• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1706-1712
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• 2006

Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

• Journal of Environmental Health Sciences
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• v.39 no.1
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• pp.71-82
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• 2013

Objectives: This study was performed in order to investigate hand hygiene practices among food-service businesses employees based on the awareness of hand-washing and load of indicator bacteria on their hands. It focused on the comparison of full-time and part-time workers in food-service workplaces. Methods: A direct-interview questionnaire survey and microbiological analysis were carried out with sixty workers each. Samples for microbiological analysis were collected through a modified glove-juice method from the hands of the food-service workers and were analyzed for aerobic plate count, total coliform, fecal coliform, Escherichia coli, Staphylococcus aureus, and Salmonella spp. Microbiological analysis was done according to the Food Code of Korea. Results: Significant differences (p<0.01) were found in the survey between the full-time and part-time workers in hand-washing frequency, use of hand-washing agents, and hand-drying methods. More full-time workers responded to washing their hands after preparing food, after visiting outside, after handling raw materials, and before putting on gloves/when changing gloves than did part-time workers (p<0.05). No remarkable difference was found in bacterial load on the hands except in the aerobic plate count between the two groups. The detection of E. coli, S. aureus, and Salmonella spp. on the hands of some food-service workers in both groups revealed poor hand hygiene practices. Conclusions: The results of this study indicate that there is a need for training programs in order to improve hand hygiene practices and strict hand hygiene compliance by food-service workers.

• Food Science of Animal Resources
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• v.34 no.5
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• pp.614-621
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• 2014

This study was conducted to isolate and characterize bacteriocin-producing bacteria against Clostridium perfringens (C. perfringens) from domestic animals to determine their usefulness as probiotics. Bacteriocin-producing bacteria were isolated from pig feces by the spot-on-lawn method. A total of 1,370 bacterial stains were isolated, and six were tentatively selected after identifying the inhibitory activity against the pathogenic indicator C. perfringens KCTC 3269 and KCTC 5100. The selected strains were identified as Enterococcus faecalis (E. faecalis) by 16s rRNA sequencing. Most of the isolated bacterial strains were resistant to 0.5% bile salts for 48 h and remained viable after 2 h at pH 3.0. Some E. faecalis also showed strong inhibitory activity against Listeria monocytogenes KCTC 3569, KCTC 3586 and KCTC 3710. In the present study, we finally selected E. faecalis AP 216 and AP 45 strain based on probiotic selection criteria such as antimicrobial activity against C. perfringens and tolerance to acid and bile salts. The bacteriocins of E. faecalis AP 216 and AP 45 strains were highly thermostable, showing anticlostridial activities even after incubation at $121^{\circ}C$ for 15 min. These bacteriocin-producing bacteria and/or bacteriocins could be used in feed manufacturing as probiotics as an alternative to antibiotics in the livestock industry.

• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1271-1278
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• 2012

Genetic fusion of two proteins frequently induces beneficial effects to the proteins, such as increased solubility, besides the combination of two protein functions. Here, we study the effects of the bacterial surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a on the folding of a C-terminally fused enhanced green fluorescent protein (EGFP) moiety. Although GFPs are generally unable to adopt a functional confirmation in the bacterial periplasm of Escherichia coli cells, we observed periplasmic fluorescence from a chimera of a 150-amino-acid N-terminal truncation of SgsE and EGFP. Based on this finding, unfolding and refolding kinetics of different S-layer-EGFP chimeras, a maltose binding protein-EGFP chimera, and sole EGFP were monitored using green fluorescence as indicator for the folded protein state. Calculated apparent rate constants for unfolding and refolding indicated different folding pathways for EGFP depending on the fusion partner used, and a clearly stabilizing effect was observed for the SgsE_C fusion moiety. Thermal stability, as determined by differential scanning calorimetry, and unfolding equilibria were found to be independent of the fused partner. We conclude that the stabilizing effect SgsE_C exerts on EGFP is due to a reduction of degrees of freedom for folding of EGFP in the fused state.

• Journal of Environmental Health Sciences
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• v.38 no.6
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• pp.520-528
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• 2012

Objective: This study was performed to assess bacterial distribution concerned in sanitation management of public lavatory in Seoul. Methods: In this research, bacteriological investigation on public lavatory was accomplished for bidet water, bidet nozzle, washbowl and lavatory stool in the 50 public facilities such as public institutions, subway stations, cinema, department stores, large-scale buildings and hospitals amount to 374 specimens. Results: The geometric mean of colony forming unit(CFU) in total aerobic colony count were analyzed as follows; $5.2{\times}10^2/100cm^2$ on lavatory stool, $7.2{\times}10^3/ea$ on bidet nozzle, $7.8{\times}10^3/ea$ on center ring of washbowl, $1.4{\times}10/mL$ in bidet water (ml) and 7.0/ea on doorknob. Opportunistically pathogenic germs such as Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa were isolated in 3.7%, 5.9%, 3.2% and 1.9% of total specimens, respectively. Conclusion: The result of this study shows that there were some facilities where the pathogenic germs were detected to may cause urological infection. And the CFU of general bacteria as the representative indicator of disinfection and lavatory cleaning were high enough to imply the improvement of sanitation management of public lavatories should be contrived.

• Journal of the East Asian Society of Dietary Life
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• v.10 no.2
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• pp.160-169
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• 2000

This study was conducted to understand the inhibitory garlic and ginger against the growth of food born pathogenic bacteria. Juice was prepared from the raw spices by using an electric homogenizer and membrane filter. Dry-powdered spices were treated with double distilled water and 70% ethanol to extract the antibacterial substances, respectively. Growth inhibitory effects of juice and extracts of the spices were monitored by using bacterial strains such as B. subtilis, L. moncytogenes, S. aureus,E. coli O157 : H7, P. aeruginosa, and S. typhimurium. On a solid medium where E. coli and S. aureus cells were grown, ginger juice formed inhibitory zone at the concentrations of 2-10% by paper disc test. The Bone formed by ginger juice was wider and more transparent than that formed by garlic juice on the same concentration.1. monocytogenes and B. subtilis were more sensitive to garlic juice than others, and stopped growing at 2% garlic juice. Ginger juice showed the growth inhibition by 30-50% at 1.0% concentration. On the contrast, P. aeruginosa which resisted to the garlic juice was the most sensitive to ginger juice. Water extract of garlic was not effective to inhibit the bacterial growth, while 2% ginger extract completely inhibited the growth of E. coli and S. aureus. Alcohol extract of ginger inhibited the growth of bacteria at the concentration of 0.3%. This growth inhibition is almost 10 times lower than that of the garlic extract. It was clear that ginger had more potential than garlic as an inhibitor to control the growth of the indicator organisms.

• Korean Journal of Plant Resources
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• v.23 no.3
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• pp.248-255
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• 2010

We previously demonstrated that root colonization of the rhizobacterium, Pseudomonas chlororaphis O6, induced expression of a galactinol synthase gene (CsGolS1), and resulting galactinol conferred induced systemic resistance (ISR) against fungal and bacterial pathogens in cucumber leaves. To examine the role of galactinol on ISR, drought or high salt stress, we obtained T-DNA insertion Arabidopsis mutants at the AtGolS1 gene, an ortholog of the CsGolS1 gene. The T-DNA insertion mutant compromised resistance induced by the O6 colonization against Erwinia carotovora. Pharmaceutical application of 0.5 - 5 mM galactinol on roots was sufficient to elicit ISR in wild-type Arabidopsis against infection with E. carotovora. The involvement of jasmonic acid (JA) signaling on the ISR was validated to detect increased expression of the indicator gene PDF1.2. The T-DNA insertion mutant also compromised tolerance by increasing galactinol content in the O6-colonized plant against drought or high salt stresses. Taken together, our results indicate that primed expression of the galactinol synthase gene AtGolS1in the O6-colonized plants can play a critical role in the ISR against infection with E. carotovora, and in the tolerance to drought or high salt stresses.

• The Korean Journal of Food & Health Convergence
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• v.5 no.3
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• pp.35-40
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• 2019

In this study, general bacterial counts and coliform counts, which are hygienic indicator microorganisms, were tested for candy, chocolate, and jelly which are easily available and enjoyed around. After dropping each sample on the desk, indoors, and outdoors, it is immediately collected, or washed and collected to confirm the myth of the 3-second rule. Immediately after removing the wrapping paper, each sample was dropped on the desk, indoors, and outdoors, and after 3 seconds from the moment of contact with the surface, and then collected in a sample bag using sterilized sanitary gloves. After the same operation, each sample was rinsed for 5 seconds using sterilized sanitary gloves and sterilized distilled water, and then collected in a sample bag. The number of bacteria detected in non-washing candies was 41 CFU/g at outdoor and the number of bacteria detected in non-washing chocolate was 76 CFU/g at outdoor. The number of bacteria detected in non-washing jellies was 79 CFU/g at outdoor. Coliform group was not detected in all samples. This showed good results at the level of m = 10,000 or less, which is an allowable value suggested in the Food Code. Also, effect of washing on contaminated food was confirmed. This result is remarkably low compared with the microorganism specimens shown in Food Code, and it is confirmed that contamination occurs but not high value. Therefore, the myth of the 3-second rule is true compared to the figures based on Food Code. However, it showed the characteristics of bacteria that could survive and cross-contaminate on dry food surfaces and emphasized the importance of hygiene through food contact to unsanitary surfaces to minimize the risk of food poisoning.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.690-695
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• 2014

Bacterial 1-aminocyclopropane-1-carboxlyate (ACC) deaminase (AcdS) is an enzyme that cleaves ACC, a precursor of the plant hormone ethylene, into ${\alpha}$-ketobutyrate and ammonia. The acdS gene was cloned from Pseudomonas fluorescens, which was capable of improving the seedling of Chinese cabbage under salinity condition. The recombinant AcdS (rAcdS) exhibited optimal activity at pH 8.5 and $30^{\circ}C$. Strong activity was sustained at up to 100 mM NaCl. The polyclonal anti-P. fluorescens AcdS antibody was produced in a rabbit that had been immunized with the purified rAcdS. This antibody successfully recognized the homologous antigens derived from the total proteins of isolated plant growth-promoting microorganisms. A statistically significant correlation was observed between the intensity of hybridization signal and AcdS activity measured by a biochemical method, suggesting its application as a useful indicator for active deaminases.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1060-1067
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• 2006

This study was undertaken to determine the effect of reservoirs on water quality and the distribution of pathogenic and indicator bacteria in a drinking water distribution system (total length 14km). Raw water, disinfected water, and water samples from the distribution system were subjected to physicochemical and microbiological analyses. Most factors encountered at each season included residual chloride, nitrate, turbidity, and phosphorus for heterotrophic bacterial distribution, and hardness, heterotrophic bacteria, sampling site, and DOC (dissolved organic carbon) for bacteria on selective media. No Salmonella or Shigella spp. were detected, but many colonies of opportunistic pathogens were found. Comparing tap water samples taken at similar distances from the water treatment plant, samples that had passed through a reservoir had a higher concentration of heterotrophic bacteria, and a higher rate of colony formation with 10 times as many bacteria on selective media. Based on the results with m-Endo agar, the water in reservoirs appeared safe; however, coliforms and opportunistic pathogenic bacteria such as Pseudomonas aeruginosa were identified on other selective media. This study illustrates that storage reservoirs in the drinking water distribution system have low microbiological water quality by opportunistic pathogens, and therefore, water quality must be controlled.