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Pesticides are frequently used to eradicate invasive ant species, but pose ecological harm. Previous studies assessed non-target effects only in terms of the increase or decrease of abundance or species richness after pesticide applications. Positive effects of the release from pressure caused by invasive ant species have not been considered so far. To more accurately assess pesticide effects in the field, the non-target effects of pesticides should be considered separately from the positive effects of such releases. Here, we used monitoring data of ants and other invertebrates collected in a program for the eradication of the Argentine ant, Linepithema humile (Mayr), using fipronil. First, we separately assessed the effects of L. humile abundance and fipronil exposure on non-target ants and other invertebrates using generalized linear models. The abundance of L. humile and the number of pesticide treatments were negatively associated with the total number of non-target individuals and taxonomic richness. We also noted negative relationships between the number of individuals of some ant species and other invertebrate taxonomic groups. The L. humile × pesticide interaction was significant, suggesting that the abundance of L. humile affected the level of impact of pesticide treatment on non-target fauna. Second, we evaluated the dynamics of non-target ant communities for 3 years using principal response curve analyses. Non-target ant communities treated with fipronil continuously for 3 years recovered little, whereas those treated for 1 year recovered to the level of the untreated and non-invaded environment.

Species native to oceanic islands are highly sensitive to the ecological impacts of invasive alien species, especially ants. The Ogasawara (Bonin) Islands are located approximately 1000 km south of mainland Japan and have a characteristic insular ecosystem. The risk of ecological harm via human and cargo transportation has increased as the number of tourists visiting the islands has grown since their designation as a World Natural Heritage Site. To evaluate the risk of biological invasion, we surveyed the ant faunas at ports of Tokyo Bay on the Japanese mainland and the Ogasawara Islands using sticky traps baited with honey and tuna. Our findings indicated that ant species composition was remarkably different between the ports of Tokyo and Ogasawara, and potential alien species were detected in both areas. The baited traps were likely to be more attractive than the non-baited traps, but need to be improved in future research. In particular, this is the first study to detect the invasive big-headed ant Pheidole megacephala (Fabricius) in mainland Japan, which likely arrived from Ogasawara via an ocean liner. Our study demonstrates the importance of periodically monitoring around ports to detect alien ants during the early invasion stage, when control and eradication methods are more effective.

The green anole (Anolis carolinensis) is on the list of the designated invasive alien species in Japan because of its negative effects on indigenous entomofauna. Therefore, control operations of the lizard are being carried out in some areas on the Ogasawara Islands. The primary population-control measure is the use of adhesive traps on tree trunks. Identification of the characteristics of the tree trunk type that this lizard uses frequently will contribute to effective setting of the traps, thereby improving catchability. To clarify whether or not there is a perching site preference according to plant species, we evaluated the available (n=1,228) and observed (n=811) perching sites in terms of tree species. A lizard food resource survey was also conducted on four major plant species. As for the selectivity of plant species, the screw pine (Pandanus boninensis) was strongly preferred by adult and juvenile anoles (P

Our goal was to create colonization and invasion risk maps for the Argentine ant, Linepithema humile, based on occurrence data in Japan, by combining colonization- and invasion-related variables and spatial filters that alleviate spatial autocorrelation. With these data, we will be better able to implement surveillance and control programs. Species distribution models were generated, using the maximum entropy approach, from presence-only data collected from 12 locations. Colonization-related variables (e.g., temperature, precipitation) and invasion-related variables (e.g., urban area, distance from ports) were used as environmental variables and spatial filters that alleviate spatial autocorrelation were included at the same time. The high invasion risk area was restricted to coastal areas, whereas high colonization risk applied to a broader area. Elevation, minimum temperature, and flow accumulation were the most effective variables for predicting colonization risk, whereas urban area, elevation, and the port distance index were the most effective variables for predicting invasion risk. The invasion risk map had a higher level of accuracy than the colonization risk map. We identified those areas with a high risk of invasion in the early stages and strong propagule pressure with a model using both invasion-related variables and colonization-related variables to accurately estimate the initial invasion distributions. We found that high colonization risk areas were concentrated in the Okinawa and Ogasawara Islands; ecosystems with highly endemic ant species that are likely to have a high sensitivity to L. humile introduction. Our data will aid in strengthening both domestic and international quarantine systems to prevent such introductions.

In the last 30 years some limited successes in alien ant control have been documented globally, and control programs remain challenging. Moreover, the potential non-target impacts of toxicants have not been well studied. We assessed the efficacy and non-target effects of multiple products containing the active compound fipronil in the attempted control of two populations of the invasive Argentine ant Linepithema humile (Mayr) in Tokyo, Japan. Three treatments were conducted: control, low-dose treatment (0.1 g/ha per treatment), and high-dose treatment (0.2 g/ha). Treatments were applied once per month for 11 months. The abundance of L. humile declined rapidly by up to 99.8 % in treated areas, but remained at extremely high densities in the control area. The treatments had few negative non-target effects, with the abundances of native ant species and other ground-dwelling invertebrates except for cockroaches being greater in the treated areas after L. humile suppression. Thus, fipronil is an effective compound for controlling L. humile and can be used with minimal toxic effects on non-target organisms. The treatments cost approximately US$ 575/ha for the low-dose treatment and US$ 1250/ha for the high-dose treatment. Our research supports the creation of more ambitious invasive ant management projects.

In the original publication of the article, the Table 1 was incorrectly published.Tapinoma melanocephalum (TM) and Camponotus nipponicus (CN) were labeled incorrectly as “absence” and “presence” at Takeshiba, respectively. The correct version of the table is given below where TM is labelled as “presence” and CN is labelled as “absence”.

During a survey of human rotaviruses in Okayama Prefecture, Japan in the 2011-2012 rotavirus season (between September 2011 and August 2012), G3P[8] was found to be a predominant genotype overall. However, G1P[8] emerged in the latter half of the season. To clarify the genetic background of the G1P[8] strains, the VP7, VP4, VP6, NSP4, and NSP5/6 genes of the strains were sequenced and genotyped. As a result, it was demonstrated that the strains with two different genotype constellations (G1-P[8]-I1-E1-H1 and G1-P[8]-I2-E2-H2) prevailed in the season. The G1P[8] strains possessing the DS-1-like VP6, NSP4, and NSP5/6 genes (the DS-1-like G1P[8] strains), which should reveal a short electropherotype, were originated from possible intergenogroup reassortment events. The DS-1-like G1P[8] strains accounted for 74.1% of all G1P[8] strains and were detected continuously throughout the season but not in the preceding season, indicating the possibility of new introduction and rapid spreading of these strains in the 2011-2012 season. The results suggest that the intergenogroup reassortants, considered generally unstable, can spread rapidly and become relevant.

An Anaplasma species closely related to Anaplasma phagocytophilum detected in sika deer in Hokkaido, Japan was molecularly analyzed using 16S rRNA, citrate synthase (gltA), and heat-shock operon (groEL) gene sequences. Genome walking was performed to determine its complete gltA and groEL sequences (1233 bp and 1650 bp, respectively). Percent identities to the closest A. phagocytophilum sequences from the US and European strains were 98.6-98.8%, 76.5%, and 80.3-80.8% for 16S rRNA, gltA, and groEL genes, respectively. For deduced amino acid sequences, percent identities to the closest A. phagocytophilum sequences were 66.7% and 97.6% for gltA and groEL genes, respectively. Phylogenetic analyses revealed divergence from any known A. phagocytophilum strain. The lower identities and the divergent phylogenetic position of the Anaplasma sp. detected from sika deer in Japan with established A. phagocytophilum strains provide evidence of its potential novelty.

OP354-like P[8] (P[8]b subtype) species A rotaviruses (RVAs) were isolated first time in Japan during a RVA survey in Okayama Prefecture between 2006 and 2009. Two of 236 RVA-positive samples were identified as G1P[8]b by reverse transcription polymerase chain reaction. P[8]b strains (RVA/human-wt/JPN/OH1998/2008/G1P[8]b and RVA/human-wt/JPN/OH2024/2008/G1P[8]b) were isolated only in May, 2008 and both patients infected with P[8]b viruses lived in the same city, suggesting that the prevalence of P[8]b RVAs is limited considerably in Okayama Prefecture. Molecular analysis of four genes (VP4, VP6, VP7, and NSP4 genes) of Japanese P[8]b strains revealed that the VP4 genes of these strains were related closely to those of Southeast Asian and Indian P[8]b strains. In contrast, the VP6, VP7, and NSP4 genes of Japanese P[8]b strains were highly homologous to G1P[8]a strains prevalent in the same area. These results suggest that the Japanese P[8]b strain may be a result of reassortment events between Japanese G1P[8]a viruses and unidentified Asian viruses possessing the P[8]b VP4 gene. J. Med. Virol. 84:697–704, 2012. © 2011 Wiley Periodicals, Inc.

Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) and tick-borne fever in ruminants. A closely related and potentially novel Anaplasma sp. in Japan was recently characterized. The aims of the study were to provide molecular evidence for the presence of these 2 species in Japan, and to develop a reliable PCR method based on the nucleotide differences within the citrate synthase (gltA) gene. DNA samples from 182 ixodid ticks (134 Ixodes persulcatus, 35 Haemaphysalis douglasii and 13 I. ovatus) collected from 2 sites in Hokkaido, Japan, were screened for A. phagocytophilum and its closely related Anaplasma sp. (herein designated as Anaplasma sp. Japan) using 16S rRNA PCR, revealing a combined prevalence rate of 27.5% (50 samples). The positive samples were then used to evaluate a newly developed gltA-based nested PCR method. Selected positive samples were further characterized using the groEL gene for confirmation and phylogenetic analyses. Two groups of sequence results were obtained: those that had closer identities with (1) A. phagocytophilum (99.5-99.6% for 16S rRNA, 97.5% for gltA and 98.4% for groEL), and those that had closer identities with (2) Anaplasma sp. closely related to A. phagocytophilum in Japan (99.3% for 16S rRNA, 96.4-98.7% for gltA and 97.5-97.9% for groEL). The present study confirmed the distinct presence of A. phagocytophilum and its closely related Anaplasma sp. in Japan, and developed a new PCR detection method based on gltA that can distinguish the 2 organisms.

日本紅斑熱の岡山県初発例を経験した．症例は64才女性．高熱，紅斑，刺し口を有し，肝障害と血小板数の低下を認めた．皮疹が四肢末梢に強いこと，手掌にも認めたことがつつが虫病との鑑別点となった．副作用のため短期間の投与となったが，ミノサイクリンが奏功した．後日ペア血清において抗体価（免疫蛍光抗体法）の有意な上昇が認められ，痂皮のPCR法では日本紅斑熱DNAが抽出された．本疾患は4類届出感染症であり，近年報告数が増加し，新たに届出をする県が増えている．治療が遅れると致死的となることもあるため，高熱と皮疹を伴う肝障害症例を診た際には常に本症も念頭におき，疑えばすぐに治療を開始することが重要である．

In June 2007, a questionnaire survey related to the surveillance, recognition, and reporting of Tsutsugamushi disease (TD) and Japanese spotted fever (JSF) — diseases considered endemic in Miyazaki Prefecture — was distributed to general practice clinics in the prefecture. The response rate was 40.9% (232/567). While 75.5% of the responding clinics knew TD to be a notifiable disease, only 41.8% knew JSF was notifiable. The recognition level of JSF surveillance was lower in the low-incidence areas of JSF within Miyazaki Prefecture. In 2006, 25 cases were clinically suspected as TD by the responding clinics; of the 25 cases, 9 were confirmed and 8 of these were reported to the National Epidemiological Surveillance of Infectious Diseases (NESID). Only 1 of 6 clinically suspected JSF cases from the responding clinics was confirmed in 2006, and it was not reported to NESID. The clinics located in the high-incidence areas for TD tended not to perform laboratory confirmation of the clinically suspected cases of either of the diseases. Considering that NESID requires laboratory confirmation of the reported cases of these diseases, their extent may be underestimated, especially in the high-incidence areas. For clinics in Miyazaki Prefecture, we need to publicize the existence of JSF surveillance and inform clinics about the laboratories available for confirmation of JSF and TD in the prefecture.

Rickettsia, a genus that includes the causative agents for spotted fever rickettsioses and typhus fever, comprises obligate intracellular bacteria (1). The first case of Japanese spotted fever (JSF), caused by R. japonica (2), was reported in 1984 in Japan (3). According to the national surveillance system in Japan (http://idsc.nih.go.jp/idwr/ybata/report-Ea.html), JSF cases, including sporadic cases resulting in death, have been gradually increasing. Rapid diagnosis of rickettsial infections is important because rickettsioses can be cured when appropriate antimicrobial drug treatment is given during the early clinical stages of the disease. Furthermore, development of a rapid and specific diagnostic system for R. japonica is now a matter of increasing urgency (4) because JSF has also been reported in several other countries in Asia (1).

アマチュア尺八奏者でピアノの訓練経験もあったKM は，五線譜および尺八譜の読み書きが可能な二楽譜使用者であった。くも膜下出血後尺八譜の読み書き障害は軽度だったが，五線譜の読み書き能力は顕著に障害され，既知のメロディーを聴いて書譜する課題や音読課題では，五線譜と尺八譜の成績が乖離した。楽曲を正確に記譜する五線譜に対し，尺八譜は楽器の操作法を仮名文字で表記する奏法譜である。西洋音楽と異なり邦楽には，演奏する前にリズムを付けて音名を唱える「唱譜」という口伝の習得様式が存在するため，楽譜は唱譜によって暗記した演奏法を記憶から再生するための補助手段として用いられる。既知のメロディーの書譜，音読課題で尺八譜が五線譜よりも成績が良かったのは，唱譜で覚えた記憶から正答を引きだした可能性が考えられた。このような尺八譜の特異性が楽譜の読み書き課題において成績の乖離に関与したと考えられた。

Chlamydophila pneumoniae is known to be associated with atherosclerosis. Recent studies have reported that components of Chlamydophila pneumoniae (chlamydophilal antigens) induce foam cell formation in macrophages. However, the mechanism of foam cell formation induced by chlamydophilal antigens has yet to be elucidated. In this paper, we first found that mitogen-activated protein kinases including extracellular signal-regulated kinase, p38 and c-Jun NH2 terminal kinase are phosphorylated after stimulation by chlamydophilal antigens. We then showed that chlamydophilal antigens induce foam cell formation mainly via c-Jun NH2 terminal kinase. Finally, we demonstrated that foam cell formation and phosphorylation of mitogen-activated protein kinases induced by chlamydophilal antigens are mainly recognized through Toll-like receptor 2. These results collectively indicated that chlamydophilal antigens induce foam cell formation mainly via Toll-like receptor 2 and c-Jun NH2 terminal kinase.

Rickettsia was first detected in seabird soft-bodied ticks, Carios capensis and C. sawaii in Japan. According to sequence analysis, Rickettsia in Japan was identical to Rickettsia scc31 in C. capensis in the U.S.A. This suggested that an environmental circulation had consisted among microorganisms, ticks and long distance migratory seabirds around the Pacific Ocean.

Despite major improvements in the diagnosis of pathogenic organisms causing acute respiratory infections (ARIs), details of infections caused by atypical pathogens are not well understood, particularly in developing countries. This clinical and epidemiological research was conducted in Bangladesh to explore the prevalence of atypical pathogens in causing childhood pneumonia. Sixty-four children with ARI were studied at the Pediatric Outpatient Department of Dhaka Medical College Hospital, Bangladesh, during September through December 2000. In addition to clinical examination, hematological, radiological, and bacteriological examinations were performed. Antibody titers from paired sera against Mycoplasma pneumoniae and Legionella spp. in the acute and convalescent phases revealed that none of these children were infected with M. pneumoniae, while only one serum sample was positive for L. pneumophila serogroup 4. Antibody titers against Chlamydophila (Chlamydia) pneumoniae, determined by an indirect microimmunofluorescence method, and by an enzyme-linked immunosorbent assay (ELISA) kit (HITAZYME C. pneumoniae kit) indicated that 13 children (20.3%) were infected with C. pneumoniae. Our results indicate a high prevalence rate of C. pneumoniae, suggesting it is as an important causative pathogen of childhood pneumonia in Bangladesh.

A study was made to evaluate the cutoff value of indirect immunofluorescent-antibody (IFA) test for Q fever diagnosis in Japan. We used 346 sera, including 16 from confirmed Q fever cases, 304 from Japanese pneumonia patients, and 26 from negative cases. Thirteen sera from the confirmed Q fever cases with an immunoglobulin M (IgM) titer of ≥1:128 and/or IgG titer of ≥1:256 by the IFA test were positive by both enzyme-linked immunosorbent assay (ELISA) and Western blotting assay (WBA), whereas 298 sera from pneumonia patients and 26 negative sera with an IgM titer of ≤1:16 and an IgG titer of ≤1:32 by the IFA test were negative by both ELISA and WBA. In the proposed “equivocal area,” with an IgM titer of ≥1:32 and ≤1:64 and/or an IgG titer of ≥1:64 and ≤1:128, we found 9 sera, 3 from confirmed Q fever cases and 6 from Japanese pneumonia patients, by the IFA test. Three sera from the confirmed Q fever cases and one of the sera from pneumonia patients were IgM and/or IgG positive by both ELISA and WBA. These results suggest that a single cutoff value for the IFA test may cause false-positive and false-negative results. In conclusion, this study showed that an “equivocal area” should be used for the IFA test rather than a single cutoff value and that sera in the equivocal area should be tested by additional serological assays for confirmation.

Recent evidence has suggested an association between Chlamydia pneumoniae infection and coronary atherosclerosis. A significant association has also been detected between heat shock protein (HSP) 60 antibody and the severity of coronary atherosclerosis. The aim of this study was to define the relationship between instability of ischemic heart disease (IHD) and serum levels of HSP60 and C. pneumoniae antibodies. Blood samples for the measurement of serum antibody titers were obtained from 1131 patients with ischemic heart disease (65 ± 9 years; male/female, 828/303) and 127 non-IHD controls with normal coronary arteries (64 ± 9 years; male/female, 60/67) on the day of cardiac catheterization. The serum levels of anti-human HSP60 IgG antibody and anti-chlamydial IgM, but not IgG or IgA, antibody were significantly higher in ACS patients than in stable IHD patients or controls. These results suggest that acute C. pneumoniae infection with HSP60-related immunological responses may contribute to the pathophysiology of acute coronary syndromes.

To the Editor: Japanese spotted fever (JSF), caused by Rickettsia japonica, is the most prevalent tickborne infectious disease in Japan (1), occurring most frequently in central and western regions (http://idsc.nih.go.jp/idwr/CDROM/Main.html [in Japanese]). Cases of unknown fever with rickettsiosis-like symptoms not associated with JSF have been reported in JSF-endemic regions of Japan (2). Several spotted fever group (SFG) rickettsiae (R. japonica, R. heilongjiangensis, R. helvetica, R. tamurae, R. asiatica, Candidatus R. tarasevichiae) and other related Rickettsia spp. have been identified in Japan (1,3–6). Human infections with R. heilongjiangensis and R. tamurae have been confirmed (3,5), and Anaplasma phagocytophilum and Ehrlichia chaffeensis, known human pathogens, have been detected in ticks and deer in Japan. We conducted this study to determine the risk in central and western Japan for human exposure to ticks harboring SFG rickettsiae, A. phagocytophilum, or Ehrlichia spp. In 2007–2011, we collected 827 Haemaphysalis, Amblyomma, and Ixodes spp. ticks (392 adults, 435 nymphs) by flagging vegetation in the prefectures of Shizuoka, Mie, Wakayama, Kagoshima, Nagasaki (Goto Island), and Okinawa (the main island and Yonaguni Island) (Technical Appendix Figure 1). We extracted DNA from the salivary glands of each tick and performed PCR to amplify gltA, 16S rDNA, and ompA of SFG rickettsiae. To detect A. phagocytophilum and Ehrlichia spp., we performed nested PCR targeting the p44/msp2 and p28/omp-1 multigenes, respectively. PCR gltA screening revealed SFG rickettsiae in 181 (21.9%) of the 827 ticks (Table). We obtained nearly full-length (1.1-kb) gltA sequences and classified them into 5 groups by phylogenetic analyses (Technical Appendix Figure 2). Sequences for groups 1 (prevalence 1.0%) and 2 (prevalence 3.2%) were identified as R. japonica YH (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AP011533","term_id":"348592266","term_text":"AP011533"}}AP011533) and R. tamurae (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AF394896","term_id":"21360604","term_text":"AF394896"}}AF394896), respectively (Table). Group 3 (prevalence 15.1%) sequences were identical to that of Rickettsia sp. LON (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AB516964","term_id":"256353425","term_text":"AB516964"}}AB516964). The sequence for group 4 (prevalence 1.6%) was closely related to that for R. raoultii strain Khabarovsk (98.8% similarity), and a part of the sequence (342 bp) was identical to that of Rickettsia sp. Hf 151 (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"AB114815","term_id":"33146352","term_text":"AB114815"}}AB114815). Group 5 consisted of 4 newly identified rickettsiae (Technical Appendix Figure 2). Of these 4 rickettsiae, 3 (Mie311, Goto13, and Mie334) were closely related to R. raoultii strain Khabarovsk (98.0% identity) and 1 (Mie201) was similar to Candidatus R. principis (99.7% identity). Table PCR survey results for Haemaphysalis, Amblyomma, and Ixodes spp. ticks tested for rickettsiae, central and western Japan, 2007–2011* We further analyzed the 16S rDNA and ompA in gltA-positive tick samples. The 16S rDNA and ompA for group 1 samples shared 100% identity with 16S rDNA and ompA of R. japonica YH ({"type":"entrez-nucleotide","attrs":{"text":"AP011533","term_id":"348592266","term_text":"AP011533"}}AP011533). The 16S rDNA of group 2 was identical to that of R. tamurae ({"type":"entrez-nucleotide","attrs":{"text":"AY049981","term_id":"21360334","term_text":"AY049981"}}AY049981). In groups 3–5, some of the specific amplicons in 16S rDNA or ompA could be detected; their sequences were confirmed to be similar (but not identical) to those of several known rickettsial sequences. We amplified the p44/msp2 amplicons of A. phagocytophilum from 25 (3%) of 827 ticks (Table). By cloning (TA Cloning Kit; Life Technologies, Carlsbad, CA, USA) and sequencing these amplicons, we obtained and identified 60 new TA-clone sequences (366–507 bp) for p44/msp2 (GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"JQ697880-JQ697950","start_term":"JQ697880","end_term":"JQ697950","start_term_id":"383088493","end_term_id":"383088630"}}JQ697880-JQ697950); these sequences may include a potentially novel Anaplasma species. (7). Ehrlichia p28/omp-1 was detected from 2 (0.2%) of the 827 ticks. Of 5 TA-clone sequences (284–315 bp) obtained from the 2 ticks, 2 from an A. testudinarium tick (GenBank accession nos. {"type":"entrez-nucleotide","attrs":{"text":"JQ697886","term_id":"383088503","term_text":"JQ697886"}}JQ697886 and {"type":"entrez-nucleotide","attrs":{"text":"JQ697887","term_id":"383088505","term_text":"JQ697887"}}JQ697887) shared 83.3%–86.7% similarity with E. ruminantium Gardel Map-1 (GenBank accession no. YP196842), and 3 from an H. longicornis tick (GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"JQ697888-JQ697890","start_term":"JQ697888","end_term":"JQ697890","start_term_id":"383088507","end_term_id":"383088511"}}JQ697888-JQ697890) showed the closest relationship to E. ewingii omp-1–15 (67%–73% similarity; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"EF116932","term_id":"133711094","term_text":"EF116932"}}EF116932). We identified the tick species associated with R. japonica as H. formosensis, H. hystricis, and H. cornigera, and another study reported an association with Dermacentor taiwanensis, H. flava, H. longicornis, and I. ovatus (4). In our study and previous studies, the tick species associated with A. phagocytophilum in Japan were identified as H. formosensis, H. longicornis, H. megaspinosa, A. testudinarium, I. ovatus, and I. persulcatus (8). Thus, it appears that 3 tick species (H. formosensis, H. longicornis, and I. ovatus) are associated with R. japonica and A. phagocytophilum. In addition, in an H. formosensis tick, we detected an SFG rickettsia that is closely related to R. raoultii, the etiologic agent of Dermacentor-borne necrosis erythema and lymphadenopathy in Europe and Russia (9). We detected Candidatus R. principis in H. flava in Japan; this species was previously detected in H. japonica douglasi and H. danieli ticks in Russia and China, respectively, (10). And, we found a high prevalence of R. tamurae in A. testidinarium ticks; Imaoka et al. (5) recently reported that R. tamurae causes local skin inflammation without general JSP-like symptoms. We did not detect the human pathogen E. chaffeensis, but we identified 2 potentially new Ehrlichia species. Our findings contribute to the known risks for exposure to Rickettsia-related pathogens in central and western Japan. Further studies may be required for the surveillance of additional pathogens, such as Candidatus Neoehrlichia mikurensis (2), which was recently recognized as a human pathogen. Technical Appendix: Phylogenetic classification of Rickettsia spp. gltA sequences detected in ticks during 2007–2011 in central and western Japan and locations of tick collection sites. Click here to view.(154K, pdf)

Aims: To clarify the inhibitory effects of ozone on Chlamydia trachomatis and C. pneumoniae. Methods and Results: Cell culture was performed using HeLa229 cells for C. trachomatis, and Human Line cells for C. pneumoniae. C. trachomatis strain D/UW-3/Cx and C. pneumoniae strain AR-39 were used. Ozone water was generated by an ozone water dispenser and diluted to desired concentration just before each experiment. Preinoculation minimum cidal concentration (MCC) and postinoculation MCC methods were employed. In preinoculation MCC, chlamydial strains were treated with serially diluted ozone water followed by inoculation to cells. In postinoculation method, chlamydial strains were inoculated to cells and incubated for 24 h. Then infected cells were treated with ozone water, followed by additional incubation for 48 h. Complete inactivation was obtained in preinoculation MCC method at 0·5 ppm of ozone water for 30 s, or 4 ppm for 5 s. Conclusion: Ozone at a concentration of 4 ppm was enough for immediate inactivation of both C. trachomatis and C. pneumoniae. Significance and Impact of the Study: Ozone water at 4 ppm should be applicable for prevention of C. trachomatis urogenital infections.

気管支喘息の増悪に Chlamydia pneumoniae (以下 C. pneumoniae) 感染が関与することはすでに数多く報告されているが, C. pneumoniae 感染に有効な抗生剤の喘息発作への効果についての研究はほとんどない. 今回われわれは非発作時の小児気管支喘息40例を, アジスロマイシン (以下AZM) 投与群 (10mg/kg/day, 3日間) と非投与群に分けて, 投与前, 投与1か月後, 3か月後の喘息発作点数と治療点数を比較した. 36例について血清抗体価, PCR, 培養で診断すると, C. pneumoniae 感染は投与群で68%, 非投与群で41%で, 両群間の感染陽性率に有意差はなかった.喘息発作点数を2群で比較すると, 前, 1か月後では有意差がなかったが, 3か月後では, 有意差をもってAZM投与群の発作点数が低かった. 治療点数には有意差はなかった. C. pneumoniae 感染があった19例をAZM投与群12例と非投与群7例で分けて検討するとAZM投与群では, 投与前から1か月後は有意に発作点数が減少したが, 非投与群では発作点数の改善はなかった. しかし C. pneumoniae 感染がなくてもAZMを投与した群7例で1か月, 3か月後とも発作点数は有意に減少した. AZM投与は, 小児気管支喘息の発作を減少させるのに有効と考えられた.

This data paper reports census data of ground-dwelling beetle and other fauna of the forest floor environment; collections were made from a network of 22 forest sites in Japan. To our knowledge, this represents the largest dataset for long-term monitoring of a ground-dwelling beetle community and other taxa in a ground environment in forests, which covers a broad climatic range in the temperate zone and is freely available. The network forms part of the Monitoring Sites 1000 Project launched by the Ministry of the Environment, Japan. It covers subalpine, cool- and warm-temperate and subtropical climatic zones and the four major forest types of Japan. Thirty-three permanent plots usually 1 ha in size were established in old-growth, secondary natural and a few plantation forests. Censuses of the ground-dwelling beetle community were conducted using pitfall trapping and forest floor environment monitoring every year from 2004 to the present. During the initial 9 years of the census (2004–2012), 59,762 beetle individuals (including 3182 larvae) of more than 314 species were recorded. This dataset includes taxonomy and biomass of each beetle individual and each taxonomic group of other invertebrates coincidently captured in pitfall trapping. The dataset also includes data related to ground coverage by forest floor vegetation, dry mass of the accumulated organic litter layer, and carbon and nitrogen contents and cellulose decomposition rate in organic layer and surface mineral soil. The data could be used to investigate geographical patterns and intra- and inter-annual dynamics of individual body mass, populations and community structures of ground-dwelling beetles, and their relationships with the forest floor environment. Furthermore, the data could be analyzed with other open datasets related to tree community dynamics and litter fall continuously measured in the same study plots. This dataset also provides information related to the distribution and average body mass of each beetle species.

Surveillance for scrub typhus was conducted in Japan in 1998 using a questionnaire. A total of 462 cases were reported. Scrub typhus occurred in both the fall and spring in the northern part of Honshu (the main island), and in the fall in the central part of Honshu and on the island of Kyushu. The occurrence of the disease varied with age, gender, and activity. Seventy-six percent of the patients were more than 51 years old, and 36% and 16% of the patients were engaged in farm work and forestry, respectively. Fever, rash, and eschar were detected in 98%, 93%, and 97% of the patients, respectively. Elevated levels of C-reactive protein, aspartate transaminase, and alanine transaminase were detected in 96%, 87%, and 77% of the patients, respectively. Disseminated intravascular coagulation developed in 34 cases and had a unique regional distribution. This study shows the status of scrub typhus in Japan in 1998 and provides important information for diagnosis and prevention.

A 29-year-old woman with primary myelofibrosis developed severe pneumonia, and 20 days later her 31-year-old sister also contracted pneumonia. The first patient had been in contact with parakeets but the second patient had not been in contact with any birds. Psittacosis was diagnosed in both cases by microplate immunofluorescence antibody technique. Person-to-person transmission between the sisters was suspected to have taken place.

Previously we reported the development of a highly sensitive enzyme-linked immunosorbent assay specific for anti-tuberculous glycolipid (anti-TBGL) for the rapid serodiagnosis of tuberculosis. In this study, the usefulness of an anti-TBGL antibody assay kit for rapid serodiagnosis was evaluated in a controlled multicenter study. Antibody titers in sera from 318 patients with active pulmonary tuberculosis (216 positive for Mycobacterium tuberculosis in smear and/or culture tests and 102 smear and culture negative and clinically diagnosed), 58 patients with old tuberculosis, 177 patients with other respiratory diseases, 156 patients with nonrespiratory diseases, and 454 healthy subjects were examined. Sera from 256 younger healthy subjects from among the 454 healthy subjects were examined as a control. When the cutoff point of anti-TBGL antibody titer was determined as 2.0 U/ml, the sensitivity for active tuberculosis patients was 81.1% and the specificity was 95.7%. Sensitivity in patients with smear-negative and culture-negative active pulmonary tuberculosis was 73.5%. Even in patients with noncavitary minimally advanced lesions, the positivity rate (60.0%) and the antibody titer (4.6 ± 9.4 U/ml) were significantly higher than those in the healthy group. These results indicate that this assay using anti-TBGL antibody is useful for the rapid serodiagnosis of active pulmonary tuberculosis.

Clinical features of tsutsugamushi disease (scrub typhus) were analyzed, based on 416 cases reported in Japan in 1998. Three major clinical symptoms: eschar, fever and rash were found in 87%, 98% and 92% of the cases, respectively. Elevated levels of CRP, GOT, GPT and LDH were observed in 96%, 85%, 78% and 91%, respectively. These clinical and laboratory findings were observed in the majority of the cases and considered important for diagnosis. Disseminated intravascular coagulation developed in 21 cases, indicating that scrub typhus can be life threatening. Lymphadenopathy was observed in 51% of the cases. Enlarged lymph nodes were limited to the local sites in 75% of these lymphadenopathy cases and most of these sites were adjacent to eschars. Most eschars were scabbed and located in the abdomen and the lower half of the body, especially the feet. This suggests that these parts are frequently exposed to tsutsugamushi mites. Furthermore, the skin is soft in these parts and covered by cloth. These factors may make it possible for mites to keep biting without being noticed for several hours, long enough for rickettsial transmission. Interestingly, eschar and rash were absent in 14% and 8% of the cases, respectively. This result suggests that the cases without the unique symptoms may have been misdiagnosed as common cold or other febrile illnesses. One hundred and fifty-four suspected cases were not scrub typhus cases by the serological tests. The three major clinical symptoms were present in approximately a half of these negative cases, eschar being observed in approximately 70%. This may suggest the presence of new type of scrub typhus can not be diagnosed by the present laboratory tests. Clinical features of scrub typhus in Japan were well revealed, and information obtained in the present study is useful for improving clinical diagnosis. It should, however, be stressed that there were cases that could not be correctly diagnosed only by the clinical symptoms, suggesting that it is important to improve the serological tests.

Two Japanese travelers from Bali were diagnosed with murine typhus in Japan during the same period. Although one had only mild illness, the other experienced liver and kidney dysfunction. Murine typhus may be missed not only in endemic areas around the world, but also in travelers, especially those returning from marine resorts in these areas.

"HITAZYME C. pneumoniae" (or "HITAZYME CPN", for short) is a diagnostic reagent that has been recently developed by adopting an ELISA method for detection of anti-Chlamydia pneumoniae (C. pneumoniae) antibodies. When this reagent is used under a current diagnostic standard that has been set as a provisional standard, however, high antibody positive rates are often produced for both IgG and IgA even using the specimens of healthy persons. So, it is difficult to distinguish C. pneumoniae-infected patients from healthy persons. Therefore, this time, we tried to establish a new diagnostic standard by setting up of special cut-off values for a single serum and rise rates of antibody titers for paired sera to improve the accuracy for diagnosis of C. pneumoniae infection. For a single serum testing, we set a special cut-off value at ID 3.00 for both IgG and IgA, so that most healthy persons fall within the range of the "negative" zone. This value was based on the calculation of "Mean+2SD" using measurement results (or IDs) of healthy persons. When this cut-off value was applied, the rate of > or = ID 3.00 for either IgG or IgA was 7.6% for healthy persons, and 64.9% for infected patients. (The rate reached 76.4% when the highest IDs of multiple specimens taken from each patient for this test were used in calculation) As a diagnostic standard for a single serum, therefore, it was defined that: "If ID is 3.00 or greater for IgG and/or IgA, it is highly likely that the case has an acute or a present infection." Using paired sera, we could confirm almost a linear relationship between the results by HITAZYME CPN and those by micro-IF method. Under micro-If method, if the antibody titer increases four times or greater using paired sera, acute infection is diagnosed. As it was found that the four-fold increase in antibody titer corresponds to the increase of 1.35 in ID for IgG and 1.00 for IgA, we defined a diagnostic standard for paired sera as follows: "If ID increases by 1.35 or greater for IgG, and/or if ID increases by 1.00 or greater for IgA, the case may be diagnosed as acute infection."

Mycoplasma pneumoniae infection is often experienced as community-acquired pneumonia, whereas Chlamydia trachomatis pneumonia is sometimes experienced as a vertical infection from mother to child through delivery, but is rare after six months of age. We reported the first case to date of a seven-year-old boy with coinfection of C. trachomatis and M. pneumoniae. The patient, who was referred to our hospital because of suspected pneumonia, had had severe cough and pyrexia for four days before his admission. The coinfection was confirmed because the antibodies to both bacteria were significantly increased. In addition the PCR for C. trachomatis detected the presence of C. trachomatis in the throat even after treatment. The PCR from a cervix swab of his mother and from a throat swab of his father were negative, but their antibodies to C. trachomatis were positive. We think that this may have been community-acquired pneumonia or due to persistent infection after a vertical transmission.

The in vitro and in vivo activities of AM-1155, a new quinolone, against Chlamydia spp. were investigated. The MIC of AM-1155 for 10 standard strains of different Chlamydia spp. and 25 wild-type strains of Chlamydia pneumoniae isolated in Japan, which were morphologically different from clinical isolates from the United States, ranged from 0.063 to 0.125 microg/ml. Its activity was almost the same as those of sparfloxacin and tosufloxacin and was 4 and 16 times superior to those of levofloxacin and ciprofloxacin, respectively, but lower than those of clarithromycin and minocycline (range for each, 0.016 to 0.031 microg/ml). The minimal chlamydiacidal concentration of AM-1155 ranged from 0.063 to 0.125 microg/ml, while those of clarithromycin and minocycline ranged from 0.016 to 0.031 microg/ml and 0.016 to 0.063 microg/ml, respectively. The therapeutic effect of a 7-day course of AM-1155 at doses of 5 and 10 mg/kg of body weight administered orally twice daily to mice with experimental Chlamydia psittaci pneumonia was excellent, with a 100% survival rate at 21 days after infection. The efficacy was equal to those of clarithromycin and minocycline and higher than those of ciprofloxacin and ofloxacin.

Biosynthesized tea polyphenols showed antichlamydial activity against Chlamydia trachomatis D/UW-3/Cx and L 2 /434/Bu using cell culture. The most active compounds were (−)-epigallocatechin gallate and (−)-epicatechin gallate, followed by (−)-epicatechin (EC). (+)-Epicatechin and (−)-epigallocatechin were intermediate. EC was the least toxic. These results warrant evaluation of tea polyphenols as topical antichlamydial agents.