AimTo comprehend the phylogeographic patterns of genetic variation in anurans at Taiwan Island, this study attempted to examine (1) the existence of various geological barriers (Central Mountain Ranges, CMRs); and (2) the genetic variation of Bufo bankorensis using mtDNA sequences among populations located in different regions of Taiwan, characterized by different climates and existing under extreme conditions when compared available sequences of related species B. gargarizans of mainland China.Methodology/Principal FindingsPhylogenetic analyses of the dataset with mitochondrial DNA (mtDNA) D-loop gene (348 bp) recovered a close relationship between B. bankorensis and B. gargarizans, identified three distinct lineages. Furthermore, the network of mtDNA D-loop gene (564 bp) amplified (279 individuals, 27 localities) from Taiwan Island indicated three divergent clades within B. bankorensis (Clade W, E and S), corresponding to the geography, thereby verifying the importance of the CMRs and Kaoping River drainage as major biogeographic barriers. Mismatch distribution analysis, neutrality tests and Bayesian skyline plots revealed that a significant population expansion occurred for the total population and Clade W, with horizons dated to approximately 0.08 and 0.07 Mya, respectively. These results suggest that the population expansion of Taiwan Island species B. bankorensis might have resulted from the release of available habitat in post-glacial periods, the genetic variation on mtDNA showing habitat selection, subsequent population dispersal, and co-distribution among clades.ConclusionsThe multiple origins (different clades) of B. bankorensis mtDNA sequences were first evident in this study. The divergent genetic clades found within B. bankorensis could be independent colonization by previously diverged lineages; inferring B. bankorensis originated from B. gargarizans of mainland China, then dispersal followed by isolation within Taiwan Island. Highly divergent clades between W and E of B. bankorensis, implies that the CMRs serve as a genetic barrier and separated the whole island into the western and eastern phylogroups.

During the breeding season, dynamic changes in body coloration are regularly observed in the male brown tree frog Buergeria robusta. This study investigated the hypothesis that this sexual dichromatism in male B. robusta is mediated through hormonal regulation. Frogs were exogenously injected with testosterone (T) or estradiol (E2). This manipulation revealed that the body coloration (hue, brightness, and saturation) of the male frog increased significantly (i.e., the brilliant yellow color developed) in response to T but not in response to E2. Concurrently, the levels of expression of brain-derived neurotrophic factor (BDNF) and pituitary adenylate cyclase-activating polypeptide (PACAP) in the pituitary gland were reduced in frogs whose coloration was pale brown on a yellow background. In particular, the weakest expressions of BDNF, PACAP, and PACAP type II receptors (VPAC-1R) were found in male frogs with a brilliant yellow body color during the breeding season regardless of background color. These changes may decrease α-melanocyte-stimulating hormone production associated with the PACAP receptors (VPAC-1R), resulting in the aggregation of black pigment in melanophores and the production of a brilliant yellow body color. The effects of hormones on skin coloration were further examined in isolated skin in vitro. The results of this investigation showed that the dispersion of xanthophores was stimulated by T or prolactin (PRL) and that the melanophores were aggregated by melatonin (MEL) but not by E2. Furthermore, yellow pigments in the xanthophores were significantly dispersed following the PRL+T treatment. In the T+MEL, PRL+MEL, and T+PRL+MEL treatments, xanthophores were dispersed, and melanophores were aggregated and subsequently moved to the low spongiosum layer of the dorsal skin, causing the increase in yellow coloration. These results reveal that multiple hormones play major roles in the regulation of the brilliant yellow coloration of male B. robusta by high plasma T during the breeding season.

Background: Bufo bankorensis is an endemic species in Taiwan, and its populations are geographically and reproductively isolated. However, the distinction of Taiwanese B. bankorensis as a separate species from the Chinese Bufo gargarizans remains in dispute. Results: A primer set was designed to explore the mitochondrial (mt)DNA cytochrome (Cyt) b sequence (700 bp) of B. bankorensis in 148 individuals collected from 12 locations in Taiwan. After a polymerase chain reaction and sequencing, we found that the nucleotide sequence of Cyt b contained two restricted enzyme sites of BamHI and TspRI. Following BamHI enzyme digestion, samples of B. bankorensis were divided into two clades: western (which were undigested) and eastern (which were digested) clades. Additionally, Cyt b of the western clade of B. bankorensis was not cut by BamHI, while it was cut by TspRI into two sublineages. The result infers that at least two broadly divergent phylogroups of B. bankorensis exist in Taiwan and are not morphologically distinguishable. Based on the divergent sequence of Cyt b and cutting restriction enzymes, these populations were classified into three distinct phylogroups.Conclusion: Genetically, one (western group 1, uncut by BamHI and cut by TspRI) is most likely B. gargarizans, a second one (western group 2, uncut by both BamHI and TspRI) is B. bankorensis, and a third one (eastern clade, cut by BamHI but not cut by TspRI) could be a new subspecies. All three phylogroups were found in some areas, suggesting that they are sympatric, not allopatric.

Ocean litter has accumulated rapidly and is becoming a major environmental concern, yet quantitative and regular observations and exploration that track litter origins are limited. By implementing monthly sample collections over five years (2012)(2013)(2014)(2015)(2016) at Dongsha Island, a remote island in the northern South China Sea (SCS), we assessed macro ocean litter dynamics, identified source countries of individual plastic bottles, and analyzed the origins of the litter by a backward-tracking model simulation considering both the effects of current velocity and windage. The results showed that large amounts of litter, which varied monthly and annually in weight and quantity, reached the island during the study years, and there were spatial differences in accumulation patterns between the north and south coasts. Styrofoam and plastic bottles were the two primary sources of macro ocean litter both annually and monthly, and most of the litter collected on the island originated from China and Vietnam, which were collectively responsible for approximately 47.5%-63.7% per month. The simulation indicated that current advection at the near-surface depths and low windage at the sea surface showed similar patterns, while medium to high windage exhibited comparable expression patterns in response to potential source regions and drifting time experiments. At either the surface with low windage or current advection at depths of 0.5 m and 1 m, macro ocean litter in the Western Philippine Sea, i.e. through the Luzon Strait between Taiwan and the Philippines, was an important contributor to the litter bulk from October to March, whereas the litter was predicted to mainly originate from the southwestern SCS from April to September. With an increasing windage effect, litter in the Taiwan Strait was predicted to be an additional major potential source. Surprisingly, a small proportion of the macro ocean litter was predicted to continuously travel in the northern SCS for a long duration (> 2 years) before drifting onto Dongsha Island. The estimated drifting time of macro ocean litter also showed monthly and directional variability. This study demonstrated that a tremendous quantity of macro ocean litter, which may cause great damage to the marine ecosystem, drifts in the ocean surface layer and is finally pushed onto beaches. Therefore, we proposed an action plan for effective ocean litter management development at regional and global spatial scales, which is vital for improving and restoring the health and sustainability of the oceanic environment.

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