The Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University is the only organization in Japan to possess a series isolates of Coccidioides spp., which are the most virulent pathogenic fungi and whi...The Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University is the only organization in Japan to possess a series isolates of Coccidioides spp., which are the most virulent pathogenic fungi and which are treated as biosafety level 3 microorganisms. Recently, the genus Coccidioides has been classified into two species, C. immitis and C. posadasii, based on their endemic areas and genotyping; the former species is endemic to the state of California, and the latter is endemic to other parts of North and South America. We reevaluated 19 isolates of Coccidioides immitis stored in our center using a multiple gene analysis. Five isolates were identified as C. immitis and 14 as C. posadasii. Their sequence information in GenBank will help to identify the two genospecies of Coccidioides spp.
An epidemiological survey of dermatomycoses and the causative fungus flora of dermatophytoses in Japan for 2002 was made on a total number of 72,660 outpatients who visited 14 dermatological clinics throughout Japan. The...An epidemiological survey of dermatomycoses and the causative fungus flora of dermatophytoses in Japan for 2002 was made on a total number of 72,660 outpatients who visited 14 dermatological clinics throughout Japan. The results were as follows: 1) Dermatophytosis was the most prevalent cutaneous fungal infection (7,994 cases) seen in these clinics, followed by candidiasis (755 cases) and then Malassezia infections (220 cases). 2) Among dermatophytoses, tinea pedis was the most frequent (4,813 cases: male 2,439, female 2,374), then in decreasing order, tinea unguium (2,123 cases: male 1,093, female 1,030), tinea corporis (497 cases: male 281, female 216), tinea cruris (299 cases: male 249, female 50), tinea manuum (248 cases: male 144, female 104) and tinea capitis including kerion (14 cases, male 6, female 8). 3) Tinea pedis and tinea unguium are seen to increase in summer season, among the aged population and among males in each clinic. When compared to the previous surveys (1992 and 1997) by clinical form, t. unguium patients increased from 1.9% of total outpatients in 1992, to 2.0% in 1997, then to 2.9% in 2002. 4) As the causative dermatophyte species, Trichophyton rubrum was the most frequently isolated among all dermatophyte infections except tinea capitis. 5) T. rubrum was isolated from 63.3%(1,431/2,262) of tinea pedis lesions, followed by Trichophyton mentagrophytes (36.6%, 829/2,262), and also 88.8% (325/366) of t. corporis, 95.4% (185/194) of t. cruris and 85.6% (462/540) of t. unguium. 6) Cutaneous candidiasis was seen in 755 cases (1.0%) of 72,660 outpatients. Intertrigo (347 cases) was the most frequent clinical form, followed by erosio interdigitalis (103 cases) and diaper candidiasis (102 cases). It has a tendency to affect the aged being complicated with topical predisposing factors. 7) Cutaneous Malassezia infections and other superficial fungal infections are seen in 220 cases, without any characteristic features by gender or clinical form.
The patient was a 57-year-old woman who initially consulted our department on May 31, 2002 with a chief complaint of other dermatoses. The initial examination demonstrated deformation of approximately 1/3 of the inner pa...The patient was a 57-year-old woman who initially consulted our department on May 31, 2002 with a chief complaint of other dermatoses. The initial examination demonstrated deformation of approximately 1/3 of the inner part of the nail plate in the right hallux, in addition to brown discoloration and keratin hypertrophy under the deformed nail plate. KOH-prepared direct microscopy revealed the presence of large round spores and hyphae. Since colonies similar to those of Trichophyton rubrum were obtained by culture, daily administration of terbinafine (125 mg/day) was initiated from June 17, 2002 based on a suspicion of tinea unguium induced by T. rubrum. However, the morphology of the colonies began to change toward a brown powder-like configuration after 2-3 weeks of culture, and band forms were also detected by slide culture, resulting in the diagnosis of onychomycosis induced by Scopulariopsis brevicaulis. From August 6, 2002, the dose of terbinafine was increased to 250 mg/day, which was maintained until November 22, 2002. When the patient visited our department approximately 2 months later, KOH-prepared direct microscopy revealed that she was still positive for S. brevicaulis. Therefore, terbinafine (250 mg/day) was administered again for 1 month from April 25, 2003, followed by a judgment of complete healing on July 4, 2003. Thereafter, there was no recurrence of onychomycosis up until January 29, 2005.
By the combined use of agar diffusion, agar vapor and agar vapor-inhibitory assays, contribution of the vapor activity of essential oils was quantitatively estimated. The test organisms were Trichophyton mentagrophytes a...By the combined use of agar diffusion, agar vapor and agar vapor-inhibitory assays, contribution of the vapor activity of essential oils was quantitatively estimated. The test organisms were Trichophyton mentagrophytes and Aspergillus fumigatus. Agar vapor assay was used to confirm the vapor activity of the oils. The parameter delta defined as a contribution index of the vapor activity was calculated by (1 - b-c/a-c) x 100, where a is inhibitory diameter in the diffusion assay, b is inhibitory diameter in the vapor-inhibitory assay and c is diameter of the sealed ring in the vapor-inhibitory assay (21 mm). Many of the essential oils examined showed a delta value near 100, thus providing the major contribution of the vapor activity to the inhibitory diameter. Essential oils containing aldehyde as major constituent showed low delta value, indicating the major inhibition was due to agar diffusion. Major essential oil components behaved similarly; the delta value was increased in the following order: aldehyde < phenol < alcohol < ester, oxide, hydrocarbon, indicating the enhanced contribution of the vapor activity in that order.
Five aerobic actinomycete strains isolated from patients in Japan were assigned provisionally to the genus Nocardia based on morphological and physiological characteristics. The five strains, IFM 10481, IFM 0668, IFM 090...Five aerobic actinomycete strains isolated from patients in Japan were assigned provisionally to the genus Nocardia based on morphological and physiological characteristics. The five strains, IFM 10481, IFM 0668, IFM 0901, IFM 0583 and IFM 0342, were not classified into any Nocardia species reported as infectious agents in Japan. Therefore, they were studied further to determine their specific taxonomic positions. Detailed chemotaxonomic and physiologic characterization and 16S rDNA sequence data of the five strains showed that they belonged to respective species of Nocardia carnea, N. elegans, N. paucivorans, N. puris and N. takedensis. This is the first isolation report of these five Nocardia species from patients in Japan.
Dermatophytosis, cryptococcosis and sporotrichosis are dermatological zoonosis caused by fungi. Control of dermatophytosis is still difficult, although it is well known as a transmittable disease from animal to human. Ex...Dermatophytosis, cryptococcosis and sporotrichosis are dermatological zoonosis caused by fungi. Control of dermatophytosis is still difficult, although it is well known as a transmittable disease from animal to human. Exotic animals such as hamster, guinea pig and rabbit, some imported from foreign countries, have become a frequent source of human dermatophytosis, and the emergence of fungal pathogens followed. The nasal cavity of dogs and cats is an important reservoir of pathogens of cryptococcosis, in addition to bird droppings. Though there have been many reports of human sporotrichosis caused by patients' pet, especially cats in Europe and U.S., since there has been no report of zoonotic transmission of sporotrichosis, zoonosis is not believed do be present in Japan. The interrelation and cooperation of medical and veterinary doctors and all other scientists involved is therefore required to prevent fungal zoonosis, because the condition may be partly due to changes in the environments of human and animal life and the more intimate relationships between humans and animals.
The genus Malassezia is now divided into eleven species. Different species initiate or aggravate different skin diseases. In seborroheic dermatitis, M. restricta play an important role, while in atopic dermatitis, M. glo...The genus Malassezia is now divided into eleven species. Different species initiate or aggravate different skin diseases. In seborroheic dermatitis, M. restricta play an important role, while in atopic dermatitis, M. globosa and/or M. restricta are major cutaneous microflora. M. globosa is a causative species of tinea versicolor, and this species is also a causative species of malassezia folliculitis. We should therefore obtain better knowledge of the ecological and pathogenic roles of malassezia.
Recent progress in the epidemiology, diagnosis and treatment of onychomycosis is summarized. The risk factors of this condition were investigated, and the results of analyses of the systemic state of patients, shape of n...Recent progress in the epidemiology, diagnosis and treatment of onychomycosis is summarized. The risk factors of this condition were investigated, and the results of analyses of the systemic state of patients, shape of nails, blood circulation around nails, nail growth rate and causative fungi were reported. In making a diagnosis of onychomycosis, the major effect of the quality of collected nail samples on the results was reconfirmed. In addition to the KOH method and culture method, a molecular biological method was introduced. From the therapeutic perspective, 1) prevention of recurrence, 2) identification of patients with high risk of onychomycosis, and 3) attempting new treatments (development of new drugs, improvement of administration and dosage, and evaluation of combined therapy) were discussed. The effectiveness of concomitant use with nail lacquer is anticipated in combined therapy. New studies on accurate diagnosis, early detection and early stage treatment are now being undertaken.
Environments may act as reservoirs for pathogenic fungi, a determinant of the establishment of fungal infection, or an exacerbating factor of disease. In recent years, skin disease caused by geophilic fungi has been decr...Environments may act as reservoirs for pathogenic fungi, a determinant of the establishment of fungal infection, or an exacerbating factor of disease. In recent years, skin disease caused by geophilic fungi has been decreasing, while case reports of zoonoses from various animals are increasing. Outbreaks of anthropophilic T. tonsurans infection pose a problem to medical mycologists. Tinea pedis is the most common exogenous dermatomycosis in Japan. Although T. rubrum is presumed to be the dominant pathogen of this disease, T. mentagrophytes is detected more frequently from various environments, so far, the reason for this discrepancy has not been fully understood. The latest knowledge about the route of dermatophyte foot infection is as follows: (1) Dermatophyte propagules disseminated from patients may contaminate not only bath-mats but also wood floors, Japanese style mattings, concrete floors, slippers, cushions, etc., and from them adhere to healthy skin. (2) The agar stamping method can easily detect dermatophytes from the skin and the environment. (3) Propagules of T. mentagrophytes can survive for more than three months under certain conditions such as in rubber boots. (4) In order to eliminate dermatophytes gathered in socks and footwear, simple procedures (washing, bathing with hot water, or wiping with a towel) are all effective. (5) Prior application of an antifungal agent promptly eradicates dermatophyte propagules adhering to the skin from the environment. The author also mentioned the possibility of asymptomatic dermatophyte colonization, and the high prevalence of dysgeucia in oral carriers of Candida albicans.
At the 48th Annual Meeting of The Society for Japanese Medical Mycology, held in October, 2004, we reported our findings from a survey on Trichophyton tonsurans infections in the Hokuriku and Kinki regions of Japan. The...At the 48th Annual Meeting of The Society for Japanese Medical Mycology, held in October, 2004, we reported our findings from a survey on Trichophyton tonsurans infections in the Hokuriku and Kinki regions of Japan. The survey revealed that a few epidemics had occurred across these regions. In this article, we introduce our subsequent studies relating to 1) molecular epidemiology of isolates taken from people in many parts of Japan and 2) factors affecting adequate sampling of the scalp with hairbrushes, essential for surveying and monitoring the infection. In total, 198 isolates of Trichophyton tonsurans were analyzed using restriction fragment length polymorphisms of the non-transcribed spacer regions of ribosomal RNA genes. The restriction enzyme Mva I indicated two molecular types of strains, implying that the causative agents of the epidemic had different origins. None of the isolates obtained from the epidemic showed the same restriction profile as that of isolates from aged and sporadic cases. The published hairbrush method suitable for obtaining samples from the scalp of Judo trainees was reevaluated by changing several factors. We found that sampling should not be done soon after the students' physical training because other fungal elements may give a false positive, samples should not be obtained from students who have recently applied topical antimycotics, and samples should be taken under the guidance of qualified instructors familiar with the sampling method.
Visceral fungal infections are difficult to manage in patients with collagen diseases and immunocompromised hosts. In particular aspergillosis can be a life-threatening complication in these patients. Here we report that...Visceral fungal infections are difficult to manage in patients with collagen diseases and immunocompromised hosts. In particular aspergillosis can be a life-threatening complication in these patients. Here we report that combined use of two antifungal agents (micafangin and itraconazole) was effective against severe aspergillosis of the bilateral pleural cavities in a 48-year old male patient diagnosed with Wegener's granulomatosis. Immunosuppressive therapy with corticosteroids and cyclophosphamides improved his nasal and pulmonary symptoms, but inflammation of the bilateral pleural cavities caused bronchial fistulas. Aspergillus fumigatus then infected the bilateral pulmonary cavities through these fistulas. This patient was treated with combined therapy of ITCZ and MCFG was given to this patient because of the risk of renal dysfunction associated with AMPH-B. After 5 weeks of treatment his clinical findings had improved and the fungus was suppressed.
To study the changes of visceral mycoses in autopsy cases, data on visceral mycosis cases with leukemia and myelodysplastic syndrome (MDS) reported in the Annual of the Pathological Autopsy Cases in Japan in 1989, 1993,...To study the changes of visceral mycoses in autopsy cases, data on visceral mycosis cases with leukemia and myelodysplastic syndrome (MDS) reported in the Annual of the Pathological Autopsy Cases in Japan in 1989, 1993, 1997 and 2001 were analyzed. The frequency rate of visceral mycoses with leukemia and MDS was 27.9% (435/1,557) in 1989, 23.0% (319/1,388) in 1993, 22.3% (246/1,105) in 1997 and 25.1% (260/ 1,037) in 2001, which was clearly higher than the rate of cases without leukemia and MDS: 3.4%, 2.7%, 3.5% and 3.7%, respectively. Furthermore, in comparing the rate of mycoses in recipients of stem cell transplantation with that of non-recipients, that of recipients was about 10% higher. The predominant causative agents were Candida and Aspergillus, at approximately the same rate (Candida 33.6%, Aspergillus 33.3%) as in 1989. However, Aspergillus increased conspicuously in 1993 (Candida 22.3% Aspergillus 44.5%), and continued to increase (Candida 22.8%, Aspergillus 50.8% in 1997; Candida 16.9%, Aspergillus 54.2% in 2001). In aspergillosis and zygomycosis, the lung and bronchi comprised the most commonly infected organs: 74.7% and 75.6% of the total cases, respectively. Among a total of 1,260 cases with mycotic infections in the four years studied, acute lymphatic leukemia and acute myeloid leukemia were the major diseases (35.5% and 33.5%, respectively) followed by MDS (29.0%). Given these facts, we emphasize that a greater interest in mycoses should be taken by clinicians, and immunocompromised patients should be protected from opportunistic invasive fungal infections, especially aspergillosis.
BACKGROUND: To identify the pathogenic fungi of dermatophytosis, restriction fragment length polymorphism (RFLP) analysis of PCR amplified ribosomal DNA including internal transcribed spacers (ITS) has been established i...BACKGROUND: To identify the pathogenic fungi of dermatophytosis, restriction fragment length polymorphism (RFLP) analysis of PCR amplified ribosomal DNA including internal transcribed spacers (ITS) has been established in Japan. Our purpose was to evaluate the usability of PCR-RFLP analysis to identify the causative agent of tinea unguium directly from a nail sample. METHOD: Samples of tinea unguium from 100 nails were collected and cultured on Sabouraud's glucose agar and observed for 2 months. DNA was extracted from these samples, and the PCR product was digested with restriction enzymes Mva I and Hinf I. Weight of the samples was determined. RESULT: Sensitivity of PCR-RFLP analysis (73%) was higher than that of culture (20%) showing that PCR is more advantageous for identification of the causative agent of tinea unguium. Sensitivity of PCR-RFLP did not depend on weight of the nail sample.
A statistical 30-year study of dermatomycosis in Sendai National Hospital (1968-1997) revealed many changes in the prevalent diseases: Tinea pedis and tinea unguium constantly increased during this period, and the ratio...A statistical 30-year study of dermatomycosis in Sendai National Hospital (1968-1997) revealed many changes in the prevalent diseases: Tinea pedis and tinea unguium constantly increased during this period, and the ratio of the former associated with nail infection finally reached 30% of all tinea pedis patients. On the contrary, tinea corporis and cruris showed a remarkable decreasing tendency. Patient age distribution of each disease also showed distinctive changes, generally increasing in the older generation and decreasing in the younger. The number of patients with tinea pedis and unguium gradually increased among the middle and older generations, with the peak of the age-distribution curve shifting upward year after year. On the other hand, cases of tinea cruris among the younger generation were few in the latest years, and middle-age patients remained at a low number. In the first stage of this study the kinds of atiologic dermatophytes consisted of multiple species, but after middle period the isolation of Epidermaphyton floccosum decreased. Microsporum canis appeared first in 1976 but in the recent several years has completely disappeared. In the last few years of the period studied Trichophyton rubrum and Trichophyton mentagrophytes were the only isolates found from among all types of dermatophytoses. Infantile candidiasis remarkably increased in 1970-1975 but thereafter decreased rapidlly. Candidial intertrigo also increased in the same period but did not decrease as much thereafter and continued at the same intermediate level. The number of other types of candidiases were not greatly changed throughout the 30-year period. Malassezia infection also showed no remarkable changes, and only 20 cases of sporothrichosis were found. One case of the deep seated form of cutaneous aspergillosis was found, and this was also true of chromomycosis caused by Fonceaea pedrosoi.
In order to confirm the phylogenetic relationships of Histoplasma capsulatum, the partial sequences of large subunit (28S) ribosomal gene (D1/D2 region) of 49 isolates were studied. The similarity values of the 49 isolat...In order to confirm the phylogenetic relationships of Histoplasma capsulatum, the partial sequences of large subunit (28S) ribosomal gene (D1/D2 region) of 49 isolates were studied. The similarity values of the 49 isolates were more than 99.0% across 617 base pairs, however, the 49 isolates were divided into 9 groups. These 9 groups were independent of 3 varieties, var. capsulatum, var. farciminosum and var. duboisii. These results showed that analysis of the nucleotide sequence of the 28S rRNA gene was very effective for identification of H. capsulatum and that three varieties of H. capsulatum should be reclassified according to the phylogenetic relationship established from analysis of the D1/D2 region sequences.
A total of 168 patients with tinea pedis, but without onychomycosis, were treated with 1 cycle of terbinafine (TBF) (1 cycle: defined as 250 mg/day for 1 week). KOH preparation for direct microscopy was performed 4, 8 an...A total of 168 patients with tinea pedis, but without onychomycosis, were treated with 1 cycle of terbinafine (TBF) (1 cycle: defined as 250 mg/day for 1 week). KOH preparation for direct microscopy was performed 4, 8 and 12 weeks after starting therapy to determine if testing was positive for tinea. Patients with no negative results on KOH examination or no evidence of obvious clinical improvement at 8 weeks, another cycle of the therapy was prescribed. The "cure", "no cure", "dropout", and "discontinuation/unevaluable" rates were 89.3%, 4.8%, 4.8% and 1.2%, respectively. The number of cycles required for cure in the plantar type was 1 cycle in 65.9% and 2 cycles in 54.5% of cases; in the interdigital type, 1 cycle in 79.1% and 2 cycles in 20.9% of cases; and mixed type, 1 cycle in 29.1% and 2 cycles in 60.9% cases. Among patients who were followed for at least 3 years after cure, the relapse rates were about 10% each year: 1 year, 11.3%; 2 years, 8.9%; and 3 years, 11.2%. The relapse rate of about 10% each year over a 3-year period suggests that reinfection may be likely.
Six cases of Trichophyton (T.) violaceum infection seen in a nursing home are reported. A 66-year-old female (case 1) was found with tinea corporis on her face, chest and shoulder, associated with black dot ringworm. A K...Six cases of Trichophyton (T.) violaceum infection seen in a nursing home are reported. A 66-year-old female (case 1) was found with tinea corporis on her face, chest and shoulder, associated with black dot ringworm. A KOH examination of hair showed endothrix parasitism. Reddish purple colonies were isolated from the patient on Sabouraud's dextrose agar, and intercalary and terminal chlamydospores were observed on slide culture. PCR-RFLP analysis of the microorganism showed a pattern of T. violaceum type. Therefore, the isolated fungus was identified as T. violaceum, a typical anthropophilic dermatophyte which had spread among residents and staffs easily. Using a mycological method, we examined 59 persons (21 residents and 38 staff members) who had had contact with case 1. The results were as follows. An 85-year-old female (case 2) and an 83-year-old female (case 3) were carriers of T. violaceum. A 23-year-old male (case 4) had tinea corporis on his right forearm due to T. violaceum. A 24-year-old male (case 5) probably had tinea corporis on his right forearm due to T. violaceum. One year after case 1's first visit to our clinic, we observed an 88-year-old female (case 6) of tinea capitis by T. violaceum. It seems that the organism was preserved in surroundings and members of the nursing home. The contagion in our cases could either have been caused by directly touching the person or by sharing their comb. PCR-RFLP analysis was performed within a short time, so that we managed effectively to select a way of treatment and to prevent the infection from spreading.
A 74-year-old woman visited a clinic in Kisarazu, Chiba Prefecture in December 2002 complaining of itching, scale and alopecia. She had been diagnosed as having tinea capitis by a direct microscopic examination of scales...A 74-year-old woman visited a clinic in Kisarazu, Chiba Prefecture in December 2002 complaining of itching, scale and alopecia. She had been diagnosed as having tinea capitis by a direct microscopic examination of scales, and been treated with an antifungal cream and steroid lotion since 1999. The bald area spread from frontal to occipital in which multiple black dots and red papules were scattered. Abundant endothrix spores were observed in the hair shaft. A mycelial colony was isolated from the black dots. A giant colony on Sabouraud's agar was white, powdery and flattened with cottony elevation at the center in the obverse, and a reddish-brown pigmentation in the reverse. The isolate produced abundant microconidia that were round to club-and balloon-shaped with extreme swelling, while macroconidia and spiral bodies were few. Hair perforation test was negative and urease activity test was positive. ITS1-5.8S-ITS2 rDNA sequencing revealed 100% homology with T. tonsurans isolated from two old women in Niigata Prefecture. On the other hand, 3 bases were different from those of the outbreak isolates from judo and wrestling players infected through international matches. T. tonsurans has polymorphism and the present isolate might be an autochthonous genotype in Japan. This is the first time T. tonsurans was isolated in Chiba Prefecture. But this prefecture had been known as an endemic area of Trichophyton coccineum, which was very similar in morphological and physiological characteristics to those of T. tonsurans before World War II. These facts raise the question of whether T. tonsurans has existed in this prefecture before.
A 74-year-old woman presented in April, 2003, with cutaneous lesions of the face by Paecilomyces lilacinus infection. The patient had received predonisolone and azathioprine for 20 months for treatment of autoimmune hemo...A 74-year-old woman presented in April, 2003, with cutaneous lesions of the face by Paecilomyces lilacinus infection. The patient had received predonisolone and azathioprine for 20 months for treatment of autoimmune hemolytic anemia. The lesion first developed on the right lateral eyelid 1.5 years earlier, and gradually enlarged. Physical examination revealed a dark reddish or brownish plaque and scattered papules and abscesses around the plaque on right lateral and lower eyelids, and the cheek. She noted mild tenderness on pressure. Cultures obtained from pus and biopsy specimen showed moulds, and those were identified as Paecilomyces lilacinus. Griseofulvin, 500 mg per day, was not effective for the lesion, so itraconazole, 200-300 mg per day, was administered orally for 11 weeks. Since culture from pus still yielded P. lilacinus despite clinical effectiveness, itraconazole pulse therapy (400 mg daily, 7 days a month) was started. The lesion cleared after three cycles of the pulse therapy.