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In 1980 and 1981, eight and nine individual sika deer Cervus nippon were reintroduced in southern Hokkaido, Japan, respectively, to address population declines in this species during 1900s. As recent population growth has led to human-wildlife conflicts, this study investigated the responses of sika deer to resource availability and geomorphic factors during the summer and winter seasons in southern Hokkaido. Global positioning system-collared data collected from 2016 to 2018 were used to assess the home range patterns and habitat selection of 14 female sika deer located in Mount Esan and Shiriuchi. The core home range size was defined using a 50% kernel density estimation that indicated a larger home range in winter than summer for all deer. Habitat selection was assessed using generalized linear mixed models. The results showed variation in habitat selection between resident deer of Mount Esan and Shiriuchi, as well as migratory deer in Shiriuchi during summer. Resident deer in Mount Esan and Shriuchi preferred areas closer to crops during summer. Interaction effects revealed that migratory deer utilized natural grassland close to forest edge habitat in Shiriuchi. By contrast, resident deer in Shiriuchi selected forest edge habitat and natural grasslands close to crops. In winter, low elevation was the most important habitat factor for all deer across the study area. Thus, sika deer habitat selection depends on resource availability in summer and topographic factors in the winter.

We report on the discovery of a new supergiant fast X-ray transient (SFXT), MAXI J0709$-$159, and its identification with LY CMa (also known as HD 54786). On 2022 January 25, a new flaring X-ray object named MAXI J0709$-$159, was detected by Monitor of All-sky X-ray Image (MAXI). Two flaring activities were observed in the two scans of $\sim 3$ hours apart, where the 2-10 keV flux reached $5\times 10^{-9}$ erg cm$^{-2}$ s$^{-1}$. During the period, the source exhibited a large spectral change suggesting that the absorption column density $N_\mathrm{H}$ increased from $10^{22}$ cm$^{-2}$ to $10^{23}$ cm$^{-2}$. NuSTAR follow-up observation on January 29 identified a new X-ray source with a flux of $6\times 10^{-13}$ erg cm$^{-2}$ s$^{-1}$ at the position consistent with LY CMa, which has been identified as B supergiant as well as Be star, located at the 3 kpc distance. The observed X-ray activity characterized by the short ($\lesssim$ several hours) duration, the rapid ($\lesssim$ a few seconds) variabilities accompanied with spectral changes, and the large luminosity swing ($10^{32}$-$10^{37}$ erg s$^{-1}$) agree with those of SFXT. On the other hand, optical spectroscopic observations of LY CMa revealed a broad $H\alpha$ emission line, which may indicate the existence of a Be circumstellar disk. These obtained results suggest that the optical companion, LY CMa, certainly has a complex circumstellar medium including dense clumps., Comment: 13 pages, 10 figures, accepted for publication in PASJ (v2. author names in metadata are modified.)

In recent years, Arenz et al. proposed the idea of reachable set characterization based on the quantum speed limit (QSL); that is, the reachable set of the target unitary gate in a closed qubit system can be characterized by considering the QSL as the necessary condition that the control setup must satisfy in order to achieve the goal. Inspired by this idea, in this paper we characterize a general Markovian open quantum system based on the QSL derived in \cite{Kohei2}. Note that this bound is not only explicitly computable with respect to system parameters, but also tighter than the other bounds. Some examples for demonstrating this analysis will be given., Comment: 6 pages, 2 figures

Nocturnal predators of many taxa are known to come to artificial light at night for foraging on clumped food resources. Both innate and acquired light preferences seem to be possible mechanisms of light approaching behavior although empirical tests are lacking in most nocturnal predators. Here, using a Japanese gecko Gekko japonicus, we investigated whether geckos have a light preference and how foraging experiences under the light reinforce light approaching tendency. In a comparative experiment, there was no difference in light approaching behavior between urban and suburban geckos irrespective of their original light habitats. In an associative learning experiment, geckos did not significantly change light approaching behavior even after repeated opportunities to forage crickets near a lamp in the laboratory setting. These results imply that light approaching behavior of Japanese geckos may not be easily reinforced by foraging experiences under the light. Although we often witness geckos coming to artificial light at night, our findings may not suggest their light preference. Geckos may approach the light-up foraging spot based on other cues relating to the artificial light environment.

Circadian rhythm is an approximately 24 h endogenous biological rhythm. Chronic disruption of the circadian clock leads to an increased risk of diabetes, cardiovascular disease, and cancer. Hence, it is important to develop circadian clock modulators. Natural organisms are a good source of several medicines currently in use. Crude drugs used in Japanese traditional Kampo medicine or folk medicines are an excellent source for drug discovery. Furthermore, identifying new functions for existing drugs, known as the drug repositioning approach, is a popular and powerful tool. In this study, we screened 137 crude drug extracts to act as circadian clock modulators in human U2OS cells stably expressing the clock reporter Bmal1-dLuc, and approximately 12% of these modulated the circadian rhythm. We further examined the effects of several crude drugs in Rat-1 fibroblasts stably expressing Per2-luc, explant culture of lung from Per2::Luciferase knockin mice, and zebrafish larvae in vivo. Notably, more than half of the major ingredients of these crude drugs were reported to target AKT and its relevant signaling pathways. As expected, analysis of the major ingredients targeting AKT signaling confirmed the circadian clock-modulating effects. Furthermore, activator and inhibitor of AKT, and triple knockdown of AKT isoforms by siRNA also modulated the circadian rhythm. This study, by employing the drug repositioning approach, shows that Kampo medicines are a useful source for the identification of underlying mechanisms of circadian clock modulators and could potentially be used in the treatment of circadian clock disruption.

Recently, controlling cell function has attracted significant attention, with the possibility of changing various forms of cells and organs. Cell modification technology operates through various methods, such as the virus vector and lipofection methods; however, these approaches have problems in terms of efficiency and process optimization. In contrast, our photochemical cell membrane perforation method exhibits high efficiency, but requires a thermosetting silicone polymer-based cell membrane perforator as a consumable item, which requires 4 h for production. In this study, we present a process using an injection molding machine for manufacturing thermoplastic elastomer-based perforators for mass production, successfully shortening the production time to < 1 h while preserving the perforating function.

研究チームは、最小の細菌である「マイコプラズマ・モービレ」が滑走するための分子モーターの動きを検出することに世界で初めて成功しました。この発見は、ナノスケールのデバイスや医薬品の開発への応用が期待されます。, Mycoplasma mobile, a parasitic bacterium, glides on solid surfaces, such as animal cells and glass, by a special mechanism. This process is driven by the force generated through ATP hydrolysis on an internal structure. However, the spatial and temporal behaviors of the internal structures in living cells are unclear. In this study, we detected the movements of the internal structure by scanning cells immobilized on a glass substrate using high-speed atomic force microscopy (HS-AFM). By scanning the surface of a cell, we succeeded in visualizing particles, 2 nm in height and aligned mostly along the cell axis with a pitch of 31.5 nm, consistent with previously reported features based on electron microscopy. Movements of individual particles were then analyzed by HS-AFM. In the presence of sodium azide, the average speed of particle movements was reduced, suggesting that movement is linked to ATP hydrolysis. Partial inhibition of the reaction by sodium azide enabled us to analyze particle behavior in detail, showing that the particles move 9 nm right, relative to the gliding direction, and 2 nm into the cell interior in 330 ms and then return to their original position, based on ATP hydrolysis.

In this paper, we propose a concept to use a quantum speed limit (QSL) as a measure of robustness of states, defining that a state with bigger QSL is more robust. In this perspective, it is important to have an explicitly-computable QSL, because then we can formulate an engineering problem of Hamiltonian that makes a target state robust against decoherence. Hence we derive a new explicitly-computable QSL that is applicable to general Markovian open quantum systems. This QSL is tighter than another explicitly-computable QSL, in an important setup such that decoherence is small. Also the Hamiltonian engineering problem with this QSL is a quadratic convex optimization problem, and thus it is efficiently solvable. The idea of robust state characterization and the Hamiltonian engineering, in terms of QSL, is demonstrated with several examples., 7 pages, 4 figures

In this study, we aim to develop a system of remote-controlled avatar robot for elderly and disabled patients. Most of teleoperation systems have interfaces to visually present the state of the robots including feedback information and receive the control commands manually sent from the operator. However, in elderly and patients with disabilities, they might have difficulty in the manual control of the robot. This paper therefore presents a multimodal interface for remotely controlling a robotic avatar. We furthermore propose a cognitive platform for remotely controlling a robot based on a concept of perceiving-acting cycle. The platform consists of a perceptual system for incremental environment modeling and an action system for extracting patterns of operator behavior. In each system, a self-organized neural network based on unsupervised learning is used. Moreover, we use a spiking neural network for spatial-temporal modeling of interaction between an operator and environment.

We report the results from the broad-band X-ray monitoring of the new Galactic black hole candidate MAXI J1803$-$298 with the MAXI/GSC and Swift/BAT during its outburst. After the discovery on 2021 May 1, the soft X-ray flux below 10 keV rapidly increased for $\sim 10$ days and then have been gradually decreasing over 5 months. At the brightest phase, the source exhibited the state transition from the low/hard state to the high/soft state via the intermediate state. The broad-band X-ray spectrum during the outburst was well described with a disk blackbody plus its thermal or non-thermal Comptonization. Before the transition the source spectrum was described by a thermal Comptonization component with a photon index of $\sim 1.7$ and an electron temperature of $\sim 30$ keV, whereas a strong disk blackbody component was observed after the transition. The spectral properties in these periods are consistent with the low/hard state and the high/soft state, respectively. A sudden flux drop with a few days duration, unassociated with a significant change in the hardness ratio, was found in the intermediate state. A possible cause of this variation is that the mass accretion rate rapidly increased at the disk transition, which induced a strong Compton-thick outflow and scattered out the X-ray flux. Assuming a non-spinning black hole, we estimated a black hole mass of MAXI J1803$-$298 as $5.8 \pm 0.4~(\cos i/\cos 70^\circ)^{-1/2} (D/8~\mathrm{kpc})~M_\odot$ (where $i$ and $D$ are the inclination angle and the distance) from the inner disk radius obtained in the high/soft state., Comment: 13 pages, 10 figures, Accepted for publication in ApJ

Adult male brown bears (Ursus arctos Linnaeus, 1758) display tree-marking behavior to chemically signal their dominance throughout the nondenning period, and this behavior peaks during the breeding season. Within the scent-marking sequence, back rub is a core marking posture. The present study investigated (i) seasonal changes in sebaceous glands in the back skin of brown bears and (ii) the relationship between those changes and testosterone levels. Back skin tissue samples and blood were collected from captive adult intact and castrated males during prebreeding, transitional, breeding, and postbreeding seasons, which were concurrent with back skin observations. In intact males, during the transitional and breeding seasons, an oily secretion from the back skin was observed along with enlarged sebaceous glands. The plasma testosterone concentrations during the transitional and breeding seasons were increased compared with the pre- and post-breeding seasons. Secretions and enlarged sebaceous glands were not found in castrated males, and the plasma testosterone concentrations remained at baseline levels. Oily secretions of the back skin glands that appear more abundant during the breeding season are rubbed against trees. Changes in size and volume of sebaceous glands, and thus their secreting capacity, are likely testosterone-regulated.

Recently, the widespread occurrence of Sri Lankan cassava mosaic virus (SLCMV), genus Begomovirus, family Geminiviridae, which causes a mosaic disease in cassava (Manihot esculenta Crantz) in South-East Asia have, become a serious economic issue. Since cassava is propagated through vegetative cuttings, a rapid virus diagnostic method is crucial for generating virus-free planting materials. In this study, a loop-mediated isothermal amplification (LAMP) assay using six primers was developed and validated for the rapid detection of SLCMV in cassava leaves. This SLCMV assay had a detection sensitivity that was up to 10,000 times higher than that of the conventional polymerase chain reaction assay and can detect the virus from symptomless stem cutting, which is a potential long-distance spreader of the virus. Furthermore, a practical LAMP protocol using stable dried reagents from a commercial kit was established so that the assay could be performed in the field by incubating the reactions in water at 60-65 °C instead of using a thermal cycler. The primer sequences and the LAMP protocol described here should be useful for the rapid and sensitive on-site detection of SLCMV.

Quantum information technologies require careful control for generating and preserving a desired target quantum state. The biggest practical obstacle is, of course, decoherence. Therefore, the reachability analysis, which in our scenario aims to estimate the distance between the controlled state under decoherence and the target state, is of great importance to evaluate the realistic performance of those technologies. This paper presents a lower bound of the fidelity-based distance for a general open Markovian quantum system driven by the decoherence process and several types of control including feedback. The lower bound is straightforward to calculate and can be used as a guide for choosing the target state, as demonstrated in some examples. Moreover, the lower bound is applied to derive a theoretical limit in some quantum metrology problems based on a large-size atomic ensemble under control and decoherence., 10 pages, 3 figures

This paper aims to develop a measurement system for the finger joint angle by using a flexible polymer sensor. In rehabilitation, it is important to evaluate human motions quantitatively for effective treatment. In this study, we focus on hand's grasping motion, which is important motion for eating and drinking. About the flexible polymer sensor, in the previous studies, it has been elucidated that the outputs from the flexible polymer sensor are proportional to the average curvature of measuring surface. Therefore, using the sensor, finger bending motion can be measured by attaching the sensor on finger surface. Firstly, the relationship between the average curvature of finger surface and finger joint angle is theoretically investigated. Then, in order to obtain the relationship between sensor outputs and finger joint angles, sensor outputs and finger joint angles during grasping acrylic cylinders are measured. From the results, it was found that the developed system is useful to measure the finger joint angles.

Several reptiles and amphibians frequently occur in environments exposed to artificial lights and appear to effectively hunt prey animals attracted to the light. Therefore, it would be beneficial for them to use such a habitat to survive in urban areas. In this paper, we examined site fidelity of a nocturnal gecko, Gekko japonicus, to a particular artificial light environment in the field. We observed short-term dependence on the same light environment by individual geckos for four to nine consecutive nights and also examined long-term site fidelity once every 10 days from March to November. The same individual frequently came to the particular light for several consecutive nights, but day and time of appearance varied among individuals. Over several months, 20% of marked individuals were repeatedly sighted at the same place although the number of their re-sightings was low (1–5 times). These results suggest that each G. japonicus may settle down in a certain artificially lit environment and use it repeatedly, but the dependence on lights not be so strong.

Accumulation of mutations in mitochondrial DNA (mtDNA) is thought to be responsible for mitochondrial, and other, diseases and biological phenomena, such as diabetes, cancer, neurodegenerative diseases, and aging. Mouse models may elucidate the relationship between mutations in mtDNA and these abnormalities. However, because of the difficulty of mtDNA manipulation, generation of mouse models has not sufficiently progressed to enable such studies. To overcome this difficulty and to establish a source of diverse mtDNA mutations, we here generated cultured mouse cells containing mtDNA derived from an mtDNA mutator mouse that accumulates random mtDNA mutations with age. Mutation analysis of the obtained transmitochondrial cytoplasmic hybrid cells (cybrids) revealed that the cells harbored diverse mtDNA mutations occurring at a higher frequency than in mouse tissues, and exhibited severe respiration defects that would be lethal in tissues or organs. Abnormal respiratory complex formation and high stress on the mitochondrial protein quality control system appeared to be involved in these severe respiration defects. The mutation rates of the majority of highly accumulated mutations converged to either approximately 5%, 10%, or 40%, suggesting that these mutations are linked on the respective mtDNA molecules, and mtDNA in cybrid cells likely consisted of mtDNA molecules clonally expanded from the small population of introduced mtDNAs. Thus, the linked mutations in these cybrid cells cannot be evaluated individually. In addition, mtDNA mutations homologous to confirmed pathogenic mutations in human were rarely observed in our generated cybrids. However, the transmitochondrial cybrids constitute a useful tool for concentrating pathogenic mtDNA mutations and as a source of diverse mtDNA mutations to elucidate the relationship between mtDNA mutations and diseases.

1 Laboratory of Theriogenology, The Graduate School of Veterinary Sciences, Gifu University, Obihiro, Hokkaido 080-8555, Japan 2 Laboratory of Zoo and Wildlife Medicine, Faculty of Applied Biological Sciences, Gifu University, Gifu 501-1193, Japan 3 Laboratory of Theriogenology, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0808, Japan 4 Laboratory of Theriogenology, Department of Applied Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan

The recruitment of mural cells such as pericytes to patent vessels with an endothelial lumen is a key factor for the maturation of blood vessels and the prevention of hemorrhage in pathological angiogenesis. To date, our understanding of the specific trigger underlying the transition from cell growth to the maturation phase remains incomplete. Since rapid endothelial cell growth causes pericyte loss, we hypothesized that suppression of endothelial growth factors would both promote pericyte recruitment, in addition to inhibiting pathological angiogenesis. Here, we demonstrate that targeted knockdown of apelin in endothelial cells using siRNA induced the expression of monocyte chemoattractant protein-1 (MCP-1) through activation of Smad3, via suppression of the PI3K/Akt pathway. The conditioned medium of endothelial cells treated with apelin siRNA enhanced the migration of vascular smooth muscle cells, through MCP-1 and its receptor pathway. Moreover, in vivo delivery of siRNA targeting apelin, which causes exuberant endothelial cell proliferation and pathological angiogenesis through its receptor APJ, led to increased pericyte coverage and suppressed pathological angiogenesis in an oxygen-induced retinopathy model. These data demonstrate that apelin is not only a potent endothelial growth factor, but also restricts pericyte recruitment, establishing a new connection between endothelial cell proliferation signaling and a trigger of mural recruitment. Electronic supplementary material The online version of this article (doi:10.1007/s10456-013-9349-6) contains supplementary material, which is available to authorized users.

Artificially generated pancreatic β-cells from pluripotent stem cells are expected for cell replacement therapy for type 1 diabetes. Several strategies are adopted to direct pluripotent stem cells toward pancreatic differentiation. However, a standard differentiation method for clinical application has not been established. It is important to develop more effective and safer methods for generating pancreatic β-cells without toxic or mutagenic chemicals. In the present study, we screened several endogenous factors involved in organ development to identify the factor, which induced the efficiency of pancreatic differentiation and found that treatment with erythropoietin (EPO) facilitated the differentiation of mouse embryonic stem cells (ESCs) into definitive endoderm. At an early stage of differentiation, EPO treatment significantly increased Sox17 gene expression, as a marker of the definitive endoderm. Contrary to the canonical function of EPO, it did not affect the levels of phosphorylated JAK2 and STAT5, but stimulated the phosphorylation of ERK1/2 and Akt. The MEK inhibitor U0126 significantly inhibited EPO-induced Sox17 expression. The differentiation of ESCs into definitive endoderm is an important step for the differentiation into pancreatic and other endodermal lineages. This study suggests a possible role of EPO in embryonic endodermal development and a new agent for directing the differentiation into endodermal lineages like pancreatic β-cells.

When deprived of an anchorage to the extracellular matrix, fibroblasts arrest in the G1 phase with inactivation of Cdk4/6 and Cdk2 and destruction of Cdc6, the assembler of prereplicative complexes essential for S phase onset. How cellular anchorages control these kinases and Cdc6 stability is poorly understood. Here, we report that in rat embryonic fibroblasts, activation of mammalian target of rapamycin complex 1 by a Tsc2 mutation or overexpression of a constitutively active mutant Rheb overrides the absence of the anchorage and stabilizes Cdc6 at least partly via activating Cdk4/6 that induces Emi1, an APC/CCdh1 ubiquitin ligase inhibitor.Structured summaryMINT-7890626: cdc27 (uniprotkb:Q4V8A2) physically interacts (MI:0915) with Cyclin-A (uniprotkb:Q6AY13) by anti bait coimmunoprecipitation (MI:0006)

Objective: To examine the therapeutic efficacy of N-acetylglucosamine (GlcNAc) in rabbits with experimental osteoarthritis (OA). Methods: Experimental OA was induced in rabbits by anterior cruciate ligament transection (ACLT). In the first study, rabbits (six in each group) received intramuscular injections of GlcNAc or normal saline three times a week starting 1 week postoperatively. In the second study, rabbits (eight in each group) were injected intra-articularly with GlcNAc (either once or twice a week) or normal saline. In the third study, rabbits (seven in each group) were injected intra-articularly twice a week with either GlcNAc, hyaluronan, or normal saline. Animals were killed 8 weeks after ACLT for macroscopic and histological assessment of the knee joints. Results: Intramuscular administration of GlcNAc in rabbits with experimental knee OA did not show chondroprotective effects but showed mild anti-inflammatory activity. In contrast, intra-articular administration of GlcNAc twice a week reduced cartilage degradation. Additionally, intra-articular GlcNAc also suppressed synovitis. Once a week intra-articular injections of GlcNAc did not demonstrate therapeutic efficacy. The chondroprotective efficacy of GlcNAc was better than that of viscosupplementation treatment with hyaluronan. Conclusion: Intra-articular GlcNAc has chondroprotective and anti-inflammatory activity in experimental OA.

ObjectiveIntraarticular injections of sodium hyaluronate (Na-HA) appear effective in reducing subjective symptoms of osteoarthritis (OA) and may also have protective effects on the cartilage matrix. The present study analyzed the suppressive effects of Na-HA on the release and degradation of aggrecan and on levels of nitric oxide (NO) in the joint fluid of patients with knee OA.DesignSixteen OA patients with knee joint effusion were treated by 5 weekly intraarticular injections of Na-HA. Prior to each Na-HA injection, joint fluid was collected to determine the levels of chondroitin 4-sulfate (C4S) and chondroitin 6-sulfate (C6S), intact aggrecan and NO.ResultsOne week after the final injection, the joint fluid levels of C4S, C6S, and NO were significantly decreased. In contrast, the joint fluid level of intact aggrecan was stable during the series of Na-HA injections. A trend was seen for a positive correlation (P

The motor is desirable high efficiency, because it is consuming about 50% electrical energy. Therefore, Permanent Magnet Synchronous Motor (PMSM) is gathering attention and being widely applied. The vector control of PMSM must have information of magnet pole position. So, it is necessary to attach position sensor. However, several problems occurs by attaching a position sensor. For example that degradation of reliability, adaptability of the environment is reduced, and cost is high. These problems should be solved. Thus, vector control without a position sensor has attracted attention. A compressor is a load of fluctuation torque because it repeats suction and compression of refrigerant gas. When such load is applied to the sensorless control, it becomes unstable. In this paper, the effect of torque fluctuation is checking when the compressor is moved by sensorless control of PMSM.

A novel method of quantitating cell migration has been proposed for the potential utilization of tissue engineered scaffolds. Applying Alt's conservation law to describe the motion of first passage ACL and MCL cells, we have developed a quantitative method to assess innate differences in the motility of cells from these two ligamentous tissues. In this study, first passage ACL and MCL cells were cultured from four mature New Zealand white rabbits. One side of the cell monolayer was scraped completely away to create a wound model. The cell moved into the cell-free area, and cell density profiles were analyzed at 6 h and 12 h. Values of the random motility coefficient (mu) were then estimated by curve fitting the 6 h and 12 h data to a mathematical model, derived from the conservation law of cell flux. During 6 h of incubation in medium supplemented with 1% FBS, MCL cells (mu(MCL) = 4.63 +/- 0.65 X 10(-6) mm(2)/sec) were significantly (p < 0.05) more mobile than ACL cells (mu(ACL) = 2.51 +/- 0.31 X 10(-6) mm(2)/sec). At 12 h, the MCL cells also appeared to move faster (mu(ACL) = 4.39 +/- 0.63 X 10(-6) mm(2)/sec, mu(MCL) = 6.59 +/- 1.47 X 10(-6) mm(2)/sec), but the difference was not statistically significant (p = 0.18). Exposure of the cells to growth factors PDGF-BB or bFGF for 6 h had no significant effect on the migration of the ACL and MCL cells. However, exposure of the ACL cells (p < 0.05) and the MCL cells (p = 0.19) to 1 ng/mL of PDGFBB for 12 h enhanced their migration. Incubation with a high concentration (100 ng/mL) of PDGF-BB or bFGF at concentrations tested (1 or 100 ng/mL) for 12 h, produced little or no migratory stimulation on these ligament cells. Our findings support the previous qualitative observations made by numerous investigators. The novel methodology developed in this study may provide a basis for tissue engineering, and the results may be applied to tissue reconstruction techniques of the knee ligaments.

Raccoons (Procyon lotor), which are not native to Japan, have been suspected to transmit various pathogens by frequent intrusion into agricultural and residential areas. To determine influenza A virus seropositivity in raccoons in Japan, we examined a total of 634 raccoons captured in 19 towns (A–S) from 2009 to 2012. Agar gel precipitation tests showed that the antibody prevalence was 1.89% (12/634). All positive raccoons were captured in three towns (A–C) located within a radius of approximately 30 km, and 75% had antibodies to multiple subtypes (H1, H3–5, N1, N6, and N8). H3 and N8 antibodies were most frequently detected (75%). Among all the raccoons captured, 67% (8/12) were found in town A in 2009 and 2010, and all five raccoons captured in 2010 had H3 and N8 antibodies, suggesting that transmission of the subtype might occur. H5 and N1 antibodies were also detected in two raccoons captured in town A. Virus neutralization tests examining the highly pathogenic avian influenza virus (HPAIV) H5N1 subtype (four isolates of which have been detected in Japan to date) and the low PAIV (LPAIV) H5N3 subtype showed that raccoon sera highly cross-reacted with three H5N1 strains (clade 2.5: Ck/Yamaguchi/7/04; clade 2.3.2.1: Whooper swan/Hokkaido/1/08 and Whooper swan/Hamanaka/11), while they displayed a low cross-reactivity with the antisera to the clade 2.2 virus (Ck/Miyazaki/K11/07) and H5N3 LPAIV (Whistling swan/Shimane/499/83). Thus, the origin of the H5N1 virus was not clearly defined. The viral M gene was detected in four antibody-negative raccoons captured in three towns by real-time reverse transcription-polymerase chain reaction (rRT-PCR) with high Ct values, although no virus was isolated. This study is the first report showing that raccoons of Japan were infected with multiple subtypes of influenza A virus, including H5N1. It remains to be elucidated how raccoons play a role in persistence of influenza A virus in nature and if they could pose risks to animal and human health, for example, by playing a role as “mixing vessel” to generate novel strains of influenza A virus.