1886 and there were three distinct species: the common Hirudinaria manillensis, 78.2% of all collected specimens and at all 17 locations, Hirudinaria javanica at 20.3% of collected samples and from five locations and a rarer unidentified morphospecies (Hirudinaria sp.) with six samples from only two locations. The karyotypes of these three species were examined across their range in this study area for 38, 11 and 6 adult specimens of Hirudinaria manillensis, Hirudinaria javanica and Hirudinaria sp., respectively. This revealed different chromosome numbers among all three species, with Hirudinaria javanica having n = 13, 2n = 26, Hirudinaria manillensis lacked one small chromosome pair with n = 12, 2n = 24, and the unknown Hirudinaria sp. differed from any known Hirudinaria karyotypes in exhibiting a higher chromosome number (n = 14, 2n = 28) and a gradual change in size from large to small chromosomes. This suggests that the unknown Hirudinaria sp. is a new biological species. However, phylogenetic analysis based upon a 658 bp fragment of the cytochrome oxidase subunit I gene placed this unknown morphospecies within the Hirudinaria manillensis clade, perhaps then suggesting a recent sympatric speciation, although this requires further confirmation. Regardless, the chromosomes of all three species were asymmetric, most with telocentric elements. A distinct bi-armed chromosome marker was present on the first chromosome pair in Hirudinaria javanica, whilst it was on pairs 1, 2, 3 and 5 in Hirudinaria manillensis, and on pairs 3 and 5 for the unknown Hirudinaria sp.     

Illustrated type catalogue of Amphidromus Albers, 1850 in the Natural History Museum,London, and descriptions of two new species

The collection of the Southeast Asian tree snail genus Amphidromus Albers, 1850 at the Natural History Museum, London includes more than 100 lots of type specimens representing 85 name-bearing types, 9 paratypes and 6 paralectotypes, and one nomen nudum. Lectotypes are here designated for Amphidromus cambojiensis, Amphidromus perakensis globosus, Amphidromus columellaris gloriosa, Amphidromus maculiferus inflata, Amphidromus lepidus, Amphidromus sinistralis lutea, Amphidromus moniliferus, Amphidromus maculiferus obscura, Amphidromus sinistralis rosea and Amphidromus sinensi vicaria. In addition, the missing types of A.A. Gould were discovered and their type status is discussed. A complete catalogue of these types, including colour photographs is provided for the first time. After examining these type specimens, two new Amphidromus species, Amphidromus (Syndromus) globonevilli Sutcharit & Panha, sp. n. and Amphidromus (Syndromus) principalis Sutcharit & Panha, sp. n. were recognized and are described herein.     

Biochemical assessment of the taxonomic diversity of the operculate land snail, Cyclophorus fulguratus (Gastropoda: Cyclophoridae), from Thailand

Allozyme variation was examined in 223 samples of the operculate land snail Cyclophorus fulguratus from 13 localities across three regions of Thailand. Using horizontal starch gel electrophoresis, 13 allozyme presumed loci (12 polymorphic) were screened. Heterozygosity was moderate in C. fulguratus (H_exp = 0.008–0.127) with a high genetic heterogeneity among samples (F_st = 0.734). Populations showed a greater genetic differentiation in central Thailand (F_st = 0.380) than in northeastern Thailand (F_st = 0.108), suggesting frequent gene flow among populations in northeastern Thailand. C. fulguratus exhibits a strong pattern of isolation by distance over the entire tested species range in Thailand and may potentially have been involved in an extensive local fragmentation. Results of the distance analysis revealed that large genetic divergence has occurred among the central, northeastern and eastern Thailand groups [D = 0.361–0.701], strongly suggesting populations from these three geographical regions may actually represent or else be evolving into separate species.     

Molecular Evolution of the HIV-1 Thai Epidemic between the Time of RV144 Immunogen Selection to the Execution of the Vaccine Efficacy Trial

The RV144 HIV-1 vaccine trial (Thailand, 2003 to 2009), using immunogens genetically matched to the regional epidemic, demonstrated the first evidence of efficacy for an HIV-1 vaccine. Here we studied the molecular evolution of the HIV-1 epidemic from the time of immunogen selection to the execution of the efficacy trial. We studied HIV-1 genetic diversity among 390 volunteers who were deferred from enrollment in RV144 due to preexisting HIV-1 infection using a multiregion hybridization assay, full-genome sequencing, and phylogenetic analyses. The subtype distribution was 91.7% CRF01_AE, 3.5% subtype B, 4.3% B/CRF01_AE recombinants, and 0.5% dual infections. CRF01_AE strains were 31% more diverse than the ones from the 1990s Thai epidemic. Sixty-nine percent of subtype B strains clustered with the cosmopolitan Western B strains. Ninety-three percent of B/CRF01_AE recombinants were unique; recombination breakpoint analysis showed that these strains were highly embedded within the larger network that integrates recombinants from East/Southeast Asia. Compared to Thai sequences from the early 1990s, the distance to the RV144 immunogens increased 52% to 68% for CRF01_AE Env immunogens and 12% to 29% for subtype B immunogens. Forty-three percent to 48% of CRF01_AE sequences differed from the sequence of the vaccine insert in Env variable region 2 positions 169 and 181, which were implicated in vaccine sieve effects in RV144. In conclusion, compared to the molecular picture at the early stages of vaccine development, our results show an overall increase in the genetic complexity of viruses in the Thai epidemic and in the distance to vaccine immunogens, which should be considered at the time of the analysis of the trial results.     

The semi-aquatic freshwater earthworms of the genus Glyphidrilus Horst, 1889 from Thailand (Oligochaeta,Almidae) with re-descriptions of several species

The semi-aquatic freshwater earthworm genus Glyphidrilus Horst, 1889 from Thailand was investigated based on extensive recent collecting. The species in this genus were characterized by their external and internal morphological characters of the location of wings, genital openings, genital organ structures and their locations, as well as the dimensions of body length and number of segments. Several type specimens were compared with both previous and newly collected materials. Ten new species are described from several river systems in Thailand; as Glyphidrilus borealis sp. n., Glyphidrilus chaophraya sp. n., Glyphidrilus chiensis sp. n., Glyphidrilus huailuangensis sp. n., Glyphidrilus kratuensis sp. n., Glyphidrilus quadratus sp. n., Glyphidrilus trangensis sp. n., Glyphidrilus wararamensis sp. n., Glyphidrilus vangthongensis sp. n. and Glyphidrilus vesper sp. n. Each species is endemic to a single river system. All 26 previously described species are re-described, and eight lectotypes have been designated. An identification key and a morphological comparison summary are provided.     

An annotated catalogue of type specimens of the land snail genus Cyclophorus Monfort, 1810 (Caenogastropoda,Cyclophoridae) in the Natural History Museum,London

The collection of land caenogastropod snails in the genus Cyclophorus Monfort, 1810 housed in the Natural History Museum, London (NHM), includes 52 type lots. Lectotypes have been designated for 43 available species-level names to stabilize existing nomenclature, two previously designated lectotype, two holotypes, one paratype, one syntype, one possible syntype and two paralectotypes are also listed. A complete catalogue of the Cyclophorus types in NHM, London is provided for the first time.