• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Pediatric Infection and Vaccine
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• v.24 no.2
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• pp.95-101
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• 2017

Purpose: Acute bacterial gastroenteritis (ABG) can cause more severe symptoms than acute viral gastroenteritis in children. This study was aimed at determining the etiologic trends and to examine the clinical characteristics of ABG in children. Methods: We sent stool samples from the children with acute gastroenteritis who were treated at a secondary hospital located in Seoul, Korea between January 2011 and December 2014 to Seoul Metropolitan Government Research Institute of Public Health and Environment to find the causative organisms. Clinical characteristics of patient were analyzed through a medical records review. Results: Out of 664 stool samples, 183 (27.6%) yielded bacterial pathogens. Staphylococcus aureus was the most common bacterial pathogen, found in 72 cases (39.3%), even though it was only tested for since 2012. The monthly isolation rate was the highest (24.6%) in August. The isolation rate of Campylobacter spp. by patient's age group was high (16.7%) in the 12- to 18-year-age group (P=0.04). In patients with bloody stool, Campylobacter spp. was the most commonly isolated (31.0%, P=0.04). When comparing C-reactive protein, the Salmonella spp.- or Campylobacter spp.-isolated group showed higher values than the S. aureus- or pathogenic Escherichia coli-isolated group ($5.7{\pm}0.6mg/dL$ vs. $2.1{\pm}0.3mg/dL$, P<0.01). Conclusions: S. aureus, Salmonella spp., pathogenic E. coli, and Campylobacter spp. were important pathogens of ABG among children. Considering the differences in pathogens found according to age, a clinical symptom and inflammation index might be helpful in assuming the causative organism.

• Journal of Life Science
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• v.26 no.11
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• pp.1259-1268
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• 2016

In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by $C_{18}$ reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], $20.8{\mu}g/ml$), Bacillus subtilis (MEC, $0.2{\mu}g/ml$), Streptococcus mutans (MEC, $0.2{\mu}g/ml$), gram-negative bacteria including Pseudomonas aeruginosa (MEC, $5.7{\mu}g/ml$), Escherichia coli (MEC, $2.6{\mu}g/ml$) and fungi, Candida albicans (MEC, $56.3{\mu}g/ml$). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.249-254
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• 2002

To determine the abilities as both lactic starter and probiotics for fermented foods, we investigated the potency of acid production, proteolytic activity and lactose metabolism of Lactobacillus amylovorus IMC-1. And the strain was cultured with lactococci in 10% skim milk medium. It was also examined the bactericidal action of antibacterial substance, produced by the strain IMC-1, against pathogenic bacteria. L. amylovorus IMC-1 showed excellent production of acid in 10% skim milk supplemented with yeast extract, and produced 0.8 and 2.7% of acid at 12 and 72 h incubation, respectively. It was found that the activity of ${\beta}-galactosidase$, about $39\;{\mu}M/minute/dry$ cell weight (mg), was stronger than that of $phospho-{\beta}-galactosidase$ in the strain IMC-1. The strain showed weak proteolytic activity in 10% skim milk, thus it produced 6 and $69\;{\mu}g/mL$ of free tyrosine at 12 and 72 h cultivation, respectively. It was known that the strain utilized mainly ${\alpha}-casein$ than ${\beta}-casein$ from patterns of SDS-PAGE. Mixed culture produced more acid than single cultures of L. amylovorus IMC-1 and Streptococcus thermophilus NIAI 510. Single culture of Str. thermophilus and mixed culture showed increasing cheese flavor with incubation times. Optimal fermentation time of mixed culture for the acid production and flora of lactic starter was 16 and 12 h by adding 0.1 and 0.5% of yeast extract to 10% skim milk, respectively. Antibacterial substance produced by the strain IMC-1 reduced about 2 log of the viable cell counts of both Escherichia coli O157 and Shigella flexneri after 24 and 4 h incubation, and they were not detected after 48 and 6 h incubation, respectively.

• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.414-419
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• 2016

The aims of this study were to investigate microbial contamination levels and to survey sanitation management between Good Agricultural Practices (GAP) and non-GAP farms of lettuce and cucumber. The samples (lettuce, cucumber, soil, agricultural water, gloves, and packing plastic bag) were tested to analyze sanitary indicator bacteria (total aerobic bacteria, coliforms and Escherichia coli) and major pathogenic bacteria (Staphylococcus aureus, Bacillus cereus). In the lettuce farms, the contamination levels of total aerobic bacteria and coliforms in GAP farms were little lower than non-GAP farms or similar. Staphylococcus aureus and Bacillus cereus in soil and agricultural water of GAP farms were detected at higher levels than non-GAP farms in soil and agricultural water. In the case of cucumber farms, levels of total bacteria and Bacillus cereus in soil and total bacteria and coliform in gloves of GAP farms were higher than those of non-GAP farms, and other bacteria contamination levels in collected samples were similar. These results indicate that agricultural products produced from GAP farm still exhibited potential microbial risks. According to the field survey, a sanitation management in GAP farms was insufficient. These results could be useful as basic data to suggestion of plan for preventing microbial contamination and to improvement of GAP certification.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1202-1208
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• 2009

Synbiotics is the term used for a mixture of probiotics (live microbial feed additives that beneficially affects the host animal) and prebiotics (non-digestible food ingredients that beneficially affect the organism). This study investigated the effect of probiotics from anaerobic microflora with prebiotics on growth performance, nutrient digestibility, noxious gas emission and fecal microbial population in weaning pigs. 150 pigs with an initial BW of 6.80${\pm}$0.32 kg (20 d of age) were randomly assigned to 5 dietary treatments as follows: i) US, basal diet+0.15% antibiotics (0.05% oxytetracycline 200 and 0.10% tiamulin 38 g), ii) BS, basal diet+0.2% synbiotics (probiotics from bacteria), iii) YS, basal diet+0.2% synbiotics (probiotics from yeast), iv) MS, basal diet+0.2% synbiotics (probiotics from mold), v) CS, basal diet+0.2% synbiotics (from compounds of bacteria, yeast and mold). The probiotics were contained in $10^{9}$ cfu/ml, $10^{5}$ cfu/ml and $10^{3}$ tfu/ml of bacteria, yeast and molds, respectively. The same prebiotics (mannan oligosaccharide, lactose, sodium acetate and ammonium citrate) was used for all the synbiotics. Pigs were housed individually for a 16-day experimental period. Growth performance showed no significant difference between antibiotic treatments and synbiotics-added treatments. The BS treatment showed higher (p<0.05) dry matter (DM) and nitrogen digestibility while ether extract and crude fiber digestibility were not affected by the dietary treatment. Also, the BS treatment decreased (p<0.05) fecal ammonia and amine gas emissions. Hydrogen sulfide concentration was also decreased (p<0.05) in BS, YS and MS treatments compared to other treatments. Moreover, all the synbioticsadded treatments increased fecal acetic acid concentration while the CS treatment had lower propionic acid concentration than the US treatment (p<0.05) gas emissions but decreased in fecal propionate gas emissions. Total fecal bacteria and Escherichia coli populations did not differ significantly among the treatments, while the Shigella counts were decreased (p<0.05) in synbiotics-included treatment. Fecal bacteria population was higher in the YS treatment than other treatments (p<0.05). The BS treatment had higher yeast concentration than YS, MS and CS treatments, while US treatment had higher mold concentrations than MS treatment (p<0.05). Therefore, the results of the present study suggest that synbiotics are as effective as antibiotics on growth performance, nutrient digestibility and fecal microflora composition in weaning pigs. Additionally, synbiotics from anaerobic microflora can decrease fecal noxious gas emission and synbiotics can substitute for antibiotics in weaning pigs.

• Journal of Environmental Science International
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• v.25 no.6
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• pp.789-798
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• 2016

Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.5
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• pp.721-726
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• 2012

Lactic acid bacteria (LAB) are able to secrete antimicrobial peptides called bacteriocins, which inhibit other bacteria such as pathogenic microorganisms. Therefore, bacteriocin-producing starters can be used as natural biopreservatives for various foods. The objective of this study was to screen and characterize bacteriocin-producing LAB from Kimchi and to investigate their applicability as a starter in Kimchi fermentation. To screen bacteriocin-producing LAB, gram-positive and gram-negative bacteria were used as indicators. To measure the antimicrobial activities of isolates, agar well diffusion assay method was used. According to the results, bacteriocin produced by $Lb.$ $sakei$ B16 showed antimicrobial activity against $Listeria$ $monocytogenes$ ATCC 19115, $Escherichia$ $coli$ KCTC 1467, and$Lactobacillus$ $plantarum$ KTCT 3104. Furthermore, bacteriocin was very stable after treatment with high temperature and high and low pH, but its effects were inhibited by treatment with proteolytic enzymes such as trypsin, proteinase K, and ${\alpha}$-chymotrypsin, revealing their bacteriocin-like protein- based structure. These results suggest that $Lb.$ $sakei$ B16 and its bacteriocin are good candidates as a functional probiotic and natural biopreservative, respectively, in fermented foods.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.270-277
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• 2010

In order to manufacture Bacillus cereus-free fermented soybean products, an antimicrobial agentproducing isolate against B. cereus was obtained from 150 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK 121057 was most closely related to Bacillus licheniformis. The B. licheniformis isolate began to produce the antimicrobial agent after 48 h of incubation. The agent was nonproteinaceous and insensitive to heat, long term storage and protease K. Electron microscopic observation indicated that the agent attacked the membrane of B. cereus, leaving the ghost cell. The isolate inhibited growth of B. subtilis, Lactobacillus brevis and various types of pathogenic strains including Escherichia coli, E. faecalis, Micrococcus luteus, Staphylococcus aureus, Aspergillus flavus, A. ochraceus, and A. parasiticus as well as B. cereus. After coinoculation of B. licheniformis SCK 121057 and B. cereus in the ratio (as the basis of CFU/g sample) of 10 to 1 on the surface of cooked soybeans, cell numbers of B. cereus had been dramatically reduced after 31 days of incubation compared to those of single inoculation of B. cereus.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.308-312
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• 2011

Plant lactic acid bacteria were isolated from plant-associated fermentative foods and crops, and their probiotic properties were investigated. Isolates K27 and O2 were isolated from Kimchi and onion, and identified as Lactobacillus plantarum on the basis of 16S rRNA gene analysis. The two strains were highly resistant to acid (an MRS broth at pH 2.5), where the survival rates of L. plantarum K27 and L. plantarum O2 were 90.2% and 97.3%, respectively. L. plantarum K27 and L. plantarum O2 also showed high bile resistance to 0.5% oxgall, with a more than 70% survival rate. They showed an inhibitory effect against pathogenic strains of Escherichia coli KCCM 40880 and Pseudomonas aeruginosa ATCC 10145. The antibacterial effect of the two strains was probably due to the presence of lactic acid. ACE inhibitory activities of the two strains ranged from 72.8% to 80.6% in MRS broth. Notably, the two strains showed high ACE inhibitory activity (89.2~98.2%) in MRS broth containing 10% skim milk. Antioxidant activity was tested by DPPH radical scavenging activity, with antioxidant activities of the strains being in the range of 56.8~61.5%. The results obtained in this study suggest that L. plantarum K27 and L. plantarum O2 may be potential probiotic starter cultures with applications with fermentative products.