Association of Mandibular Posterior Anatomic Limit with Skeletal Patterns and Root Morphology Using Three-Dimensional Cone Beam Computed Tomography Comprehensive Analysis.

This study aimed to clarify the relationship between the mandibular posterior anatomic limit (MPAL) and skeletal anteroposterior and vertical skeletal patterns, with consideration of factors that may be related. In total, 230 people were included: 49 Japanese, 122 Egyptian, and 59 Korean people. The MPAL was measured at 0, 2, 4, and 6 mm from the root furcation along the sagittal and cuspal lines at the distance from the distal root of the mandibular right second molar to the mandibular cortex of the lingual bone. Eight different MPALs were evaluated using multiple regression analysis with explanatory variables for anteroposterior and vertical skeletal patterns and qualitative variables for age, sex, population, the presence of third molars, number of roots, presence of C-shaped roots, and Angle malocclusion classification. The MPAL was significantly larger as the mandibular plane angle decreased. The MPAL near the root apex was significantly larger as the A-nasion-point B angle increased, and the MPAL near the root apex measured at the cuspal line was significantly larger for C-type roots. The present study showed that a C-shaped root affected the MPAL in addition to the anteroposterior and vertical skeletal patterns.

Current Genetic Status of Nagaremon-charr, a Threatened Morphotype of Salvelinus leucomaenis in the Ane River, Lake Biwa System, Central Japan, With Comments on Its Conservation.

Stream-dwelling white-spotted charr, Salvelinus leucomaenis, populations tend to have unique color and spot patterns in different regions and may even display stream-specific patterns. An extreme edge of such diversity, found in individuals with atypical body color patterns (so-called nagaremon-type charr, a rare morphotype of Salvelinus leucomaenis [hereafter termed nagaremon-charr]), sympatrically occurring with normal-charr, has been reported from only six small isolated populations in Japan. Based on morphological and ecological perspectives, nagaremon-charr has been considered as an intraspecific color variant of white-spotted charr, although the genetic status of nagaremon-charr has not been determined. In this study, genetic diversity and population structure of the nagaremon-charr in a tributary of the Ane River (Lake Biwa system) were investigated through microsatellite and mtDNA analyses. Nagaremon-charr and sympatric normal-charr in the tributary shared the mtDNA haplotypes and were assigned to the same cluster in the STRUCTURE analysis and discriminant analysis of principal components (DAPC). These results suggested that nagaremon-charr in the Ane River is an intra-populational specific color variant of white-spotted charr. Above a waterfall, nagaremon-charr specimens exhibited extremely reduced genetic diversity, indicating that genetic drift may account for the fixation of the nagaremon-morphotype. Normal-charr below the waterfall clustered separately from hatchery-reared charr, indicative of native status of the former. Thus, both nagaremon-charr and normal-charr in the entire Ane River tributary should be conserved.

Prevalence of saliva immunoglobulin A antibodies reactive with severe acute respiratory syndrome coronavirus 2 among Japanese people unexposed to the virus.

While the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a threat to public health as the number of cases and COVID-19-related deaths are increasing worldwide, the incidence of the virus infection is extremely low in Japan compared with many other countries. To explain this uncommon phenomenon, we investigated the prevalence of naturally occurring ("natural") antibodies, focusing on those of the secretory immunoglobulin A (sIgA) form, reactive with SARS-CoV-2 among Japanese people. One hundred and eighty healthy Japanese volunteers of a wide range of age who had been considered to be unexposed to SARS-CoV-2 participated in this study. Saliva samples and blood samples were collected from all of the 180 participants and 139 adults (aged ≥ 20 years) included therein, respectively. The determination of saliva IgA antibodies, mostly comprising sIgA antibodies, as well as serum IgA and immunoglobulin G antibodies, reactive with the receptor binding domain of the SARS-CoV-2 spike-1 subunit proteins was conducted using an enzyme-linked immunosorbent assay. The major findings were that 52.78% (95% confidence interval, 45.21%-60.25%) of the individuals who had not been exposed to SARS-CoV-2 were positive for saliva IgA antibodies with a wide range of levels between 0.002 and 3.272 ng/mL, and that there may be a negative trend in positivity for the antibodies according to age. As we had expected, a frequent occurrence of assumable "natural" sIgA antibodies reactive with SARS-CoV-2 among the studied Japanese participant population was observed.

Interploidy gene flow involving the sexual-asexual cycle facilitates the diversification of gynogenetic triploid Carassius fish.

Asexual vertebrates are rare and at risk of extinction due to their restricted adaptability through the loss of genetic recombination. We explore the mechanisms behind the generation and maintenance of genetic diversity in triploid asexual (gynogenetic) Carassius auratus fish, which is widespread in East Asian fresh waters and exhibits one of the most extensive distribution among asexual vertebrates despite its dependence on host sperm. Our analyses of genetic composition using dozens of genetic markers and genome-wide transcriptome sequencing uncover admixed genetic composition of Japanese asexual triploid Carassius consisting of both the diverged Japanese and Eurasian alleles, suggesting the involvement of Eurasian lineages in its origin. However, coexisting sexual diploid relatives and asexual triploids in Japan show regional genetic similarity in both mitochondrial and nuclear markers. These results are attributed to a unique unidirectional gene flow from diploids to sympatric triploids, with the involvement of occasional sexual reproduction. Additionally, the asexual triploid shows a weaker population structure than the sexual diploid, and multiple triploid lineages coexist in most Japanese rivers. The generated diversity via repeated interploidy gene flow as well as an increased establishment of immigrants is assumed to offset the cost of asexual reproduction and might contribute to the successful broad distribution of this asexual vertebrate.

Facultative amphidromy involving estuaries in an annual amphidromous fish from a subtropical marginal range.

Variation in the life cycle of diadromous fishes can be explained by differential food availability between marine and freshwater habitats, since migration is often interpreted as a mechanism for exploiting food resources. Theoretically, a migration pattern of mainly remaining in freshwater occurs in tropical and subtropical habitats where fluvial productivity possibly exceeds marine productivity. However, in Yakugachi River, Amami-Oshima Island, southern Japan, low nutrient concentrations in the river suggest that food availability is limited for the subtropical Ryukyu-ayu Plecoglossus altivelis ryukyuensis. Since Ryukyu-ayu is an amphidromous fish that mainly grows in rivers after spending 2 months in the sea, limited food availability in rivers would force this species to migrate to other habitats with better food availability. Otolith increment and Sr:Ca analyses of 48 adult Ryukyu-ayu collected from the Yakugachi River revealed that all individuals visited estuaries more than three times after moving upstream. Although the specific growth rates of this species in the river had no correlation with the salinity profile in the fluvial period, this movement may be an adaptive choice because the salinity profile significantly affected the body size at maturity. Our results highlighted individual-based variations in amphidromous migration for utilizing estuaries, which could be explained by relatively higher productivity in estuarine than in freshwater and marine habitats.

Using dense locality sampling resolves the subtle genetic population structure of the dispersive fish species Plecoglossus altivelis.

In dispersive species with continuous distributions, genetic differentiation between local populations is often absent or subtle and thus difficult to detect. To incorporate such subtle differentiation into management plans, it may be essential to analyse many samples from many localities using adequate numbers of high-resolution genetic markers. Here, we evaluated the usefulness of dense locality sampling in resolving genetic population structure in the ayu (Plecoglossus altivelis), a dispersive fish important in Japanese inland fisheries. Genetic variability in, and differentiation between, ayu populations around the Japan-Ryukyu Archipelago were investigated in 4746 individuals collected from 120 localities by genotyping 12 microsatellite markers. These individuals represented the two subspecies of ayu, namely the Ryukyuan subspecies (Plecoglossus altivelis ryukyuensis) and both amphidromous and landlocked forms of the nominotypical subspecies (P. a. altivelis) along the archipelago. We successfully detected an absence of genetic differentiation within the landlocked form and subtle but significant differentiation and clear geographic patterns of genetic variation among populations of the amphidromous form, which had been considered genetically homogeneous. This suggests that dense locality sampling effectively resolves subtle differences in genetic population structure, reducing stochastic deviation in the detection of genetic differentiation and geographic patterns in local populations of this dispersive species. Resampling analyses based on empirical data sets clearly demonstrate the effectiveness of increasing the number of locality samples for stable and reliable estimations of genetic fixation indices. The genetic population structure observed within the amphidromous form provides useful information for identifying management or conservation units in ayu.

Exposure of a herbivorous fish to ¹³4Cs and ¹³7Cs from the riverbed following the Fukushima disaster.

Ayu Plecoglossus altivelis, a herbivorous fish, is an important fishery resource and key component of the foodweb in many Japanese streams. Radionuclide contamination of this species is likely transferred to higher trophic levels, include humans, in the food chain. After the Fukushima accident in March 2011, ayu were exposed to highly contaminated silt while feeding on algae attached to the riverbed stones. To understand the route by which herbivorous fish are exposed to radionuclides, the activity concentrations of sum of (134)Cs and (137)Cs (radiocesium) were analyzed in riverbed samples (algae and silt) and in the internal organs and the muscle of ayu in five river systems in the Fukushima Prefecture between summer 2011 and autumn 2013. Although there was a positive correlation between the radiocesium activity concentrations in the muscle and the internal organs of ayu, the median activity concentration in the muscle was much lower than those in the internal organs. The activity concentrations of radiocesium in the riverbed samples and the internal organs and the muscle of ayu were correlated with contamination levels in soil samples taken from the watershed upstream of the sample sites. The results of the generalized linear mixed models suggest that the activity concentrations in both the internal organs and the muscle of ayu declined over time. Additionally, the activity concentrations in the internal organs were correlated with those in the riverbed samples that were collected around the same time as the ayu. The activity concentrations in the muscle were correlated with ayu body size. Our results suggest that ayu ingest (134)Cs and (137)Cs while grazing silt and algae from the riverbed, and a part of the (134)Cs and (137)Cs is assimilated into the muscle of the fish.

Prolactin and upstream migration of the amphidromous teleost, ayu Plecoglossus altivelis.

Changes in mRNA levels of prolactin (PRL) during the upstream migration were examined in fry of the amphidromous fish, ayu Plecoglossus altivelis. Quantification of mRNA has been done with real-time PCR and expressed as whole body or pituitary contents depending the body size of fry. PRL mRNA levels of ayu caught in seawater of the coastal area remained low during early spring. Prior to the start of the upstream migration, the fish caught in the coastal area in mid spring showed increased levels of PRL mRNA. There were further increases in PRL levels in the fish caught in the river. Analysis of proportions revealed that there were significant differences among PRL mRNA in the fish caught in different environmental salinities. Body weight showed a positive relation with PRL mRNA in ayu caught in seawater. A landlocked population of ayu, which migrates from lake to river, showed no significant change in PRL mRNA levels before and after upstream migration. Results in this study indicate the importance of up-regulation of PRL gene expression of ayu during the upstream migration from seawater to fresh water. There is a possible relationship between body size and PRL in the early developmental stage of ayu in seawater, but not in the fish in fresh water.

Historical effect in the territoriality of ayu fish.

Ayu fish form algae-feeding territories in a river during a non-breeding (growing) season. We build a cost-benefit theory to describe the breakdown and formation of territory. In the early stage of a growing season, all fish hold territories at low densities. Once all territory sites are occupied, excess fish become floaters. When fish density further increases, a phase transition occurs: all the territories suddenly break down and fish form a school. In contrast, when the fish density is decreased, territories are suddenly formed from the school. Both theory and experiments demonstrate that ayu should exhibit a historical effect: the breakdown and formation processes of territory are largely different. In particular, the theory in formation process predicts a specific fish behavior: an "attempted territory holder" that tries to have a small territory emerges just before the formation of territory.

Changes in prolactin mRNA levels during downstream migration of the amphidromous teleost, ayu Plecoglossus altivelis.

Changes in mRNA levels of prolactin (PRL) during seaward migration and after experimental transfer from fresh water (FW) to seawater (SW) were examined in larvae of the amphidromous fish, ayu Plecoglossus altivelis. In the field study, ayu larvae caught in the surf zone showed lower levels of PRL mRNA than those in the river, while growth hormone (GH) levels showed no significant change. Decrease in PRL gene transcription was also observed 24h after direct transfer from FW to SW, whereas there was no significant influence of water temperature. On the other hand, there was no significant change in GH mRNA levels in relation to SW transfer or environmental temperature. In a raceway with a vertical salinity gradient, PRL mRNA levels of ayu larvae showed a significant reduction during spontaneous migration from FW to SW, which mimicked the results from the field observation and the transfer experiment, and then a gradual increase during the course of development. Whole body water and sodium contents of larvae in a salinity gradient were stable during migration to SW. Results in this study indicated the importance of regulation of PRL gene expression in the downstream migration and acclimation to SW during the early development of ayu.

Biogeography and evolution of the Carassius auratus-complex in East Asia.

BACKGROUND: Carassius auratus is a primary freshwater fish with bisexual diploid and unisexual gynogenetic triploid lineages. It is distributed widely in Eurasia and is especially common in East Asia. Although several genetic studies have been conducted on C. auratus, they have not provided clear phylogenetic and evolutionary descriptions of this fish, probably due to selection bias in sampling sites and the DNA regions analysed. As the first step in clarifying the evolutionary entity of the world's Carassius fishes, we attempted to clarify the phylogeny of C. auratus populations distributed in East Asia. RESULTS: We conducted a detailed analysis of a large dataset of mitochondrial gene sequences [CR, 323 bp, 672 sequences (528 sequenced + 144 downloaded); CR + ND4 + ND5 + cyt b, 4669 bp in total, 53 sequences] obtained from C. auratus in East Asia. Our phylogeographic analysis revealed two superlineages, one distributed mainly among the Japanese main islands and the other in various regions in and around the Eurasian continent, including the Ryukyus and Taiwan. The two superlineages include seven lineages with high regional specificity that are composed of endemic populations indigenous to each region. The divergence time of the seven lineages was estimated to be 0.2 million years ago (Mya) by a fossil-based method and 1.0-1.9 Mya by the molecular clock method. The antiquity and endemism of these lineages suggest that they are native to their respective regions, although some seem to have been affected by the artificial introduction of C. auratus belonging to other lineages. Triploids of C. auratus did not form a monophyletic lineage but were clustered mostly with sympatric diploids. CONCLUSIONS: The results of the present study revealed the existence of two superlineages of C. auratus in East Asia that include seven lineages endemic to each of the seven regions examined. The lack of substantial genetic separation between triploids and diploids indicates that triploids are not composed of a single independent lineage. The ancient origins and evolutionary uniqueness of the seven lineages warrant their conservation. An overall phylogenetic framework obtained from the present study will be of use for estimating the phylogenetic relationships of Carassius fishes on the Eurasian continent.

Mitochondrial DNA population structure of the white-spotted Charr (Salvelinus leucomaenis) in the Lake Biwa water system.

A phylogeographic analysis of mitochondrial DNA sequences was performed in order to elucidate the origin, dispersal process, and genetic structure of white-spotted charr in the Lake Biwa water system. Two haplotypes were most common in the Lake Biwa water system, and were also common in the adjacent inlet rivers of the Sea of Japan. These results suggest that in the glacial periods of the Pleistocene, white-spotted charr dispersed into the northern inlet rivers of Lake Biwa from adjacent inlet rivers of the Sea of Japan by watershed exchanges, colonizing the whole of the Lake Biwa water system. Mitochondrial DNA diversity contrasted sharply between the western and eastern parts of the system, suggesting that the populations in the western part might be more reduced than those in the eastern part in relation to the smaller habitat size. The high overall FST estimate (0.50), together with pairwise comparisons of FST, indicated significant genetic divergence between populations due to isolation and small population size. Hierarchical analysis (AMOVA) also showed that genetic variation was more pronounced among regions (28.39%) and among populations within regions (47.24%) than within populations (24.37%). This suggests that each population in and around the Lake Biwa water system should be treated as a significant unit for conservation and management.

Unexpected ceiling of genetic differentiation in the control region of the mitochondrial DNA between different subspecies of the ayu Plecoglossus altivelis.

Sequence analyses of the non-coding, control region (CR) and coding region of the ND4-tRNA(Ser) genes in the mitochondrial DNA (mtDNA) were conducted for populations of the ayu Plecoglossus altivelis altivelis and the Ryukyu-ayu P. a. ryukyuensis. The level of genetic differentiation between the two subspecies evaluated from the CR data was substantially low, when comparing with that estimated from ND4-tRNA(Ser) gene region data, as well as those from nuclear genome data sets. By contrast, the differentiation between subspecies in the ND4-tRNA(Ser) gene region was substantial, being consistent with the results from the previous nuclear genome analyses. Results of UPGMA and minimum spanning network analyses also implied the unexpected ceiling of genetic differentiation in the CR. These results suggest that the CR does not reflect accurately the level of overall genetic differentiation between the populations of the ayu, but other coding regions of the mtDNA do reflect it so that the mtDNA on the whole may function as a rich source of useful markers for genetic assessment of populations of this species.