One of the glycoprotein toxins (Fr II-2) isolated from a Candida albicans strain is descrived with complement-activation capacity both in vivo and in vitro. Fr II-2 was not found to convert C3 into its faster electrophoretic form when incubated in normal human serum. EDTA and EGTA completely inhibited the complement-activating activity of the toxin, suggesting that this complement activation would be entirely accomplished via the classical pathway. The toxin decreased the number of peripheral lymphocytes and increased that of neutrophiles when intravenously injected into rabbits. The same effects on peripheral leukocytes were also attained by intravenous injection of cellular mannan, formol-killed cells or living cells of C. albicans. It seems, therefore, that the glycoprotein toxins would play an important part in the course of C. albicans infection.

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Three strains of Exophiala jeanselmei were isolated from 177 natural samples which were collected in rural regions of both Chiba and Ibaragi prefectures, Japan. Two of them were isolated from rotting wood and one from pine (Pinus densiflora) bark (Table 1). The colonies of these isolates on Sabouraud's dextrose agar at 27°C were blackish in color with a velvety appearance or with a yeast-like one. These isolates produced annellated conidiogenous cells (annellides) and annelloconidia (Fig. 1, 2). Conidiogenous cells of these isolates were cylindrical, obclavate or lageniform, and conidia were non-septate, subglobose, ellipsoidal to cylindrical and hyaline. Toruloid hyphae and free conidiogenous cells were present (Fig. 1-c). These morphological features of the natural isolates were almost similar to those of clinical isolates of E. jeanselmei (Fig. 3). Furthermore, there were no distinct differences between both the natural and clinical isolates in physiological characteristics concerning thermotolerance and decomposition of starch, skim milk and hypoxanthine (Table 2). One or 2 out of 3 rats, which received subcutaneous injection of 10⁶ conidial cells of each natural isolate of E. jeanselmei, produced nodules at the sites of the injected skin (Table 3). Histopathologically, these lesions were granulomas in the lower dermis (Fig. 4-a). Fungus elements were present in all of the granulomas (Table 3, Fig. 4-b, c). The recovery of a culture from the tissues was positive in 2 of 4 animals which produced nodular lesions and in one of 5 animals which looked apparently normal at the sites (Tabel 3). These results of pathogenicity test suggest that the natural isolates of E. jeanselmei might be pathogenic for rats, even though their pathogenicity is relatively weak one. The present paper is the first report on the isolation of E. jeanselmei from nature in Japan.

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We obtained Candida albicans cells grown predominantly in the yeast-like (Y) and mycelial (M) forms, which developed in Sabouraud's glucose medium and in synthetic media containing methionine, respectively. Protoplasts were prepared from the two forms by using a cell wall lytic enzyme (Zymolyase) from Arthrobacter luteus and plasma membranes were isolated by hypotonic disruption of protoplasts and differential centrifugation. The comparative lipid analysis of plasma membranes thus obtained showed that sterol content of the M-form cells was approximately three times greater than that of the Y-form cells. Major polar lipids were phosphatidylcoline and phosphatidylethanolamine, and their fatty acids were mainly palmitic, palmitoleic, oleic and linoleic acids. No marked difference was observed in the phospholipid class composition between two forms, whereas there was a greater proportion of oleic acid in the Y-form cells. Data of ESR spectra obtained by using a spin probe (5 SAL) demonstrated that the order parameter of plasma membrane lipids from the M-form cells was greater than that from the Y-form cells. It implies that the physical state of the M-form plasma membrane is different from that of the Y-form plasma membrane, the former being less fluid. The activity of chitin synthetase (EC: 2.4.1.16) which is associated with plasma membranes was approximately three to five times greater in the M-form than in the Y-form. These results provide evidence which suggests that the lipid composition of plasma membranes would be somehow related with the dimorphic morphogenesis of Candida albicans.

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Several reliable methods for bioassay for miconazole in serum or other clinical specimens employing Candida albicans as the indicator organism are described. The well agar plate method with Yeast Morphology Agar was sensitive to 1μg of drug per ml and linear from 1 to equal to or greater than 16μg per ml. The disc agar plate method was much more sensitive, by which 0.06μg of miconazole per disc was quantitatively measurable. The bioautographic method was the most sensitive; the lower limit of the assay was as low as 0.002μg of drug per sample applied. In the latter two methods, Yeast Morphology Agar was used as the assay medium and serum specimens were extracted with cyclohexane or ethyl ether for assay.

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Several sensitive methods for quantitative microbial assay for the miconazole content of blood were tested and results from volunteers who received miconazole vaginal tablets were presented. The sensitivity limit of bioassay performed by the well agar plate method using Candida pseudotropicalis MTU 12040 and Blastomyces dermatitidis MTU 17008 as the indicator organism was 0.16 and 0.12μg of drug per ml, respectively. The disc agar plate method with B. dermatitidis was much more sensitive, the assay limit being 0.004μg of drug per disc. The microbial assay for miconazole was performed with blood specimens from 9 volunteers who were treated by the daily insertion of one 100mg tablet for successive 14 days, but none of specimens tested contained any detectable level (above 0.004μg/ml) of miconazole.
The minimum inhibitory concentration and the minimum cidal concentration of miconazole against several typical Lactobacillus members of human vaginal flora were determined. Results of the sensitivity testing with 6 species and 12 strains of Lactobacillus using the agar dilution method showed that all the strains are virtually insensitive to the drug.

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The effectiveness of 31 antimicrobial agents against 74 strains of Nocardia, Actinomadura, Streptomyces and Rhodococcus were determined with use of plate dilution agar method. Of 31 antimicrobial agents tested, sulfa drugs, minocycline and gentamicin were most bioactive, and ketoconazole, streptomycin and fusidic acid were moderately active. The sulfa drugs and gentamicin showed higher activity against Nocardia brasiliensis than Nocardia asteroides, whereas minocycline and doxycycline showed similar activity against both strains of N. asteroides and N. brasiliensis. The mean MIC's (Minimum inhibitory concentration) of sulfadimethoxin (a sulfa drug), gentamicin and minocycline against N. brasiliensis and N. asteroides were 9.5μg/ml and 68.5μg/ml, 16.5μg/ml and 98.2μg/ml, and 4.9μg/ml and 5.9μg/ml, respectively. β-Lactam antibiotics including ampicillin and cephaloridine were inactive and the mean MIC's of ampicillin and cephaloridine against N. asteroides were 132.6μg/ml and 120.1μg/ml, respectively.

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