• Journal of Microbiology and Biotechnology
• /
• 제26권3호
• /
• pp.521-529
• /
• 2016

Extensive use of antibiotics over recent decades has led to bacterial resistance against antibiotics, including gentamicin, one of the most effective aminoglycosides. The emergence of resistance is problematic for hospitals, since gentamicin is an important broad-spectrum antibiotic for the control of bacterial pathogens in the clinic. Previous study to identify gentamicin resistance genes from environmental samples have been conducted using culture-dependent screening methods. To overcome these limitations, we employed a metagenome-based culture-independent protocol to identify gentamicin resistance genes. Through functional screening of metagenome libraries derived from soil samples, a fosmid clone was selected as it conferred strong gentamicin resistance. To identify a specific functioning gene conferring gentamicin resistance from a selected fosmid clone (35-40 kb), a shot-gun library was constructed and four shot-gun clones (2-3 kb) were selected. Further characterization of these clones revealed that they contained sequences similar to that of the RNA ligase, T4 rnlA that is known as a toxin gene. The overexpression of the rnlA-like gene in Escherichia coli increased gentamicin resistance, indicating that this toxin gene modulates this trait. The results of our metagenome library analysis suggest that the rnlA-like gene may represent a new class of gentamicin resistance genes in pathogenic bacteria. In addition, we demonstrate that the soil metagenome can provide an important resource for the identification of antibiotic resistance genes, which are valuable molecular targets in efforts to overcome antibiotic resistance.

• 미생물학회지
• /
• 제33권3호
• /
• pp.165-169
• /
• 1997

병원하수에서 분리한 gentamicin 저항성 세균으로부터 aacC2 유전자를 가지고 있는 R 플라스미드 pGM5와 pGM6를 선발하였다. 이들 플라스미드에서 gentamicin 저항성 유전자를 포함하는 부분을 pUC18의 BamHI 자리에 클로닝하여 재조합 플라스미드 pSY5와 pSY6를 각각 얻었다. 재조합 플라스미드의 삽입된 부분에 대한 제한효소 지도를 통해 Tn3 염기서열이 aacC2 유전자의 상류부위에 위치하는 것을 알았다. 재조합 플라스미드의 gentamicin에 대한 민감성의 비교를 통해 Tn3의 bla 유전자 부분과 3'역행중복 부위의 염기서열이 gentamicin 저항성 유전자의 발현에 중요한 역할을 담당하고 있었다.

• 미생물학회지
• /
• 제17권3호
• /
• pp.152-159
• /
• 1979

Using Micromonospora strain, gentamicin was produced by fermentation. The increase in gentamicin productivity was studied by strain improvement and systematic optimization of fermentation process variables. The productivity of parent strain of M.purpurea (ATCC15835) was improved by selection of superior mutant after U.V. irradiation and induction of genetamicin resistance. Potato starch and soy bean meal were the best carbon and nitrogen sources for gentamicin fermentation, respectively. The optimum stimulating concentration of Co ion for gentamicin production was 0.006g $CoCl_2$ per liter of broth. Oxygen ws found to be an important factor for gentamicin yield. The optimum pH for the cell growth and gentamicin production were 7.2 and 6.8 respectively. By controlling the pH, oxygen, and other conditions found in this study at the optimal conditions for cell growth and gentamicin production, the total productivity of gentamicin was increased significantly.

• 대한약침학회지
• /
• 제19권4호
• /
• pp.312-318
• /
• 2016

Objectives: Bacterial resistant infections have become a global health challenge and threaten the society's health. Thus, an urgent need exists to find ways to combat resistant pathogens. One promising approach to overcoming bacterial resistance is the use of herbal products. Green tea catechins, the major green tea polyphenols, show antimicrobial activity against resistant pathogens. The present study aimed to investigate the effect of catechins, green tea extract, and methylxanthines in combination with gentamicin against standard and clinical isolates of Staphylococcus aureus (S. aureus) and the standard strain of Pseudomonas aeruginosa (P. aeruginosa). Methods: The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) values of different agents against bacterial strains were determined. The interactions of green tea extract, epigallate catechin, epigallocatechin gallate, two types of methylxanthine, caffeine, and theophylline with gentamicin were studied in vitro by using a checkerboard method and calculating the fraction inhibitory concentration index (FICI). Results: The MICs of gentamicin against bacterial strains were in the range of $0.312-320{\mu}g/mL$. The MIC values of both types of catechins were $62.5-250{\mu}g/mL$. Green tea extract showed insufficient antibacterial activity when used alone. Methylxanthines had no intrinsic inhibitory activity against any of the bacterial strains tested. When green tea extract and catechins were combined with gentamicin, the MIC values of gentamicin against the standard strains and a clinical isolate were reduced, and synergistic activities were observed (FICI < 1). A combination of caffeine with gentamicin did not alter the MIC values of gentamicin. Conclusion: The results of the present study revealed that green tea extract and catechins potentiated the antimicrobial action of gentamicin against some clinical isolates of S. aureus and standard P. aeruginosa strains. Therefore, combinations of gentamicin with these natural compounds might be a promising approach to combat microbial resistance.

• 미생물학회지
• /
• 제40권1호
• /
• pp.60-64
• /
• 2004

Gentamicin에 저항성을 나타내는 세균을 병원 하수로부터 분리하여 aminoglycoside-(3)- N-acetyltransferase를 암호화하는 유전자인 aac(3)II의 존재여부를 dot-blot hybridization으로 조사하였다. aac(3)II유전자의 일부를 탐침으로 사용한 결과 gentamicin저항성 세균의 41% (39/95)가 이 유전자를 가지고 있었다. aac(3)II와 Tn3에서 각각 설계된 primer를 사용한 PCR 결과 aac(3)II를 가지고 있는 39개 균주 중 13개 균주가 TnS-aac(3)II구조를 가지고 있음을 알 수 있었다. aac(3)II의 상류에 Tn3를 가지고 있는 13개 균주의 gentamicin에 대한 최소억제농도는 가지고 있지 않은 균주에 비해 상대적으로 높았다. 13개 균주를 동정한 결과 5개는 Eschelichia coli, 3개는 Acinetobacter johnsonii, Enterobacter agglomerans와 Micrococcus luteus가 각각 2개, 그리고 1개의 Pseudomonas facilis로 동정되었다. 이러한 결과들은 Tn3-aac(3)II구조가 gentamicin 저항성 세균들에서 널리 분포하고 있음을 말해준다.

• 한국가금학회지
• /
• 제7권1호
• /
• pp.53-64
• /
• 1980

A total of 1503 specimens were submitted to the Poultry Disease Diagnostic Service Laboratory during the year 1966 and 1978. The most frequently diagnosed diseases in order of prevalence were avian mycoplasmosis, staphylococcosis, colibacillosis, salmonellosis and pullorum disease, the percentages of the conditions being 24.6%, 20.0%, 18.0%, 12.6% and 6.4%, respectively, The drug resistance of pathogenic mirnoorganisms isolated during the year 1978 from chicken with colicabacillosis, staphylococcosis or salmonellosis were investigated by the use of disc diffusion technique, the results being as follow. 1) Drug resistance of 63 strains of Escherichia coli More than 95% of the strains tested were sensitive to colistin and gentamicin. The percentages of strains sensitive to kanamycin, chloramphenicol, ampicillin and nitrofurantoin were 66.7%, 60.3%, 60.3% and 47.6%, respectively. Majority of the strains were highly resistant to streptomycin and tetracyline. All the strains were resisistant to bacitracin lincomycin, oleandomycin, penicillin and erythromycin. All the strains tested were resistant to more than two among 10 drugs in common use such as penicillin, erythromycin, streptomycin, tetracycline, neomycin, chloramphenicol, kanamycin, ampicillin and gentamicin, and 27 different resistance patterns were noted. The most frequent multiple resistance pattern was PC, EM, SM and TC (11.1%). 2) Drug resistance of 48 strains of Salmonella More than 95% of the strains tested were sensitive to colistin, gentamicin ana ampicillin. The percentages of st rains sensitive to kanamycin, tetracycline, neomycin and nitrofurantoin were 81,3%, 79%, 72.9%, and 68.0% respectively. None of them was sensitive to streptomycin, oleandomycin, erythromycin, lincomycin and bacitracin. All the strains were resistant to more than one among 7 drugs in common use such as streptomycin, erythromycin, neomycin, tetracycline, kanamycin, ampicillin and gentamicin. The most frequent resistance pattern was SM and EM(66.7%). 3) Drug resistance of 54 strains of Staphylococci All the strains tested were sensitive to gentmaicin, kanamycin and cephalothin. Majority of them were highly sensitive to bacitracin, methicillin, nitrofurantoin and chloramphenicol. The Percentages of strains sensitive to streptomycin, ampicillin, lincomycin and tetracycline were 66.7%, 55.6%, 44.4% and 27.8%, respectively. Among them, 51 strains were resistant to more than one among 11 drugs in common use such as tetracycline, lincomycin, ampicillin, penicillin, streptomycin, erythromycin, neomycin, oleandomycin, chloramphenicol, methicillin and bacitracin, and thirty one different resistance patterns were noted.

• 약학회지
• /
• 제30권2호
• /
• pp.55-61
• /
• 1986

A total of 130 of Staphylococcus strains isolated from various clinical specimens of admitted patients of a university hospital with systemic or severe cases of infection. All of these were tested for the antimicrobial susceptibility to 11 drugs of common use. The hospital strains isolated showed higher frequency of resistance against four drugs including gentamicin, penicillin, erythromycin, kanamycin but amikacin, cephalothin, streptomycin were effective. And also 47.7% of methicillin-resistant Staphylococcus were isolated from the clinical patients. However, isolated Escherichia coli strains showed higher frequency of resistance, but two drugs, tobramycin and gentamicin were effective to them.

• 대한약침학회지
• /
• 제22권1호
• /
• pp.49-54
• /
• 2019

Objective: Antibiotic resistance is a global health problem and threatens health of societies. These problems have led to a search for alternative approaches such as combination therapy. The aim of the present study was to investigate the effect of caffeine and omeprazole in combination with gentamicin or ciprofloxacin against standard and clinically resistant isolates of Staphylococcus aureus and Escherichia coli. Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of different agents against bacterial strains were determined. The interaction of non- antibiotic drugs with gentamicin and ciprofloxacin was studied in vitro using a checkerboard method and calculating fraction inhibitory concentration index (FICI). Verapamil as efflux pump inhibitor was used to evaluate the possible mechanism of bacterial resistance to antibiotics. Results: The MIC and MBC values of gentamicin against bacterial strains were in the range of $20-80{\mu}g/ml$ and $40-200{\mu}g/ml$, respectively. Caffeine and omeprazole had no intrinsic inhibitory activity against tested microorganisms. However, upon combination of caffeine with antibiotics, the synergistic effects were observed. Verapamil was able to reduce the MIC values of gentamicin (4 folds) only in some bacterial strains. Conclusion: These findings indicated that caffeine was effective in removing bacterial infection caused by S. aureus and E. coli. The relevant mechanisms of antibiotic resistance were not related to the drug efflux.

• 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.

• 한국동물위생학회지
• /
• 제23권2호
• /
• pp.153-163
• /
• 2000

This study was carried out the antimicrobial resistance of staphylococcus aureus isolated from bovine mastitic milk, chickens, korean native cattle, korean native goats, pigs, dog and mice in northern area of kyongbuk. The result were summarised as follows ; A total of 149 S aureus were isolated from bovine mastitic milk, chickens, korean native cattle, korean native goats, pigs, dog and mice. In 80 isolates of S aureus from bovine mastitic milk, 60% of isolates revealed resistance to penicillin and ampicillin, 19% to gentamicin, 6% to tetracycline. In 36 isolates of S aureus from chickens, 72% of isolates revealed resistance to tetracycline, 58% to penicillin and ampicillin, 42% to streptomycin, 31% to lincomycin, 25% to norfloxacin, 22% to gentamicin. In 17 isolates of S aureus from korean native cattle, 100% of isolates revealed resistance to penicillin and ampicillin, 88% to lincomycin, 76% to tetracycline. 2 MRSA were isolated from the isolates of S aureus from bovine mastitic milk and revealed multi-drug resistance.