• Korean Journal of Microbiology
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• v.26 no.2
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• pp.82-87
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• 1988

The genetic analysis and characterization of protoplast fusion hybrids between complementary auxotrophic mutants of Candida pseudotropicalis were carried out. Nuclear fusion appeared to occur in fusion hybrids (e.g., F15 and F33), as strongly suggested by isolation of recombinants after mitotic segregation of parental genetic markers. This was confirmed by KNA content, nuclear staining and comparison of survival rate to UV light. After keeping fusion hybrids for approximately one year, the frequency of spontaneous mitotic segregation was $3.0\times 10^{-4}$ - $8.1\times 10^{-4}$ while that of induced mitotic segregation was $1.4\times 10^{-3}$- $1.7\times 10^{-3}$. These results suggested that they maintained stable karyogamy state. It was also found that the production of $\beta$-D-galactosidase from F15, F33 and F158 was somewhat increased when compared with that from either auxotrophic parents or wild type.

• Korean Journal of Veterinary Research
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• v.28 no.2
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• pp.361-363
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• 1988

The prevalence of yeasts in mammary glands of dairy cows and teat cups of milking machines was studied in Chinju area. The rate of subclinical yeast infection in 330 quarters was 3.6%. Of 12 isolates from the milk, 4 Candida pseudotropicalis, 3 C tropicalis, 2 C krusei, 2 C albicans and 1 Rhodotorula spp were identified. The 91.7% of the isolates belonged to the genus Candida and C pseudotropicalis was the predominant species. From 20.5% of 200 teat cups tested, 51 strains of yeasts were isolated. These were 13 C pseudotropicalis, 9 C guilliermondii, 7 C tropicalis, 5 C krusei, 5 C parapsilosis, 3 C albicans, 2 Torulopsis glabrata, 2 Geotrichum candidum and 5 unidentified yeasts. C pseudotropicalis was most frequently encountcred.

• Korean Journal of Microbiology
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• v.22 no.1
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• pp.49-56
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• 1984

Formation and regeneration of protoplasts by Novozym 234 from Kluyveromyces fragilis N100 and Candida pseudotropicalis CBS607 were studied. This enzyme was more effective on cells grown at exponential phase than those at stationary one to convert intact cells into protoplasts. As osmotic stabilizer, ammonium sulphate was suitable for not only protoplast formation but also regeneration in K. fragilis as well as in C. pseudotropicalis. Optimal enzyme concentration was 3mg per ml in K. fragilis and 1~3mg per ml in C. pseudotropicalis, respectively. After the exposure of K. fragilis cells to 3mg per ml of enzyme for 3hr at 30$^{\circ}C$ , approximately 95% of protoplast formation of all observed cells was obgained, while about 100% from C. pseudotropicalis under the same condition was produced. The regeneration frequency of protoplasts by this enzyme was much lower than that by snail enzyme(Glusulase) although Novozym 234 converted cells from above two species into protoplasts free of cell debris effectively, compared with Glusulase. Novozym 234 appears to be suitable for subcellular fractionation to obtain nuclei or other organelles rather than protoplast regeneration.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.188-196
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• 1988

Intergeneric or intraspecific protoplast fusion between Saccharomyces cerevisiae X-2180-1A, Candida pseudotropicalis ATCC 8619 and CBS 607 was attempted to produce ethanol from lactose containing materials. Teh intergeneric fusion frequency between Saccharomyces cerevisiae X-2180-1A (ade rho) and Candida pseudotropicalis CBS 607 (his met) was $1.0\times 10^{-5}$. These values exhibited approximately 2-3.5 fold increase when compared with fusion frequency obtained without the treatment of bovine serum albumin, myoinositol and ergosterol, suggesting that these compounds may improve intergeneric of intraspecific protoplast fusion. Nuclear fusion appears to occur in fusants between intergenera(S. cerevisiae+C. pseudotropicalis) and intraspecies (C. pseudotropicalis strains) as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and isolation of recombinants after mitotic segragation. It was also found that alcohol production from intraspecific hybrids was somewhat increased when compared with that from their parents.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.274-281
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• 1987

The effects of bovine serum albumin, myoinositol and ergosterol on protoplast formation, regeneration and fusion from auxotrophic mutants of Candida pseudotropicalis were examined. Frequency of protoplast formation ranged from 48 to 98% depending on auxotrophic types. When myoinositol (0.5mg/ml) and ergosterol (0.1mg/ml) were supplemented in the medium of cell growth, and bovine serum albumin (4mg/ml)was added to protoplasting buffer, 50-100% of cells were converted to protoplasts. Such a treatment of three additives improved 2.2-3.0 fold of regeneration rate of protoplasts. The fusion frequencies between complementary auxotrophs ranged from $7.0\times 10^{-4}$ to $1.5\times 10^{-3}$ in the optimal conditions. These values showed 1.9-2.3 fold increase when compared with fusion frequencies obtained without the treatment of additives. These results suggested that these comsion frequencies obtained without the treatment of additives. These results suggested that these xompounds may improve protoplast regeneration and fusion between complementary auxotrophs used in this study.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.243-250
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• 1986

Protoplast formation and regeneration from wild-type and auxotrophic mutants of Candida pseudotropicalis CBS 607 as well as fusion between complementary mutants were carried out. Frequencies of protoplast formation from wild-type and histidine or adenine requiring mutants ranged from 96 to 100% whereas those from methionine or tryptophan auxotrophs were 52 and 72%, respectively. When bovine serum albumin(4mg/ml, BSA) was added to protoplasting buffer for cells of methionine or tryptophan auxotrophs grown in a medium supplemented with myoinositol(0.5mg/ml), 96-99 % of cells were converted to protoplasts. Protoplasts were regenerated at the frequencies ranging from 18 to 20%. However, the addition of BSA to protoplasting buffer and the supplement of myoinositol to a medium of cell growth doubled the regeneration rate except adenine auxotroph in which such an improvement was not observed. It was found that optimal concentrations of polyethylene glycol and $CaCl_2$ are 20% and 100mM while optimal pH and exposure time are 6.0 and 30min. The fusion frequencies between complementary mutants ranged from $1.5{\times}10^{-3}\;to\;8.8{\times}10^{-3}$ and were enhanced by the improvement in the rate of protoplast regeneration. When histidine auxotroph was fused with tryptophan mutant, several fusion products were obtained which were found to be in the state of aneuploid or diploid, judging from DNA content and the presence of a large nucleus in the products.

• The Journal of the Korean dental association
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• v.11 no.10
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• pp.671-674
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• 1973

From September to November 1967, Isolation of Candida species from oral cabity was attempted with 270 cases who made visit at outpatient Oral Surgery Department at Soo Do Military Hospital. 1. Out of 270, yeast like organisms were isolated in 108 cases. 2. Among these yeast like organism, 87 proved to be candida species and 21 to be undetermined group. 3. Eighty seven Candida species are consisted of 26 of C. albicans, 31 of C. krusei, 6 of C. Stellatoidea, 9 of C. tropicalis, 4 of C. guilliermondii, 6 of C. parapsilosis. and 5 of C. Pseudotropicalis.

• Korean Journal of Veterinary Research
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• v.22 no.2
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• pp.121-138
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• 1982

The present study was conducted in order to investigate the role of yeast-like fungi in bovine mastitis. Attempts were made to isolate and identify yeast-like fungi from the milk from normal udders and those with clinical or subclinical mastitis and from feces. Also incuded in the study were trials for the pathogenicity of the isolates for laboratory animals and efficacy of an anti-fungal drug for the treatment of mastitis. A total of 133 isolates of yeast-like fungi was made from milk and feces and they were identified as Candida (C.) albicans (5 isolates), C. krusei (63 isolates), C. tropicalis (27 isolates), Torulopsis (T.) glabrata (10 isolates), Rhodotourla sp. (6 isolates), Hansenula sp. (6 isolates) and Pichia sp. (1 isolate). Sixty seven strains of yeast-like fungi were isolated from the milk of 64 quarters (4.3% of quarters examined) of 55 cows (14.3% of cows examined). C. krusei, C. tropicalis, C. pseudotropicalis, C. parapsilosis and T. glabrata were isolated as the causative agents from 20 quarters (1.3% of quarters examined) with clinical mastitis. C. krusei, C. tropicalis, C. albicans, C. pseudotropicalis, T. glabrata, Rhodotorula sp. and Hansenula sp. were isolated as the causative agents from 22 quarters (1.5% quarters examined) with subclinical mastitis. C. tropicalis, C. krusei, T. glabrata and Rhodotorula sp. were isolated as the contaminants from 22 normal quarters (1.5% of quarters examined). C. krusei, C albicans, C. tropicalis, C. pseudotropicalis, C. parapsilosis, T. glabrata, Hansenula sp., Rhodotorula sp. and Pichia sp. were isolated as the contaminants from feces and all of the species except Pichia sp. were isolated from milk of the same cows at the some time. Intramammary infusion of nystatin was effective for the treatment of mastitis caused by C. albicans, C. krusei, C. tropicalis, C. pseudotoropicalis, C. parapsilosis, T. glabrata and Rhodotorula sp. C. albicans, C. krusei, C. tropicalis, C. pseudotropicalis, T. glabrata, Hamsenula sp. and Pichia sp. were pathogenic for rat but C. parapsilosis and Rhodotorula sp. were not.

• YAKHAK HOEJI
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• v.30 no.2
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• pp.79-86
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• 1986

In order to study 2-formylpyridine dithiocarbohydrazones (DTCH's) and their Cu(II) chelates as potential effective antimicrobial agents, twelve new compounds of six DTCH's and their Cu(II) chelates were synthesized. The compositions of Cu(II) chelates were determined on the basis of the data obtained from elemental analysis, electronic and IR spectrophotometry, and other method. They were tested for antimicrobial activity in vitro against a gram-positive, seven gram-negative bacterial species and three fungal species. DTCH's exhibited high antibacterial activity against the gram-positive Staphylococcus aureus, but low activity against the various gram negative bacterial species. In contrast to DTCH'S, their Cu(II) chelates exhibited higher antibacterial activity against St. aureus, but weaker, if any, activity against the gram-negative bacterial species. Both DTCH's and their Cu(II) chelates showed relatively potent antifungal activity against Cryptococcus neoformans, but weak activity against Candida albicans and Candida pseudotropicalis.

• Korean Journal of Veterinary Research
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• v.29 no.1
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• pp.69-73
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• 1989

In vitro antifungal susceptibility test was carried out on 53 strains of Candida spp isolated from milk of dairy cows with subclinical mastitis and teat cups of milking machines, Nystatin, clotrimazole, miconazole, econazole, 5-fluorocytosine, cycloheximide, haloprogin and griseofulvin were tested by the agar dilution method. The 84.8% to 98.2% of Candida strains were inhibited by clotrimazole, econazole and miconazole at $${\leq_-}25{\mu}g/ml$$, and clotrimazole was most active. Interspecies differences of antifungal susceptibility were recognized and these were as follows. C albicans was most sensitive to clotrimazole (GM-MIC, $5.49{\mu}g/ml$) followed by 5-fluorocytosine, econazole and miconazole. C pseudotropicalis and C guilliermondii were notably sensitive to haloprogin, clotrimazole, miconazole, cconazole, 5-fluorocytosine, and haloprogin (GM-MIC, $0.17{\sim}0.19{\mu}g/ml$) was most active. C krusei was most sensitive to cycloheximide (GM-MIC, $0.54{\mu}g/ml$) followed by clotrimazole, haloprogin, miconazole and econazole. C parapsilosis was somewhat sensitive to econazole, cycloheximide, clotrimazole, and econazole (GM-MIC, $7.26{\mu}g/ml$) was most active. C tropicalis showed very low sensitivity to all tested drugs (GM-MIC, $${\geq_-}20.32{\mu}g/ml$$).