A new “bead method” for Candida mannan detection developed recently was compared with avidin-biotin enzyme-linked immunosorbent assay (AB-ELISA) using the serum of mice infected with Candida albicans.
BALB/c male mice (normal mice) and those treated with cyclophosphamide (CP-treated mice) were the experimental animals used. These mice were infected with C. albicans and sacrificed under ether anesthesia every day following inoculation, and the brain, heart, lung, liver, spleen, kidney, and serum were obtained from them. C. albicans was recovered from some organs of both mouse groups during the experimental period.
Histopathologically, in the normal mice the kidney and brain were more affected than the other organs. In the CP-treated mice, many lesions with massive fungal growth were observed in the kidney, heart and lung.
The maximum values of Candida mannan in the normal mouse serum were obtained two days after inoculation, 33.3ng/ml by the bead method and 21.3ng/ml by AB-ELISA. In the CP-treated mice, the maximum values were obtained three days after inoculation, 166.6ng/ml by the bead method and 100ng/ml by AB-ELISA.
The detection limit of Candida mannan by both the bead method and AB-ELISA was 0.8ng/ml.
The bead method is thus useful for detecting Candida mannan.

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Dermatophytes were isolated from the house dust of three homes where patients with tinea pedis were living. The isolates of three patients were all identified as Trichophyton mentagrophytes. Initial examination of house dust demonstrated many colonies of T. mentagrophytes in the house of case 1, and a few colonies of T. mentagrophytes and T. rubrum in those of case 2 and case 3. When the house dust was cultured after lesions and fungi were no longer visible on the patients, many colonies of T. mentagrophytes were still isolated from the houses of case 1 and 2, and a few colonies of T. rubrum from that of case 3. This suggested that other family members were probably also infected with dermatophytes. We found T. mentagrophytes infection on the wife of case 1, T. mentagrophytes and T. rubrum infection on the husband of case 2, and T. rubrum infection on the father-in-law of case 3. The number of colonies of dermatophyte from the house dust gradually decreased with the treatment of the family member. It was concluded that identification of dermatophytes in house dust not only suggests the presence of the infection in more than one family member, but also is an important indication of the efficacy of the treatment of dermatophyte infection.

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Malassezia furfur has recently attention as a pathogen of seborrheic dermatitis. The purpose of this investigation was to develop an experimental model for seborrheic dermatitis with M. furfur.
Materials and methods: Sixteen male guinea pigs, 5 C3H mice and 5 unde mice were used. M. furfur was inoculated without occlusion on the back and the ear. Clinical isolates of M. furfur were cultured in Dixon's broth.
Results: Seborrheic dermatitis-like lesions were observed on the back and the ear in some guinea pigs after the 7 days. Erythema appeared on 9/12 backs and 10/12 ears on the 25th day. The lesions appeared in the group given repeated applications group faster than in the group given a single application. Ebelacton B, an esterase inhibitor, inhibited the development of these lesions. In nude mice, only pity scale appeared on the 22nd day. These results suggest that certain reactions with fungal elements and/or metabolic products of M. furfur are responsible for the development of seborrheic dermatitis.

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Some antifungal agents are known to work more effectively in vivo than expected by their MIC level measured in vitro. To determine whether this enhanced antifungal effect in vivo is caused by the synergy of these agents with the host defense mechanism, we investigated the synergistic effect of flucytosine against Candida krusei with murine peritoneal macrophages. C. krusei were added to thioglycollate-elicited murine peritoneal macrophages and were incubated with or without flucytosine. After 24 hours macrophages were lysed and the number of viable yeasts was measured by counting CFU to determine the antifungal activity. The results showed that: 1) the macrophages were fungistatic against C. krusei; 2) at the concentration of 0.1-1μg/ml, flucytosine had no effect against C. krusei when given to the yeasts by itself, and showed antifungal activity at 10μg/ml; 3) when added together with macrophages, flucytosine showed antifungal activity at as low as 0.1μg/ml. Thus macrophages can work synergistically with flucytosine against C. krusei. This synergy can also be seen in other antifungal agents, causing the discrepancy between MIC and the clinical efficacy.

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We report the case of a 38-year-old man with superficial hyperkeratotic candidiasis of palm with nail involvement. He visited our hospital because he developed scaly erythema with hyperkeratosis on his left palm with pachonychia of his left thumb and middle finger. Direct examination of the nail using the Parker-KOH method revealed numerous fungal elements, including both hyphae and grape-shaped spores, however, examination of the palm showed numerous hyphae but no spores. Candida tropicalis and C. parapsilosis were isolated and identified from both palm and nail. One week after treatment with oral itraconazole and topical antifungal reagent, same positive results were obtained with the Parker-KOH method. Cultures of the nail with Sabouraud dextrose agar were positive in both candida species, but those of scales from palm were negative. Observations of the culture with BHI broth with thiamin and inositol were the same as the culture with Sabouraud dextrose agar. However, Candida-elements on the palm were detectable by both PAP method with polyclonal anti-candida antibodies and PAS staining.

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A preliminary in vitro study was performed for the purpose of detecting immunoreactive proteinase. A polyclonal antibody against S. schenckii proteinase (II) was prepared in a rabbit. A highly reactive lot was isolated and its sensitivity and specificity examined by enzyme-linked immunosorbent assay (ELISA). Using this antibody, 0.1μg/ml proteinase was detected without any cross-reactivity with Candida albicans proteinase. Proteinase activity and the amount of immunoreactive proteinase were well correlated with fungal growth in a liquid culture medium containing albumin as the sole nitrogen source. Maximal amount of immunoreactive proteinase was observed at a level of 25μg/ml on the 6th culture day. This antibody did not detect immunoreactive proteinase from C. albicans though high proteinase activity was observed. These in vitro data suggest that detection of immunoreactive proteinase in the serum of sporothricosis patients is possible.

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A 46-year-old Japanese man visited our clinic because of a persistent subcutaneous nodule of the right foot. The lesion was 2×1.5cm in size and had no sinus. Multiple hard black granules Ca. 1mm in size were found in the fibrous hard dermal tissue at the time of biopsy. The histological examination revealed inflammatory granulomatous reaction in both the dermis and subcutaneous fat tissue. Hyphal and chlamydospore-like elements were observed in biopsied specimens stained with PAS and GMS. A black colony was developed on Sabouraud's dextrose agar from these black granules and identified as Madurella grisea. From these findings, the lesion was diagnosed as mycetoma. The infection route is unknown.
Total excision was performed. To our knowledge, this is the first case of mycetoma pedis caused by Madurella grisea in Japan.

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