• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.801-806
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• 2002

A multiplex competitive polymerase chain reaction (PCR) method was developed for the rapid identification and quantification of Leuconostoc mesnteroides and Lactobacillus plantarum populations which are the key microorganisms in kimchi fermentation. The strain-specific primers were designed to selectively amplify the target genes encoding 165 rRNA of L. plantarum and dextransucrase of L. mesenteroides. There was a linear relationship between the band intensity of PCR products and the number of colony forming units of each model organism. The PCR quantification method was compared with a traditional plate-counting method f3r the enumeration of the two lactic acid bacteria in a mixed suspension culture and also applied to a real food system, namely, watery kimchi. The population dynamics of the two model organisms in the mixed culture were reliably predictable by the competitive PCR analysis.

• Journal of Ginseng Research
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• 제34권4호
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• pp.321-326
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• 2010

Enzymatic treatment conditions for red ginseng residue (RGR) were investigated to apply RGR as a microbial medium. Polysaccharide hydrolyase and protease were screened to obtain high solid and carbohydrate yields, and a good degree of carbohydrate hydrolysis. The optimal dosage and reaction time for Viscozyme, the chosen polysaccharide hydrolyase, were found to be 1.0% (w/w) and 3 h, respectively. Of the tested proteases, Flavourzyme, whose optimal dosage was 0.5% (w/w), was selected. Co-treatment with the optimal dosages of Flavourzyme and Viscozyme increased solid yield, carbohydrate yield, and degree of carbohydrate hydrolysis by 76%, 65%, and 1,865%, respectively, over levels in non-treated RGR. The culture characteristics of Leuconostoc mesenteroides strain KACC 91459P grown in enzymatically hydrolyzed red ginseng residue (ERGR) and RGR suspensions were compared. After cultivation for 6 h, the viable cell counts of both cell suspensions rapidly increased to $1.3{\times}10^9$ colony-forming units (CFU)/g. Moreover, while the viable cell population drastically decreased to $2.4{\times}10^6\;CFU/g$ for cells grown in RGR medium, it was maintained in cells fermented in ERGR medium for 24 h.

• International Journal of Oral Biology
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• 제40권3호
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• pp.143-146
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• 2015

The purpose of this study was to compare the effect of different methods of hand washing by counting the number of bacteria on the hand surface. Eighteen clinicians were chosen and divided into three groups, consisting of six clinicians each. Culturing of the right raw palms of all individuals was performed. Individuals in the control group washed hands for 5 seconds with antimicrobial soap. Group 1 washed their hands for 10 seconds with antimicrobial soap. Group 2 washed with an instant alcohol-based hand sanitizer. After the respective washes, re-culturing of the right raw palm was done for each member of all groups. The colony-forming units (CFU) were calculated at each time point, and the reduction rate of CFU among the three groups were statistically evaluated using student t-test. All groups showed a significant decrease in CFU, according to the time applied (P<0.01). In addition, the reduction rate of CFU between the groups were statistically evaluated with ANOVA (P<0.01). It showed statistically difference between the control group and group 1, control group and group 2. The present study confirmed that the hand washing method with antimicrobial soap for 10 seconds and hand sanitizer, including alcohol, were excellent for decreasing the number of bacteria on the hand surface.

• Preventive Nutrition and Food Science
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• 제8권2호
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• pp.196-199
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• 2003

We investigated the antagonistic effects of two lactococcal bacteriocins, nisin or lacticin NK24, on the growth and the survival of Listeria monocytogenes in vacuum or modified atmosphere-packaged ground beef, Ground beef was inoculated with approximately 3 log colony-forming units (CFU) of L. monocytogenes ATCC 15313 culture per gram of ground beef. Inoculated samples were blended with/without 100 AU/g nisin or lacticin NK24, and subsequently vacuum or modified atmosphere packed at 4$^{\circ}C$. Listeria in the bacteriocin-treated and control samples was subsequently isolated from both vacuum and modified atmosphere packs and enumerated as CFU on Listeria Isolation Agar medium. Microbial counts in ground beef treated with bacteriocin declined steadily, while those of non-treated beef samples increased steadily. The results obtained demonstrate that nisin inhibits the growth of L. monocytogenes more effectively than lacticin NK24 at 100 AU/g. The use of lactococcal bacteriocins, such as nisin or lacticin NK24, in vacuum or modified atmosphere packaging offers a promising approach for eliminating or reducing the risk of L monocytogenes contamination in ground beef.

• 약학회지
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• 제57권6호
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• pp.383-387
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• 2013

In the present study, we determined the antifungal effect of obacunone isolated from Phellodendri Cortex against Candida ablicans, a pathogenic fungus. The antifungal effect was analyzed by an in-vitro susceptibility test and in a murine model of disseminated candidiasis. Possible mechanism of the antifungal activity was also examined. Analyses of data resulting from the susceptibility test revealed that the compound inhibited C. albicans growth. At 25 ${\mu}g$ obacunone/ml, there was app. 45% reduction of CFUs (colony forming units) as compared to obacunone-untreated C. albicans yeast cells (P<0.01). In the murine model of disseminated candidiasis due to C. albicans, obacunone enhanced resistance of mice against disseminated candidiasis. During an entire period of 30-day observation, control animals all died within 14 days, whereas 60% of obacunone-treated mice survived (P<0.05). In addition, obacunone inhibited the hyphal production, a major virulence factor of C. albicans, from the blastoconidial form. Thus, obacunone appears to have antifungal activity for C. albicans infection. This may possibly be mediated by the blockage of hyphal production.

• 한국식물병리학회지
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• 제14권2호
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• pp.164-170
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• 1998

This study was carried out to develop DNA probe for specific detection of Erwinia carotovora subsp. carotovora. Universal rice primer (URP, 20 mer) developed from repetitive sequence of rice was applied for producing PCR DNA fingerprints of Erwinis spp. In E. carotovora subsp. carotovora strains, primer URP2F amplyfied polymorphic bands which are distinguisable from other Erwinia spp. A PCR band of 0.6 kb selected from PCr polymorphic bands of E. carotovora subsp. carotovora strains was cloned and evaluated as a diagnostic DNA probe. Among 28 bacterial strains including 22 Erwinia spp, the probe (pECC2F) only hybridized to total DNAs from e. carotovora subsp. carotovora strains and E. carotovora subsp. wasabiae, but sizes of hybridized bands were different between these subspecies, 10.0 kb and 3.5 kb respectively. In dot blot assays using probe pECC2F, as few as 103 colony forming units (CFU) of E. carotovora subsp. carotovora could be detected in a suspension containing about 1$\times$103 CFU of soil bacteria.

• Food Science and Biotechnology
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• 제14권3호
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• pp.387-391
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• 2005

Competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Listeria monocytogenes in pork meat. Pork meat was artificially inoculated with L. monocytogenes and DNA was extracted using guanidine thiocyanate-phenol-chloroform and subjected to PCR amplification. Sixteen primer sets for L. monocytogenes hlyA gene were tested for sensitive detection and the DG69/DG74 primer set was selected. The detection limit achieved with this primer set was as low as 860 colony-forming units (cfu) per 0.1 g of pork meat. When the samples were cultured at $30^{\circ}C$ for 16 hr in Brain Heart Infusion (BHI) medium, even a single bacterium could be detected with this primer set by PCR. For cPCR, the hlyA gene, which features a 148 bp-deletion, was cloned in the pGEM-4Z vector. A known amount of competitor DNA which has the same primer binding sites was co-amplified with L. monocytogenes total DNA from the artificially inoculated pork meat. The cell-number determined by cPCR was approximately equal to cfu from the Most Probable Number (MPN) method. The whole procedure took only 5 hr.

• Food Science and Biotechnology
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• 제15권3호
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• pp.458-461
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• 2006

To determine whether the toxicity of Bacillus cereus would be seen in human cell lines and mice, we screened B. cereus B-38B, B. cereus B-50B, and B. cereus KCCM40935 for genes that coded for 5 enterotoxins using the polymerase chain reaction and cultivated them for 17 hr, by whose time they had grown to $10^7-10^8$ colony-forming units (CFU) per milliliter. Cell-free supernatant was added to make up 1% of the total reaction solution. Human cells from normal lung, lung carcinoma, embryonic kidney, and cervical adenocarcinoma cell lines were grown in culture. The cytotoxicity induced by adding the reaction solution was indicated by cell death rates of 0 to 70%, depending on the bacterial strain involved and the cell line. A lethality of 20% was observed when B. cereus cultures containing $10^7-10^8$ viable cells were administrated orally to mice. Therefore, the culture of B. cereus containing $10^7-10^8$ viable cells seems to have high cytotoxicity on human cell lines and lethality on mice.

• 한국환경과학회지
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• 제30권5호
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• pp.369-378
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• 2021

In this study, we investigated the effects of indole on biofilm formation inhibition in Pantoea agglomerans (P. agglomerans). In the biofilm growth assay, indole inhibited biofilm formation across all the growth time. Depending on biofilm growth stage, indole exhibited biofilm inhibition and anti-bacterial effects on planktonic cells. Through the analysis of the proportion rate between biofilm and Colony Forming Units (CFU) and inhibition rate of indole, we confirmed that depending on the biofilm stage of P. agglomerans, indole treatment timing was more important than the treatment duration. By comparing gene expression rates through rt-qPCR P.agglomerans affected by indole was found to significantly change quorum sensing (pagI/R) and indole transportation (bssS) gene expressions. Throughout all, indole exhibited both antimicrobial and anti-biofilm effects on P. agglomerans. In addition, we confirmed the anti-biofilm effects of indole on mature biofilm. In conclusion, indole as a signal molecule, can exhibit anti-biofilm effects through bacterial quorum sensing inhibition and indole affects. Therefore, indole can regulate biofilm bacteria especially gram-negative opportunistic pathogens.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.429-438
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• 2020

The emergence of multi-drug resistant, pathogenic methicillin-resistant Staphylococcus aureus (MRSA) is a threat to global health and has created a need for novel functional therapeutic agents. In this study, we evaluated the underlying mechanisms of the anti-MRSA effect of an azaphilone pigment, sclerotiorin (SCL) from Penicillium sclerotiorum. The antimicrobial activity of SCL was evaluated using agar disc diffusion, broth microdilution, time-kill assays and biophysical studies. SCL exhibits selective activity against Gram positive bacteria including MRSA (range, MIC = 128-1028 ㎍/ml) and exhibited rapid bactericidal action against MRSA with a > 4 log reduction in colony forming units within three hours of administration. Biophysical studies, using fluorescent probes and laser or electron microscopy, demonstrated a SCL dose-dependent alternation in membrane potential (62.6 ± 5.0.4% inhibition) and integrity (> 95 ± 2.3%), and the release of UV260 absorbing materials within 60 min (up to 3.2 fold increase, p < 0.01) of exposure. Further, SCL localized to the cytoplasm and hydrolyzed plasmid DNA. While in vitro checkerboard studies revealed that SCL potentiated the antimicrobial activity of topical antimicrobials such as polymixin, neomycin, and bacitracin (Fractional Inhibitory Concentration Index range, 0.26-0.37). Taken together these results suggest that SCL targets the membrane and DNA of MRSA to facilitate its anti-MRSA antimicrobial effect.