Uncovering overlooked diversity using molecular phylogenetic approach: A case of Japanese sphaeriid clams (Bivalvia: Sphaeriidae).

Revealing the species and lineage diversity of a taxon is important for many biological studies of wildlife. In recent decades, DNA-based approaches have been widely utilised to elucidate the diversity of taxa, especially those that are difficult to distinguish based on morphological traits. This study focused on freshwater clams (Sphaeriidae) in Japan, a biodiversity hotspot of freshwater molluscs. Molecular phylogenetic approaches, including divergence time estimation, species delimitation, rarefaction, and biogeographic area estimation, were used to reveal the nature of the species diversity and its formation process, which are largely unknown. Our delimitation and rarefaction analyses suggest that Japanese sphaeriid clams consist of at least 18 delimitated lineages. This lineage diversity is relatively high compared to other Japanese freshwater molluscs, and in addition, the majority of the Japanese lineage appears to have high endemicity despite the possibility of long-distance dispersal in sphaeriid clams. Our biogeographical analyses suggest that this diversity may be due to the combination of colonisation, during the period when Japan was connected to the continent, and the relatively recent dispersal. Our results highlight the overlooked biodiversity of Japan and provide a basis for further Japanese sphaeriid research, including conservation perspectives.

A complete dietary review of Japanese birds with special focus on molluscs.

Birds often hold important positions in the food webs of ecosystems. As a result, interactions between birds and their prey have attracted attention not only in ecology, but also in fields like agriculture and conservation. Avian food resources are well researched in Japan, however there is no database critically reviewing molluscs as a food resource for birds. Here, we present a new database reviewing dietary information for all Japanese bird species. In addition to addressing general diet categories and specific food habits for each bird, we include detailed data on the molluscan prey observed for all species that consume them. The information within this database was collected through intense literary review to provide a complete look at bird species historically present around the country. We also include new information on snail species found in the upper digestive tract of harvested wild birds. This database is publicly available in the Zenodo repository. The information should aid research around the Japanese archipelago, especially projects involving birds or molluscs.

A comprehensive phylogeography of the widespread pond snail genus Radix revealed restricted colonization due to niche conservatism.

To clarify the effect of niche conservatism on evolutionary history, we focused on freshwater snails, which have different ecological and phylogenetic properties from previously tested taxa. We conducted a phylogenetic analysis using 750 lymnaeid individuals from 357 sites of eleven Radix species. Then, we estimated the ancestral distribution using the geographic coordinates and colonization routes. In addition, a statistical test of the colonization distances in the latitudinal and longitudinal directions was performed. We also conducted ecological niche modeling for two widely distributed species using climatic data. Ancestral geographic reconstruction estimated the origin of the genus to be around the Indian subcontinental region and showed that latitudinal immigration distances were shorter than longitudinal immigration distances in the diversification process. Ecological niche models suggested that the current distribution was restricted by climate, with annual mean temperature and precipitation of the driest month as particularly strong factors. Niche conservatism to the climate can affect the diversification of freshwater snails.

Distribution records of three species of Leucochloridium (Trematoda: Leucochloridiidae) in Japan, with comments on their microtaxonomy and ecology.

Insectivorous birds serve as definitive hosts for trematodes of the genus Leucochloridium. The parasites exclusively use amber snails of the family Succineidae as intermediate hosts. A pulsating and colorful display of the larval broodsac in the snail's eyestalk seems to be a caterpillar mimic for attracting birds. A colored design of the broodsac is very useful for parasite identification. In Japan, characteristic broodsacs from amber snails have been recorded from 1980's, but their taxonomic discrimination from Asian, European, and North American species has not been achieved. In this study, old scientific records, sighting information on broodsacs from the general public, and direct molecular evidence by DNA barcoding clearly showed that at least three species of Leucochloridium are distributed in Japan. A vertical-striped broodsac found from Succinea sp. in Okinawa, the subtropical island of Japan, were treated as Leucochloridium sp., but being almost identical to that of Leucochloridium passeri in neighboring Taiwan. The European species of Leucochloridium perturbatum and Leucochloridium paradoxum were frequently detected from Succinea lauta in Hokkaido, the northernmost island of Japan. The former species was common in inland areas of Hokkaido, whereas the latter species was frequently seen in the coastal areas. A possible explanation for the parasite distribution pattern is that principal definitive hosts (migratory or resident birds) differ in each parasite. The conspecificity of Leucochloridium variae in North America and L. perturbatum in Europe and the Far East is also discussed.

Phylogeography of freshwater planorbid snails reveals diversification patterns in Eurasian continental islands.

BACKGROUND: Islands have traditionally been the centre of evolutionary biological research, but the dynamics of immigration and differentiation at continental islands have not been well studied. Therefore, we focused on the Japanese archipelago, the continental islands located at the eastern end of the Eurasian continent. While the Japanese archipelago is characterised by high biodiversity and rich freshwater habitats, the origin and formation mechanisms of its freshwater organisms are not clear. In order to clarify the history of the planorbid gastropod fauna, we conducted phylogenetic analysis, divergence time estimation, ancestral state reconstruction, and lineage diversity estimations. RESULTS: Our analyses revealed the formation process of the planorbid fauna in the Japanese archipelago. Most lineages in the Japanese archipelago have closely related lineages on the continent, and the divergence within the Japanese lineages presumably occurred after the late Pliocene. In addition, each lineage is characterised by different phylogeographical patterns, suggesting that immigration routes from the continent to the Japanese archipelago differ among lineages. Furthermore, a regional lineage diversity plot showed that the present diversity in the Japanese archipelago potentially reflects the differentiation of lineages within the islands after the development of the Japanese archipelago. CONCLUSIONS: Although additional taxon sampling and genetic analysis focused on each lineage are needed, our results suggest that immigration from multiple routes just prior to the development of the Japanese archipelago and subsequent diversification within the islands are major causes of the present-day diversity of the Japanese planorbid fauna.

Activity of invasive slug Limax maximus in relation to climate conditions based on citizen's observations and novel regularization based statistical approaches.

Citizen science is a powerful tool that can be used to resolve the problems of introduced species. An amateur naturalist and author of this paper, S. Watanabe, recorded the total number of Limax maximus (Limacidae, Pulmonata) individuals along a fixed census route almost every day for two years on Hokkaido Island, Japan. L. maximus is an invasive slug considered a pest species of horticultural and agricultural crops. We investigated how weather conditions were correlated to the intensity of slug activity using for the first time in ecology the recently developed statistical analyses, Bayesian regularization regression with comparisons among Laplace, Horseshoe and Horseshoe+ priors for the first time in ecology. The slug counts were compared with meteorological data from 5:00 in the morning on the day of observation (OT- and OD-models) and the day before observation (DBOD-models). The OT- and OD-models were more supported than the DBOD-models based on the WAIC scores, and the meteorological predictors selected in the OT-, OD- and DBOD-models were different. The probability of slug appearance was increased on mornings with higher than 20-year-average humidity (%) and lower than average wind velocity (m/s) and precipitation (mm) values in the OT-models. OD-models showed a pattern similar to OT-models in the probability of slug appearance, but also suggested other meteorological predictors for slug activities; positive effect of solar radiation (MJ) for example. Five meteorological predictors, mean and highest temperature (°C), wind velocity (m/s), precipitation amount (mm) and atmospheric pressure (hPa), were selected as the effective factors for the counts in the DBOD-models. Therefore, the DBOD-models will be valuable for the prediction of slug activity in the future, much like a weather forecast.

Do Native Rodents Prey on Land Snails? An Experimental and Quantitative Study in Hokkaido, Japan.

Predator-prey interaction is one of the most important and pervasive pressures in the ecology and evolution of prey species. However, accurate description of the food web is sometimes extremely difficult as there are many predator-prey interactions in the wild are obscure. Recent studies have reported that two closely related land snails, Karaftohelix editha and K. gainesi, on Hokkaido Island, Japan, were diversified due to predation, probably by carabid beetles. However, it is unclear 1) whether native rodents prey upon land snails on Hokkaido Island, and 2) how frequently land snails are preyed upon by rodents in this region, although it has been reported that several species of rodents are predators of land snails in many regions. Thus, we investigated these issues in this study by captive feeding trials and field observations. No rodent species other than the Siberian chipmunk, Tamias sibiricus, were found to prey upon land snails around the research site on Hokkaido Island. In addition, the population density of T. sibiricus was lower than those of other rodent species, and it has been reported that T. sibiricus is omnivorous and preys upon snails considerably less than on other food sources. Overall, these findings suggest that T. sibiricus is not an important predator of Karaftohelix land snails in Hokkaido.

Single-gene speciation: Mating and gene flow between mirror-image snails.

Variation in the shell coiling, or chirality, of land snails provides an opportunity to investigate the potential for "single-gene" speciation, because mating between individuals of opposite chirality is believed not possible if the snails mate in a face-to-face position. However, the evidence in support of single-gene speciation is sparse, mostly based upon single-gene mitochondrial studies and patterns of chiral variation between species. Previously, we used a theoretical model to show that as the chiral phenotype of offspring is determined by the maternal genotype, occasional chiral reversals may take place and enable gene flow between mirror image morphs, preventing speciation. Here, we show empirically that there is recent or ongoing gene flow between the different chiral types of Japanese Euhadra species. We also report evidence of mating between mirror-image morphs, directly showing the potential for gene flow. Thus, theoretical models are suggestive of gene flow between oppositely coiled snails, and our empirical study shows that they can mate and that there is gene flow in Euhadra. More than a single gene is required before chiral variation in shell coiling can be considered to have created a new species.

Parallel evolution of passive and active defence in land snails.

Predator-prey interactions are major processes promoting phenotypic evolution. However, it remains unclear how predation causes morphological and behavioural diversity in prey species and how it might lead to speciation. Here, we show that substantial divergence in the phenotypic traits of prey species has occurred among closely related land snails as a result of adaptation to predator attacks. This caused the divergence of defensive strategies into two alternatives: passive defence and active defence. Phenotypic traits of the subarctic Karaftohelix land snail have undergone radiation in northeast Asia, and distinctive morphotypes generally coexist in the same regions. In these land snails, we documented two alternative defence behaviours against predation by malacophagous beetles. Furthermore, the behaviours are potentially associated with differences in shell morphology. In addition, molecular phylogenetic analyses indicated that these alternative strategies against predation arose independently on the islands and on the continent suggesting that anti-predator adaptation is a major cause of phenotypic diversity in these snails. Finally, we suggest the potential speciation of Karaftohelix snails as a result of the divergence of defensive strategies into passive and active behaviours and the possibility of species radiation due to anti-predatory adaptations.

Complete mitochondrial genome of an enigmatic dragonfly, Epiophlebia superstes (Odonata, Epiophlebiidae).

This study reported the 15,435 bp-long complete mitochondrial genome of the relict Epiophlebia superstes (Odonata, Epiophlebiidae), an enigmatic dragonfly of the paraphyletic 'Anisozygoptera' possessing characteristics similar to members of both extant odonate suborders, the Zygoptera and the Anisoptera. This mitogenome comprises the common set of 37 genes and an A + T-rich control region, and has a gene arrangement identical to those of all available odonates. The genome contains three non-coding inter-genic spacers (s1-s3), which occurs in all of other known odonates, but it lacks the inter-genic spacer s5 typically found in the Anisoptera. This result suggests that E. superstes possesses a mitogenmic organization more closely related to that of the Zygoptera than that of the Anizoptera.

Feasibility study of a combined treatment of electromyography-triggered neuromuscular stimulation and mirror therapy in stroke patients: a randomized crossover trial.

BACKGROUND: Mirror therapy (MT) and electromyography-triggered neuromuscular stimulation (ETMS) are both effective treatments for impaired upper limbs following stroke. A combination of these two treatments (ETMS-MT) may result in greater gain than either treatment alone. OBJECTIVES: The feasibility and possible effects of ETMS-MT upon upper extremity function were investigated in stroke patients. METHODS: Thirteen post-acute stroke patients were randomly assigned to an immediate ETMS-MT group or a delayed ETMS-MT group and then underwent an 8-week training program. The immediate ETMS-MT group received ETMS-MT in addition to physical and occupational therapy (PT+OT) for 4 weeks. They then received only PT+OT for the next 4 weeks. In the delayed ETMS-MT group, interventions were provided in the reverse order. The main outcome measure was the Fugl-Meyer Assessment (FMA). RESULTS: The immediate ETMS-MT group showed significantly greater gain in FMA in the first 4 weeks. The delayed ETMS-MT group showed significantly greater gain in active range of motion during the latter 4 weeks. No adverse effects were reported following ETMS-MT. CONCLUSION: ETMS-MT might be as effective as independent MT or ETMS without causing any side effects. Future research should focus upon the direct comparisons between independent and combined interventions.