Predicting ecosystem changes by a new model of ecosystem evolution.

In recent years, computer simulation has been increasingly used to predict changes in actual ecosystems. In these studies, snapshots of ecosystems at certain points in time were instantly constructed without considering their evolutionary histories. However, it may not be possible to correctly predict future events unless their evolutionary processes are considered. In this study, we developed a new ecosystem model for reproducing the evolutionary process on an oceanic island, targeting Nakoudojima Island of the Ogasawara Islands. This model successfully reproduced the primitive ecosystem (the entire island covered with forest) prior to the invasion of alien species. Also, by adding multiple alien species to this ecosystem, we were able to reproduce temporal changes in the ecosystem of Nakoudojima Island after invasion of alien species. Then, we performed simulations in which feral goats were eradicated, as had actually been done on the island; these suggested that after the eradication of feral goats, forests were unlikely to be restored. In the ecosystems in which forests were not restored, arboreous plants with a high growth rate colonized during the early stage of evolution. As arboreous plants with a high growth rate consume a large amount of nutrient in soil, creating an oligotrophic state. As a result, plants cannot grow, and animal species that rely on plants cannot maintain their biomass. Consequently, many animals and plants become extinct as they cannot endure disturbances by alien species, and the ecosystem loses its resilience. Therefore, even if feral goats are eradicated, forests are not restored. Thus, the founder effect from the distant past influences future ecosystem changes. Our findings show that it is useful to consider the evolutionary process of an ecosystem in predicting its future events.

An EBSD camera as a tool to characterise in-plane magnetisation vectors on Fe-Si (001) surface.

Previous studies have shown that type-II magnetic-domain contrasts are caused by differences in the backscattering yields of magnetic domains of opposite magnetisation. Imaging the magnetic domains when the magnetisation vectors in the opposite-magnetisation domains are perpendicular to the tilt axis of the specimen has been considered difficult, because of the lack of change in the backscattering yields between the domains. An alternative way to obtain the type-II magnetic-domain contrasts is to utilise the difference in the exit angular distribution of the backscattered electrons from different magnetic domains. In this study, it is found that an electron backscatter diffraction (EBSD) camera can be used to obtain the type-II magnetic-domain contrasts caused by the above two mechanisms simultaneously. We verify this by distinguishing all four possible in-plane magnetisation vectors on a Fe-Si (001) surface without a sample rotation, using an EBSD detector as an array of electron detectors. The change in contrast between the magnetic domains, with respect to the location of a virtual electron detector, can provide information on the directions of the magnetisation vectors. A method to suppress the topographic contrast superimposed on the magnetic-domain contrast is also demonstrated.

Contrasting patterns of population structure of Bulwer's petrel (Bulweria bulwerii) between oceans revealed by statistical phylogeography.

The patterns of population divergence of mid-latitude marine birds are impacted by only a few biogeographic barriers to dispersal and the effect of intrinsic factors, such as fidelity to natal colonies or wintering grounds, may become more conspicuous. Here we describe, for the first time, the phylogeographic patterns and historical demography of Bulwer's petrel Bulweria bulwerii and provide new insights regarding the drivers of species diversification in the marine environment. We sampled Bulwer's petrels from the main breeding colonies and used a statistical phylogeography approach based on surveying nuclear and mitochondrial loci (~ 9100 bp) to study its mechanisms of global diversification. We uncovered three highly differentiated groups including the Western Pacific, the Central Pacific and the Atlantic. The older divergence occurred within the Pacific Ocean, ca. 850,000 ya, and since then the W Pacific group has been evolving in isolation. Conversely, divergence between the Central Pacific and Atlantic populations occurred within the last 200,000 years. While the Isthmus of Panama is important in restricting gene flow between oceans in Bulwer's petrels, the deepest phylogeographic break is within the Pacific Ocean, where oceanographic barriers are key in driving and maintaining the remarkable structure found in this highly mobile seabird. This is in contrast with the Atlantic, where no structure was detected. Further data will provide insights regarding the extent of lineage divergence of Bulwer's petrels in the Western Pacific.

A widespread survey of avian haemosporidia in deceased wild birds of Japan: the hidden value of personally collected samples.

Widespread surveys of avian haemosporidia (Plasmodium, Haemoproteus, and Leucocytozoon) in wild birds have substantially advanced information on the haemosporidian fauna of Japan. However, many areas and bird species remain insufficiently investigated. Bird carcasses collected for personal specimen collection seldom reach academic audience particularly in the veterinary field. The presence of avian haemosporidia was investigated in these personally collected bird carcasses, in order to better understand the avian haemosporidian fauna in Japan. Bird carcasses were donated through personal contact upon approval of the study. Tissue samples were collected from the birds and examined for haemosporidian parasites using nested-PCR targeting the cytochrome b gene. One hundred and forty-three birds of 85 species were donated, including 34 species and two subspecies that were molecularly or collectively investigated for the first time in Japan. Avian haemosporidian DNA was detected from 37 of the 134 tested birds (27.61%). In 8 bird species, avian haemosporidia was detected for the first time. Twenty-nine lineages were detected, including 8 novel and 9 known lineages detected in Japan for the first time. Furthermore, 16 lineages were detected from novel host species. While information that could be drawn was limited and risk management of zoonotic diseases needs re-consideration, these findings expanded information on the host range and distribution of several lineages. Collectively, this method of investigation using personally collected bird samples can provide important additions to more fully understand the avian haemosporidian fauna of Japan, as well as other areas with limited investigations.

Quantitative Analysis of Nitrogen by Atom Probe Tomography Using Stoichiometric γ'-Fe4N Consisting of 15N Isotope.

The nitrogen deficiency in steels measured by atom probe tomography (APT) is considered to arise from the obscurement of singly charged dimer nitrogen ions (N2+) by the iron-dominant peak (56Fe2+) at 28 Da. To verify this by quantifying the amount of N2+ ions, γ'-Fe4N consisting of the 15N isotope was prepared on iron substrates by plasma nitriding using a nitrogen isotopic gas (15N2). Although considerable amounts of 15N2+ were observed at 30 Da without overlap with any iron peak, the observed nitrogen concentrations of γ'-Fe4N were clearly lower than the stoichiometric composition (19­20 at%), using both pulsed voltage and pulsed laser atom probes. The origin of the missing nitrogen, excluding nitrogen obscured by other ion species, was predicted to be the occurrence of neutral nitrogen or nitrogen gas molecules in field evaporation. The generation rate of iron nitride ions (FeN2+) for 15N was significantly lower than that for 14N in γ'-Fe4N, which affected the amount of the missing nitrogen. The isotope effect suggests that the isotopic ratio cannot always be determined from only one ion species among the multiple species observed in the APT analysis. We discuss the mechanism of the isotope effect in FeN2+ formation by field evaporation.

Determination of the Chemical Compositions of Fine titanium Carbide and Niobium Carbide Precipitates in Isothermally Aged Ferritic Steel by Atom Probe Tomography Analysis.

The carbon (C) ratios, namely the atomic ratios of C/(C + M), in nano-sized coherent MC precipitates (M = Ti, Nb) with the NaCl-type (B1) structure in ferritic steels, which had been isothermally aged at 580 °C, were investigated using atom probe tomography (APT). Considering the influences of the trajectory aberration, detection loss, and peak overlap, we determined the C ratios to be ~0.40 and ~0.45 for an equivalent volume diameter of 1.5­5 nm and 1­5 nm for the TiC and NbC precipitates, respectively, suggesting that there is a considerable fraction of C vacancies in both nano-sized precipitates. The apparent C ratios show significant scatter with decreasing particle size, while the apparent mean C ratios of very fine TiC particles, smaller than 1.5 nm, decreased with decreasing particle size. With the use of one of the latest APT instruments with a high detection efficiency, the scattering in the apparent C ratios was reduced because the counting statistics were improved; however, the artificial enrichment of C atoms to particular crystallographic directions of ferrite hindered the determination of the C ratio for very fine TiC particles smaller than 1.5 nm.

Cryptic Speciation of the Oriental Greenfinch Chloris sinica on Oceanic Islands.

The Oriental greenfinch, Chloris sinica, is a small seed-eating finch that breeds in the eastern Palearctic region, an area that spans from Russia in the east to China, Korea, and Japan in the south and southwest. Several subspecies have been described based on subtle morphological characteristics, although the taxonomy varies among different authors. Although many ecological studies have been performed, there has been no phylogenetic study that encompasses the species' entire geographical range. We used four regions of mitochondrial DNA to analyze the intraspecies genetic phylogeny and diversity of the Oriental greenfinch. In addition, we performed morphometric analyses using museum specimens. Genetic analysis identified two clades that diverged approximately 1.06 million years ago. These were a population from the Ogasawara Islands, Japan (subspecies kittlitzi, Clade B), and the other populations (Clade A, which could not be subdivided according to geographic context). Morphometric analyses showed that the population on the Kuril Islands (subspecies kawarahiba) had the longest mean wing length, whereas C. s. kittlitzi had the shortest wings. Chloris s. kittlitzi also had the longest mean bill length, probably because it has adapted to feeding on the Ogasawara Islands. Based on molecular phylogeny and morphology analyses, we recommend that C. s. kittlitzi should be treated as a completely distinct species, called the Ogasawara greenfinch, Chloris kittlitzi. It is critically endangered and needs to be specially protected.

Study on Quantitative Analysis of Carbon and Nitrogen in Stoichiometric θ-Fe3C and γ'-Fe4N by Atom Probe Tomography.

The quantitative analysis performance of carbon and nitrogen was investigated using stoichiometric θ-Fe3C (25 at% C) and γ'-Fe4N (~20 at% N) precipitates in pulsed voltage and pulsed laser atom probes. The dependencies of specimen temperature, pulse fraction, and laser pulse energy on the apparent concentrations of carbon and nitrogen were measured. Good coincidence with 25 at% carbon concentration in θ-Fe3C was obtained for the pulsed voltage atom probe by considering the mean number of carbon atoms per ion at 24 Da and the detection loss of iron, while better coincidence was obtained for the pulsed laser atom probe by considering only the mean number of carbon at 24 Da. On the other hand, a lack of nitrogen concentration in γ'-Fe4N was observed for the two atom probes. In particular, the pulsed laser atom probe showed a significant lack of nitrogen concentration. This implies that a large amount of 14N2+ was obscured by the main iron peak of 56Fe2+ at 28 Da in the mass-to-charge spectrum. Regarding preferential evaporation or retention, carbon in θ-Fe3C exhibited little of either, but nitrogen in γ'-Fe4N exhibited definite preferential retention. This result can be explained by the large difference in ionization energy between carbon and nitrogen.

Genetic Structure and Diversity of Two Populations of the Eastern Buzzard (Buteo japonicus japonicus and B. j. toyoshimai) in Japan.

The nominotypical subspecies of the Eastern buzzard (Buteo japonicus japonicus; BJJ) is a common raptor inhabiting East Asia and Japan. Another subspecies, B. j. toyoshimai (BJT), inhabits only the Bonin Islands of the Ogasawara Islands, where there are only an estimated 85 breeding pairs. Because of this low population size, this subspecies is classified as endangered (class IB) in Japan. The aims of the present study were to examine genetic differences between BJJ and BJT, determine the genetic structure of the Eastern Buzzard, and assess genetic diversity within each subspecies. We sequenced 1526 bp within the control region of the mtDNA of 10 BJJ individuals during the breeding season in four sites; similarly, we sequenced 23 BJJ individuals during winter in three sites. We detected 24 haplotypes among the 33 individuals. In a similar analysis performed with 12 BJT individuals, three haplotypes were detected. The phylogenetic analysis showed that BJJ and BJT have diverged into distinct clades, supporting the genetic differentiation between the subspecies. Network and mismatch distribution analyses indicated that BJJ may have experienced population expansion. In addition, comparisons with other raptors revealed a high degree of genetic diversity in the BJJ population. In contrast, the genetic diversity of the BJT population is lower than that in other raptors. Our results indicated that it is necessary to protect BJT to prevent the reduction in its genetic diversity.

Origin of Japanese White-Eyes and Brown-Eared Bulbuls on the Volcano Islands.

The Ogasawara Archipelago comprises two groups of oceanic islands: the Bonin Islands, formed in the Paleogene, and the Volcano Islands, formed in the Quaternary. These groups are located within a moderate distance (ca. 160-270 km) of one another; thus, most land bird species are not distinguished as different subspecies. Two land birds, however, show unusual distribution. The Japanese white-eyes Zosterops japonicus originally inhabited only the Volcano Islands, but has been introduced to the Bonin Islands. The brown-eared bulbuls Hypsipetes amaurotis are distributed as a different subspecies. We investigated their genetic differences and divergences in the Ogasawara Archipelago using mitochondria DNA. The Volcano population of white-eyes had four endemic haplotypes that were divergent from one another, except for the Bonin population, which shared three haplotypes with the Volcano, Izu, and Ryukyu Islands and did not have any endemic haplotype. This is the first genetic suggestion that the Bonin population is a hybrid of introduced populations. With respect to bulbuls, the Volcano and Bonin Islands each had a single endemic haplotype. The Volcano haplotype is closest to a haplotype shared with Izu, the Japanese mainland, Daito and Ryukyu, whereas the Bonin haplotype is closest to one endemic to the south Ryukyu Islands. This indicates that the sources of the two bulbul populations can be geologically and temporally distinguished. The populations of the two species in the Ogasawara Archipelago are irreplaceable, owing to their genetic differences and should be regarded as evolutionarily significant units. In order to prevent introgression between the two populations, we must restrict interisland transfers.

Increases in soil water content after the mortality of non-native trees in oceanic island forest ecosystems are due to reduced water loss during dry periods.

The control of dominant, non-native trees can alter the water balance of soils in forest ecosystems via hydrological processes, which results in changes in soil water environments. To test this idea, we evaluated the effects of the mortality of an invasive tree, Casuarina equisetifolia Forst., on the water content of surface soils on the Ogasawara Islands, subtropical islands in the northwestern Pacific Ocean, using a manipulative herbicide experiment. Temporal changes in volumetric water content of surface soils at 6 cm depth at sites where all trees of C. equisetifolia were killed by herbicide were compared with those of adjacent control sites before and after their mortality with consideration of the amount of precipitation. In addition, the rate of decrease in the soil water content during dry periods and the rate of increase in the soil water content during rainfall periods were compared between herbicide and control sites. Soil water content at sites treated with herbicide was significantly higher after treatment than soil water content at control sites during the same period. Differences between initial and minimum values of soil water content at the herbicide sites during the drying events were significantly lower than the corresponding differences in the control quadrats. During rainfall periods, both initial and maximum values of soil water contents in the herbicided quadrats were higher, and differences between the maximum and initial values did not differ between the herbicided and control quadrats. Our results indicated that the mortality of non-native trees from forest ecosystems increased water content of surface soils, due primarily to a slower rate of decrease in soil water content during dry periods.

In situ determination of misorientation angle of grain boundary by field ion microscopy analysis.

We proposed an advanced analysis technique for characterizing a grain boundary using field ion microscopy (FIM) for atom probe analysis. The technique enables quick and precise estimation of the misorientation angle of the grain boundary by matching the calculated crystallographic pole positions with the actual FIM image including the grain boundary. We investigated the accuracy in estimation of the misorientation angle using target grain boundaries which had been analyzed by electron backscatter diffraction pattern (EBSD) analysis. From the comparison between EBSD and FIM analyses, we found that the technique enables the determination of the misorientation angle with a high accuracy of ± 0.4°, which is comparable with that achieved by EBSD.

Limited overwater dispersal and genetic differentiation of the snake-eyed skink (Cryptoblepharus nigropunctatus) in the Oceanic Ogasawara Islands, Japan.

The genetic differentiation and speciation of lizards on oceanic islands may be affected by their rate of overwater dispersal. Cryptoblepharus is one of the most geographically widespread scincid lizards throughout the Indo-Pacific and Australian regions. Cryptoblepharus nigropunctatus is the northernmost species of the genus, dwelling on several small Pacific islands. To examine the colonization history of this lizard, mitochondrial 16S rDNA and D-loop sequences were compared among populations of the Ogasawara Islands consisting of four island groups (the Muko-jima, Chichi-jima, Haha-jima, and Kazan groups), and an isolated island, Minamitori-shima (Marcus Island). These four groups and Minamitori-shima have not been connected to each other because each is surrounded by deep sea (>100 m). DNA analyses showed that the lizard populations on individual islands had each representative haplotypes. The ancestors of C. nigropunctatus probably arrived on the islands from the southern Pacific Ocean via wave dispersal and differentiated to produce the present state. They appear to have dispersed from their origin along two independent pathways: one between Kitaiwo-to (Kazan group) and the Muko-jima and Chichi-jima groups, and the other among the Minamitori-shima, Minamiiwo-to (Kazan group), and Haha-jima groups. Limited long-distance overwater dispersal may be responsible for the genetic structure of the C. nigropunctatus populations on these oceanic islands. However, among the small islands within the same island group, D-loop haplotypes were shared and the local genetic diversity was usually high, suggesting frequent gene flow across the same group of islands.

Three-dimensional atom probe analysis of heavily drawn steel wires by probing perpendicular to the pearlitic lamellae.

Three-dimensional atom probe analysis of heavily drawn pearlitic steel wires was conducted by probing in the direction perpendicular to the pearlitic lamellae. A needle tip perpendicularly intersecting the lamellar structure was prepared using focused ion beam (FIB) milling combined with the lift-out method. The specimen preparation technique enabled the analysis of many lamellae and their interfaces with higher depth spatial resolution, which was suitable for drawn pearlitic steel wires having inhomogeneous and fine lamellar structure. Carbon concentration peaks at the cementite lamellae appeared higher and narrower than those obtained by probing parallel to the lamellae, which implies that the conventional analysis overestimated the extent of cementite decomposition in drawn pearlitic steel wires.

Genetic and morphological differences among populations of the Bonin Islands White-eye in Japan.

The Bonin Islands White-eye, Apalopteron familiare , is the sole endemic avian species surviving on the Bonin Islands. The current distribution of this species is limited to only three islands of the Hahajima Island group: Hahajima, Imotojima, and Mukohjima. Imotojima and Mukohjima, which are small satellite islands of Hahajima, are about 3.6 km and 5.5 km, respectively, from the larger island. To investigate genetic and morphological differences among A. familiare populations on these islands, we assayed 634 bp of mitochondrial control region sequence for 132 birds from five locations among the three islands. We detected five haplotypes: two endemic haplotypes each on Hahajima and Imotojima and one on Mukohjima. Principal component analysis based on eight morphological characters of 162 birds from the three island populations revealed that birds from the small satellite islands had significantly different beak morphological characters. Our findings indicate that over-sea dispersal is rare in A. familiare , even when islands are separated by only a few kilometers, and suggest that little movement occurs on Hahajima. Thus, each population on each island should be conserved as an evolutionarily significant unit. The low dispersal ability of this species suggests that it is vulnerable to habitat fragmentation. Although the populations are currently stable and do not require rapid countermeasures, they should be monitored, especially those on the smaller islands.

Hopping and climbing gait of Japanese Pygmy Woodpeckers (Picoides kizuki).

Single cycles of hopping and climbing were investigated in Japanese Pygmy Woodpeckers Picoides kizuki using motion analyses on video. Body movements on substrate angled from 0-90 degrees were compared for every 10 degrees. The body was inclined forward during stance phase for both small and large substrate angles, and the inclination amplitude increased when the substrate angle increased. The tail was bent ventrally almost simultaneously to this body inclination, and its amplitude was apparently high at large substrate angles. Most of the gait parameters changed when the stride length increased. The minimum body-tail angle and most of the parameters representing body movements during stance phase changed when the substrate angle increased, probably because gravity pulled the birds further backward when they were moving on a steeper slope. These parameters showed a clear difference between the data on substrate steeper than 40 degrees and lower than 30 degrees. The abrupt changes in these parameters most likely mean that the motor pattern changed from hopping to climbing between these angles.

Development of atom probe specimen preparation techniques for specific regions in steel materials.

More elaborated specimen preparation techniques for atom probe analysis were developed using a focused ion beam with a sample lift-out system so as to expand the application field in steel materials. The techniques enable atom probe analysis of sample steel at site-specific regions of interest. The preferable form of the needle specimen was provided by electrostatic field calculation using a finite element method. The new techniques were applied to the observation of a bainite-ferrite interface in a low carbon steel, and atomic-scale partitioning and segregation of alloying elements at the phase interface were directly observed in three dimensions.