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Aspergillus fumigatus is a pathogenic fungus with a global distribution. The emergence of azole-resistant A. fumigatus (ARAf) other than the TR-mutants is a problem in Japan. Additionally, the genetic diversity of A. fumigatus strains in Japan remains relatively unknown. Here we show the diversity in the A. fumigatus strains isolated in Japan as well as the complexity in the global distribution of the pathogenic strains. First, we analyzed the genome sequences of 171 strains from Japan as well as the antifungal susceptibility of these strains. Next, we conducted a population analysis of 876 strains by combining the available genomic data for strains isolated worldwide, which were grouped in six clusters. Finally, a genome-wide association study identified the genomic loci associated with ARAf strains, but not the TR-mutants. These results highlight the complexity of the genomic mechanism underlying the emergence of ARAf strains other than the TR-mutants. Whole-genome analysis of 876 Aspergillus fumigatus strains including 171 strains from Japan highlights the diversity in the A. fumigatus strains isolated in Japan as well as the complexity in the global distribution of the pathogenic strains. [ABSTRACT FROM AUTHOR]

Background: High‐attenuation mucus (HAM) is a specific manifestation of allergic bronchopulmonary mycosis (ABPM) on chest computed tomography (CT). Objectives: To compare the diagnostic accuracy of the two definitions of HAM and to clarify the clinical and radiographic characteristics of HAM‐positive and HAM‐negative ABPM. Methods: CT images at the diagnosis of ABPM using Asano's criteria were retrospectively analysed. In Study #1, radiographic data obtained using the same CT apparatus in a single institute were analysed to determine the agreement between the two definitions of HAM: a mucus plug that is visually denser than the paraspinal muscles or that with a radiodensity ≥70 Hounsfield units. In Study #2, HAM was diagnosed by comparison with the paraspinal muscles in patients with ABPM reporting to 14 medical institutes in Japan. Results: In Study #1, 93 mucus plugs from 26 patients were analysed. A substantial agreement for HAM diagnosis was observed between the two methods, with a κ coefficient of 0.72. In Study #2, 60 cases of ABPM were analysed; mucus plugs were present in all cases and HAM was diagnosed in 45 (75%) cases. The median A. fumigatus‐specific IgE titre was significantly lower in HAM‐positive patients than in HAM‐negative patients (2.5 vs. 24.3 UA/mL, p =.004). Nodular shadows were observed more frequently in the airways distal to HAM than in those distal to non‐HAM mucus plugs (59% vs. 32%, p <.001). Conclusion: In conclusion, agreement between the two methods to diagnose HAM was substantial. HAM was associated with some immunological and radiographic characteristics, including lower levels of sensitization to A. fumigatus and the presence of distal airway lesions. [ABSTRACT FROM AUTHOR]

Background: Allergic bronchopulmonary mycosis (ABPM) is an allergic disease caused by type I and type III hypersensitivity to environmental fungi. Schizophyllum commune, a basidiomycete fungus, is one of the most common fungi that causes non‐Aspergillus ABPM. Objective: Herein, we attempted to clarify the clinical characteristics of ABPM caused by S. commune (ABPM‐Sc) compared with those of allergic bronchopulmonary aspergillosis (ABPA). Methods: Patients with ABPM‐Sc or ABPA were recruited from a nationwide survey in Japan, a multicenter cohort, and a fungal database at the Medical Mycology Research Center of Chiba University. The definition of culture‐positive ABPM‐Sc/ABPA is as follows: (1) fulfills five or more of the 10 diagnostic criteria for ABPM proposed by Asano et al., and (2) positive culture of S. commune/Aspergillus spp. in sputum, bronchial lavage fluid, or mucus plugs in the bronchi. Results: Thirty patients with ABPM‐Sc and 46 with ABPA were recruited. Patients with ABPM‐Sc exhibited less severe asthma and presented with better pulmonary function than those with ABPA (p = 0.008–0.03). Central bronchiectasis was more common in ABPM‐Sc than that in ABPA, whereas peripheral lung lesions, including infiltrates/ground‐glass opacities or fibrotic/cystic changes, were less frequent in ABPM‐Sc. Aspergillus fumigatus‐specific immunoglobulin (Ig)E was negative in 10 patients (34%) with ABPM‐Sc, who demonstrated a lower prevalence of asthma and levels of total serum IgE than those with ABPM‐Sc positive for A. fumigatus‐specific IgE or ABPA. Conclusions: Clinical characteristics of ABPM‐Sc, especially those negative for A. fumigatus‐specific IgE, differed from those of ABPA. [ABSTRACT FROM AUTHOR]

The taxon Aspergillus section Fumigati comprises several causative agents of aspergillosis. Here, the distribution of Aspergillus sect. Fumigati in outdoor environments of Izu and Ogasawara Islands was investigated. Different strains were isolated from soil samples collected from 68 sites on 9 islands (Izu-oshima, Toshima, Shikinejima, Kozushima, Miyakejima, Hachijojima, Mukojima, Chichijima, and Hahajima), including different landscapes, and identified using morphological characteristics and calmodulin (CaM) sequences. Seven Aspergillus sect. Fumigati species were identified. The occurrence frequency of Aspergillus fumigatus was higher in forest sites on the islands, except for Ogasawara Islands, whereas that of species other than A. fumigatus was higher in bare land and grassland sites on all islands. The occurrence frequency of A. fumigatus was more than 50% on islands between Izu-oshima and Toshima, decreased on islands between Shikinejima and Hachijojima, and was zero on Ogasawara Islands. Considering other Aspergillus species, Aspergillus felis showed high occurrence frequency on islands between Izu-oshima and Shikinejima, Aspergillus pseudoviridinutans on islands between Kozushima and Hachijojima, and Aspergillus udagawae on Ogasawara Islands. At two study sites (grassland and forest sites), the soil was sampled throughout the year to evaluate whether the occurrence frequency of each fungal species was affected by sampling season. At the grassland site, A. pseudoviridinutans was isolated at more than 90% frequency, regardless of the sampling season. A. fumigatus occurrence frequency at the forest site ranged from 0% to 60% and greatly varied among sampling seasons. Thus, differences in island location and landscape affected the distribution of Aspergillus sect. Fumigati. [ABSTRACT FROM AUTHOR]

Aspergillus section Nigri, a group of black Aspergillus, has several cryptic species that were recently discovered to be intrinsically resistant to azole antifungals. In this study, susceptibility testing of 35 clinical isolates of Aspergillus tubingensis and Aspergillus welwitschiae in Japan was carried out using microdilution method. Strains tolerant to itraconazole in A.tubingensis (14/17 strains) and A.welwitschiae (6/18 strains) were discovered with hyphal growth and conidial germination above the minimal inhibitory concentration by microscopic observation, while no resistant strain was observed macroscopically. In contrast, no strain with reduced susceptibility to voriconazole, posaconazole, and amphotericin-B was found. Further examination may be required to determine the susceptibility of cryptic species in Aspergillus section Nigri to antifungals. [ABSTRACT FROM AUTHOR]

The white koji fungus, Aspergillus luchuensis mut. kawachii, is used in the production of shochu, a traditional Japanese distilled spirit.White koji fungus plays an important role in the shochu production process by supplying amylolytic enzymes such as α-amylase and glucoamylase. These enzymes convert starch contained in primary ingredients such as rice, barley, buckwheat, and sweet potato into glucose, which is subsequently utilized by the yeast Saccharomyces cerevisiae to produce ethanol.White koji fungus also secretes large amounts of citric acid, which lowers the pH of the shochu mash, thereby preventing the growth of undesired microbes and enabling stable production of shochu in relatively warm regions of Japan. This review describes the historical background, research tools, and recent advances in studies of the mechanism of citric acid production by white koji fungus. [ABSTRACT FROM AUTHOR]

Pulmonary oxalosis can be fatal, and Aspergillus tubingensis is commonly resistant to azoles in Japan. We report a case of bronchopulmonary oxalosis caused by A. tubingensis in a non-neutropenic patient who was successfully treated with voriconazole monotherapy. The susceptibility of the isolates to voriconazole and the effective elimination of contagious necrotic tissue by expectoration seemed to be two major factors contributing to the patient's survival. According to the literature review, pulmonary oxalosis is associated with a high mortality rate over a short term. An exploration of detailed information about the genomic characteristics and drug susceptibility of Aspergillus isolates is important for the development of treatment strategies for this life-threatening disease. [ABSTRACT FROM AUTHOR]

Koji mold, which belongs to the Aspergillus section Nigri, is used in the production of shochu. The section Nigri is composed of very morphologically similar members that in some cases produce mycotoxins, which rises concerns as to whether the presence of mycotoxin-producing fungi in shochu producing sites can compromise consumer safety. Thus, we examined the presence of mycotoxin-producing sec. Nigri fungi in six shochu factories (named A–F) in Japan. Airborne fungal levels in the factories were determined, and a traditional koji called “kona-koji” made from the mold naturally present in factory C (Aogashima village) was analyzed. Isolates of sec. Nigri fungi were identified morphologically and confirmed via cytochrome b gene analysis. In factory A (Nago city), airborne fungal levels of sec. Nigri were 4,000 and 100 cfu/m³ in the koji-making and fermentation rooms, respectively. In factories B, C, and D, the levels were 40, >104 cfu/m³, and 100 cfu/m³, respectively. In factory F (Iki city), there were high levels of airborne white-koji mold (a white mutant of Asp. luchuensis). The most dominant fungal species of sec. Nigri was isolated and identified as Asp. luchuensis via genetic analysis. This is likely to have originated from the commercial fermentation culture used. Asp. niger and Asp. luchuensis were isolated from kona-koji. Mycotoxin production (ochratoxin and fumonisin B2) by Asp. luchuensis (eight strains) and Asp. niger (three strains) was virtually inexistent; only one strain of Asp. niger was positive for fumonisin B2. This study clearly shows that mycotoxin-producing fungi are not dominant in the fungal flora present in the shochu factories examined and therefore, that the liquor can be safely fermented.Implications: In this study, we examined the presence of mycotoxin-producing Aspergillus sec. Nigri fungi in six shochu (Japanese distilled beverage) factories. The most dominant fungal species of sec. Nigri was isolated and identified as Aspergillus luchuensis (black-koji mold). The proportion of mycotoxin-producing Aspergillus niger and Aspergillus carbonarius was very small. In addition, the Asp. niger isolated from koji mold did not have the ability to produce ochratoxins or fumonisin B. This study clearly shows that shochu can be safely fermented. [ABSTRACT FROM AUTHOR]

Invasive fungal disease (IFD) in patients with haematological disorders is a fatal disease, making rapid identification and treatment crucial. However, the identification of the causative fungus is often difficult, sometimes even impossible. There have been few reports concerning the causative species of IFD. This study aimed to investigate the epidemiology and causative organism of IFD in patients with haematological diseases in Japan. We analyzed the IFD cases among the patients with haematological malignancies identified at the Medical Mycological Research Center, Chiba University, between 2013 and 2019. The most common underlying disease was acute myeloid leukaemia (34.3%). Forty-six point one percent of IFD patients received haematopoietic stem cell transplantation (HSCT). The major pathogens were Aspergillus, Candida, and Fusarium. Aspergillus fumigatus was the most common Aspergillus species, and Candida fermentati and Fusarium petroliphilum were the most common Candida and Fusarium species, respectively, in this analysis. Furthermore, various cryptic species and non-albicans Candida were identified. The drug susceptibility of such relatively rare strains suggests that analysis of the causative fungi should provide valuable information for therapeutic options. Therefore, our study indicated that it is clinically significant to identify the organism in as much detail as possible. [ABSTRACT FROM AUTHOR]

Aspergillus fumigatus-related species are responsible for causing aspergillosis, which is a fatal infectious disease. Recently, there has been a series of reports of A. fumigatus-related species that are resistant to azole drugs used in clinical practice for the treatment of fungal infections. Some of these species have been isolated from outdoor environments. Testing the drug susceptibility of the strains from outdoor environments, therefore, is important. In this study, we isolated and cultured 72 strains of A. fumigatus-related species from the outdoor environment in Japan. The isolates identified via morphological observation and molecular phylogenetic analysis were Aspergillus felis, Aspergillus lentulus, Aspergillus pseudoviridinutans, Aspergillus udagawae, and Aspergillus wyomingensis. The results of the drug susceptibility testing revealed that A. felis (6 of 14 strains) and A. pseudoviridinutans (13 of 17 strains) were resistant to itraconazole (ITCZ), with 4 mg/L or higher minimum inhibitory concentrations (MICs). The voriconazole (VRCZ)-resistant strains with 4 mg/L or higher MICs were A. felis (14 of 14), A. lentulus (4 of 4), A. pseudoviridinutans (15 of 17), A. udagawae (23 of 34), A. wyomingensis (1 of 3), and A. pseudoviridinutans (1 of 3). Among them, A. felis (1 of 14) and A. pseudoviridinutans (7 of 17) demonstrated 8 mg/L or higher MICs for ITCZ and VRCZ. These results indicate that A. fumigatus-related species resistant to ITCZ and VRCZ are widely distributed in outdoor environments in Japan. [ABSTRACT FROM AUTHOR]

The black koji mold, Aspergillus luchuensis , which belongs to Aspergillus section Nigri , is used for the production of traditional Japanese spirits (shochu) mainly in the southern districts of Japan. This mold is known to produce amylolytic enzymes essential for shochu production; however, mechanisms regulating amylolytic gene expression in A. luchuensis have not been studied in as much detail as those in the yellow koji mold, Aspergillus oryzae. Here, we examined the gene expression profiles of deletion mutants of transcription factors orthologous to A. oryzae AmyR and CreA in A. luchuensis. A. luchuensis produces acid-unstable (AmyA) and acid-stable (AsaA) α-amylases. AmyA production and amyA gene expression were not influenced by amyR or creA deletion, indicating that amyA was constitutively expressed. In contrast, asaA gene expression was significantly down- and upregulated upon deletion of amyR and creA , respectively. Furthermore, the glaA and agdA genes (encoding glucoamylase and α-glucosidase, respectively) showed expression profiles similar to those of asaA. Thus, genes that play pivotal roles in starch saccharification, asaA , glaA , and agdA , were found to be regulated by AmyR and CreA. Moreover, despite previous reports on AsaA being only produced in solid-state culture, deletion of the ortholog of A. oryzae flbC, which is involved in the expression of the solid-state culture-specific genes, did not affect AsaA α-amylase activity, suggesting that FlbC was not associated with asaA expression. [ABSTRACT FROM AUTHOR]

1-Octen-3-ol is a major aroma component of awamori, a traditional distilled liquor produced in Okinawa Prefecture, Japan. As 1-octen-3-ol is thought to affect the sensory properties of awamori, it is important to fully characterize the compound's biosynthetic pathway and control mechanism. We previously reported that the fatty acid oxygenase ppoC (ppo: psi-produced oxygenase) of Aspergillus luchuensis is directly involved in the production of 1-octen-3-ol in rice koji (Kataoka et al., J. Biosci. Bioeng., 129, 192–198, 2020). In the present study, we constructed A. luchuensis ppoD disruptants to characterize the role of ppo genes in 1-octen-3-ol biosynthesis. A small-scale awamori fermentation test was performed using ppoA , ppoC , and ppoD single disruptants (Δ ppoA , Δ ppoC , and Δ ppoD , respectively), along with the parent strain, Δ ligD. 1-Octen-3-ol was not detected in the distillate prepared using the Δ ppoC strain. We conclude that A. luchuensis ppoC is the only 1-octen-3-ol–producing factor in the awamori brewing process. Because Δ ppoA and Δ ppoD slightly enhanced 1-octen-3-ol productivity, these two genes may play a role in negatively controlling 1-octen-3-ol biosynthesis. [ABSTRACT FROM AUTHOR]

The incidence of non-dermatophyte mould onychomycosis has been increasing worldwide for the past several decades, but it is not well recognized in Japan. Recent molecular techniques and phylogenetic analyses contributed to the identification of uncommon and emerging species. We came across 13 (0.5%) cases of non-dermatophyte onychomycosis among a total of 2,591 onychomycosis cases in 106,703 outpatients during the past 5 years (January 2015-December 2019). The cases included 5 patients with Aspergillus species, 4 patients with Fusarium species, and one patient each with Scopulariopsis brevicaulis and Botryosphaeria dothidea, respectively. Botryosphaeria dothidea is closely related phylogenetically to Neoscytalidium dimidiatum. In Japan, at the time of writing this report, there are 26 reported cases of ungual aspergillosis and 18 cases of hyalohyphomycosis caused by Fusarium species. We summarize these cases and report the symptoms and mycological features. [ABSTRACT FROM AUTHOR]

Awamori is a distilled spirit produced in Okinawa Prefecture, in southern Japan. Awamori contains the volatile organic compound 1-octen-3-ol, an important flavor component. Here, using solid-phase microextraction–gas chromatography–mass spectrometry (SPME-GCMS), we demonstrate that the black koji mold Aspergillus luchuensis produces 1-octen-3-ol in rice koji. To examine the role of the fatty acid oxygenase genes ppoA and ppoC in 1-octen-3-ol biosynthesis by A. luchuensis , we constructed ppoA and ppoC disruptants, Δ ppoA and Δ ppoC , respectively, via protoplast-PEG transformation. No clear differences in growth and conidiation were observed between the transformants and the parent strain. Volatile compounds in rice koji prepared using these gene disruptants were analyzed by SPME-GCMS. The amount of 1-octen-3-ol contained in koji produced by the Δ ppoA strain was the same as that produced by the parental strain. In contrast, although the Δ ppoC strain grew on the rice koji, 1-octen-3-ol was not detected. These results indicate that ppoC is involved in 1-octen-3-ol biosynthesis in A. luchuensis. [ABSTRACT FROM AUTHOR]

Aspergillus fumigatus is the most prevalent species that causes aspergillosis. A. fumigatus strains with tandem repeats in the cyp51A promoter have emerged in the environment. Aspergillus species other than A. fumigatus have also been recognized as causative agents of aspergillosis; however, they show lower susceptibility to antifungals compared with A. fumigatus. Therefore, it is important to precisely identify Aspergillus species and determine their antifungal susceptibility. Herein, we collected 119 mold strains isolated from clinical specimens collected at a hospital between November 2013 and December 2018. The collected strains were identified by sequencing several regions, including internal transcribed spacers, and determined their susceptibility to the antifungals itraconazole, voriconazole, and amphotericin B. Of 119 strains, 107 were Aspergillus species, which were identified as A. fumigatus (67), Aspergillus section Nigri (21), A. flavus (7), A. terreus (6), and A. nidulans (6). In Aspergillus section Nigri , the number of A. niger was less than the number of A. welwitschiae and A. tubingensis. Two azole-resistant A. fumigatus samples were included among the isolates. Four of the eight A. tubingensis isolates showed less susceptibility to voriconazole; however, all isolates of A. niger and A. welwitschiae were susceptible to itraconazole and voriconazole. Because of lack of susceptibility data for non-fumigatus Aspergillus and an increasing frequency of antifungal resistance among A. fumigatus , our data along with further surveillance may contribute to determining the frequency and susceptibility of Aspergillus spp. clinical isolates in Japan. [ABSTRACT FROM AUTHOR]

Background: Aspergillus is one of the important pathogens that contribute to high mortality in patients with coronavirus disease 2019 (COVID-19) in intensive care units (ICUs). Although incidence rates of Aspergillus coinfection are high globally, a Japanese national survey reported a low incidence. This study aimed to describe the clinical characteristics of patients with COVID-19-associated pulmonary aspergillosis at our institute., Methods: We identified patients with microbiologically confirmed COVID-19 on mechanical ventilation in the ICU. Of these patients, we identified patients in whom Aspergillus was cultured from the respiratory specimen., Results: Of a total of 169 patients, seven had aspergillosis (4.1%), which included three patients, three patients, and one patient with possible, probable, and proven aspergillosis, respectively, according to the criteria of the European Confederation of Medical Mycology International Society. All patients received systemic steroid therapy. Two patients (one each with proven and probable aspergillosis) had tracheobronchitis diagnosed by bronchoscopy. All patients in whom Aspergillus was repeatedly isolated from samples died. The mortality rates for all cases and probable and proven cases were 57% (4/7) and 75% (3/4), respectively., Conclusions: The incidence rate of aspergillosis in patients with COVID-19 in the ICU was higher in our institute than that reported by a Japanese national survey (4.1% vs. 0.5%). Repeated detection of Aspergillus might suggest a true Aspergillus infection, such as chronic aspergillosis, rather than colonization. In patients with severe COVID-19 patients, it is important to always keep CAPA in mind., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: The Japanese traditional cuisine, Washoku, considered to be responsible for increased longevity among the Japanese, comprises various foods fermented with the non-pathogenic fungus Aspergillus oryzae (koji). We have recently revealed that koji contains an abundant amount of glycosylceramide. Intestinal microbes have significant effect on health. However, the effects of koji glycosylceramide on intestinal microbes have not been studied. Materials and methods: Glycosylceramide was extracted and purified from koji. C57BL/6N mice were fed a diet containing 1 % purified koji glycosylceramide for 1 week. Nutritional parameters and faecal lipid constituents were analyzed. The intestinal microbial flora of mice on this diet was investigated. Results: Ingested koji glycosylceramide was neither digested by intestinal enzymes nor was it detected in the faeces, suggesting that koji glycosylceramide was digested by the intestinal microbial flora. Intestinal microbial flora that digested koji glycosylceramide had an increased ratio of Blautia coccoides. Stimulation of B. coccoides growth by pure koji glycosylceramide was confirmed in vitro. Conclusions: Koji functions as a prebiotic for B. coccoides through glycosylceramide. Since there are many reports of the effects of B. coccoides on health, an increase in intestinal B. coccoides by koji glycosylceramide might be the connection between Japanese cuisine, intestinal microbial flora, and longevity. [ABSTRACT FROM AUTHOR]

Three strains of a xerophilic Aspergillus species were isolated from house dust and honey in Japan. A molecular phylogenetic analysis based on the combined dataset for four regions (internal transcribed spacer rDNA, calmodulin, β-tubulin, and RNA polymerase II second largest subunit) revealed that the strains formed an independent lineage, sister to Aspergillus halophilicus classified in section Restricti . Morphological comparisons show that the strains differ from A. halophilicus in three aspects: (i) the size of cleistothecia, as well as the surface structure and size of ascospores, (ii) the ability to grow on Harrold's agar and dichloran 18 % glycerol agar, and (iii) the lack of conidiophore formation on potato dextrose agar +20 % NaCl. These strains could be clearly distinguished from all known Aspergillus section Restricti species. Therefore, we consider it to be a novel species and propose the name Aspergillus verrucosus sp. nov. (NBRC 115547 T ).

Many survivors of the tsunami that occurred following the Great East Japan Earthquake on March 11, 2011, contracted a systemic disorder called "tsunami lung," a series of severe systemic infections following aspiration pneumonia caused by near drowning in the tsunami. Generally, the cause of aspiration pneumonia is polymicrobial, including fungi and aerobic and anaerobic bacteria, but Aspergillus infection is rarely reported. Here we report a case of tsunami lung complicated by disseminated aspergillosis, as diagnosed during autopsy. [ABSTRACT FROM AUTHOR]

Abstract: The industrial fungus Aspergillus luchuensis is used to produce a distilled spirit in Okinawa Island, Japan. Recently, the genome sequence of A. luchuensis RIB2604 (Aspergillus awamori NBRC 4314) was revealed and many functional genes are now expected to be analyzed. Gene targeting is necessary for analyzing the function of a gene; however, gene targeting frequencies in A. luchuensis are very low. To develop a highly efficient gene-targeting system for A. luchuensis, we disrupted A. luchuensis ligD (ALligD) encoding the human DNA ligase IV (ligIV) homologue using an Agrobacterium mediated gene transformation method. Deletion of ALligD dramatically improved homologous recombination efficiency (reached 100%) compared to that in the wild-type strain (0.8%), when 1000-bp homologous flanking regions were used. The ALligD disruptant showed no apparent defect in vegetative growth, and it exhibited increased sensitivity to phleomycin and high methyl methanesulphonate concentrations compared to the wild-type strain. Furthermore, using this ALligD disruptant, we disrupted ALpksP encoding an Aspergillus fumigatus polyketide synthase P (alb1/pksP) orthologue. The ALpksP disruptant displayed a decolourized conidial phenotype. This result indicated that ALpksP is a key factor for conidial black pigmentation in A. luchuensis. Our results indicate that the ALligD mutant is an efficient host for targeted gene disruption in A. luchuensis. [Copyright &y& Elsevier]

The data on visceral mycoses reported in the "Annual of Pathological Autopsy Cases in Japan" were analyzed epidemiologically every four years from 1989 to 2005, and in 2007. The frequency rates of visceral mycoses dropped sharply between 1989 (4.5%) and 1994 (3.2%), but by 2001 had risen again and have remained (4.4-4.6%) generally stable since then. The predominant causative agents were Candida and Aspergillus. Although the rate of candidosis showed a gradual decrease, the rate of aspergillosis showed an increase by degrees. Furthermore, the rate of aspergillosis exceeded that of candidosis in 1994, and the difference in the rates between the two conditions apparently further increased until 2001. After 2005, however no changes in this difference were observed. For complicated infections, the incidence of coinfection with Aspergillus and Candida showed a decreasing, and that with Aspergillus and Zygomycetes showed an increasing tendency. Severe infections with Zygomycetes showed a clear increase from 57.4% in 1989 to 88.9% in 2007. Comparing underlying diseases with mycoses in 1989 and 2007, leukemia (including myelodysplastic syndrome) decreased from 26.1% to 18.8% and bacterial infections (including interstitial pneumonia) increased from 11.1% to 22.1%. By age, the highest frequency rate of mycoses was observed in the range of 60-79 years, and the frequency rate of exogenous fungal infections such as aspergillosis, cryptococcosis, zygomycosis and trichosporonosis showed an increasing trend in the less than one-year old group. [ABSTRACT FROM AUTHOR]

As there is not yet a standardized in vitro susceptibility test of micafungin (MCFG), we evaluated the methods of such testing, focusing on the judgment method of MIC, based on the National Committee for Clinical Laboratory Standards (NCCLS) M27-A2, M38-A, the proposed standards of The Japanese Society for Medical Mycology (JSMM) for yeast (JSMM-Y) and for filamentous fungi (JSMM-F) against Candida spp. and Aspergillus spp. The judgment of MIC value was performed spectrophotometrically and visually in both (NCCLS and JSMM) assays. Only the spectrophotometric MIC judgment against Aspergillus spp. in the NCCLS assay used two end points: 80% inhibitory concentration (IC80) of the growth control and 50% inhibitory concentration (IC50). The end point for the visual judgment against Aspergillus spp. in the NCCLS assay was determined to be no growth from the small clumps of altered hyphae in the microtiter plate. The other MIC judgments used an IC80 end point. The MICs of MCFG for Candida spp. were ≤0.0039-1 μg/ml in both judgments by the two assays. In contrast, there was a large difference of the MIC values against Aspergillus spp. between the NCCLS assay results using the IC80 end point and the visual judgment (>4 μg/ml and 0.0078-0.0313/μg/ml). However, the MICs using the IC50 end point and those by JSMM assay agreed with the result of the visual assessment. Therefore, we recommend the JSMM assay, the NCCLS assay using the IC50 end point or the novel visual judgment for the susceptibility testing of MCFG against Aspergillus spp. [ABSTRACT FROM AUTHOR]

The highly humid climate of Japan facilitates the growth of various molds. Among these molds, Aspergillus oryzae is the most important and popular in Japan, and has been used as yellow-koji in producing many traditional fermented beverages and foods, such as Japanese sake, and soy sauce. Taka-amylase A (TAA), a major enzyme produced by the mold, is well known worldwide to be a leading enzyme for industrial utilization and academic study, since many extensive studies have been carried out with TAA. In southern Kyushu, the other koji’s of citric acid-producing molds have often been used, such as in the production of a traditional distilled liquor of shochu. The koji molds black-koji and white-koji produce two types of α-amylase, namely, acid-stable (AA) and common neutral (NA). The latter enzyme is enzymatically and genetically similar to TAA. In this review, we investigate AA from three molds, Aspergillus niger, A. kawachii and A. awamori, and the yeast Cryptococcus sp. regarding the distinguishable properties between AA and NA. (i) The N-terminus amino acid sequences of AA determined by molecular cloning started with the sequence of L-S-A-, whereas those of NA started with A-T-P-. (ii) Most of the full sequences of AA were composed of, besides a core catalytic domain, an extra domain of a hinge region and a carbohydrate binding domain, which could be responsible for raw-starch-digestibility. The AA from A. niger has no exceptionally extra domain, similarly to NA. (iii) Simple methods for distinguishing AA from NA using CNP-α-G3 and G5 as substrates were developed by our group. (iv) The number of subsite in AA on the basis of its cleavage pattern of maltooligosaccharides was estimated to be five, which differs from that of TAA, 7–9. AA has many advantages in industrial applications, such as its acid-stability, thermostability, and raw-starch digesting properties. [Copyright &y& Elsevier]

No abstract available. [ABSTRACT FROM AUTHOR]

The genome of Aspergillus oryzae, a fungus important for the production of traditional fermented foods and beverages in Japan, has been sequenced. The ability to secrete large amounts of proteins and the development of a transformation system have facilitated the use of A. oryzae in modern biotechnology. Although both A. oryzae and Aspergillus flavus belong to the section Flavi of the subgenus Circumdati of Aspergillus, A. oryzae, unlike A. flavus, does not produce aflatoxin, and its long history of use in the food industry has proved its safety. Here we show that the 37-megabase (Mb) genome of A. oryzae contains 12,074 genes and is expanded by 7–9 Mb in comparison with the genomes of Aspergillus nidulans and Aspergillus fumigatus. Comparison of the three aspergilli species revealed the presence of syntenic blocks and A. oryzae-specific blocks (lacking synteny with A. nidulans and A. fumigatus) in a mosaic manner throughout the genome of A. oryzae. The blocks of A. oryzae-specific sequence are enriched for genes involved in metabolism, particularly those for the synthesis of secondary metabolites. Specific expansion of genes for secretory hydrolytic enzymes, amino acid metabolism and amino acid/sugar uptake transporters supports the idea that A. oryzae is an ideal microorganism for fermentation. [ABSTRACT FROM AUTHOR]

The article discusses the result of the study on the cloning and characterization of saponin hydrolases from Aspergillus oryzae and Eupenicillium brefeldianum, in Japan. Glycoproteins were confirmed to be the enzymes that come from these hydrolases. Each enzyme's deduced amino acid sequences from the cloned genes showed a homology with that of the saponin hydrolase Sdn1 from Neocosmospora vasinfecta var. vasinfecta. The soyasaponin I to soyasapogel B and triose were hydrolyzed with recombinant enzymes.

The article discusses the result of the study regarding the contribution of a single free cysteine residue and disulfide bond to the thermostability of Aspergillus saitoi 1,2-α-mannosidase, in Japan. A. saitoi 1,2-α-mannosidase contains three conserved cysteine residues, which were not essential for the activity analyzed by site-directed mutagenesis and kinetics. The destabilization of the enzyme was due to the substitution at each cysteine residue. Thus, the hydrophilic and large side chain causes the destabilization.

The article discusses the result regarding the study of Taka-amylase A (TAA) of conidia from Aspergillus oryzae RIB40, in Japan. The TAA not only exists in germinated conidia but also in conidia, based from the results. The activity of TAA was detected in cell extracts of conidia using 0.5M acetate buffer. Investigators conclude that the TAA might belong to a set of basal proteins in the conidia that facilitates the imbibition and germination of conidia.

The diversity of sporocarp-inhabiting fungi (SCIF) was examined using six samples of xylarialean fungi from two different forests in Ibaraki Prefecture, Japan: a moist forest in the Sakuragawa area and an urban dry forest in the Tsukuba area. These fungi were enumerated using direct observation and dilution plate methods. We obtained 44 isolates, and careful morphological and molecular phylogenetic studies of these isolates revealed that approximately 30% of the operating taxonomic units were undescribed or cryptic species related to known fungi. Although typical mycoparasitic fungi, such as helotialean fungi and Trichoderma spp., were not isolated, the genera Acremonium, Acrodontium, and Simplicillium were detected. Comparisons of SCIF communities between the two forests suggested that the number of isolated species in the Sakuragawa area was lower than that in the Tsukuba area. Soil-borne fungi, such as Aspergillus, Beauveria, Penicillium, and Talaromyces, or polypores/corticioid mushrooms, are frequently detected in the Tsukuba area. Factors affecting SCIF communities in the two forests are discussed. Some noteworthy fungi are briefly described with notes on taxonomy, ecology, and molecular phylogeny. [ABSTRACT FROM AUTHOR]

• Aspergillus luchuensis albino strains have other mutations than pigment deficiency. • Passaged albino strains are genetically distinct from A. luchuensis and A. kawachii. • Albino strains with isocitrate lyase mutation showed no growth with acetate. • Passage may influence the mutation of genes for stress responses in albino strains. Black Aspergillus luchuensis and its white albino mutant are essential fungi for making alcoholic beverages in Japan. A large number of industrial strains have been created using novel isolation or gene/genome mutation techniques. Such mutations influence metabolic and phenotypic characteristics in industrial strains, but few comparative studies of inter-strain mutation have been conducted. We carried out comparative genome analyses of 8 industrial strains of A. luchuensis and A. kawachii IFO 4308, the latter being the first albino strain to be isolated. Phylogenetic analysis based on 8938 concatenated genes exposed the diversity of black koji strains and uniformity among albino industrial strains, suggesting that passaged industrial albino strains have more genetic mutations compared with strain IFO 4308 and black koji strains. Comparative analysis showed that the albino strains had mutations in genes not only for conidial pigmentation but also in those that encode N-terminal acetyltransferase A and annexin XIV-like protein. The results also suggest that some mutations may have emerged through subculturing of albino strains. For example, mutations in the genes for isocitrate lyase and sugar transporters were observed only in industrial albino strains. This implies that selective pressure for increasing enzyme activity or secondary metabolites may have influenced the mutation of genes associated with environmental stress responses in A. luchuensis albino strains. Our study clarifies hitherto unknown genetic and metabolic characteristics of A. luchuensis industrial strains and provides potential applications for comparative genome analysis for breeding koji strains. [ABSTRACT FROM AUTHOR]

Invasive fungal infections (IFIs) are serious infections that develop in conjunction with neutropenia after chemotherapy for acute leukemia or with hematopoietic stem cell transplantation. Conventionally, empirical antifungal therapy was recommended to treat IFIs for patient safety despite a lack of evidence of fungal infections. However, many studies have indicated that antifungals were not necessary for over half of patients, and several detriments of empirical therapy were noted, e.g., antifungals caused adverse reactions, an increase in drug-resistant fungi was a possibility, and medical costs soared. β-D-glucan (BDG) is a component of clinically important fungi such as Aspergillus and Candida. The G-test was developed in Japan as a way to measure BDG in serum using a coagulation factor from the blood of the horseshoe crab. Pre-emptive antifungal therapy based upon serodiagnosis with a BDG or galactomannan assay and CT imaging has been introduced. With pre-emptive antifungal therapy, the prognosis is equivalent to that with empirical therapy, and the dose of the antifungal has been successfully reduced. Measurement of BDG has been adopted widely as a method of diagnosing IFIs and is listed in the key guidelines for fungal infections and febrile neutropenia. [ABSTRACT FROM AUTHOR]

To understand fungal contamination in the indoor environments of the disaster region, surveys were conducted to detect mycoflora in temporary shelters, prefabricated temporary housing, private housing, and rented apartments in areas affected by the Great East Japan Earthquake. The results from the surveys of temporary shelters indicated that the indoor-air fungal counts at all sampling points were less than 1000 colony forming units (cfu)/m 3 , which is the recommended limit for fungal contamination in indoor air. However, the Aspergillus counts were high compared to the indoor environments of typical housing. Since Aspergillus is a known allergenic genus, careful attention should be paid to residents' health. The results of the surveys of private housing and rented apartments also indicated that fungal counts were highest during the rainy season throughout the summer. In contrast, temporary housing had a maximum fungal count in the winter. The extremely high level of fungal condensation in indoor air may have been due to the high relative humidity and loss of heat insulation in the buildings' attics. It is thought that these problems happen most commonly in colder regions, such as the entire area affected by the Great East Japan Earthquake. The case of a patient with allergic bronchopulmonary mycosis caused by a large amount of Eurotium herbariorum mold in his temporary housing was reported to demonstrate the health risks posed by fungi in this disaster region.

Aspergillus section Nigri, a group of black Aspergillus, has several cryptic species that were recently discovered to be intrinsically resistant to azole antifungals. In this study, susceptibility testing of 35 clinical isolates of Aspergillus tubingensis and Aspergillus welwitschiae in Japan was carried out using microdilution method. Strains tolerant to itraconazole in A. tubingensis (14/17 strains) and A. welwitschiae (6/18 strains) were discovered with hyphal growth and conidial germination above the minimal inhibitory concentration by microscopic observation, while no resistant strain was observed macroscopically. In contrast, no strain with reduced susceptibility to voriconazole, posaconazole, and amphotericin-B was found. Further examination may be required to determine the susceptibility of cryptic species in Aspergillus section Nigri to antifungals.

Abstract Purpose: To analyze the incidence and causes of fatality of patients with chronic granulomatous disease (CGD) in Japan and to provide an opportunity for comparison with reports from other countries. Methods: The Study Group of Phagocytic Disorders in Japan conducted a questionnaire survey on CGD patients in Japan, results of which formed the basis of the study. Results: Clinical details of 221 patients were analyzed: 194 male and 27 female (ratio: 7.2/1), 152 living, 51 dead and 18 unknown. The prevalence of CGD was estimated to be 1/287,709 live births. The fatality rate was 23.1%. The mean age of the surviving patients increased from 8 years 4 months in 1985 to 16 years 0 months in 1998. Although the mean age of death was advanced by 4 years 11 months during the same period of time, the fatality rate has remained practically unchanged during the past 13 years. The number of living adult patients has tended to increase (32.1%). At the time of study, approximately 90% of patients had been placed on sulfamethoxazole-trimethoprim prophylactically or therapeutically, either singly or in combination with other modalities, including interferon-γ, antifungal agents and various antibiotics. Pulmonary infections were responsible for 58.3% of fatalities. Conclusions: With early diagnosis and prompt institution of appropriate therapy, the mean age of CGD patients in Japan has been increasing, but the fatality rate has remained practically unchanged during the last 13 years, mostly due to the fungal infections. Key words Aspergillus infection, chronic granulomatous disease, questionnaire survey. [ABSTRACT FROM AUTHOR]

The International Space Station (ISS) is a closed facility that orbits the earth carrying not only its crew but also microorganisms. We have participated in microbiota analysis projects for the Japanese Experiment Module KIBO (ISS; operations nomenclature: Microbe-I, II, III, and IV) and were in charge of fungal screening. The interior of KIBO was sampled using swabs and microbe detection sheets (MDSs) for fungal detection. The dominant genera obtained by culture were Aspergillus and Penicillium. DNA analyses of the fungal biota using a clone library showed that KIBO was dominated by Malassezia, a fungal inhabitant of human skin. Three fungal species, Aspergillus sydowii, Penicillium palitans, and Rhodotorula mucilaginosa, which grew under microgravity in KIBO were observed under a field emission-scanning electron microscope on the ground. No novel phenotypic characteristics were noted. The results of antifungal susceptibility testing of all isolates did not differ significantly from previous reports of corresponding fungi. In Microbe-I (August 2009), MDSs were culture negative, while in the next stages the CFU of MDSs were 10 for Microbe-II (February 2011), 24 for Microbe-III (October 2012), and 151 for Microbe-IV (February 2015). These results indicated that fungi inside KIBO are increasing and expanding over time, and therefore continuous surveillance is crucial., (© 2021 The Societies and John Wiley & Sons Australia, Ltd.)

Objectives: To verify the effectiveness of our infection control measures based on the infection control risk assessment (ICRA) to minimize the risk of Aspergillus dispersion before, during, and after demolition work in a university hospital., Background: It is widely accepted that invasive aspergillosis is associated with construction, renovation, and demolition activities within or close to hospital sites. However, the risk is underestimated, and only limited preventive measures are taken in Japanese hospitals., Method: The demolition process, carried out in July 2014, was supervised by our facility management in collaboration with the infection prevention team and followed an adapted ICRA tool. Dust containment measures were implemented to reduce the risk of airborne Aspergillus contamination. Air sampling was performed at four wards in the adjacent hospital buildings to assess the containment measures' effectiveness., Results: A high, undetermined number of colonies of bacteria and molds were detected on all outside balconies before demolition. During demolition, Aspergillus spp. was detected only in the ward closest to the demolition site. However, no case of aspergillosis was reported. The difference-in-difference analysis revealed that the interaction between the demolition activity, height of the ward, and distance of the air intake to the demolition activities resulted in a significant increase in the numbers of Aspergillus spp., Conclusions: When large-scale demolition work occurs in hospital premises, Aspergillus spp. may increase in the ward where the vertical and horizontal distance of air intake from the demolition site is close, even though infection control measures based on the ICRA are implemented.

Presents abstracts of studies presented at the symposium on frontier of the molecular mechanism of plant microtubule organization during the 69th annual meeting of the Botanical Society of Japan. "Microtubule Organization in Physcomitrella Patens," by Yoshikatsu Sato; "Essential Role of Microtubule in Polarized Hyphal Growth of Aspergillus Nidulans," by Tetsuya Horio and Berl R. Oakley; "The Microtubule Organizing Mechanism by Histone H1-Tubulin Complex on the Nuclear Surface," by Takashi Hotta and Koichi Mizuno.

Background: There are several clinical diagnostic criteria for allergic bronchopulmonary aspergillosis (ABPA). However, these criteria have not been validated in detail, and no criteria for allergic bronchopulmonary mycosis (ABPM) are currently available., Objective: This study proposes new diagnostic criteria for ABPA/ABPM, consisting of 10 components, and compares its sensitivity and specificity to existing methods., Methods: Rosenberg-Patterson criteria proposed in 1977, the International Society for Human and Animal Mycology (ISHAM) criteria proposed in 2013, and this new criteria were applied to 79 cases with pathological ABPM and the control population with allergic mucin in the absence of fungal hyphae (n = 37), chronic eosinophilic pneumonia (n = 64), Aspergillus-sensitized severe asthma (n = 26), or chronic pulmonary aspergillosis (n = 24). These criteria were also applied to the 179 cases with physician-diagnosed ABPA/ABPM in a nationwide Japanese survey., Results: The sensitivity for pathological ABPM with Rosenberg-Patterson criteria, ISHAM criteria, and this new criteria were 25.3%, 77.2%, and 96.2%, respectively. The sensitivity for physician-diagnosed ABPA/ABPM were 49.2%, 82.7%, and 94.4%, respectively. The areas under the curve for the receiver-operating characteristic curves were 0.85, 0.90, and 0.98, respectively. The sensitivity for ABPM cases that were culture-positive for non-Aspergillus fungi were 13.0%, 47.8%, and 91.3%, respectively., Conclusions: The new diagnostic criteria, compared with existing criteria, showed better sensitivity and specificity for diagnosing ABPA/ABPM., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

To understand fungal contamination in the indoor environment of the disaster region, a field survey was performed to measure the number of fungal counts and identify isolates in the indoor air of prefabricated temporary housing, privately independent-housing, and rented apartments flooded by the East Japan Great Earthquake disaster tsunami. As a result, the period with the highest detected fungal count was from the rainy season to summer in independent-housing and rented apartments. Moreover, in the temporary housing, the fungal number increased further in winter as indicated by the maximum fungal-number throughout the measurement period. The detection frequency of Aspergillus species was relatively higher in the indoor air of temporary housing than in typical housing in the non-disaster area. Since Aspergillus is known as an allergenic genus, it requires careful attention to the health risk for residents. The extremely high level of fungal condensation in indoor air possibly occurred due to high relative humidity and loss of heat insulation in the building attics. It is suggested that this problem commonly happened in the cold region including the entire disaster region of the East Japan Great Earthquake.

Pulmonary mycosis is a fungal disease that commonly affects bottlenose dolphins (Tursiops truncatus) and is generally treated by the oral administration of azoles, such as itraconazole (ITZ) and voriconazole (VRZ). However, antifungal susceptibility testing of clinical isolates has not been well performed as a routine clinical examination in aquaria. In this study, we collected fungal species from the blowholes of 14 bottlenose dolphins, of which 12 were treated with ITZ or VRZ. All dolphins were housed in the Port of Nagoya Public Aquarium. The fungal species Candida albicans, C. tropicalis, C. glabrata, Aspergillus fumigatus, and A. niger were isolated. E-tests were performed to determine the minimum inhibitory concentrations (MICs) of ITZ and VRZ on these isolates. VRZ-resistant C. tropicalis (MIC: >32 µg/ml) and A. niger (MIC: >32 µg/ml) were isolated from three dolphins treated with ITZ or VRZ. Additionally, azole-resistant isolates of C. albicans and C. glabrata were collected from two dolphins that had never received azole therapy. To the best of our knowledge, our study is the first to report the isolation of VRZ-resistant C. albicans, C. tropicalis, and A. niger from the blowholes of bottlenose dolphins. Thus, antifungal susceptibility testing is a crucial strategy for selecting antifungal agents to treat respiratory fungal infections in bottlenose dolphins in aquaria.

Background: Allergic bronchopulmonary aspergillosis (ABPA) is an allergic pulmonary disease characterized by a hypersensitivity reaction to Aspergillus species colonizing the airways. The clinical characteristics of ABPA may differ depending on genetic and environmental background. We performed a nationwide survey to determine the clinical characteristics of ABPA in Japan., Methods: In 2013, a questionnaire on physician-diagnosed ABPA/allergic bronchopulmonary mycosis was sent to 903 medical centers specializing in respiratory or allergic diseases. Cases fulfilling the following criteria were categorized as possible ABPA-central bronchiectasis (ABPA-CB): 1) presence of specific serum immunoglobulin E (IgE) antibodies or a positive skin reaction to Aspergillus, and 2) bronchiectasis or mucoid impaction in the central bronchi., Results: Of 499 physician-diagnosed cases reported by 132 clinical centers, 358 cases met the criteria for possible ABPA-CB. Median age of ABPA-CB onset was 57 (interquartile range, 44-68) years; later-onset disease, developing ≥50 years of age, accounted for 66% of the cases and was associated with female sex, delayed onset of asthma, and lower levels of serum IgE. A third of the patients (120 patients, 34%) exhibited low levels of serum total IgE (<1000 IU/mL). Aspergillus species were isolated from sputum in 126/213 cases (59%), and Schizophyllum commune was identified in 12 (6%) patients. During the course of the treatment, ABPA recurred in 169 (48%) cases., Conclusions: This nationwide survey identified several unique clinical characteristics of ABPA in Japan, such as late-onset, relatively lower serum IgE levels, and frequent recurrences/flares., (Copyright © 2017 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.)

Presents a case of allergic bronchopulmonary aspergillosis (ABPA) in a boy without signs of cystic fibrosis. Overview of ABPA; Initial treatment of the patient; Diagnosis of the boy's condition.

Objective:The purpose of this study was to evaluate the effectiveness of the combination of inferior and middle meatal antrostomies for treatment of a maxillary sinus fungus ball by functional endoscopic sinus surgery.Methods:A retrospective analysis including 28 patients with non-invasive fungal maxillary sinusitis was performed. Fourteen patients underwent FESS with both middle and inferior meatal antrostomies (combined group). The remaining 14 patients were treated with FESS through only the middle meatal antrostomy (control group).Results:Post-operative computed tomography showed normal maxillary sinuses in all patients in the combined group. In contrast, in the control group, five patients (36 per cent) exhibited a normal maxillary sinus, seven (50 per cent) showed maxillary mucosal thickening and two (14 per cent) had persistent fungus balls in the maxillary sinus.Conclusion:FESS with a combination of middle and inferior meatal antrostomies proved more effective for treating fungal maxillary sinusitis. [ABSTRACT FROM PUBLISHER]

Horikoshi, Koki (The Institute of Physical and Chemical Research, Tokyo, Japan), Shigeji Iida, and Yonosuke Ikeda. Mannitol and mannitol dehydrogenases in conidia of Aspergillus oryzae. J. Bacteriol. 89:326-330. 1965.-A sugar alcohol was isolated from the conidia of Aspergillus oryzae and identified as d-mannitol. Two types of d-mannitol dehydrogenases, nicotinamide adenine dinucleotide phosphate-linked and nicotinamide adenine dinucleotide-linked, were found in the conidia. Substrate specificities, pH optima, Michaelis-Menton constants, and the effects of inhibitors were studied. d-Mannitol was converted to fructose by the dehydrogenases. Synthesis of d-mannitol dehydrogenases was not observed during germination; the content of d-mannitol decreased at an early stage of germination. It was assumed, therefore, that d-mannitol might be used as the source of endogenous respiration and provide energy for the germination.