포도구균에서의 teicoplanin 내성 빈도와 디스크 확산법에 의한 내성 검출의 평가

김선무,김병기,박연준,김명신,심상인 대한감염학회 1998 감염 Vol.30 No.2

목 적 : Teicoplanin은 methicillin 내성 포도구균에 주로 사용되는 glycopeptide이나 최근 glycopeptide에 내성을 보이는 포도구균이 증가하고 있다. 또한 teicoplanin은 배지에 확산이 잘 되지 않아 디스크 확산법에 의한 감수성 결과와 항생제 최소발육저지농도(MIC)가 차리를 보이는 것으로 알려져 있다. 본 연구에서는 포도구균의 teicoplanin에 대한 감수성 검사를 디스크 확산법과 한천희석법으로 시행하여 MIC이 분포와 두 방법간의 차이를 알아보았다. 방 법 : 290균주{coagulase negative staphylococci(CNS) 77균주, Staphylococcus aureus 213균주}를 대상으로 teicoplanin에 대하여 디스크 확산법과 한천희석법으로 항생제 감수성 검사를 시행하여 MIC의 분포를 알아보고, 두 방법에 의한 감수성 검사 결과를 비교하였다. 결 과 : Teicoplanin에 대한 한천 희석법 시험 결과 CNS 77균주 중 teicoplanin에 감수성인 균주가 75균주(97.4%), 중간인 균주가 2균주(2.6%)였고, S. aureus는 213균주 모두 teicoplanin에 감수성을 보였다. 디스크 확산법에 의한 억제대의 크기와 MIC는 낮은 상관관계를 보였으나(r=0.50), 디스크 확산법에 의한 항생제 감수성 결과와 한천희석법에 의한 감수성 판정 결과 차이를 보인 균주는 모두 8균주(2.8%)로 이중 major error가 0.4%, minor error가 2.4%였다. 결 론 : CNS 77균주 중에서는 teicoplanin에 중간인 균주가 2균주(2.6%)였고, S. aureus는 모두 감수성을 보였다. 또한, 디스크 확산법만으로 teicoplanin 내성 포도구균을 검출하는 것은 불충분하므로, 이의 검출을 위한 보다 정확하고 간편한 방법이 필요할 것으로 생각된다. Background : Teicoplanin is a glycopeptide antimicrobial agent effective against methicillin-resistant staphylococci. Decreased susceptibility of staphylococci to glycopeptide has been increasing. Teicoplanin diffuses poorly in agar and therefore the correlation between the inhibition zone diameter and the minimal inhibitory concentration(MIC) is rather poor. The purpose of this study was to evaluate the prevalence of teicoplanin-resistant stapylococci and to assess the reliability of inhibition zone diameters for determining the susceptibility of staphylococci to teicoplanin by comparing the results of the agar dilution MICs. Methods : From June to August 1997, 290 clinical isolates of staphylococci(77 coagulase negative staphylococci(CNS), 213 Staphylococcus aureus) were collected. The antimicrobial susdeptibilities to teicoplanin were determined by inhibition zone diameter and the results were compared with the MICs determined by the agar dilution method. Results : Among 77 CNS strains, 75(97.4%) were susceptible and 2(2.6%) were intermediate by agar dilution method and all 213 strains of S. aureus were susceptible to teicoplanin. There was a poor correlation(r=0.50) between the zone diameters of inhibition and agar dilution MICs. In comparison with the results of disk diffusion test and agar dilution MIC, eight (2.8%) out of 290 isolates showed discrepancies (major error rates : 0.3%, minor error rates: 2.4%). Conclusions : Two(2.6%) out of 77 strains of CNS and none of 213 S. aureus strains revealed decreased susceptibility to teicoplanin. And the inhibition zone diameter was less reliable in determining the susceptibility of staphylococci than MICs. Therefore, the more effective and convenient method is needed.

96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사

이무웅,김종철,최종수,김기홍 영남대학교 의과대학 1992 Yeungnam University Journal of Medicine Vol.9 No.2

저자들은 Granade와 Artis의 방법에 따라 96-well microplate와 24-well macroplate를 이용하여 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℃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℃와 25℃에서 각각 배양시 배양온도에 따른 MIC의 차이는 96-well microplate를 사용하여 37℃에서 배양시 ketoconazole에 대한 MIC는 0.006이하-0.04㎍/ml, itraconazole에 대한 MIC는 0.006이하-0.04㎍/ml였으며 25℃에서의 ketoconazole에 대한 MIC는 0.08-5.68㎍/ml, itraconazole에 대한 MIC는 0.06-0.71㎍/ml로 37℃에서의 MIC는 25℃에서의 MIC에 비해 현저히 낮았다. 3. 24-well microplate와 96-well microplate에서 각각 배양시 배양용기의 크기에 따른 판독시기는 96-well microplate액체배지에서는 4-6일로 24-well macroplate액체배지에서의 8-12일에 비해 판독 시기가 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㎍/ml, itraconazole에 대한 MIC는 0.006이하-5.68㎍/ml로 고체배지에서의 MIC가 다소 높게 측정되었다. 5. 이상의 결과를 종합하여 볼때 96-well microplate를 사용하여, 흡광도 1.0의 균농도로 접종하여, 25℃에서 배양 후 5-6일째 육안으로 판독하는 것이 항진균제 감수성 검사를 빠르고 간편하게 실시 할 수 있는 방법이다. 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 possiblity 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 (aborbance 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℃, MIC were below 0.006-0.04㎍/ml to ketokckonazole and below 0.006 -0.04㎍/ml to itraconazole while at 25℃ 0.08-5.68㎍/ml to ketoconazole and 0.006-0.71㎍/ml to itraconazole. Significant reduction of MIC was observed at 37℃ compared to 25℃. 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㎍/ml to ketoconazole, below 0.006-0.36㎍/ml to intraconzole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006 - 5.68㎍/ml to ketoconzole, below 0.006-5.68㎍/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.

2019-2022년 국내에서 분리한 Erwinia amylovora균의 옥시테트라사이클린과 옥솔린산 항생제에 대한 저항성 평가

함현희,오가람,이방울,이용환,이용훈 한국농약과학회 2023 농약과학회지 Vol.27 No.4

Fire blight, a destructive plant disease causing great threats to the apple and pear industries, was first occurred in South Korea in 2015. Various pesticides have been registered and used for the control of fire blight. Among them, antibiotics are known as very effective agents, but developed countries including Europe have been avoiding their use due to concerns about the emergence of antibiotic resistance and ecosystem disturbance. Antibiotics have been used for fire blight control since 2015 in Korea, however, antibiotic resistance of the pathogen has not been systematically investigated yet. Therefore, in this study, we surveyed the minimal inhibition concentration (MIC) of Erwinia amylovora, a causal agent of fire blight, against the oxytetracycline and oxolinic acid using a total of 258 isolates that are collected from overall areas of South Korea from 2019 to 2022 to assess the presence of antibiotic resistance. The MIC of oxytetracycline was in a range of 0.25-2 µg/ml, and that of oxolinic acid was less than 0.25 µg/ml. In addition, the MIC of oxytetracycline was increased in 2022 compared to 2019. However, no significant differences in MIC existed between isolated regions or hosts. Taken together, the results indicate that oxytetracycline and oxolinic acid resistance may not occur in E. amylovora of Korea. This study also provides fundamental information to the assessment of resistance against antibiotics registered for the control of fire blight in Korea.

Differential Inactivation of Food Poisoning Bacteria and Lactobacillus sp. by Mandelic Acid

전정민,Hae-In Lee,Seong Geun Kim,Sang-Hyun Han,Jae-Seong So 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.3

Microbicides are used to eliminate pathogenic bacteria, but they can also eliminate beneficial and normal bacteria. This study provides antimicrobial information of mandelic acid (MA) prior to its widespread use. The MA susceptibilities of Lactobacillus sp. and pathogens were determined by dilution method based on NCCLS protocol. The concentration of MA at 5.0 mg/mL inhibited 90% of the Lactobacillus sp., whereas a 1.25 mg/mL concentration inhibited the following pathogenic bacteria strains: Listeria monocytogenes KCTC3710, Yersinia enterocolitica ATCC23715, and Shigella sonnei ATCC9290. The minimal inhibitory concentration (MIC) for Escherichia coli O157:H7, Salmonella enteritidis KCTC12400,Salmonella gallinarum ATCC9148, Salmonella typhimurium ATCC29631, and Staphylococcus aureus KCCM40881was at 2.5 mg/mL. These pathogens were also inhibited by addition of 1.0 M NaCl in 2 mg/mL MA solution (pH 5.0),while lactobacilli were not affected. These results indicate that MA can prevent disease and food poisoning when applied as a disinfectant in food industry, hospitals, and public places.

신개발 세족기 및 세족액의 젖소 적응효과

백광수,김병학,박수봉,박성재,김현섭,이왕식,기광석,전병순,안병석,강석진,서국현,Baek, K.S.,Kim, B.H.,Park, S.B.,Park, S.J.,Kim, H.S.,Lee, W.S.,Ki, K.S.,Jeon, B.S.,Ahn, B.S.,Kang, S.J.,Suh, G.H. 한국축산환경학회 2006 축산시설환경학회지 Vol.12 No.3

본 연구는 젖소에 있어서 세족기 및 신개발 세족액의 적용효과를 구명하기 위하여 2004년 축산연구소 및 전문경영체농장 2개소에서 사육중인 홀스타인 착유우를 대상으로 신개박 세족기 및 세족액을 적용하여 다음과 같은 결과를 얻었다. 1. 유산동의 MIC와 MBC는 E. coli 및 Bacillus에 대해서 공히 0.31%로 나타났다. 신개발 세족액의 경우 E. coli에 대해서는 MBC가 5%로 나타났고 Bacillus에 대해서는 2.50%로 나타났다. 2. E. coli의 경우 5% 및 16% 농도에서 각각 95.69% 및 95.87%의 감소율을 나타내었고 Bacillus의 경우 5% 및 16% 농도에서 각각 99.5% 및 99.89%의 감소율을 나타내었다. 3. 30일동안 사용하였을 때 보행자세지수(locomotion score ; LS) 2, LS3, LS4 및 LS5의 경우 100%, 43.8%, 14.3% 및 0%의 정상회복율을 나타내었고 누적하여 60일동안 사용하였을 때 LS2, LS3, LS4 및 LS5의 경우 100%, 81.3%, 71.4% 및 50.0%의 정상회복율을 나타내었다. 4. 신개발 세족액을 사용하기 전과 후의 발굽장애 발생율에 있어서 사용 전에는 18.8%의 발생율을 보였으나 사용 후에는 1.5%의 발생율을 나타내었다. This study was carried out to investigate the effect of new foot-bath facility and detergent solution (sodium molylbdenate, citrate, potassium nitrate, tataric acid, sodium hypo-cholorite, and zinc sulfate) on claw health in lactating dairy cows. Minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) of copper sulphate were 0.31% for E. coli and Bacillus isolated from cows claw. The MIC and MBC of new detergent for E. coli were 1.25% and 5%, respectively, however their respectively values for Bacillus were noticed 0.63% and 2.5%. Both 5E. coli and Bacillus populations in petri-dishes were significantly reduced (more than 95%) with the application of new detergent solution (5% or 16%). Locomotion score (LS 1-5; very good to severely bad) of lactating cows were significantly improved with in 30 days with the use of new detergent solution in foot bath. The LS2 (n=16), LS3 (n=16), and LS4 (n=7) were shown 100%, 43.8%, and 14.3% recovery rate within 30 days with the use of new detergent solution. However, LS5 (n=2) were not recovered to normal claw health and locomotion score within 30 days of new detergent application. Usage of new detergent solution for 60 days in a foot bath have shown 81.3%, 71.4% and 50.0% recovery rate in cows with LS3, LS4 and LS5, respectively. Abnormal claw incidence was reduced from 18.8% to 1.5% in overall herd (n=80) with the use of new detergent solution (16%) in a foot bath for 90 days. In conclusion, usage of 16% of our detergent solution for 60 days in a foot bath can significantly improve the cow claw health and thus mitigate the negative effects of abnormal claw on productivity of cows and dairy farm income.

진피산(秦皮散)이 Staphylococcus aureus와 Staphylococcus epidermidis에 미치는 항균효과에 대한 실험적 연구

서형식,Seo, Hyung-Sik 대한한방안이비인후피부과학회 2007 한방안이비인후피부과학회지 Vol.20 No.1

Objective : This experimental study was performed to investigate the effect of Jinpi-san on Staphylococcus aureus(S. aureus) and Staphylococcus epidermidis(S. epidermidis) that induce keratitis. Methods : Minimal inhibitory concentration(MIC) was measured by dropping to $50{\mu}l$ according to density Jinpi-san(100%, 10%, 1%, 0.1 %). Anti-bacterial potency was measured by the size of inhibition zone with change of volume. Results : 1. MIC on S. aureus in Jinpi-san was $40{\mu}l$ undiluted solution. 2. MIC on S. epidermidis in Jinpi-san was $20{\mu}l$ undiluted solution. Conclusions : These results indicate that Jinpi-san can be used to cure S. aureus, S. epidermidis that induce eye disease(keratitis). If further study is performed, the use of eye drops will be valuable and beneficial in the clinical medicines.

백년초 줄기 추출물의 화장품용 천연 방부제로서의 활용성

임미혜,홍세정 한국피부과학연구원 2016 아시안뷰티화장품학술지 Vol.14 No.4

Purpose: This study was carried out to develop natural cosmetic preservatives in replace of the chemical preservatives. Methods: The extracts of Opuntia ficus-indica var. saboten stem (OFS), which is known for its excellent anti-oxidant properties, were obtained by reflux extraction with 80% ethanol. Its anti-microbial activity was tested using minimal inhibition concentration (MIC) test and its preservative activity was tested by challenge test in mud pack. Results: As a result, the MIC of OFS for each strain represented 12.5% of Pseudomonas aeruginosa (P. aeruginosa), 12.5% of Staphylococcus aureus (S. aureus), 1.5% of Escherichia coli (E. coli), 20% of Candida albicans (C. albicans), and 20% of Aspergillus niger (A. niger). At this time, MIC of 1,2-hexanediol showed 1% of P. aeruginosa, 2.5% of S. aureus, 1.5% of E. coli, 1% of C. albicans, and 5% of A. niger. In the containing 20% of OFS mud pack, P. aeruginosa, S. aureus, and E. coli were decreased approximately 99.9999% within 2nd day and this trend was maintained up to 28th day. C. albicans was decreased approximately 99.99% within 14th day and this trend was also maintained up to 28th day. A. niger, on the other hand, was rapidly decreased within the 21st and the 28th day. The results demonstrated that containing 20% of OFS mud pack has a preservative activity in accordance to the standards of European Pharmacopoeia (EP), United States Pharmacopoeia (USP) and Cosmetic, Toiletry, and Fragrance Association (CTFA). Conclusion: Based on the result, if OFSs were to be applied to cosmetics as natural preservatives in the future, it would provide a high commercial value. 목적: 본 연구는 화장품 천연 방부제 개발을 위한 시도의 일환으로 항산화능이 뛰어난 것으로 알려진 백년초 줄기 추출물을 가지고 기존 화장품 제조분야에서 사용되는 화학적 방부제의 대체 가능성을 알아보았다. 방법: 백년초 줄기를 80% 에탄올로 환류추출하여 추출물을 얻었으며, minimal inhibition concentration (MIC) test를 이용하여 항균성을 확인하였고 백년초 줄기 추출물로 머드팩을 만들어 방부력 시험을 시행하였다.결 과: 항균력 시험결과 백년초 줄기 추출물의 최소생육저지농도는P seudomonas aeruginosa (P. aeruginosa) 12.5%, Staphylococcus aureus (S. aureus) 12.5%, Escherichia coli (E. coli) 1.5%, Candida albicans (C. albicans) 20% 및 Aspergillus niger (A. niger) 20%를 나타내었다. 이때 1,2-hexanediol의 최소생육저지농도는 P. aeruginosa 1%, S. aureus 2.5%, E. coli 1.5%, C. albicans 1% 및 A. niger 5%를 나타냈다. 20% 백년초 줄기 추출물은 머드팩내에서 P. aeruginosa, S. aureus, E. coli에 대하여 2일차에 초기 균수보다 99.9999%이상 감소하였고 28일차까지 감소세를 유지하였으며, C. albicans는 14일차에 초기 균수보다 99.99%이상 감소하였고 28일까지 감소세를 유지하였다. A. niger는 21일차 및28일차에 급격히 감소하여 EP, USP 및 CTFA의 기준에 부합하는 정도의 방부력을 지님을 알 수 있었다. 결론: 이러한 결과를 토대로 백년초 줄기 추출물을 향후 화장품용 천연 보존제 및 천연 방부제로 사용한다면 매우 안정성 높은 원료로서의 가치가 있을것으로 사료된다.

에센셜오일의 Candia albicans에 대한 항생효과

함주옥 ( Joo-ok Ham ),임미혜 ( Mi-hye Lim ) 한국미용학회 2016 한국미용학회지 Vol.22 No.6

This study was conducted to analyse antibiotic activity of essential oils to Candida albicans which is causative agent in fungal vaginitis. Antibiotic activity of 56 essential oils collected in Korea was tested against Candida albicans KCTC 7965 by disc diffusion method, broth dilution method and MTT metabolic assay. Among the 56 well known essential oils that can be easily collected, only 41 showed a positive result on antibiotic activity, whereas the other 15 types did not show any results at all. A further classification of antibiotic activity for the 41 essential oils by diameter of inhibitory zone, showed that: 12 types of the 56 fall into a range of IZ ≤ 1 cm (+); 11 types a category of 1 cm< IZ ≤ 1.5 cm(++); 9 types a category of 1.5 cm< IZ ≤ 2 cm(+++), 4 types a category of 2 cm< IZ ≤ 2.5 cm (++++); and finally, 5 types a category of IZ >2.5 cm(+++++), respectively. In order to measure the antibiotic strength on those 5 most effective essential oils(+++++) against Candida albicans, we adopted the broth microdilution method as well as MTT metabolic activity assay, and compared with the commercially available antibiotic, Amphotericin B. After comparing and matching the MIC values of each essential oil and Amphotericin B, the result showed that the essential oils ranged from 0.0225-0.12% with the MIC50, whereas Amphotericin B with the same MIC value fell into a category of 0.3-0.7%. With the value of MIC0, essential oils showed a ranging 0.06-0.2%, compared to Amphotericin B`s 3-6%. As a result of those conducts, it is not hard to say that those 5 types of natural essential oils outperformed than those commercially available Amphotericin B, alluding the possible use as an antibiotics for Candidiasis in the future.

Antimicrobial activity of Mongolian medicinal plants

Gonchig, Enkhmaa,Erdenebat, Sarnaizul,Togtoo, Ouyntsetseg,Bataa, Sukhkhuu,Gendaram, Odontuya,Kim, Young-Sup,Ryu, Shi-Yong The Korean Society of Pharmacognosy 2008 Natural Product Sciences Vol.14 No.1

The antimicrobial activity of seventy five ethanol extracts obtained from 67 different kinds of plant species of the Mongolian flora were evaluated by means of the disc diffusion method against five species of microorganisms, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Micrococcus luteus and Pseudomonas aeruginosa. Among the plant extracts examined, 34 kinds of extracts demonstrated significant antibacterial activity against one or more species of microorganisms, respectively. Especially, the root extract of Paeonia anomala, the whole herb extract of Myricaria alopecuroides, the whole herb extract of comarum zalesovianum, the whole herb extract of Agrimonia pilosa and some other plant extracts demonstrated a particularly potent antimicrobial activity. The ethylacetate fractions obtained from the whole herb extract of Myricaria alopecuroides and from those of Sedum aizoon, Paeonia anomala, Sedum hybridum and Dasiphora fruticosa exhibited a particularly potent antibacterial activity especially against Staphylococcus aureus and Micrococcus luteus.