• 한국식품위생안전성학회지
• /
• 제35권1호
• /
• pp.6-12
• /
• 2020

본 연구에서는 비브리오 패혈증 예방과 치료에 유용한 항생제를 제시하기 위해 V. vulnificus의 독소유전자 분포와 항생제 내성을 분석하였다. 2015년부터 2017년까지 3년간 전남지역에서 발생한 비브리오 패혈증 환자로부터 분리되어 보관된 18균주와 전남지역에서 채취된 어패류 및 횟집 수족관수에서 분리된 5균주, 총 23균주를 대상으로 하였다. 실험에 사용된 V. vulnificus 23균주 모두 V. vulnificus로 재확인되었다. V. vulnificus 균주의 독소유전자를 분석한 결과, 23균주 중 19균주(82.6%)에서 RtxA 독소유전자가 확인되었고, 23균주 모두에서 viuB와 vvhA 독소유전자가 검출되었다. 이러한 결과는 독소유전자의 검출율이 기존 보고에 비해 높은 것이며, 실험에 사용한 모든 V. vulnificus 균주가 1개 이상의 독소유전자를 보유한 것으로 생선회 섭취와 상처를 통한 비브리오 패혈증 감염의 위험성이 상존하고 있었다. 따라서 횟집 종사자 등에 대한 비브리오 패혈증 예방 교육이 필요한 것으로 판단되었다. V. vulnificus에 대한 항생제 내성 실험결과 cefoxitin 항생제에 94.4%가 내성을 나타내었고, chloramphenicol과 tetracycline 등 14종의 항생제에 감수성을 나타내었다. 비브리오 패혈증 치료에 chloramphenicol과 tetracycline 항생제를 사용하는 현 치료법이 유용한 것으로 판단되었다.

• Fisheries and Aquatic Sciences
• /
• 제25권4호
• /
• pp.202-213
• /
• 2022

Antibiotic resistant Escherichia coli cases are increasing high especially in Southeast Asia. Illegal use of the antibiotic in the aquaculture farming may become the culprit of the outbreak and spread into environmental source. A study was conducted to: 1) detect the chloramphenicol (CAL)-resistant gene in E. coli isolated from three aquaculture farms and six rivers of northwestern Borneo and 2) investigate the correlation between cat gene with five common antibiotics used. Isolation of E. coli was done on Eosin methylene blue agar and characterized using indole, methyl red, Voges-Proskauer, citrate tests. E. coli isolates were subsequently tested for their susceptibility to five antibiotics commonly used in aqua-farming. The CAL-resistant E. coli were further analyzed for the presence of resistant genes (cat I, cat II, cat III, cat IV) using multiplex polymerase chain reaction. 42 bacterial colonies were isolated from a total of 80 individual water samples, 34 of which were identified as E. coli. Result showed 85.3% of the E. coli isolates were resistant to amoxicillin, 35.3% were resistant to tetracycline, 29.4% were resistant to CAL, 17.6% were resistant to nitrofurantoin and 8.8% were resistant to nalidixic acid. All of the 10 CAL resistant E. coli isolateswere detected with cat II genes; five isolates detected with cat IV genes; three isolates detected with cat III genes; and another two detected with cat I genes. Pearson correlation coefficient shows highly significant relationship between resistance pattern of CAL with amoxicillin; and CAL with tetracycline. Our findings provide the supplementary information of the CAL resistance gene distribution, thereby improving our understanding of the potential risk of antibiotic resistance underlying within this microbial ecosystem.

• 한국동물위생학회지
• /
• 제34권1호
• /
• pp.45-54
• /
• 2011

This study was conducted to investigate the distribution of Salmonella spp. from pigs and cattle in slaughterhouse, the antimicrobial resistance pattern and the prevalence of resistance genes of isolates. A total of 640 fecal samples from pigs and cattle in slaughterhouse were collected for isolation of Salmonella spp.. Isolation rate was revealed as 15% in pigs and 1.6% in cattle. As result of serotyping, group B (56.6%) were identified as most common in pigs and cattle isolates, in order of group C (24.5%) and group E (15.1%). S. Typhimurium (50.9%) was most common serotype. The major serotypes were in order of S. Rissen and S. London (11.3%) and S. Riggil (7.6%). In antimicrobial test, all isolates were demonstrates susceptibility to nitrofurantoin. But isolates were revealed resistance other antibiotics in order of tetracycline (64.6%), streptomycin (68.3%), ampicillin and amoxicillin (56.3%) and spectinomycin (47.9%). With polymerase chain reaction, antimicrobial resistance gene strA (75.0%) and aadA1 (3.1%) were detected in streptomycin resistance isolates and tetA (94.3%) and tetB (11.3%) gene were detected in tetracycline resistant isolates, but tetG was not detected. Class 1 integron gene was detected in all Salmonella isolates.

• 한국동물위생학회지
• /
• 제36권3호
• /
• pp.171-180
• /
• 2013

This study was carried out to investigate the antimicrobial resistance pattern and distribution of resistance gene in 44 Enterobacteriaceae and 21 Pseudomonas (P) aeruginosa isolated from hospitalized dogs and cats in animal hospital from 2010 to 2011 in Daegu. Among Enterobacteriaceae, Escherichia (E) coli was highly resistant to ampicillin (56.7%), followed by tetracycline (53.3%), cephalothin, streptomycine, sulfamethoxazole/trimethoprim, gentamicin and norfloxacin (40.0~43.3%). The remaining isolates of Enterobacteriaceae had high resistance to ampicillin (64.3%) and streptomycin (42.9%). Whereas, P. aeruginosa was low resistant to all antimicrobials tested (less than 15%). int I 1 gene was detected in 20 (57.1%) of 35 antimicrobial resistant Enterobacteriaceae and 2 (9.5%) of 21 P. aeruginosa., but int I 2 gene was not detected in all isolates. The eight resistance genes were found either alone or combination with other gene (s): $bla_{TEM}$, aadA, strA-strB, clmA, tetA, tetB, sul I and sul II. About 78% of integron-positive isolates were resistance to more than four antimicrobial agents. The findings suggest that class I integrons are widely distributed in E. coli among Enterobacteriaceae from dogs and cats and multi-drug resistance related to the presence of class I integrons. The prudent use of antimicrobials and continuous monitoring for companion animals are required.

• 미생물학회지
• /
• 제51권1호
• /
• pp.39-47
• /
• 2015

방선균이 생산하는 이차대사산물은 자기조절인자(${\gamma}$-butyrolactone autoregulator)라고 불리는 저분자의 신호전달물질과 이에 특이적으로 결합하는 autoregulator receptor protein의 상호작용에 의해 조절되는 것으로 알려져 있다. 그러므로 non-host에 autoregulator receptor 혹은 pleiotropic regulator의 발현은 이차대사산물 혹은 새로운 대사화합물의 효율적인 생산을 유도할 것으로 기대된다. 희소방선균 Saccharopolyspora erythreae으로부터 receptor (seaR) 유전자의 기능을 연구하기 위해 다른 속의 균주인 Streptomyces coelicolor A3(2)로 seaR 유전자를 삽입하여 형질전환하였다. S. coelicolor A3(2)의 형질전환은 oriT, attP, $ermEp^*$과 seaR gene 단편을 가지고 있는 ${\Phi}C31$ 유래의 integration vector인 pEV615 (6.6 kb)를 이용하여 Escherichia coli ET12567/pUZ8002를 DNA 공여체로 이용한 접합전달법을 사용하여 확립하였다. seaR 유전자의 삽입 유무는 PCR방법으로 확인하였고, seaR 유전자의 전사 발현은 RT-PCR방법으로 확인하였다. S. coelicolor A3(2)의 경우 표현형 microarray 실험을 통하여 seaR 유전자의 발현에 따른 표현형의 변화를 확인하였다. 특히, 표현형 microarray 실험에 나타난 tetracycline 항생제 기질에 대하여 wild type이 transformant에 비해 빠르게 성장하는 것은 항균제 감수성 검사와 일치하였다. 이는 tetracycline 생합성 유전자 및 내성 유전자의 발현 억제에 따른 변화라고 예상할 수 있으며 이를 위하여 tetracycline 생합성 관련 유전자 및 내성 유전자의 발현 패턴 분석등과 같은 분자 수준에서의 연구가 필요할 것으로 생각된다.

• 수산해양교육연구
• /
• 제29권3호
• /
• pp.834-846
• /
• 2017

Aquaculture practices to ensure greater production, such as high density breeding and excessive feeding, are become stressors that raise the prevalence of diseases. Accordingly, increasingly large volumes of antibiotics are used more frequently each year. Long term use antibiotics can generate resistant bacteria, which interrupt treatments and cause a potential transfer to human bodies. Thus, antibiotic resistance is of importance in public health. Tetracycline (Tc) is one of the typical medicines used in the aquaculture drugs, which has a wide range of application including gram-positive and gram-negative bacteria. In the examination of 153 strains isolated from olive flounder (Paralichthys olivaceus) farms located in Jeju in 2016, it turned out that a total of 84 strains were resistant to Tc or oxytetracycline (OTC). The extent to which the strains are resistant to Tc and OTC was confirmed through MIC test, mostly within the range of 25 to $100{\mu}g/m{\ell}$. Twelve different types of tet genes were detected using single and multiplex PCR in the 84 Tc-resistant strains. The PCR was used to find tet(K), tet(M), tet(O), and tet(S), which are known to exist primarily in gram positive strains. According to the results, - tet(S) is the most dominant gene in 49 strains of Streptococcus parauberis, accounting for 63.2%. And there were two strains that have two different types of resistant genes. The multiplex PCR was used to detect tet(A), tet(B), tet(C), tet(D), tet(E), and tet(G), which are commonly found in gram-negative strains. Each of tet(B), tet(D), and tet(B)&(M) was found in a strain presumed to be Vibrio sp., and only tet(D) was found in 10 Edwardsiella tarda strains.

• 미생물학회지
• /
• 제49권2호
• /
• pp.126-130
• /
• 2013

원유시료에서 분리한 장알균속 세균 245균주의 항생제 내성에 관여하는 다중약물 유출 펌프 유전자 분포도와 항생제 내성 패턴을 연구하였다. 그 결과 245 장알균속 균주의 ampicillin에 대한 내성률은 44.1%, erythromycin에 대한 내성률은 79.2%, tetracycline에 대한 내성률은 76.3%, chloramphenicol에 대한 내성률은 36.3%였으며 vancomycin과 ciprofloxacin에 대해서는 모두 감수성임을 알 수 있었다. 내성 관련 유전자 중 MFS타입의 eme(A)는 82.1%의 장알균에 분포하였으며, ABC 타입의 유전자인 efr(A)는 72.7%, efr(B)는 77.1%, lsa는 71.8%의 장알균에 분포하였다. 특히 이러한 유전자의 기원 세균인 Enterococcus faecalis의 경우 eme(A)는 92.5%, efr(A)는 87.4%, efr(B)는 88.4%, lsa는 88.4%의 분포도를 나타내었다. 한편 동일한 장알균속이지만 종이 다른 장알균에서 eme(A)는 E. faecium 4균주, E. avium 7균주, E. durans 4균주 및 E. raffinosus 2균주에 분포하고 있었다. efr(A)는 E. faecium 2균주와 E. durans 2균주에 분포하였으며, efr(B)는 E. faecium 4균주, E. avium 5균주 및 E. durans 4균주에 분포하였다. 본 연구는 우리나라 원유시료에서 분리한 장알균속의 여러 종의 세균에서 동일한 다중약물 유출 펌프(multidrug efflux pump)의 분포에 대한 첫 번째 보고라 사료되며 E. faecalis 이외의 장알균속에서 이러한 유전자의 분포는 서로 다른 종간의 유전자의 수평적인 이동의 가능성을 시사한다.

• Journal of Microbiology and Biotechnology
• /
• 제33권4호
• /
• pp.500-510
• /
• 2023

In this study, lactic acid bacteria were isolated from 21 top-selling probiotic products on Korean market and their antimicrobial resistance were analyzed. A total 152 strains were claimed to be contained in these products and 70 isolates belonging to three genera (Bifidobacterium, Lactobacillus, and Lactococcus) were obtained from these products. RAPD-PCR showed diversity among isolates of the same species except for two isolates of Lacticaibacillus rhamnosus from two different products. The agar dilution method and the broth dilution method produced different MICs for several antimicrobials. With the agar dilution method, five isolates (three isolates of Bifidobacterium animalis subsp. lactis, one isolate of B. breve, one isolate of B. longum) were susceptible to all nine antimicrobials and 15 isolates were multi-drug resistant. With the broth microdilution method, only two isolates (one isolate of B. breve and one isolate of B. longum) were susceptible while 16 isolates were multi-drug resistant. In this study, only two AMR genes were detected: 1) lnu(A) in one isolate of clindamycin-susceptible and lincomycin-resistant Limosilactobacillus reuteri; and 2) tet(W) in one tetracycline-susceptible isolate of B. longum B1-1 and two tetracycline-susceptible isolates and three tetracycline resistant isolates of B. animalis subsp. lactis. Transfer of these two genes via conjugation with a filter mating technique was not observed. These results suggest a need to monitor antimicrobial resistance in newly registered probiotics as well as probiotics with a long history of use.

• 생명과학회지
• /
• 제21권2호
• /
• pp.257-264
• /
• 2011

2009년 7월부터 12월까지 순천 소재 한 병원에 내원한 환자의 검체로부터 methicillin 내성 Staphylococcus aureus (MRSA) 75균주와 methicillin 감수성 S. aureus (MSSA) 24균주를 분리하였다. 분리균의 항생제 감수성 조사는 디스크 확산법을 사용하여 측정하였다. 분리균의 독소 유전자 보유는 multiplex PCR을 이용하여 장독소(enterotoxin; SE), 독성 쇼크 증상 독소 1(toxic shock syndrome toxin-1; TSST-1), 피부박탈성 독소(exfoliative toxin; ET) 및 백혈구 용해 독소(Panton-Valentine leukocidin; PVL) 유전자를 검출하였다. 분리된 MRSA 60개 균주는 1개 혹은 2개의 독소 유전자를 가지고 있으며, 22.7%의 균주가 seb, sec, seg, sei와 tst 유전자를 동시에 보유하고 있었으며 18.7%는 sec, seg, sei와 tst 유전자를 동시에 보유하고 있었다. 백혈구 용해독소를 암호하는 pvl 유전자는 검출되지 않았다. MRSA는 sec, seg, sei와 tst 유전자 보유에 높은 상관성을 보였다. MRSA 균주들은 erythromycin (분리균의 89%), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%)과 tetracycline (58.7%)에 내성이 높은 반면, MSSA 균주들은 erythromycin를 제외한 다른 항생제에는 민감하였다. 독소 유전자 seb, sec와 tst는 tetracycline 내성과 높은 상관관계가 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제31권2호
• /
• pp.250-258
• /
• 2021

Among various species of marine bacteria, those belonging to the genus Halomonas have several promising applications and have been studied well. However, not much information has been available on their antibiotic resistance. In our efforts to learn about the antibiotic resistance of strain Halomonas socia CKY01, which showed production of various hydrolases and growth promotion by osmolytes in previous study, we found that it exhibited resistance to multiple antibiotics including kanamycin, ampicillin, oxacillin, carbenicillin, gentamicin, apramycin, tetracycline, and spectinomycin. However, the H. socia CKY01 resistance pattern to kanamycin, gentamicin, apramycin, tetracycline, and spectinomycin differed in the presence of 10% NaCl and 1% NaCl in the culture medium. To determine the mechanism underlying this NaCl concentration-dependent antibiotic resistance, we compared four aminoglycoside resistance genes under different salt conditions while also performing time-dependent reverse transcription PCR. We found that the aph2 gene encoding aminoglycoside phosphotransferase showed increased expression under the 10% rather than 1% NaCl conditions. When these genes were overexpressed in an Escherichia coli strain, pETDuet-1::aph2 showed a smaller inhibition zone in the presence of kanamycin, gentamicin, and apramycin than the respective control, suggesting aph2 was involved in aminoglycoside resistance. Our results demonstrated a more direct link between NaCl and aminoglycoside resistance exhibited by the H. socia CKY01 strain.