Sequence information of 25S rRNA gene was useful for the genotype determination in Candida albicans. Genotypes of 301 C. albicans strains by this single PCR method were determined, and out of them, two strains of new genotype (genotype E) which are closely related to C. dubliniensis in its intron structure were discovered. Analysis of internal transcribed spacer (ITS) region sequence including 5.8S rRNA region in three varieties of C. neoformans was found to be an useful method for genotype determination, and a new genotype (Africa genotype) of var. gattii was discovered. In comparison with other traditional taxonomic methods in pathogenic fungi, usefulness of these genotype determination methods in their epidemiological studies was discussed. New PCR identification systems which were developed based on RAPD band pattern analyses for Histoplasma capsulatum, Paracoccidioides brasiliensis and Penicillium marneffei were also introduced. These genetic studies lead to a development of new DNA microarray identification method, and their usefulness was discussed.

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The incidence of invasive aspergillosis (IA) , which is commonly caused by Aspergillus fumigatus, has increased recently in immunocompromised patients and has become the common cause of death in these patients. Antifungal resistance is one of the reasons for treatment failure. Since the first itraconazole-resistant A. fumigatus was reported in 1997, the reports on clinical strains of triazole-resistant A. fumigatus have increased, as well as studies on the resistant mechanisms. In this paper, the known molecular mechanisms of antifungal resistance in Aspergillus, especially in A. fumigatus, are reviewed.

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In recent years fungi have been flourishing in immunocompromised patients of tertiary care centers. The data on the burden of opportunistic mycoses in India is not clear though the climate in this country is well suited for a wide variety of fungal infections. There are very few good diagnostic mycology laboratories and clinicians are still not aware of the emerging trends. Within the limited data available, an increased incidence of invasive candidiasis, aspergillosis, and zygomycosis are reported. The emergence of fungal rhinosinusitis, penicilliosis marneffei and zygomycosis due to Apophysomyces elegans is unique in the Indian scenario. Invasive candidiasis is the most common opportunistic mycosis. The global change in spectrum of Candida species is also observed in India; however, the higher prevalence of candidemia due to Candida tropicalis instead of C. glabrata or C. parapsilosis is interesting. Invasive aspergillosis is the second contender. Though due to difficulty in antemortem diagnosis the exact prevalence of this disease is not known, high prevalence is expected in Indian hospitals where construction activities continue in the hospital vicinity without a proper impervious barrier. The other opportunistic mycosis, invasive zygomycosis is an important concern as the world's highest number of cases of this disease is reported from India. The infection is commonly observed in patients with uncontrolled diabetes mellitus. Though antiretroviral therapy in AIDS patients has been introduced in most Indian hospitals, no decline in the incidence of cryptococcosis and penicilliosis has yet been observed. Thus there is need of good diagnostic mycology laboratories, rapid diagnosis, and refinement of antifungal strategies in India.

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We provide foot and nail care services to outpatients at our dermatology clinic in Saiseikai Kawaguchi General Hospital. Patients with dermatological foot problems such as onychogryphosis, onychomycosis, and ingrown nails are recommended by dermatologists to obtain foot and nail care services performed by a specially trained nurse. These services include a footbath, foot massage, nail clipping, and corn and nail filing. If ingrown nails exist, a small piece of cloth is placed under the nail edge.

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Fungal keratitis is one of the most challenging types of microbial keratitis for the ophthalmologist to diagnose and treat. Fungi causing human keratitis take the form of either yeasts or mold. Candida, the major pathogenic yeasts, can be detected in the normal ocular surface flora. Preceding ocular surface disorder, the wearing of contact lenses and the use of antibiotic/steroid eye drops may lead to candida keratitis. Infectious focus caused by Candida tends to melt the corneal stroma. Keratitis caused by mold often develops after an injury caused by soil and/or a plant. Mold can reach the anterior chamber without destroying the stromal layer of the cornea, which results in distinctive clinical features such as endothelial plaque and hyphate ulcer.
Fungal keratitis needs to be managed by antifungal agents, most of which must be prepared by ourselves to apply to the ocular surface. Candida keratitis should be managed with azoles. If the infection seems to be caused by mold, several antifungal drugs including pimaricin, which is the only agent officially applicable to the eye, should be used. Some cases of mold keratitis need to have therapeutic penetrating keratoplasty because of their lack of response to intensive medication.
Mold causing keratitis is variegated. Fusarium and Aspergillus can reach the intraocular space rapidly. Alternaria and some other unclassified molds remain in the superficial layer of the cornea for a long time. Our experiments indicate that the progress of focus in the cornea is regulated by the receptiveness of mold against temperature.

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In July 2007, information on the current status of mycological examinations in Japanese university clinics and requests for a university network system was gathered by a questionnaire completed by 98 of 112 professors or directors of those clinics. A summary of the findings follows: only 9% of the hospitals performed fungal culture studies for all or most cases, indicating a drop of 7% from 2000 (16% in 2000, reported by Kasai) . Also, just 55% of relevant departments maintained the ability to identify most or all clinical isolates in-house, which down 27% from 2000 (83% in 2000, reported by Kasai) . These findings indicate that mycological diagnoses by many departments have been rapidly decreasing. Eighty-nine percent of respondents indicated a desire for some help from the university network including: basic training in medical mycology of young doctors (senior residents in university hospitals) ; aid in diagnosing rare fungal infections; and support in obtaining knowledge and new technologies. The basic training of senior residents in each department is not possible in the network system so each department is required to take responsibility for training their own. However, a project of an advanced course at the leader level, which educates the person in charge of each department, may be possible in the network system. Employing this would make it realistic for the educated leaders to train the senior residents in their departments.

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Due to the serious financial condition of the health care insurance system in Japan, many clinical microbiology laboratories in hospitals have been forced to close or downsize, and therefore identification of pathogenic fungi isolated in these hospitals has become more and more difficult. This problem becomes even more serious when rare but clinically important fungi are the causative agents. For the smooth and accurate identification of the fungi, formation of a collaborative network among hospital laboratories, private clinical laboratory test centers and university research laboratories is now required.
In contrast, the culture collection system of pathogenic fungi for deposit and distribution has been significantly improved in the past few years largely due to the support of the National BioResource Program. The most important part of this kind of system is its longevity, and further improvement is warranted to keep the system viable even after the end of the Program.

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Trichophyton tonsurans infection was found first in autumn 2000 in Gifu prefecture and spread rapidly in the Tokai region. Not only direct KOH examination but also culture is necessary to diagnose this disease. In order to collect a specimen, I recommend cellophane adhesive tape. During the past 5 years, dermatologists in the Tokai area have sent me specimens in an envelope for mycological examination. Hyphae were found in all 75 cases in the scales examined. Fungal culture revealed 61 cases to be T. tonsurans infection; the male : female ratio was 54 : 7. By age distribution, high school students accounted for 46 (75%) , elderly patients 9 (15%) and lower age 6 (10%) . Judo players accounted for 32 (52%) , wrestlers for 24 (39%) and others for 5 (8%) . Most had lesions on the face, neck, head or arm. One wrestler had a nail involvement. In some specimens from tinea corporis , hyphae in the hair shaft were observed.
This sort of tinea epidemic probably occurs more often than is reported. Therefore we have begun to cooperate with a medical department member of the Gifu Judo Society to prevent of T. tonsurans infection.

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It has been seven years since an outbreak of Trichophyton tonsurans infection occurred in Japan. We have examined the state of T. tonsurans infection by a hairbrush test of athletes and a questionnaire survey of doctors’ experience in its treatment, but have not completely grasped the actual state of infection at the sites of judo matches and its treatment or application of preventive measures. Subjects and Methods: The questionnaire was distributed to the leaders of all judo clubs (10,077 clubs) registered under the All Japan Judo Federation, and responses recovered from 1,199 clubs (11.9%) were analyzed. These leaders were asked about (1) members of the club, (2) recognition of the infection, (3) experience of the breakout of infection at present and in the past, (4) present state of preventive measures, and (5) their opinions and requests. Results: A higher percentage of the leaders of junior high school physical education judo clubs and primary school judo clubs responded they “did not know” about the infection, as compared with leaders of the other age groups. The answer regarding the “experience of the outbreak of infection” was “yes” in responses from 371 clubs (30.9%) , with a significantly higher percentage of leaders of senior high school judo clubs replying in the affirmative. Concerning preventive measures such as providing shower rooms, 540 clubs (45.1%) answered “no measures taken” , which was more frequent in junior high school and primary school judo clubs. Since T. tonsurans infection is expected to spread to younger age groups in the future, a nationwide campaign by the All Japan Judo Federation for the education of risk of infection is urgently needed.

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Penicillium marneffei was isolated from three blood cultures of a Thai woman with AIDS and then identified as such. The patient, 41 a year-old female from northeast Thailand came to Japan 10 years ago and married a Japanese man. She was reportedly the third patient infected with this fungal species in Japan, and considered to be the first case from whom the causative fungus was successfully cultured, which led to the diagnosis of penicilliosis marneffei. The colony of the isolate, which was cultured on Sabouraud dextrose agar at 25-27ºC, was initially white and pannose, gradually turned in color from yellow to yellow-green, and diffused a deep red pigment into the medium. Conidial heads were divergent, and chains of conidia were formed from phialides. Colonies of the isolate, which was cultured on brain-heart infusion agar at 35ºC, had a grayish white, membranous yeast-like form with fine plicae and microscopically consisted of short hyphae. Furthermore, 560 bases of the internal transcribed spacer (ITS) region of the ribosomal RNA gene including the 5.8S region (ITS1-5.8S-ITS2) (DDBJ accession number AB298970) were sequenced and allowed an unequivocal species identification.

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We report a case of inflammatory tinea corporis by Epidermophyton ( E. ) floccosum. A 73-year-old man visited our hospital with an eruption on his left arm where he wore his wristwatch. He had treated himself with a topical steroid. On KOH preparation, many hyphal elements were observed in the scale. Histological findings revealed granulomatous inflammation around hair follicles and positive fungal elements by PAS and Grocott staining in these follicles. Topical luliconazole and oral terbinafin were effective clinically and mycologically. Epidemiological survey in Japan has shown the number of cases of tinea caused by E. floccosum to be on the decrease recently. The anthropophilic dermatophyte, E. floccosum is known usually not to invade the hair follicle, and no case of inflammatory tinea corporis by this species has been reported in Japan. We consider this case of inflammatory tinea corporis due to E. floccosum to be very rare.

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We report a case of nail candidiasis with severe deformities. The patient was a 71-year-old woman who initially consulted our department on April 5, 2006. She had diabetes, chronic rheumatoid arthritis and multiple liver metastasis of unknown origin. She had taken prednisolone for treatment of chronic rheumatoid arthritis for a long period. The initial examination demonstrated deformation of 1/3 of the inner part of the nail plate in both the third and fourth fingers, with apparent hyperkeratosis under the deformed nail plates. KOH-prepared direct microscopy revealed the presence of numerous spores and pseudohyphae. Numerous fungal elements were detected by Grocott staining and PAS staining. Candida albicans was isolated and identified by cultivation on the ATG agar and PCR-RFLP. Fluconazole (100 mg/day) was administered from April 8, 2006. After 14 weeks of treatment her clinical findings had improved, however she died of multiple organ failure on July 25, 2006.

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