• 약학회지
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• 제59권6호
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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.

• 보존과학회지
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• 제31권3호
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• pp.227-241
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• 2015

공주 송산리 6호분에서 분리한 미생물의 배양조건에 따른 생화학적 특성을 확인하고 천연물 유효성분인 anethole과 eugenol의 항진균력을 평가하여 고분 또는 벽화에 대한 적용 가능성을 확인하였다. 송산리 6호분 내부 공기중 및 벽체에서 포집된 26종의 곰팡이는 자낭균류 15종, 접합균류 2종, 담자균류 1종으로 동정되었다. 분리된 곰팡이의 전분과 젤라틴에 대한 분해도 평가를 실시한 결과 SY-18, SY-23, SY-25 등에서 전분에 대한 분해능이 높게 나타났으며, SY-18, SY-21, SY-23 등에서 높은 젤라틴 분해능을 보였다. 각 기질의 분해능이 우수한 균주 3종을 액체배양하여 생화학적 특성을 확인하였다. 천연물 유효성분인 anethole과 eugenol의 항진균력 평가 결과, anethole과 eugenol의 혼합액(1:2)에서 곰팡이의 생장 억제력이 가장 크게 나타났다. 국내외에 동굴 또는 고분 벽화에 발생 가능한 곰팡이에 대해 강력한 항진균력을 보이는 천연물 유효성분을 이용하여 미생물의 발생을 억제할 수 있을 것으로 기대된다.

• 한국미생물·생명공학회지
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• 제50권4호
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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.

• 대한수의학회지
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• 제29권1호
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• pp.69-73
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• 1989

준임상형 유방염 이환 젖소의 유즙과 유두컵으로부터 분리된 Candida속 균의 8종 항진균성약제에 대한 감수성을 조사하였던 결과는 다음과 같다. 총 53주의 Candida속균 중 98.2%, 94.4%, 84.8%의 균주가 각각 clotrimazole, econazole, miconazole의 농도 $25{\mu}g/ml$이하에서 발육이 억제됨으로서, 이들 약제의 항균력이 타 약제에 비하여 우수하였다. 균종별로 보면 C albicans는 clotrimazole에 대한 감수성이 가장 높았으며, 기타 비교적 감수성이 높았던 약제는 5-fluorocytosine, econazole, miconazole의 순이었다. C pseudotropicalis와 C guilliermondii는 haloprogin, clotrimazole, miconazole, econazole, 5-fluorocytosine등에 높은 감수성을 나타내었으며, haloprogin의 평균 MIC가 $0.17{\sim}0.19{\mu}g/ml$로 가장 우수하였다. C krusei는 cycloheximide에 대한 감수성이 가장 높았으며, 그의 비교적 감수성이 높았던 약제는 clotrimazole, haloprogin, miconazole, econazole의 순이었다. 한편 C parapsilosis는 econazole, cycloheximide, clotrimazole등에 다소 감수성을 나타내었으나, 이들 약제의 MIC는 econazole(기하평균 MIC $7.26{\mu}g/ml$)을 제외하고는 타균종에서 보다 현저히 낮았다. 또한 C tropicalis는 전 공시약제에 대하여 감수성이 낮은 경향이었다.

• 약학회지
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• 제55권3호
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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.

• 대한치과기공학회지
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• 제19권1호
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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.

• Journal of Yeungnam Medical Science
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• 제9권2호
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• pp.396-406
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• 1992

저자들은 Granade와 Artis의 방법에 따라 96-well microplate와 24-well macroplate를 이 용하여 T. rubrum 9주를 대상으로 경구용 항진균제인 ketoconazole과 itraconazole에 대한 MIC를 측정하여 실제 임상사용 가능성을 알아보고 배양온도, 배양용기의 크기, 배지의 종류를 달리하여 MIC에 영향을 줄 수 있는 요소를 점검하여 다음과 같은 결론을 얻었다. 1. 96-well microplate를 사용하여 $25^{\circ}C$에서 배양시 균농도에 따른 판독시기의 차이는 높은 균농도(흡광도 2.0, 1.0)에서는 4일만에, 낮은 균농도(흡광도 0.5, 0.25)에서는 6-8일만에 판독할 수 있었고, MIC는 높은 균농도에서 높았으나 시간이 경과시 점차 차이가 줄어들었다. 2. $37^{\circ}C$와 $25^{\circ}C$에서 각각 배양시 배양온도에 따른 MIC의 차이는 96-well microplate를 사용하여 $37^{\circ}C$에서 배양시 ketoconazole에 대한 MIC는 0.006이하-$0.04{\mu}g/ml$, itraconazole에 대한 MIC는 0.006이하-$0.04{\mu}g/ml$였으며 $25^{\circ}C$에서의 ketoconazole에 대한 MIC는 0.08-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006-$0.71{\mu}g/ml$로 $37^{\circ}C$에서의 MIC는 $25^{\circ}C$에서의 MIC에 비해 현저히 낮았다. 3. 24-well microplate와 96-well microplate에서 각각 배양시 배양용기의 크기에 따른 판독시기는 96-well microplate 액체배지에서는 4-6일로 24-well macroplate 액체배지에서의 8-12일에 비해 판독시기가 빨랐으나, MIC의 차이는 없었다. 4. 액체배지와 고체배지에서 배양시 배지종류에 따른 MIC의 차이는 액체배지를 함유한 24-well macroplate를 이용한 경우 ketoconazole에 대한 MIC는 0.006이하-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006 이하-$0.36{\mu}g/ml$였고 고체배지에서는 ketoconazole에 대한 MIC는 0.006이하-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006 이하-$5.68{\mu}g/ml$로 고체배지에서의 MIC가 다소 높게 측정되었다. 5. 이상의 결과를 종합하여 볼때 96-well microplate를 사용하여, 흡광도 1.0의 균농도로 접종하여, $25^{\circ}C$에서 배양 후 5-6일째 육안으로 판독하는 것이 항진균제 감수성 검사를 빠르고 간편하게 실시 할 수 있는 방법이다.