• Title/Summary/Keyword: antifungal susceptibility test

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Synergistic Antifungal Activity against Disseminated Candidiasis by Combination Therapy of Crataegi Fructus Extract and Fluconazole (전신성 캔디다증에 대한 산사자 추출물과 Fluconazole의 병용요법에 의한 항진균 상승효과)

  • Han, Yongmoon
    • YAKHAK HOEJI
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    • v.59 no.6
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    • pp.259-265
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    • 2015
  • In recent, there are increasing reports about pharmacological activities of Crataegi Fructus which has been used for many centuries as medicinal and food sources in East Asia. However, its antifungal efficacy needs to be investigated. Thus, in the current study, we determined synergistic antifungal activity of the Crataegi Fructus extract (CFE) when combined with fluconazole (FLC) against disseminated candidiasis due to Candida albicans. This fungus is one of the most problematic fungal pathogens. Data resulting from a microdilution susceptibility test showed that CFE had a dose-dependent antifungal activity. When the extract was combined with FLC, the activity was synergistic. For example, the antifungal activity by the combination of CFE at $20{\mu}g/ml$ plus FLC at $0.1{\mu}g/ml$ was 4 times more effective than antifungal activity by FLC alone at the same concentration (P<0.05). In the murine model of disseminated candidiasis, the combination therapy potentiated resistance of mice, resulting in 80% of C. albicans-infected animals surviving the entire period of 40 days observation, whereas mice given CFE alone or FLC alone all died with 17 and 23 days, respectively, although they survived longer than the untreated control animals (P<0.05). The CFE's antifungal activity seemed to be related to the blockage of hyphal production of C. albicans yeast cells. In summary, CFE has a synergistic antifungal activity, which can be produced by combining CFE with FLC. Thus, our data strongly indicate that CFE may be a potential candidate as an antifungal agent for combination therapy.

Biochemical Characteristics and Growth Control for fungi isolated from mural painting of Tomb No.6 at Songsan-ri, Gongju (공주 송산리 6호분에서 분리한 진균의 생화학적 특성 분석 및 생장제어 연구)

  • Lee, Min Young;Park, Hee Moon;Chung, Yong Jae
    • Journal of Conservation Science
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    • v.31 no.3
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    • pp.227-241
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    • 2015
  • Fungi were isolated from mural painting in tomb no.6 at songsan-ri, Gong-ju. Antifungal susceptibility of essential oils extracted from natural medicine was tested and it confirmed applicability for mural painting in tombs. 26 species of fungi collected from air-borne and wall surfaces were identified to 15 species of Ascomycetes, 2 species of Zygomycetes, 1 of Basidiomycetes. Wheat starch and gelatin degradability were evaluated as isolated fungi. SY-18, SY-23, SY-25 showed high degradability of wheat starch. SY-18, SY-21, SY-23 were decomposed into gelatin. Biochemical characteristics of decomposing fungi to wheat starch glue and cowhide glue were analyzed by using ${\alpha}-amylase$ and gelatinase activity. An Antifungal test was conducted in Anethole and Eugenol. Anethole and Eugenol mixture(1:2) showed high antifungal susceptibility. Natural adhesives help microbial growth and can cause structural damage in mural painting. The expectation of this study is the possibility to control microbial growth in wall painting using natural essential oils. It can be used as a data for conservation method to control microbial damages.

Production and Identification of Secondary Metabolite Gliotoxin-Like Substance Using Clinical Isolates of Candida spp.

  • Noorulhuda Ojaimi Mahdi, Al-Dahlaki;Safaa Al-Deen Ahmed Shanter, Al-Qaysi
    • Microbiology and Biotechnology Letters
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    • v.50 no.4
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    • pp.488-500
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    • 2022
  • Most fungal infections by opportunistic yeast pathogens such as Candida spp. are the major causes of morbidity and mortality in patients with lowered immune. Previous studies have reported that some strains of Candida secret secondary metabolites play an important role in the decreasing of immunity in the infected patient. In this study, 110 Candida spp. were isolated from different clinical specimens from Baghdad hospitals. Candida isolates were identified by conventional methods, they were processed for Candida speciation on CHROMagar. The results of identification were confirmed by internal transcribed spacer (ITS) sequencing. Phylogenetic trees were analyzed with reference strains deposited in GenBank. Antifungal susceptibility testing was evaluated by the disc diffusion method and performed as recommended by the Clinical and Laboratory Standard Institute (CLSI) M44-A document. Candida isolates investigated produce secondary metabolites gliotoxin with HPLC technique and quantification. Out of 110 Candida isolates, C. albicans (66.36%) was the most frequent isolate, followed by the isolates of C. tropicalis (10.9%) and C. glabrata (6.36%) respectively. Concerning the antifungal susceptibility test, Candida isolates showed a high level of susceptibility to Miconazole (70.9%), Itraconazole (68.2%), and Nystatine (64.5%). The ability of obtained isolates of Candida spp. to produce gliotoxin on RPMI medium was investigated, only 28 isolates had the ability to secret this toxin in culture filtrates. The highest concentrations were detected in C. albicans (1.048 ㎍/ml). Gliotoxin productivity of other Candida species was significantly lower. The retention time for gliotoxin was approximately 5.08 min.

Antifungal susceptibility of Candida spp isolated from bovine mammary glands and teat cups of milking machines (Candida속 균의 항진균성약제에 대한 감수성)

  • Yeo, Sang-geon;Chung, Kyu-young;Cho, Hee-tack
    • 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, $${\geq_-}20.32{\mu}g/ml$$).

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Effect of Antimicrobial Peptide from Coptidis Rhizoma on Candida albicans Infection (황련 유래 Antimicrobial Peptide의 Candida albicans 감염 억제효과)

  • Lee, Jue-Hee
    • YAKHAK HOEJI
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    • v.55 no.3
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    • pp.227-233
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    • 2011
  • We previously reported the protein isolated from Coptidis Rhizoma (CRP), which has antifungal activity against a fungal pathogen, Candida albicans. In the current study, we investigated what portion in the CRP is responsible for the antifungal activity. For the investigation, the CRP was fractionated on a Shepadex G-50 column. Data resulting from the fractionation, seven fractions were obtained. Fractions (Fr.) I, II, and III eluted initially from the column showed no inhibitory effect on the growth of C. albicans, whereas Fr. IV, V, and VI eluted later revealed inhibition of the growth, and Fr. IV and VI showed potent antifungal activity by broth susceptibility analysis. However, Fr. VI was contained in the CRP more than Fr. IV, which led us to select the VI for the following experiments. In a murine model of a subcutaneous candidiasis caused by C. albicans, the Fr. VI displayed a therapeutic effect on nude mice pretreated with anti-neutrophil monoclonal antibody (RB68C5) and then infected subcutaneously with live C. albicans. At day 16, these mice were healed almost up to 78% of the infected area when compared to infected area of control nude mice that received diluent (Dulbecco's Phosphate-Buffered Saline; DPBS), instead of the Fr. VI (P<0.01). The Fr. VI blocked hyphal formation from blastoconidial form of C. albicans (P<0.01), which might prevent penetration of hyphae to the deeper site of skin and thus helps the healing. In the ionic strength test, the effect of Fr. was influenced by $Ca^{2+}$ ion just like other known antimicrobial peptides, but the influence was affected at an extremely high concentration such as 500 mM. Thus, such ion-concentration is considered to be meaningless in the clinical situation. Considering all data together, Coptidis Rhizoma is appeared to produce an antimicrobial peptide that has therapeutic effect on subcutaneous infection caused by C. albicans.

Isolation of Candida albicans from Denture Patients and in Vitro Activities of Amphotericin B and Miconzole (의치환자에서 Candida albicans분리와 amphotericin B 및 miconazole에 시험관내 감수성)

  • Shin, Moo-Hak;Kim, Shin-Moo
    • Journal of Technologic Dentistry
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    • v.19 no.1
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    • pp.113-120
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    • 1997
  • Candida albicans is now well recognized among the denture stomatitis patients. The broth macrodilution test is the most widely used technique for antifungal susceptibility testing. The purpose of this study was to determine the C. albicans carrier rate of the denture patients in Iksan, chonbuk. To determine the C. albicans carrier rate of denture patients, culture were made from 227 sample taken in Iksan, Chonbuk during July 1997 to August 1997. Also activities of amphotericin B and miconzole against isolates of denture patients of C. albicans were tested by broth macrodilution test using RPMI medium 1640. The results were as follows : First C. ablicans was isolated from 6.6% of denture patients samples and the frequency of isolation fo C. albicans was highest(50%) in the age group of 71-year-old to 80-year-old denture patients. Second, against C. albicans, the MIC range of amphotericn B was $0.06{\sim}0.25{\mu}g/ml$. MIC50 and MIC90 were $0.13{\mu}g/ml$ and $0.25{\mu}g/ml$, respectively. Third, the MIC range of miconazole was $10-{\ge}20{\mu}g/ml$ and MIC50 and MIC90 were $20{\mu}g/ml$ and ${\ge}20{\mu}g/ml$, respectively. It was concluded from this study that C. albicans acrriages from healthy denture individuals only over 60-year-old ages were isolated, they remain susceptible to amphotericin B and not rarely resistant to miconzole.

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Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method (96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사)

  • Lee, Moo-Woong;Kim, Jong-Chul;Choi, Jong-Soo;Kim, Ki-Hong
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.396-406
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    • 1992
  • Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

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