• Korean Journal of Food Science and Technology
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• v.41 no.5
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• pp.515-521
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• 2009

The DEFT (direct epifluorescent filter technique)/APC (aerobic plate count) test was utilized to screen powdered Ramen soup samples (RS-1, RS-2) whether or not they have been microbial-decontaminated. The initial microbial loads of commercially-packaged samples were log DEFT 6.46 (RS-1) and 7.05 (RS-2), but the viable bacterial counts were log APC 2.74 (RS-1) and 1.95 (RS-2), respectively; this finding showed that they have been already decontaminated by methods other than irradiation. The same samples were then subjected to gamma irradiation at 0, 5 and 10 kGy in order to evaluate the microbial and physicochemical changes during post-irradiation storage for 6 months under room conditions ($10{\pm}3^{\circ}C$). The DEFT count was constant in irradiated samples even at different doses, but APC decreased with dose increases; this implies that the log DEFT/APC increased in a linear fashion with dose. No coliforms, yeasts and molds were detected in any of the samples, whereas the initially detected aerobic bacteria ($5.49{\times}10^2CFU/g$) were inactivated by 5 kGy or more and no growth was observed during the subsequent storage period. The pH of RS-1 was reduced by irradiation (p<0.01), but increased (p<0.01) with increasing storage time. Irradiation induced a reduction in volatile basic nitrogen (VBN), whereas an increase in thiobarbituric acid (TBA) values was observed. The storage time proved more influential than irradiation up to 10 kGy in terms of changes in the VBN and Hunter’s color values of powdered Ramen soups.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1201-1209
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• 2006

The purpose of this study was to isolate probiotic lactic acid bacteria (LAB) that produce bile salt hydrolase (BSH), and to evaluate its effects on serum cholesterol level. One-hundred-twenty bacterial colonies were initially isolated from human feces, and five strains were selected after screening based on their resistance to acids, tolerance against bile salts, and inhibitory activity on Escherichia coli. The Lactobacillus plantarum strain with the highest level of BSH activity was identified using 16S rRNA sequences, and was named L. plantarum CK 102. L. plantarum CK 102 at a level of 1.36$\times$10$^8$cfu/ml survived in pH 2 buffer for 6 h and exhibited excellent tolerance for bile salt. Coculturing the strain with E. coli in MRS broth resulted in strong inhibition against growth of E. coli at 18 h. Furthermore, the potential effect of CK 102 on serum cholesterol level was evaluated in rats. Thirty-two rats [Sprague-Dawley (SD) male, 129$\pm$l g, 5 weeks old] were divided into four groups of eight each. For six weeks, Group 1 was fed a normal diet (negative control); Group 2 was fed a cholesterol-enriched diet (positive control); Group 3 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 1.0$\times$10$^7$cfu/ml; and Group 4 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 5.0$\times$10$^7$cfu/ml. Blood samples were collected, serum lipids were analyzed, and weights of the organs were measured. Total blood cholesterol level, triglyceride, LDL-cholesterol, and free-cholesterol values were lower in rats that were fed 1. plantarum CK 102 than in those not fed L. plantarum CK 102. This cholesterol lowering effect implies that L. plantarum CK 102 could be utilized as an additive for health-assistance foods. In conclusion, these results suggest that the 1. plantarum CK 102 isolated could be used commercially as a probiotic.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.26-33
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• 2016

Dominant lactic acid bacteria (LAB) strains were isolated from traditional fermented foods to obtain rare ${\gamma}$-aminobutyric acid (GABA)-producing LAB. Out of 147 isolates, 23 strains that could produce GABA with 1% (w/v) L-monosodium glutamate (MSG) were first isolated. After further screening of these rare GABA-producing LAB by analysis of the glutamate decarboxylase and 16S rRNA gene sequences, Enterococcus faecium JK29 was isolated, and 1.56 mM of GABA was produced after 48 h cultivation in basic de Man, Rogosa, and Sharpe (MRS) medium. To enhance GABA production by E. faecium JK29, the culture conditions were optimized. When E. faecium JK29 was cultivated in optimized MRS medium containing 0.5% (w/v) sucrose and 2% (w/v) yeast extract with 0.5% (w/v) MSG, GABA production reached 14.86 mM after 48 h cultivation at initial conditions of pH 7.5 and $30^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.389-395
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• 2012

T-RFLP (terminal restriction fragment length polymorphism) analysis, one of the most highly adopted culture-independent microbial community analysis methods, was applied to evaluate the colonization of probiotics in experimental animal gut. Lactic acid bacteria that exhibited cinnamoyl esterase activity were isolated from Korean fermented vegetables and identified by 16S ribosomal RNA sequence analysis. Lactobacillus plantarum KK3, which demonstrated high chlorogenic acid hydrolysis by cinnamoyl esterase activity, and acid/bile salt resistances, was cultured, freeze-dried, and fed to mice and the microbiota in their feces were monitored by T-RFLP analysis. The T-RF of L. plantarum was detected in the feces of mice after the start of administration and lasted at least 31 days after the initial 7 day feeding. T-RFLP analysis was considered a useful tool to evaluate the gut colonization of probiotic L. plantarum. In order to prove that L. plantarum was from viable cells, we reisolated L. plantarum in the feces using cinnamoyl esterase activity media as the screening step. The colonization of L. plantarum KK3 in the mouse gut was confirmed by this research.

• KSBB Journal
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• v.26 no.6
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• pp.517-522
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• 2011

In our previous study, we isolated and characterized a 1-deoxynojirimycin (DNJ)-producing bacterium, Bacillus subtilis MORI, from chungkookjang, a Korean traditional food. B. subtilis MORI was subjected to ${\gamma}$-irradiation and the resulting bacteria were screened for increased DNJ production. A mutant was identified that produced 7.6 times more DNJ and named B. subtilis MORI 3K-85. In this study, the protein profiles of both strains were compared by one-dimensional and two-dimensional gel electrophoresis (1-DE and 2-DE, respectively) under both native and denaturing conditions. The 1-DE native-PAGE and 1-DE SDS-PAGE analyses identified 5 and 7 bands, respectively, that were found at higher concentrations in B. subtilis MORI 3K-85 than in B. subtilis MORI. Similarly, 2-DE analyses identified 20 protein spots which were found at higher concentrations in B. subtilis MORI 3K-85. The peptide mass profiles of these 20 proteins were analyzed by MALDI-TOF and compared with peptide sequences of B. subtilis and B. amyloliquefaciens in the MASCOT database. This screening suggested that three dehydrogenases, an aldolase, a synthetase, an isomerase, a reductase, and a peroxidase are elevated in B. subtilis MORI 3K-85. Based on this data, one or more of the elevated 8 enzymes might be related to the DNJ biosynthetic pathway.

• Journal of Periodontal and Implant Science
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• v.46 no.5
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• pp.320-328
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• 2016

Purpose: Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods: This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student's t-test. Results: Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434.10,139 ng/mL) and 8,598 ng/mL (range, 7,421.9,877 ng/mL), respectively. The Student's t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions: The presence of a significant difference in CSP1 levels between healthy subjects and periodontal patients suggests that CSP1 may be a potential biomarker for the detection or screening of periodontitis patients.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

• Journal of Animal Science and Technology
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• v.54 no.1
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• pp.59-63
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• 2012

Bovine serum contains various nutrients and growth factors that can be potentially used in biological experiments, drug manufacturing process and food industry. However, almost all the bovine blood has been wasted during slaughter process in Korea, thus there is a high demand for alternative uses of the wasted sera. In order to produce high quality and safe sera, it is necessary to screen zoonotic pathogens as well as other microbial contaminants to prevent any downstream contamination. The present research has been undertaken to assess biological safety of Hanwoo sera by determining microbiological contamination during slaughtering and handling processes. Serological tests have been performed to detect bacteria, mycoplasma and virus contamination in total of 52 Hanwoo sera. No sera were found to be contaminated with mycoplasma or virus, but only two sera were found to be contaminated with Bacillus thuringiensis. The present result shows that Hanwoo sera obtained from slaughtering process are biologically safe and have potentials to be developed as a biological reagent. Moreover, the methods employed in our study may provide basic standard for microbiological screening methods once wasted Hanwoo sera gain industrial values.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.94-101
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• 2016

Acinetobacter species isolates are important opportunistic pathogens and commonly implicated in nosocomial infections. The therapeutic options for treatment of the bacterial infections are limited because the bacteria isolates are usually multidrug resistant (MDR). In the current study, we investigated various carbapenemase genes in 68 Acinetobacter species isolates. Antimicrobial susceptibilities were tested using the disk diffusion method. Screening of carbapenemase genes was performed via multiplex PCR. In addition, PCR and DNA sequencing were used to identify the carbapenemase genes. Repetitive extragenic palindromic-PCR (REP-PCR) was also performed to assess the clonality of isolates. In our study, A. baumannii isolates were highly resistant to all agents tested while all non-A. baumannii isolates were susceptible to all agents tested, with the exception of aztreonam and cefotaxime. All 51 A. baumannii isolates contained the $bla_{OXA-51}$ gene and 37 (72.5%) isolates also harbored the $bla_{OXA-23}$ gene. In addition, 39 MDR A. baumannii isolates were identified in our study and 37 isolates contained the $bla_{OXA-23}$ gene. The 37 MDR strains harboring $bla_{OXA-23}$ showed type I (n=22) or type II (n=15) banding patterns on their REP-PCR profiles. Our results suggest clonal relation and horizontal spreading of MDR A. baumannii isolates containing the $bla_{OXA-23}$ gene at the hospital located in Daejeon. Continuous investigation of antimicrobial resistant determinants and monitoring emergence and dissemination of MDR isolates is required to prevent and control infection and colonization of MDR A. baumannii isolates.

• Korean Journal of Food Science and Technology
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• v.25 no.5
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• pp.582-588
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• 1993

In order to investigate the effects of food materials toward the growth of Bifidobacterium spp. and Clostridium perfringens which have great influences on the intestinal physiology of human, 162 kinds of foodstuffs and foods were collected. Among their extracts, 31 samples showed the inhibitory effects against the growth of B. bifidum and C. perfringens by agar diffusion method. Especially, the methanol extracts of Caltha palustris, Deonjang, onion, mustard and potato inhibited the growth of C. perfringens, while they did not remarkably inhibit other intestinal bacteria including Bifidobacterium spp. By the cultivation of faecal inoculum in the 1 %(v/v) extract broths of Caltha palustris, onion and mustard, population of Bifidobacterium spp. increased by 10 order and that of C. perfringens decreased. ${\beta}$-glucuronidase activities and indole amounts in the cultures of onion and mustard extracts were lower than those of the control culture and ${\beta}-glucosidase$ activities were not detected in the cultures of onion and Doenjang extracts.