• 한국환경성돌연변이발암원학회지
• /
• 제25권4호
• /
• pp.157-160
• /
• 2005

Biofilm 내부의 항생제 내성 박테리아의 성장형태는 만성감염과 질병을 발생한다. 인위적 형성한 biofilm의 체외실험 모델 시스템 통한 항생제 침투 실험을 수행하였다. 항생제 내성 균주 (E. coli, S. aureus)는 항생제 내성균주 은행으로부터 획득하였다 Ca-alginate bead를 인위적 biofilm으로 사용하였고, 세포 생존률을 향상시키기 위해 공기 압축을 이용한 세포 포획 실험도 측정되었다. biofilm의 항생제 감수성은 항생제의 농도 따라 최저 저해 농도 (MIC)를 이용하여 측정되었다. bead생성에 따른 안정성은 bead를 형성하지 않은 세포와 비교하여 감소하였다. 항생제에 민감한 E. coli의 경우 시간이 지남에 따라 균체수도 감소하였으나, 항생제 내성 E. coli는 일정한 균체수를 유지하였다. bead형성 후 항생제 투여 효과는 항생제 내성이 있고, 낮은 농도의 항생제를 처리할수록 더 높은 생존률을 보였다.

• Archives of Pharmacal Research
• /
• 제19권5호
• /
• pp.353-358
• /
• 1996

E. coli 11 and E. coli 101, clinical isolates of Escherichia coli were resistant to various quinolones, especially MICs to norfloxacin of both strains were higher than 100 mg/ml. In the presence of carbonyl cyanide m-chlorophenylhydrazone, a proton gradient uncoupler, norfloxacin uptake in both strains was increased, suggesting that an efflux system play an important role in the norfloxacin resistance. Outer membrane proteins of the susceptible and resistant strains which could affect the route of norfloxacin entry into cells were different. When quinolone resistance determining region(QRDR) of gyrA was amplified using PCR and cut with Hinf I, QRDR in the susceptible strain yielded two fragments while QRDRs in E. coli 11 and E. coli 101 yielded only one uncut fragment. When DNA sequence of QRDR was analyzed, there were two mutations as Ser-83 and Asp-87 in both resistant strains. these residues were changed to Leu-83 and Asn-87, respectively. These results showed that the norfloxacin resistance of E. coli 11 and E. coli 101 was resulted from multiple changes-an altered DNA gyrase A subunit, a change in route of drug entry, and reduction in quinolone concentration inside cells due to an efflux system.

• Environmental Engineering Research
• /
• 제14권1호
• /
• pp.63-67
• /
• 2009

Escherichia coli K-12 (E. coli K-12) is a representative indicator globally used for distinguishing and monitoring dynamic fates of pathogenic microorganisms in the environment. This study investigated how to most critically quantify lacZ ($\beta$-galactosidase) gene in E. coli K-12 by two different real-time polymerase chain reaction (real-time PCR) in association with three different DNA extraction practices. Three DNA extractions, i.e., sodium dodecyl sulfate (SDS)/proteinase K, magnetic beads and guanidium thiocyanate (GTC)/silica matrix were each compared for extracting total genomic DNA from E. coli K-12. Among them, GTC/silica matrix and magnetic beads beating similarly worked out to have the highest (22-23 ng/${\mu}L$) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/${\mu}L$) concentration of DNA retrieved. There were no significant differences in the quantification of the copy numbers of lacZ gene between SYBR Green I qPCR and QProbe-qPCR. However, SYBR Green I qPCR obtained somewhat higher copy number as $1{\times}10^8$ copies. It was decided that GTC/silica matrix extraction or magnetic beads beating in combination with SYBR Green I qPCR can be preferably applied for more effectively quantifying specific gene from a pure culture of microorganism.

• 한국식품영양과학회지
• /
• 제24권2호
• /
• pp.280-285
• /
• 1995

To develop natural food preservatives, antimicrobial effect of the ethanol extract of leaf mustard against E. coli and S. aureus were examined in terms of compositions and leakage of cellular materials in the microorganisms treated with the extract. No effect of the concentration of ethanol extract on the fatty acid composition of E. coli and S. aureus at logarithmic phase was showen, but the content of palmitic and palmitoleic acid of E. coli slightly increased and decreased, respectively, and the content of palmitic and margaric acid of S. aureus slightly increased, when compared to each control. Ethanol extract did not affect most of the amino acids E. coli and S. aureus at logarithmic phase ; however, some of them(proline, glycine, valine and histidine of E. coli and proline, methionine and histidine of s. aureus) were elevated and some other amino acid(aspartic acid, glutamic acid, tyrosine and arginine of E. coli and aspartic acid, glutamic acid, glycine, alanine and lysine of Staph. aureus) found to be decreased. The amount of cell body protein leaked from E. coli and S. aureus increased to 1.02 and 0.22mg/g cell weight, respectively, as compared to controls. Similarly, the substances with absorbance at 260 nm from E. coli and s. aureus increased to 0.12 and 0.06mg/g cell weight, respectively.

• 한국식품조리과학회지
• /
• 제13권4호
• /
• pp.440-447
• /
• 1997

향신료인 oregano(Origanum vulgare L.)를 액체배지에 첨가하여 2종류의 식중독세균(Escherichia coli O157:H7과 Staphylococcus aureus 196E)의 증식과 저온저장중 생존에 미치는 항균효과를 조사하였다. 저농도(0~2%, w/v)의 oregano를 액체배지(TSB)에 첨가하여 E. coli와 S. aureus를 $10^{6}$~$10^{7}$ cells/$m\ell$가 되게 접종한 후 35$^{\circ}C$에서의 증식과 냉장(5$^{\circ}C$)및 냉동(-2$0^{\circ}C$) 저장중 생존억제효과를 생균수의 변화로서 조사하였다. 35$^{\circ}C$에서의 E. coli의 증식은 1% 이내의 향신료 농도에서는 증식이 저해되지 않았으나, 2%의 oregano존재하에서는 짧은 유도기를 거친 후에 증식이 시작되었다. 정상기 이후의 사멸기에서는 첨가한 oregano의 농도가 높을수록 빠른 사멸속도를 나타내었다. 35$^{\circ}C$에서의 S. aureus의 증식은 0.1%의 경우에는 증식이 저해되지 않았으며 0.3%와 0.5%의 oregano 존재하에서는 긴 유도기를 거친 후에 증식이 시작되었으나 1~2%의 경우에는 계속적으로 생균수가 감소되었다. 5$^{\circ}C$에서 냉장한 경우에는 0.5%이상의 oregano에 의해 E. coli와 S. aureus 모두 우수한 항균효과를 나타내어 생존이 크게 억제되고 저장말기에 사멸하였다. -2$0^{\circ}C$에 동결저장하였을 때, E. coli는 저장기간이 길어질수록, 첨가한 향신료의 농도가 높을수록 생존억제효과가 증대되었으나 S. aureus는 저장초기에 급격히 감소한 후 거의 일정수준을 유지하였으며 첨가한 향신료의 농도에 따른 생균수의 차이는 나타나지 않았다. 동결저장에서 0.1%의 oregano를 첨가했을 때 E. coli는 대조구의 생균수에 비해 1/3~l/7로 생균수가 감소하였으나 S. aureu는 대조구에 비해 1/18~l/46로 감소하여 S. aureus에서 더 우수한 항균효과를 나타내었다.

• Journal of Microbiology
• /
• 제39권1호
• /
• pp.62-66
• /
• 2001

Unlike eukaryotic efflux pumps energized by ATPase bacterial efflux pumps are energized by the proton motive force. That is the reason why CCCP, an inhibitor of proton motive forcer is widely used to study the bacterial efflux pump. In many cases, efflux systems have been observed only in quinolone-resistant bacteria. Most of the quinolone-susceptible strains have been found to maintain little efflux pump. However some susceptible bacteria skewed the increased intracellular quinolone concentration only at a low concentration (0.01 or 0.1 mM) but net at a high concentration (1 mM) of CCCP. If bacterial cells were killed at high concentrations of CCCP and lost the integrity of their membranes, the intracellular quinolone would leak out from cells with no efflux system. The efflux pump system in the quinolone-susceptible strains could net be detected at the same concentration used for resistant bacteria. To test this hypothesist the intracellular quinolone concentration in the quinolone-susceptible and -resistant strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus was assayed at various concentrations of CCCP. Since the effect of CCCP is very rapid, the survival of bacteria was observed by assaying the DNA synthesis in 5 min. In the case of E. coli, but not P. aeruginosa or S. aureus, the quinolone susceptible strain was more susceptible to CCCP than the quinolone resistant ones, especially when the incubation with CCCP was extended. Decrease of the intracellular quinolone concentration resulted in a false result-no or weak efflux system in the quinolone susceptible strains. Results suggested that the concentration of CCCP should be optimized in order to detect the efflux system in the quinolone susceptible strains of E. coli.

• Journal of Microbiology and Biotechnology
• /
• 제11권4호
• /
• pp.720-723
• /
• 2001

The effect of long-term storage on garlic antibacterial activity was investigated. A concentration of 5% or more garlic was found to be necessary to completely inhibit Eschrichia coli growth in tryptic soy broth. This value is substantially higher than the minimum inhibitory concentration of 1% for E. coli reported previously. pH-modified garlic extract was stored at different temperatures to investigate the impact of storage conditions (i.e., temperature, pH, period of storage) on the stability of the antibacterial activity of the garlic extract used against E. coli B34. The antibacterial effectiveness of the garlic extract against E. coli remained stable when both the storage temperature and the pH of the extract were kept low. When the garlic extract was stored at $40^{\circ}C and above, most or all of the garlic antibacterial activity disappeared after a 24-h storage period, regardless of the storage pH. The antibacterial activity was weakened when the pH of the garlic extract was adjusted to 8, and at low temperatures.

• 대한의생명과학회지
• /
• 제29권3호
• /
• pp.130-136
• /
• 2023

In this study, the anti-bacterial activity of three medicinal herbs such as Dioscorea batatas, Morus alba, and Tagetes erecta, have been applied in oriental medicine against Escherichia coli and Staphylococcus aureus. The antibacterial activity of Dioscorea batatas ethanol extract (DBEE), Morus alba ethanol extract (MAEE), and Tagetes erecta ethanol extract (TEEE) was analyzed by disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. As a result, three medicinal herbs exhibited antibacterial activity in a dose-dependent manner as well as MAEE and TEEE showed the most potent antibacterial activity followed by disk diffusion assay against E. coli and S. aureus, respectively. MIC against E. coli and S. aureus was 1.6 mg/mL of MAEE and TEEE as well as 0.2 mg/mL of TEEE. MBC against both strains was 5 mg/mL of DBEE and 2.5 mg/mL of TEEE. Consequently, three medicinal herbs in this study showed potent antibacterial activity through the inhibited growth of E. coli and S. aureus, which is considered as a potential candidate for cosmeceutic agents to attenuate the pruritus. In addition, antibacterial and antihistamine effects followed by the extraction solvents should be analyzed in future studies.

• Membrane and Water Treatment
• /
• 제10권3호
• /
• pp.231-237
• /
• 2019

Peroxymonosulfate (PMS) in combination with Cu(II) was examined to inactivate E. coli and MS2 coliphage in natural water. The combined system (i.e., the Cu(II)/PMS system) caused a synergistic inactivation of E. coli and MS2, in contrast with either Cu(II) or PMS alone. Increasing the concentration of PMS enhanced the inactivation of E. coli and MS2, but after a certain point, it decreased the efficacy of the microbial inactivation. In the Cu(II)/PMS system, adding reactive oxidant scavengers marginally affected the E. coli inactivation, but the inhibitory effects of copper-chelating agents were significant. Fluorescent assays indicated that the Cu(II)/PMS system greatly increased the level of reactive oxidants inside the E. coli cells. The sequential addition of Cu(II) and PMS inactivated more E. coli than did adding the two simultaneously; in particular, the inactivation efficacy was much higher when Cu(II) was added first. The observations from the study collectively showed that the microbial inactivation by the Cu(II)/PMS system could be attributed to the toxicity of Cu(I) as well as the intracellular oxidative stress induced by Cu(III) or radical species.

• Journal of Microbiology and Biotechnology
• /
• 제16권9호
• /
• pp.1448-1452
• /
• 2006

The effects of amplifying the gluconeogenic phosphoenolpyruvate carboxykinase of Escherichia coli ($pck_{Ec}$) on succinic acid production in E. coli were examined under anaerobic condition. No significant increase in succinic acid production was observed in E. coli overexpressing the $pck_{Ec}$ gene without supplementing $NaHCO_{3}$ or $MgCO_{3}$. On the other hand, succinic acid production was enhanced as the $NaHCO_{3}$ concentration was increased. When 20 g/l of $NaHCO_{3}$ was added, succinic acid production in recombinant E. coli overexpressing PCK was 2.2-fold higher than that observed in the wild-type strain. It was concluded that the gluconeogenic $pck_{Ec}$ overexpression enabled E. coli to enhance succinic acid production only under the high bicarbonate supplementation condition.