Contributions of a small collection of terrestrial microsnails (Pupilloidea, Hypselostomatidae) from Myanmar with description of three new species.

Land snails were collected for the project 'Conserving Myanmar's Karst Biodiversity' from the limestone karsts in Mon, Kayin, and Shan states and in the regions of Tanintharyi and Mandalay between 2015 and 2017, through cooperation with Fauna and Flora International (FFI) and the Forestry Department of Myanmar. Here, we report on a portion of the collection, and list 17 species from seven genera of the Hypselostomatidae microsnails. Three new species from two genera are described as Bensonellataiyaiorum Tongkerd & Panha, sp. nov., B.lophiodera Tongkerd & Panha, sp. nov., and Gyliotrachelaaunglini Tongkerd & Panha, sp. nov. All new species are known only from the type locality in Shan State (Bensonella) and Kayin State (Gyliotrachela). A new combination of Acinolaemusdayanum and three newly recorded species, namely A.cryptidentatus, B.anguloobtusa and G.hungerfordiana are discussed. The low morphological variability of the widely distributed G.hungerfordiana is discussed, and two species are proposed for formal synonymisation. Constituting the first records for Myanmar, five species of Bensonella and two species of Acinolaemus were collected.

Global species delimitation of the cosmopolitan marine littoral earthworm Pontodrilus litoralis (Grube, 1855).

The marine littoral earthworm Pontodrilus litoralis (Grube, 1855) is widely distributed and is reported as a single species. This study utilized an integrative taxonomic approach based upon morphological examination, phylogenetic reconstruction, and molecular species delimitation, to test whether the taxon is a single species or a species complex. For this, a total of 114 P. litoralis specimens collected from North America, Africa, Australia and Oceania, Europe and Asia were used. The phylogenetic analyses revealed deeply divergent mitochondrial lineages and a high level of genetic diversity among P. litoralis populations. Both single and multi-locus species delimitation analyses yielded several molecular operational taxonomic units. Therefore, due to the homogeneity of morphological characteristics, it is likely that the morphospecies P. litoralis is a complex of four or more cryptic species, suggesting that more sampling is required and that the population structure genetic data and gene flow need to be investigated.

Four new terrestrial earthworm species from the northeast Thailand (Oligochaeta, Megascolecidae).

Earthworm specimens collected from Sakon Nakhon and Nakhon Phanom, northeast Thailand, were found to contain four new species in the family Megascolecidae, with one species in the genus Metaphire Sims & Easton, 1972, and the other three in the genus Amynthas Kinberg, 1867. These are herein named Metaphiresongkhramensis Chanabun & Panha, sp. nov. in the sexthecal houlleti species group, and Amynthassakonnakhonensis Chanabun & Panha, sp. nov., A.auriculus Chanabun & Panha, sp. nov., and A.bantanensis Chanabun & Panha, sp. nov. in the sexthecal aelianus species group. Metaphiresongkhramensis Chanabun & Panha, sp. nov. occurs in dark clay soil of the oxbow lake of the river, Amynthassakonnakhonensis Chanabun & Panha, sp. nov. occurs in wetland area, A.auriculus Chanabun & Panha, sp. nov. occurs in dark sandy loam habitats of mixed deciduous forest while the following species, A.bantanensis Chanabun & Panha, sp. nov. occurs in sandy loam habitats of paddy fields. Descriptions of the new species, including illustrations of the external and internal morphological characteristics, are provided.

A new discovery of the bioluminescent terrestrial snail genus Phuphania (Gastropoda: Dyakiidae).

The mysterious world of the bioluminescent molluscs in terrestrial ecosystems is mesmerizing, but Quantula striata was previously the only terrestrial mollusc known to be luminescent. Here, we document the new discovery of bioluminescence in four land snails, namely Phuphania crossei, P. globosa, P. carinata, and P. costata. Our observations establish clearly that these four species of Phuphania produce a continuous greenish light from the light-emitting cells located within the mantle and the foot, and that its bright luminescence is intracellular and is not due to any luminous secretion. Although both Quantula and Phuphania can produce a green light, the luminescence patterns are different. The luminescence displayed by Quantula is rhythmical blinking or flashing, while Phuphania glows continuously. In addition, the bioluminescence in Q. weinkauffiana is confirmed, which is similar to that in the related species, Q. striata.

Review of the pill millipede genus Hyperglomeris Silvestri, 1917 (Diplopoda, Glomerida, Glomeridae) with description of two new species from Laos.

The pill millipede genus Hyperglomeris Silvestri, 1917 is reported from Laos for the first time. Two new species, namely H.bicaudata Likhitrakarn, sp. nov. and H.inkhavilayi Likhitrakarn, sp. nov., from Houaphanh and Khammouane provinces, northern Laos, are described and illustrated based on morphological characters and molecular analyses. Sequences of COI gene were used as DNA barcoding markers, and successfully supported the accurate identification of other Glomeridae species. Interspecific divergence of the COI uncorrected p-distance between these new species and other Hyperglomeris species ranged from 7.84-13.07%, while the intraspecific divergence was 0.45% in H.inkhavilayisp. nov. and 5.3% in H.bicaudatasp. nov. The updated status of Hyperglomeris, a map of its distribution, and identification keys for all species are given.

Systematic revision of the snorkel snail genus Rhiostoma Benson, 1860 (Gastropoda, Caenogastropoda, Cyclophoridae) with descriptions of new species.

The snorkel snail genus Rhiostoma Benson, 1860 is comprised of terrestrial cyclophorid snails with wide-ranging species diversity and radiation in Southeast Asia. The typical characters of the genus are a depressed shell, a detached and descending portion of the last whorl with a distinctive peristomal breathing device attached, and a calcareous cup-shaped operculum. Herein, we have revised the systematics of extant species based on shell morphology combined with COI barcoding. From these thirty recognised species, twelve are described as new to science: R. ? amarapuraensesp. nov., R.anceyisp. nov., R.breviocollarsp. nov., R.ebenozosterasp. nov., R.cheliopegmasp. nov., R.furfurosumsp. nov., R.gnomus, sp. nov., R.lannaensesp. nov., R.laoensesp. nov., R.platymorphasp. nov., R.rhothonotaphrosasp. nov., and R.tigrinasp. nov. All conchological characters are provided via illustrations of type specimens and living snails, and descriptions of the shells and radulae. Phylogenetic analysis based on partial COI gene sequences strongly supports the designated morphospecies and a monophyletic Rhiostoma, confirming that all pterocyclinid snails with a calcareous, cup-shaped operculum belong to the same clade. A high intra-specific divergence was observed in R.jalorensis and R.housei populations from locations in close proximity, suggesting a lower dispersal and higher level of isolation. The low inter-specific divergence found in R.hainesi, R.samuiense, R.asiphon, and R.rhothonotaphrosasp. nov. supports their recent diversification and local adaptation, and is congruent with their marked morphological differences. Finally, nine formerly Rhiostoma-placed species were reclassified into either the genus Cyclotus or the genus Opisthoporus.

Further notes on the taxonomy of the land snail family Clausiliidae Gray, 1855 (Stylommatophora, Helicina) from Myanmar with description of two new species.

This study presents a complete species list of the door snails inhabiting Myanmar, updated to now include 33 taxa, and provides taxonomic notes together with a re-description of the shell, radula, and genitalia for 13 species and subspecies, including Oospiraphilippiana, the type species of the genus Oospira. The snails previously treated as subspecies or synonyms of Oospiragracilior and Oospiramagna are reclassified and recognized as distinct species. The lectotype of Oospirainsignis has been clarified and an illustration of the original type specimen provided. A long-overlooked species, Oospiraandersoniana, has been collected and redescribed herein. Two new species from the limestone karsts in the Salween River Basin are introduced: Oospiraluneainopsis Man & Panha, sp. nov. and Oospirazediopsis Man & Panha, sp. nov. A synoptic view of all clausiliid taxa known from Myanmar is presented along with taxonomic information and distributional records. Photographs of the type materials for all taxa are provided for further comparison or, if unavailable, photographs of the examined specimens or the original figure from the literature.

Ethogram and classification of the mating and egg-laying behaviour of the Southeast Asian apple snail Pilavirescens (Deshayes, 1824) (Mollusca, Gastropoda, Caenogastropoda, Ampullariidae).

The status of the indigenous Southeast Asian apple snails belonging to the genus Pila is of concern due to their fast rate of population decline, possibly as a result of multiple factors including habitat loss or disturbance and the introduction of globally-invasive apple snails, Pomacea spp. Conservation actions, including captive breeding of the native Pila species, have been suggested as urgent remedial practices, but the lack of knowledge regarding the fundamental reproductive biology of indigenous Pila spp. makes such practices difficult. In the present study, observations on the mating and egg-laying behaviour of an economic valuable apple snail native to Southeast Asia, P.virescens, were conducted using video recording to examine and describe their reproductive behaviour under a laboratory condition. A total of 15 types of mating and seven egg-laying behaviour were recorded. The mating sequence which subsequently resulted in egg laying was comprised of seven types of major sequential behaviour: mate probing, mounting, shell circling, positioning, insemination posture, sheath withdrawal and dismounting. Rejection of mating attempts by the female was frequently observed. Egg laying occurred during either day or night. A sequence of seven distinct types of behaviour were performed during oviposition: climbing, positioning, forming a temporary tube, mucous secreting, egg depositing, leaving and resting. Overall, these results provide an understanding of the egg-laying behavioural process and highlight its complexity in P.virescens. In addition, detailed ethograms of mating and egg-laying behaviour were derived. These will promote further systematic comparative studies of the reproductive behaviour of apple snails.

Responses to salinity in the littoral earthworm genus Pontodrilus.

The cosmopolitan littoral earthworm Pontodrilus litoralis is distributed in tropical and sub-tropical coastal habitats, whereas P. longissimus is reported only in the Thai-Malay coastal line. In the present study, we examined the difference in salinity effect on the survival rate, wet weight (hereafter weight) change, behaviour, and osmolality of these two Pontodrilus species. A 28 d exposure to varying salinity concentration (0-50 ppt) revealed that P. litoralis is able to survive over a wide salinity range than P. longissimus, with the latter species exhibiting a low survival rate over the same salinity range. During short-term exposure (0-96 h) to a salinity of less than 30 ppt, P. litoralis exhibited weight gain and this was significant in the first 12 h of exposure. However, P. longissimus gained weight when exposed to salinity at under 10 ppt in the first 72 h of exposure. The two species of Pontodrilus behaved differently when exposed to different salinities. The coelomic fluid osmolarity of Pontodrilus was related to the exposure medium and was mostly maintained as hyperosmotic to the external medium over the range of salinities tested. This study shows how two different species of the littoral earthworm genus Pontodrilus respond to a change in salinity, which may explain their dispersal pattern and shape their distribution pattern throughout Southeast Asia.

The Oriental millipede genus Nepalella Shear, 1979, with the description of a new species from Thailand and an updated key (Diplopoda, Chordeumatida, Megalotylidae).

The Oriental genus Nepalella is reviewed, rediagnosed and shown to comprise 28 species, including N.siamensis sp. nov. from southeastern Thailand. All Nepalella species are keyed, and their distributions mapped, being highly localized and mainly allopatric. Unlike most congeners, which are largely confined to subtropical environments (including montane to high-montane conditions, up to 3800 m a.s.l.) or karst caves (eight species, all in southern China alone), the new species is the southernmost in the distribution area of the entire genus, also being among the very few (four) that are restricted to lowland, purely tropical habitats.

Uncovering local endemism from southeastern Myanmar: description of the new karst-associated terrestrial snail genus Burmochlamys (Eupulmonata, Helicarionidae).

Salween River basin's karst ecosystems in southeastern Myanmar remain largely unexplored and are likely to harbour a high terrestrial snail diversity that are often associated with high levels of snail endemism. Here, an outstanding group of new karst-associated terrestrial snails, Burmochlamys gen. nov., are discovered. A study of the comparative morphological and anatomical data reveals that the reproductive tract and radula of this new genus are closely related to the helicarionid genus Sophina Benson, 1859 but shell morphology (shape, size, and sculpture) and mantle extensions are distinct from the latter genus. Burmochlamys gen. nov. now consists of four known nominal species, B.cassidula comb. nov., B.cauisa comb. nov., B.perpaula comb. nov., and B.poongee comb. nov., and five new species; B.albida sp. nov., B.fasciola sp. nov., B.moulmeinica sp. nov., B.versicolor sp. nov., and B.whitteni sp. nov. The highlight is that the members of the new genus show site-specific endemism, being restricted to karstic habitat islands of the Salween River basin. In addition, the discovery supports that the unique and complex structure of Salween River basin's karst ecosystems are habitats in which the terrestrial malacofauna have speciated and become endemic.

Revision of the carnivorous land snail family Streptaxidae (Stylommatophora, Achatinina) in Myanmar, with description of four new species.

The carnivorous terrestrial snail family Streptaxidae, recognized by having a regular to eccentric shell with complex apertural dentition, remains little-known and largely unexplored in Myanmar. This article presents historically recorded species and provides new data on this family. A total of eighteen species in five genera, namely Carinartemis, Discartemon, Haploptychius, Oophana, and Perrottetia from the southeastern and eastern parts of Myanmar, is examined herein. Among these, Haploptychius is the most diverse with eight species, while the remaining genera are comprised of fewer than five species each. Streptaxisbirmanica and Streptaxisblanfordianus are herein synonymized with Haploptychiusblanfordi, while Streptaxishanleyanus is synonymized with Carinartemissankeyi. Furthermore, the first genitalia and radula descriptions for three previously known species, D.tonywhitteni, C.exacutus, and C.sankeyi, are provided. Using comparative morphological and anatomical approaches, four new species are described: D.paurodeviatus sp. nov., H.heliakosus sp. nov., H.tenasserimicus sp. nov., and H.karenorum sp. nov. This present study enhances the understanding of the land snail fauna in Myanmar, specifically the streptaxids, and highlights that limestone areas are important for biodiversity conservation.

Annotated checklist of the operculated land snails from Thailand (Mollusca, Gastropoda, Caenogastropoda): the family Pupinidae, with descriptions of several new species and subspecies, and notes on classification of Pupina Vignard, 1829 and Pupinella Gray, 1850 from mainland Southeast Asia.

Thailand is located at the crossroads of several biogeographical regions, and boasts a high level of biodiversity, especially among the malacofauna. The most recent checklist of land snail species in Thailand was compiled more than twenty years ago, and so this checklist needs revision and the addition of newly discovered taxa. This study updates the taxonomy and species list of the operculated land snail family Pupinidae from Thailand. This snail family is diverse and abundant, and can be found in various natural habitats in Southeast Asia. Although the taxonomy of some Southeast Asian pupinid genera has been reviewed, studies of Pupina Vignard, 1829, which contains the highest number of species, and a lesser-known genus Pupinella Gray, 1850 are still lacking. Herein we present an annotated checklist with an up-to-date systematic framework of the Pupinidae in Thailand based on both field investigations and literature surveys, and include the taxonomic treatment of all Pupina and Pupinella species from mainland Southeast Asia. This annotated checklist contains 30 nominal species and two subspecies from seven genera currently known to occur in Thailand. We describe two species of Pseudopomatias (P.doiangkhangensis Jirapatrasilp, sp. nov. and P.pallgergelyi Jirapatrasilp, sp. nov.), five species and one subspecies of Pupina (P.bensoni Jirapatrasilp, sp. nov., P.bilabiata Jirapatrasilp, sp. nov., P.godwinausteni Jirapatrasilp, sp. nov., P.latisulci Jirapatrasilp, sp. nov., P.stoliczkai Jirapatrasilp, sp. nov., and P.dorriisanensis Jirapatrasilp, ssp. nov.) as new to science. New records of Coptocheilussumatranus, Pupinellamansuyi, and Rhaphaulustonkinensis are also reported from Thailand. The mainland Southeast Asian Pupina species are classified into three species groups (Pupinaartata group, Pupinaarula group, and Pupinaaureola group) based on the distinction of shell teeth and canals, and operculum. Three species formerly in Pupina from Vietnam are allocated to Pupinella (P.illustris comb. nov., P.sonlaensis comb. nov., and P.thaitranbaii comb. nov.) due to the presence of a funnel-like anterior canal.

Biodiversity of the Buffalo Leeches Genus Hirudinaria (Arhynchobdellida, Hirudinidae) in Southern Thailand Revealed from DNA Barcoding.

Leeches in the genus Hirudinaria Whitman, 1886, also known as buffalo leeches, are blood-sucking ectoparasites of vertebrates. Although they are widely distributed in Asia and had been highly abundant in the past, studies on diversity and taxonomy of this genus are still scarce. There is probably a large amount of cryptic diversity yet to be discovered, particularly from mainland Southeast Asia. In this study, we used morphology and DNA barcoding with a COI gene fragment to explored the diversity of Hirudinaria leeches in the southern region of Thailand, where a unique geographic feature could have led to the diversification of freshwater biota. Molecular phylogenetic analyses and species delimitation approaches (ABGD, bPTP, GMYC, and BOLD) revealed the presence of four putative species of Hirudinaria leeches from southern Thailand, including H. bpling, H. thailandica, and two morphologically cryptic lineages of H. manillensis. Compared to other leech genera, genetic distances of Hirudinaria leeches were relatively low (0.11-0.65% within species; 3.72-14.36% between species) and barcoding gaps were very narrow (1.54-2.88%). The species diversity, distribution pattern, and a phenomenon of low genetic divergence of Hirudinaria leeches in southern Thailand could be explained by an ancient seaway, paleo-drainage, and anthropogenic activities.

Systematic review of the dextral Hemiplecta Albers, 1850 (Eupulmonata, Ariophantidae) from Thailand with description of a new species and list of all the Indochinese species.

The genus Hemiplecta is a group of large-sized land snails which have long been used as a food resource by Indochinese people. There are five dextral and four sinistral species currently recognized from Thailand. The dextral group is comprised of two previously recorded species (H. humphreysiana and H. distincta), two newly recorded species (H. funerea and H. esculenta), and one new species (H. nemorosa sp. nov.) from northern Thailand is being proposed. We reassessed the diagnostic characters of the genitalia, mantle edge, and radula. Specimens were classified into the genus Hemiplecta on the basis of the penial verge and shell lobe, and on the characters of a bulbous gametolytic sac without a gametolytic duct. A complete species list, together with photographs of the name-bearing types or authenticated specimens and the taxonomic status of Hemiplecta s.l. that are known from Indochina including Peninsular Malaysia and Myanmar, is provided for the first time. In total, this species list contains 39 available nominal species names described from this area. Type or authentic specimens can be located for 37 nominal species names, of which 25 are illustrated herein and the other 12 were recently illustrated. However, two available species-level names could not be traced to any type specimens. In addition, lectotypes of H. funerea and H. pluto are designated herein to stabilize the names.

Clarification on the name-bearing type designation of several cyclophorid species (Mollusca, Gastropoda) by H. H. Godwin-Austen (1915).

The type series boundary and the name-bearing type designation of each cyclophorid taxon originally described by Godwin-Austen are clarified based on an interpretation that complies with the ICZN. Previous statuses of type specimens designated by previous authors are reconsidered. Lectotypes of Spiraculum oakesi Godwin-Austen, 1915, Spiraculum kempi Godwin-Austen, 1915, Pterocyclos aborensis Godwin-Austen, 1915, Pterocyclos miriensis Godwin-Austen, 1915, Pterocyclos brahmakundensis Godwin-Austen, 1915, Spiraculum luyorensis Godwin-Austen, 1915, Spiraculum putaoensis Godwin-Austen, 1915, and Theobaldius oakesi Godwin-Austen, 1915 are here designated to stabilize the existing nomenclature. In addition, the type specimens of Pterocyclos miriensis and Theobaldius oakesi are photographed and figured for the first time.

Two new species of the millipede genus Tylopus Jeekel, 1968 from Shan State, Myanmar (Diplopoda, Polydesmida, Paradoxosomatidae).

The predominantly Indochinese to southern Chinese millipede genus Tylopus presently comprises 76 described species, including two new, T. monticola sp. nov. and T. sutchariti sp. nov., both described and illustrated based on material from a limestone mountain in Taunggyi District, Shan State, Myanmar. Both new species have been found to occur syntopically near limestone caves and are assumed to be narrowly endemic to the Taunggyi Mountains, southwestern Shan State, Myanmar. A key to all six Tylopus species known to occur in Myanmar is provided, and their distributions are also mapped.

Evidence of cryptic diversity in freshwater Macrobrachium prawns from Indochinese riverine systems revealed by DNA barcode, species delimitation and phylogenetic approaches.

The diversity of Indochinese prawns in genus Macrobrachium is enormous due to the habitat diversification and broad tributary networks of two river basins: the Chao Phraya and the Mekong. Despite long-standing interest in SE-Asian decapod diversity, the subregional Macrobrachium fauna is still not yet comprehensively clarified in terms of taxonomic identification or genetic diversification. In this study, integrative taxonomic approaches including morphological examination, DNA barcoding, and molecular species delimitation were used to emphasize the broad scale systematics of Macrobrachium prawns in Indochina. Twenty-seven nominal species were successfully re-verified by traditional and molecular taxonomy. Barcode gap analysis supported broad overlapping of species boundaries. Taxonomic ambiguity of several deposited samples in the public database is related to inter- and intraspecific genetic divergence as indicated by BOLD discordance. Diagnostic nucleotide positions were found in six Macrobrachium species. Eighteen additional putative lineages are herein assigned using the consensus of species delimitation methods. Genetic divergence indicates the possible existence of cryptic species in four morphologically complex and wide-ranging species: M. lanchesteri, M. niphanae, M. sintangense, and some members of the M. pilimanus group. The geographical distribution of some species supports the connections and barriers attributed to paleo-historical events of SE-Asian rivers and land masses. Results of this study show explicitly the importance of freshwater ecosystems in Indochinese subregions, especially for the Mekong River Basin due to its high genetic diversity and species composition found throughout its tributaries.

Occurrence of bioluminescent and nonbioluminescent species in the littoral earthworm genus Pontodrilus.

Pontodrilus litoralis is a cosmopolitan littoral earthworm known to exhibit bioluminescence. Recently, a congeneric species, Pontodrilus longissimus, from Thailand was described. These species are sympatric, but their burrowing depths on Thai beaches are different. In this study, we examined the in vivo and in vitro bioluminescent properties of P. longissimus and P. litoralis. Mechanical stimulation induced in vivo luminescence in P. litoralis, as reported previously, but not in P. longissimus. In vitro cross-reaction tests between these species revealed the absence of luciferin and luciferase activities in P. longissimus. The coelomic fluid of P. litoralis had strong fluorescence that matched the spectral maximum of its bioluminescence, but the same result was not observed for P. longissimus. These results suggest that P. litoralis has luminescence abilities due to the creation of bioluminescent components (i.e., luciferin, luciferase, and light emitters). The presence of both luminous and nonluminous species in a single genus is likely widespread, but only a few examples have been confirmed. Our findings provide insight into the possible functions of bioluminescence in earthworms, such as avoiding predation by littoral earwigs.

Snail mucus from the mantle and foot of two land snails, Lissachatina fulica and Hemiplecta distincta, exhibits different protein profile and biological activity.

OBJECTIVE: Snails secrete different types of mucus that serve several functions, and are increasingly being exploited for medical and cosmetic applications. In this study, we explored the protein pattern and compared the biological properties of the mucus secreted from the mantle collar and foot of two snail species, Lissachatina fulica and Hemiplecta distincta. RESULT: Protein profile showed a different pattern between the two species and between the two secretory parts. The mantle-specific protein bands were further characterized and among them was an antibacterial protein, achacin. Accordingly, the mucus from the mantle exhibited the higher antibacterial activity than that from the foot in both snail species. The mucus from H. distincta, first reported here, also showed antibacterial properties, but with a lower activity compared to that for L. fulica. Snail mucus also exhibited anti-tyrosinase activity and antioxidant activity but with no significant difference between the foot and mantle mucus. These results indicate some different protein compositions and biological activities of snail slime from the mantle and foot, which might be associated with their specific functions in the animal and are useful for medical applications.