콩발효식품에서의 장구균과 항생제 내성특성

강태미(Tae-Mi Kang),박종현(Jong-Hyun Park) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.5

콩 발효식품인 청국장과 된장에서 분리한 Enterococcus의 항생제 내성을 평가하고자 31개의 청국장과 17개의 된장으로부터 Enterococcus를 분리, 동정한 후 항생제 내성특성을 분석하였다. 모든 청국장 시료에서 119균주, 된장시료에서 4균주, 총 123개의 Enterococcus를 분리하였고 가장 많이 분리된 종은 청국장에서는 E. faecium 69균주, E. faecalis 20균주 등이며 된장시료에서 분리한 4균주는 모두 E. faecium으로 동정되었다. 총 123개의 Enterococcus는 ampicillin, chloramphenicol, penicillin, tetracycline에 내성이 낮게 나타났다. 그리고 erythromycin, ripampin, streptomycin 등에게는 내성이 높은 것으로부터 민감성이 있는 것까지 다양하게 분포하였다. 청국장에서 분리된 Enterococcus의 vancomycin MIC 범위는 0.25~8 ㎍/mL로 나타났으며, MIC 8 ㎍/mL 정도의 낮은 내성균 17균주가 E. faecalis, E.faecium, E. gallinarum, E. casselifalvus 등에 고르게 분포되어 있음을 알 수가 있었다. 모든 분리균주에서 고농도 vancomycin 내성유전자인 vanA, vanB는 검출되지 않았다. 본 연구의 결과 Enterococcus는 자연환경에 널리 분포하고 있는 만큼 가공식품에는 항생제 내성균주와 vancomycinresistant Enterococcus가 존재할 수 있으나 이들 청국장, 된장 등의 콩 발효식품 Enterococcus에는 주요 항생제에 내성정도가 높지 않아 항생제 안전성이 있는 것으로 보인다. To evaluate the antibiotic risk of Enterococcus in fermented soy paste, Enterococcus spp. were isolated and identified from 31 Cheongkukjang and 17 Doenjang samples. Exactly 123 Enterococcus spp., 119 from Cheongkukjang and four from Doenjang, were ultimately isolated. The most frequently collected Enterococcus isolates in Cheongkukjang were 69 strains of E. faecium and 20 strains of E. faecalis. All four Enterococcus spp. from Deonjang were identified as E. faecium. All isolates were sensitive to ampicillin, chloramphenicol, penicillin, and tetracycline. However, they showed broad spectra from sensitivity to resistance to erythromycin, ripampin, and streptomycin. Vancomycin minimum inhibition concentration (MIC) of Enterococcus spp. from Cheongkukjang ranged from 0.25 to 8 ㎍/mL. Almost all strains were sensitive to vancomycin, but eight strains showed intermediate resistance to vancomycin. Seventeen strains showing the highest MIC of 8 ㎍/mL among all isolates were evenly distributed among E. faecalis, E. faecium, E. gallinarum, and E. casselifalvus, in which the strong resistant genes of vanA and vanB for vancomycin were not detected. Overall antibiotic resistance of Enterococcus isolates was relatively low and particularly low vancomycin resistance was similar to those of Enterococcus isolates obtained from other foods. Therefore, the antibiotics resistance of Enterococcus and especially vancomycin-resistant Enterococcus spp. from Cheongkukjang and Doenjang is not hazardous.

MALDI-TOF Mass Spectrometry as a Useful Tool for Identification of Enterococcus spp. from Wild Birds and Differentiation of Closely Related Species

( Dagmara Stepien-pysniak ),( Tomasz Hauschild ),( Pawel Rozanski ),( Agnieszka Marek ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.6

The aim of this study was to explore the accuracy and feasibility of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying bacteria from environmental sources, as compared with rpoA gene sequencing, and to evaluate the occurrence of bacteria of the genus Enterococcus in wild birds. In addition, a phyloproteomic analysis of certain Enterococcus species with spectral relationships was performed. The enterococci were isolated from 25 species of wild birds in central Europe (Poland). Proteomic (MALDI-TOF MS) and genomic (rpoA gene sequencing) methods were used to identify all the isolates. Using MALDI-TOF MS, all 54 (100%) isolates were identified as Enterococcus spp. Among these, 51 (94.4%) isolates were identified to the species level (log(score) ≥2.0), and three isolates (5.6%) were identified at a level of probable genus identification (log(score) 1.88-1.927). Phylogenetic analysis based on rpoA sequences confirmed that all enterococci had been correctly identified. Enterococcus faecalis was the most prevalent enterococcal species (50%) and Enterococcus faecium (33.3%) the second most frequent species, followed by Enterococcus hirae (9.3%), Enterococcus durans (3.7%), and Enterococcus casseliflavus (3.7%). The phyloproteomic analysis of the spectral profiles of the isolates showed that MALDI-TOF MS is able to differentiate among similar species of the genus Enterococcus.

국내 신선 편이식품으로부터 분리한 Enterococcus의 항생제 저항성

김현정,김승민 한국식품과학회 2018 한국식품과학회지 Vol.50 No.6

Enterococcus spp. have been considered major indicator organisms for antibiotic resistance due to their ability to easily acquire and to harbor antibiotic resistance. In this study, Enterococcus spp. were isolated from 174 retail freshcut products (fresh vegetable salads, microgreens, and sprouts) in Korea. Among the 20 Enterococcus isolates obtained, 18 (90.0%) were Enterococcus faecalis and 2 (10.0%) were Enterococcus faecium. The patterns of antibiotic resistance against nine antimicrobials were analyzed. Most of the isolates (85.0%) were resistant to quinupristin/dalfopristin, 40.0% and 50.0% of the isolates showed intermediate resistance to two critically important antimicrobials for human medicine, ciprofloxacin and erythromycin, respectively. Vancomycin-resistant enterococci were not detected in this study. Given the importance of antimicrobial resistance of enterococci in food safety as well as in public health, our results regarding the occurrence (level of contamination) and antimicrobial resistance of Enterococcus spp. could provide useful information that aids the risk analysis of antibiotic resistance.

The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model

Jeong Hyun Chang(장정현),EunJu Yang(양은주),Sun Nyoung Yu(유선녕),Soon-Cheol Ahn(안순철) 한국생명과학회 2017 생명과학회지 Vol.27 No.10

프로바이오틱스(probiotics)는 숙주에 이로운 작용을 하는 장내세균으로 주로 알려져 있으며 꾸준히 염증치료의 새로운 치료물질로 주목 받고 있다. 그러나 Lactobacillus나 Bifidobacterium과 같은 전형적인 프로바이오틱스는 장내에서 꾸준히 생존하기 어렵고 그로 인해 매개할 수 있는 다양한 효능이 줄어든다. 반면, Enterococcus는 다양한 물질을 분비하고 생물학적 활성을 지니고 있는 프로바이오틱스이긴 하나 동시에 기회감염균으로 항생제 내성능을 쉽게 획득할 수도 있어 사용을 자제할 것을 권고하고 있으며, Vancomycin 내성인자를 원래부터 가지고 있어서 이 인자의 발현과 함께 타 독성균주에 전달할 가능성이 있다. 그러나 본 연구에서는 이러한 Enterococcus의 추출물을 이용하여 항염증 물질을 확인해보고자 하였다. 특히, 호흡기계 질환인 천식의 치료효능을 보기 위해 human monocytic cell line인 THP-1 세포를 이용한 in vitro test와 천식 유발 BALB/c mice를 이용한 in vivo test를 시행하였다. 천식은 사이토카인과 IgE 등 세포 염증성 물질을 분비하고 호산구, 호중구, 호염기구, 비만세포, 단구, T 세포 등 다양한 염증 세포가 폐 부위로 침윤되는 복합적인 염증 질환으로 잘 알려져 있다. 본 연구에서는 우선 Enterococcus faecalis 에서 초음파 처리를 통해 균체성분을 분해한 다음 동결건조하여 추출물을 획득하였다. E. faecalis에서 추출한 추출물(Ef extract)을 PBS에 부유시킨 다음, 염증유발을 위해 ovalbumin (OVA)를 처리한 THP-1 세포에 처리한 결과, OVA에 의해 증가한 THP-1 세포의 생존율이 감소하였다. 또한 OVA에 의해 천식이 유발된 BALB/c mice에서 염증부위인 폐의 세척액을 채취하여 폐 조직 내 세포 변화를 관찰하였다. 그 결과, Ef extract에 의해 호산구와 호중구가 Ef extract 농도에 따라 점차 감소하였고 혈액 내에서도 Ef extract에 의해 호산구, 호중구, 호염기구가 감소하였다. 이는 Ef extract에 의해 염증세포로부터 IL-5의 분비를 억제하고 IgE의 양을 감소시켜 이와 관련된 염증세포의 침윤 및 증가를 억제한 것으로 사료된다. 이를 통해 Ef extract 가 천식 동물모델에서 항염증효과가 있음을 관찰하였지만 Ef extract의 구성 물질 분석 및 작용 메커니즘에 대한 추가연구를 통해 천식 치료물질로의 가능성을 규명할 것이다. Probiotics are usually defined as intestinal bacteria that provide healthy benefit to the host and may offer new therapeutic materials for the treatment of inflammatory diseases. Lactobacillus, Bifidobacterium and Enterococcus are known as typical probiotics. But, these bacteria have mostly a weak viability and thus decreased probiotics-mediated effects in the intestinal tract. Asthma is an inflammatory airway disease, which is characterized by the releases of inflammatory mediators including cytokine and IgE. They are mainly associated with the recruitment, activation and disregulation of specific inflammatory cells, especially mast cells, monocytes, T cells, eosinophils and neutrophils in asthma. We performed these studies as in vitro and in vivo test the human inflammatory cell lines and ovalbumin (OVA)-induced asthma mouse model. And then the inhibitory effects of Enterococcus faecalis whole extract on inflammatory responses were examined. For our examinations, the E. faecalis whole extract (Ef extract) was acquired from whole bacteria of E. faecalis using freeze/thawing after ultrasonication method. As results, OVA-mediated THP-1 cell viability was decreased by the treatment of Ef extract. In the asthmatic mouse model, Ef extract inhibited the infiltration of inflammatory cells into the inflammatory sites and blood. This whole extract may have anti-asthmatic effects associated with the regulation of IL-5 and IgE expression. It may also be a promising candidate in anti-allergic medicine for the treatment of asthma.

중환자실 간호사의 반코마이신 내성 장구균 감염관리에 대한 인지도, 이행도 및 관련요인

박영미(Park Young Mi),박형숙(Park Hyoung-Sook),박경연(Park Kyung-Yeon) 기본간호학회 2008 기본간호학회지 Vol.15 No.4

Purpose: The purpose of this study was to identify factors influencing Vancomycin-resistant enterococcus infection control among nurses in intensive care units. Method: Data were collected from August 15 to October 14, 2007 from 188 nurses working in intensive care units. The nurses answered a 26 item-questionnaire, which included management of the cohort (14 items), hand washing (6 items) and management of the environment (6 items). Descriptive statistics, t or F test, ANOVA, and mu ltiple regression analysis were used with SPSS PC+ 14.0 to analyze the data. Results: The participant's level of awareness of Vancomycin-resistant enterococcus infection control was 3.87; that of compliance was 3.74. Significant factors influencing the level of compliance with Vancomycin-resistant enterococcus infection control were ‘the level of the awareness’ and ‘the type of intensive care unit’. These two variables accounted for 21.0% of variance for compliance with the Vancomycin-resistant enterococcus infection control among the participants. Conclusion: In order to develop a strategy to increase the compliance with Vancomycin-resistant enterococcus infection control, it is necessary to be concerned about ‘the level of the awareness’, ‘the type of intensive care unit’, and ‘experiences of caring for patients with Vancomycin-resistant enterococcus’.

Enterococcus의 Aminoglycoside에 대한 고도내성

김미나,배직현,송재훈 대한임상병리학회 1992 대한임상병리학회지 Vol.12 No.3

Characteristic and role of chromosomal type II toxin-antitoxin systems locus in Enterococcus faecalis ATCC29212

Li Zhen,Shi Chao,Gao Shanjun,Zhang Xiulei,Lu Di,Liu Guangzhi 한국미생물학회 2020 The journal of microbiology Vol.58 No.12

The Gram-positive bacterium Enterococcus faecalis is currently one of the major pathogens of nosocomial infections. The lifestyle of E. faecalis relies primarily on its remarkable capacity to face and survive in harsh environmental conditions. Toxin-antitoxin (TA) systems have been linked to the growth control of bacteria in response to adverse environments but have rarely been reported in Enterococcus. Three functional type II TA systems were identified among the 10 putative TA systems encoded by E. faecalis ATCC29212. These toxin genes have conserved domains homologous to MazF (DR75_ 1948) and ImmA/IrrE family metallo-endopeptidases (DR75_ 1673 and DR75_2160). Overexpression of toxin genes could inhibit the growth of Escherichia coli. However, the toxin DR75_1673 could not inhibit bacterial growth, and the bacteriostatic effect occurred only when it was coexpressed with the antitoxin DR75_1672. DR75_1948–DR75_1949 and DR75_ 160–DR75_2161 could maintain the stable inheritance of the unstable plasmid pLMO12102 in E. coli. Moreover, the transcription levels of these TAs showed significant differences when cultivated under normal conditions and with different temperatures, antibiotics, anaerobic agents and H2O2. When DR75_2161 was knocked out, the growth of the mutant strain at high temperature and oxidative stress was limited. The experimental characterization of these TAs loci might be helpful to investigate the key roles of type II TA systems in the physiology and environmental stress responses of Enterococcus.

CASE REPORT : Refractory Clostridium difficile Infection Cured With Fecal Microbiota Transplantation in Vancomycin-Resistant Enterococcus Colonized Patient

( Mi Ok Jang ),( Jun Hwan An ),( Sook In Jung ),( Kyung Hwa Park ) 대한장연구학회 2015 Intestinal Research Vol.13 No.1

The rates and severity of Clostridium difficile infections, including pseudomembranous colitis, have increased markedly. How-ever, there are few effective treatments for refractory or recurrent C. difficile infections and the outcomes are poor. Fecal micro-biota transplantation is becoming increasingly accepted as an effective and safe intervention in patients with recurrent disease, likely due to the restoration of a disrupted microbiome. Cure rates of >90% are being consistently reported from multiple cen-ters. We cured a case of severe refractory C. difficile infection with fecal microbiota transplantation in a patient colonized by vancomycin-resistant enterococcus. (Intest Res 2015;13:80-84)

샐러드와 새싹채소에서 Enterococcus의 분리와 항생제 내성특성

강태미 ( Tae Mi Kang ),조선경 ( Sun Kyung Cho ),박종현 ( Jong Hyun Park ) 한국미생물 · 생명공학회 2008 한국미생물·생명공학회지 Vol.36 No.2

장구균 검출 배지 개발

장동호,윤준범,이근헌,박경량,Chang, Dong-Ho,Yoon, Jun-Beom,Lee, Keun Heon,Park, Kyeong Ryang 한국미생물학회 2016 미생물학회지 Vol.52 No.1

분변오염의 지표가 되는 장구균을 검출하기 위하여 장구균 선택 배지를 개발하였다. 기존에 알려진 여러 장구균 검출배지 중 총 9종류(Enterococci Confirmatory agar, Azide dextrose agar, Bromocresol-purple azide agar, Esculin bile agar, Citrate azide tween carbonate agar, KF Streptococcus agar, BROLACIN agar, Kanamycin esculin azide agar, Membrane filter Enterococcus selective agar)의 배지를 선별하였고, 배지 성분을 적절히 조합하여 새로운 배지 Enterococcus Mixed medium을 개발하였다. 배지의 효과를 검증하기 위하여 장구균 6종류(Enterococcus avium KACC 10788, Enterococcus faecium KACC 11954, Enterococcus saccharolyticus KACC 10783, Enterococcus durans KACC 10787, Enterococcus faecalis KACC 11304, Enterococcus hirae KACC 10779)와 비장구균 3종류(Escherichia coli KACC 10005, Staphylococcus aureus subsp. aureus KACC 10768, Bacillus subtilis KACC 10111)를 사용하였다. 그 결과 새롭게 개발된 장구균 검출배지에서 비장구균의 생장이 억제된 반면에 장구균 6종류는 모두 생장하는 것을 확인하였다. 또 환경시료에서 장구균을 분리할 수 있는지 확인하기 위해 하천과 바다에서 채취해온 시료를 이용하여 실험한 결과 장구균의 생장을 확인하였다. 따라서 이 배지를 이용하면 수질 분변오염의 지표가 되는 장구균을 효과적으로 검출할 수 있음을 확인하였다. An enterococci selective medium was developed to detect the presence of enterococci for use as a fecal contamination indicator. Among several media which have been known to detect enterococci, the following 9 different kinds of media were selected: Enterococci Confirmatory agar, Azide dextrose agar, Bromocresol-purple azide agar, Esculin bile agar, Citrate azide tween carbonate agar, KF Streptococcus agar, BROLACIN agar, Kanamycin esculin azide agar, and Membrane filter Enterococcus selective agar. Various components from the nine media were mixed to develop a more effective enterococcus selective medium. The newly developed medium named as 'Enterococcus Mixed medium' was more effective than the previous 9 media. Enterococci strains (Enterococcus avium KACC 10788, Enterococcus faecium KACC 11954, Enterococcus saccharolyticus KACC 10783, Enterococcus durans KACC 10787, Enterococcus faecalis KACC 11304, and Enterococcus hirae KACC 10779) and non-enterococci strains (Escherichia coli KACC 10005, Staphylococcus aureus subsp. aureus KACC 10768, and Bacillus subtilis KACC 10111) were used to test the new medium. As a result, the enterococci strains grew well on the Enterococcus Mixed medium whereas the non-enterococci strains did not grow well on it. Additionally, growth of enterococci with freshwater and seawater samples was observed to be good on the Enterococcus Mixed medium. The result of this study confirmed that the Enterococcus Mixed medium was effective in detecting the target enterococci.