• Journal of Food Hygiene and Safety
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• v.20 no.1
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• pp.48-52
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• 2005

This study was investigated to determine the antimicrobial activity of ethanol extract of green tea, and of solvent fractionated ethanol extract on Escherichia coli O157:H7. MIC value of ethanol extracts and solvent fractionated ethanol extract (hexane, ethyl-acetate, chloroform and water) were $2000{\mu}g/ml,\;0,\;500{\mu}g/ml,\;2000{\mu}g/ml,\;1000{\mu}g/ml$, respectively. The antimicrobial activity of ethyl acetate fraction against E. coli O157 :H7 ranged from $250-2000{\mu}g/disk$. The antimicrobial activity was significantly increased as concentration dependent. When E. coli O157:H7 was pH-adjusted TSB containing $1000{\mu}g/ml$ ethyl acetate, E. coli O157:H7 was significantly inhibited at initial pH of 10, whereas the pathogen grew well in the presence of pH 4.5-pH 9. The inhibitory effect of the ethyl acetate fraction on the growth of E. coli O157 :H7 was investigated. Growth of both strain in the tested conditions were rapidly occurred up to 12 h, but no growth was occurred in the presence of $250-1000{\mu}g/mL$ of ethyl acetate fraction for 72 h.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.109-117
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• 2020

Cre recombinase is widely used to manipulate DNA sequences for both in vitro and in vivo research. Attachment of a trans-activator of transcription (TAT) sequence to Cre allows TAT-Cre to penetrate the cell membrane, and the addition of a nuclear localization signal (NLS) helps the enzyme to translocate into the nucleus. Since the yield of recombinant TAT-Cre is limited by formation of inclusion bodies, we hypothesized that the positively charged arginine-rich TAT sequence causes the inclusion body formation, whereas its neutralization by the addition of a negatively charged sequence improves solubility of the protein. To prove this, we neutralized the positively charged TAT sequence by proximally attaching a negatively charged poly-glutamate (E12) sequence. We found that the E12 tag improved the solubility and yield of E12-TAT-NLS-Cre (E12-TAT-Cre) compared with those of TAT-NLS-Cre (TAT-Cre) when expressed in E. coli. Furthermore, the growth of cells expressing E12-TAT-Cre was increased compared with that of the cells expressing TAT-Cre. Efficacy of the purified TAT-Cre was confirmed by a recombination test on a floxed plasmid in a cell-free system and 293 FT cells. Taken together, our results suggest that attachment of the E12 sequence to TAT-Cre improves its solubility during expression in E. coli (possibly by neutralizing the ionic-charge effects of the TAT sequence) and consequently increases the yield. This method can be applied to the production of transducible proteins for research and therapeutic purposes.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.191-194
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• 2008

The antagonistic activity of probiotic strains (Bifidobacterium animalis BB-12, Bifidobacterium bifidum A, Bifidobacterium longum B6, Lactobacillus acidophilus ADH, Lactobacillus paracasei ATCC 25598, and Lactobacillus rhamnosus GG) against nalidixic acid resistant ($NA^R$) Escherichia coli O157:H7 MF1847, E. coli O157:H7 H2439, E. coli O157:H7 ATCC 43894, and E. coli O157:H7 C7927 was investigated using the agar-overlay, well diffusion, and broth culture tests. L. paracasei ATCC 25598 was the most effective probiotic strain in terms of in vitro antagonistic activity against $NA^R$ E. coli O157:H7, followed by L. rhamnosus GG, B. longum B6, and L. acidophilus ADH. The use of selected probiotic strains could be an effective pre-harvest intervention strategy to reduce the risk of $NA^R$ E. coli O157:H7 by maintaining a balanced microflora in animals and might provide many potential benefits in lieu of using antimicrobials.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.238-245
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• 2009

Pure culture experiments were conducted to assess the bacteriostatic and bactericidal effects of phlorotannins (PT) isolated from Ascophyllum nodosum (brown seaweed) on Escherichia coli O157:H7. In Exp. 1, one non-O157:H7 strain (25922) and three strains of E. coli O157:H7 (3081, EDL933 and E318N) were cultured in M9 medium with PT included at 0 (control), 25, 50 or $100{\mu}g/ml$ (n = 3). Bacterial growth was monitored by $OD_{600}$ at 0, 4, 6, 12 and 24 h, and by dilution plating at 0, 4, 6 and 24 h. All strains were inhibited (p<0.001) by PT to varying degrees. At 50 or $100{\mu}g/ml$, PT prevented growth of all four strains. At $25{\mu}g\;PT/ml$, growth of 25922, 3081, E318N and EDL933 was inhibited for 6, 12 and 24 h, respectively, but 25922 and 3081 resumed growth by 12 and 24 h. Direct plating confirmed bactericidal effects of PT on all four strains at $100{\mu}g/ml$, and on EDL933 and E318N at $50{\mu}g/ml$. In Exp. 2, strains 25922 and 3081 were incubated with no tannins or with $50{\mu}g/ml$ of PT, purified condensed tannins (CT) from Quebracho (Schinopsis balansaei), or purified tannic acid from Rhus semialata (Anacardiaceae) as hydrolysable tannins (HT). Strain 3081 was unaffected by HT or CT, but was completely inhibited (p<0.001) by PT at 4, 6 and 24 h. Strain 25922 was unaffected by HT, slightly inhibited by CT, and almost eradicated by PT at 4 and 6 h. Transmission electron microscopy revealed tannin-mediated alterations to bacterial cell walls. Phlorotannins from A. nodosum exhibit growth-inhibiting and bactericidal effects in vitro against the strains of E. coli O157:H7 investigated. Anti-E. coli efficacy of A. nodosum PT is superior to that of terrestrial tannins purified from Quebracho and from Rhus semialata.

• The Korean Journal of Community Living Science
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• v.18 no.2
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• pp.293-300
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• 2007

The antimicrobial activity of 10 different traditional medicinal herbs extracts against E. coli was determined using paper disc method. Caesalpinia sappan and Rhus javanica, extracts in 5 mg/ml, and Paeonia suffruticosa and Seutellaria baicalensis extracts in 10 mg/ml showed significant antimicrobial activity against E. coli. Minimum inhibitory concentrations of medicinal herbs extracts were in the range of $1.4{\sim}8mg/ml\;and\;1.2{\sim}12mg/ml$, for MeOH extracts and EtOH extracts, respectively. In addition, the antimicrobial activity of each solvent fraction was most significant with the EtOAc layer. Optical density at 620nm after 24 hours incubation of E. coli in the presence of 100, 300 or 500 ppm of Caesalpinia sappan extract ranged from 0.02 to 0.1 compared to 0.6 in the absence of Caesalpinia sappan extract, indicating that growth of E. coli was significantly inhibited within 24 hours by the addition of at least 300 ppm of Caesalpinia sappan extract. Optical density at 620 nm after 24 hours incubation of E. coli in the presence of 300 ppm of Rhus javanica extract ranged from 0.02 to 0.2 compared to 0.5 in the absence of Rhus javanica extract, indicating that growth of E. coli was also significantly inhibited within 24 hours by the addition of at least 500 ppm of Rhus javanica extract. In conclusion, these findings suggest that extracts from medicinal herbs may play important roles in antimicrobial activities against E. coli.

• Food Science of Animal Resources
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• v.32 no.1
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• pp.62-67
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• 2012

This study calculated kinetic parameters of Escherichia coli O157:H7 and developed a probabilistic model to estimate growth probabilities of E. coli O157:H7 on polyethylene cutting boards as a function of temperature and time. The surfaces of polyethylene coupons ($3{\times}5$ cm) were inoculated with E. coli O157:H7 NCCP11142 at 4 Log $CFU/cm^2$. The coupons were stored at 13 to $35^{\circ}C$ for 12 h, and cell counts of E. coli O157:H7 were enumerated on McConkey II with sorbitol agar every 2 h. Kinetic parameters (maximum specific growth rate, Log $CFU/cm^2/h$; lag phase duration, h; lower asymptote, Log $CFU/cm^2$; upper asymptote, Log $CFU/cm^2$) were calculated with the modified Gompertz model. Of 56 combinations (temperature${\times}$time), the combinations that had ${\geq}$0.5 Log $CFU/cm^2$ of bacterial growth were designated with the value of 1, and the combinations that had increases of <0.5 Log $CFU/cm^2$ were given the value 0. These growth response data were fitted to the logistic regression to develop the model predicting probabilities of E. coli O157:H7 growth. Specific growth rate and growth data showed that E. coli O157:H7 cells were grown at $28-35^{\circ}C$, but there were no obvious growth of the pathogen below $25^{\circ}C$. Moreover, the developed probabilistic model showed acceptable performance to calculate growth probability of E. coli O157:H7. Therefore, the results should be useful in determining upper limits of working temperature and time, inhibiting E. coli O157:H7 growth on polyethylene cutting board.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2060-2065
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• 2016

Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at $70^{\circ}C$, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

• Journal of Life Science
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• v.21 no.12
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• pp.1710-1715
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• 2011

The bactericidal effect of nano plasma ion (NPi) which was generated by NPi was analyzed using different kinds of microorganisms, exposure times, chamber sizes, ion amounts and distance. As the result of Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Staphylococcus aureus and Bacillus subtilis were shown different in decrement. Gram-negative bacteria E. coli showed the highest percentage (96.57%) and Gram-positive bacteria B. subtilis which produced spore has the lowest percentage (57.41%). From the exposure time of NPi most of the microorganisms were extinct at an early stage. According to the size of the chamber we compared the loss of E.coli and the experiment result shown, analyzed NPi using 5 chambers $0.005m^3$ to $30m^3$ for 2 hr, that when volume of the chamber increased, saturation ion and bactericidal effect was decreased. In addition, an NPi generator installed in the $1m^3$ chamber investigated the decrement of E. coli. Saturation ion concentration increased with decrement. Finally, E. coli showed a similar reduction according to the distance from NPi generator.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.593-598
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• 2004

The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K. oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arse) kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced amino acid sequences of the K. oxytoca Dl2 Ars proteins were compared with the amino acid sequences of the E. coli R773 Ars proteins, a significant amino acid similarity was observed (87.9％ for ArsR, 89.2％ for ArsD, 83.2％ for ArsA, 92.6％ for ArsB, and 91.3％ for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R773 and K. oxytoca Dl2.

• Environmental Engineering Research
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• v.12 no.2
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• pp.64-71
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• 2007

The necessity of disinfecting playground soil is an important issue, because pathogenic protozoa, bacteria, and parasite eggs remain viable for several months and can infect children. UV irradiation has been used to decontaminate water but its effectiveness on soil is unclear. We determined the efficacy of UV radiation for inactivation of an indicator bacteria, E. coli (strain ATCC 8739), on playground soil. While 99% inactivation of E. coli in the soil was readily achieved by UV radiation within 55 min at $0.4\;mW\;cm^{-2}$, complete inactivation was not achieved, even after prolonged treatment at $4\;mW\;cm^{-2}$. This was attributed to the irregular surface of the soil. A small number of E. coli escaped the UV radiation because they were situated in indentations or under small particles on the soil surface. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) confirmed that the surface characteristics of the soil is the major limiting factor in the inactivation of E. coli by UV radiation. Thus UV treatment may not be adequate for disinfecting some soils and should be carefully evaluated before being used on playground soils.