• Journal of Food Hygiene and Safety
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• v.23 no.3
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• pp.248-256
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• 2008

To investigate the epidemiological characteristics of 68 Listeria monocytogenes isolates, including 11 reference strains and 57 isolates from imported US beef, domestic meats(beef, pork, chicken meat), raw milk, and milk plants. L. monocytogenes was to evaluate the production of virulence proteins, such as hemolysin(LLO) and lecithinase(LCP), the adsorption of Congo red(CRA), and to detect virulence genes using the polymerase chain reaction(PCR). In the study of virulence protein production, 68(100%), 62(91.2%), and 54(79.4%) of the 68 L. monocytogenes strains were positive for LLO production, the LCP test, and the CRA test, respectively, while strains of other species, such as L. innocua, L. gray, L. murrayi, and L. welshimeri, were not. There were no significant differences between L. monocytogenes serotypes and the ability to produce LLO or LCP. L. monocytogenesstrains had very high hemolytic titers(2 to 16 fold), while the other Listeria species, other than L. ivanovii and L. seeligeri, did not. The hemolysin activities of L. monocytogenes, L. ivanovii, and L. seeligeri usually exceeded 1.0 HU/mg, while those of other Listeria spp. were less than 0.04 HU/mg. In the PCR assay, all of the L. monocytogenes strains contained the hlyA, plcA, plcB, inlA, and inlB virulence genes and produced a product of the expected size. In the PCR of the actA gene, the expected 385-bp product was seen in 39(57.4%) L. monocytogenesstrains, while an unexpected 268-bp product was seen in 29(42.6%) strains. Most L. monocytogenes strains isolated from Hanwoo beef produced the 385-bp actA gene product, while strains of imported US beef usually produced the 268-bp actA gene product. By contrast, no virulence gene products were amplified in the other Listeria spp.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1276-1284
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• 2006

Listeria monocytogenes is a well-known high-risk foodborne pathogen that grows at refrigeration temperature and is responsible for outbreaks of listeriosis. We report here the incidence of L. monocytogenes in fresh chicken carcasses and present genetic diversity of L. monocytogenes isolates. In this study, 25 g of chicken carcasses from markets in Korea were examined according to the FDA method, and presumptive isolates were confirmed by multiplex PCR assay. L. monocytogenes isolates were analyzed by Pulsed-Field Gel Electrophoresis using restriction enzymes, ApaI and AscI, to obtain strain-specific DNA fragments profiles. Antimicrobial resistance of L. monocytogenes strains against generally used antibiotics (Penicillin G, Kanamycin, Tetracycline, Vancomycin, Cephalothin, Rifampicin, Erythromycin, Ampicillin, Gentamicin, Streptomycin, and Chloramphenicol) were analyzed by NCCLS protocols to examine the presence of antimicrobial resistance in natural L. monocytogenes. Of a total 274 chicken samples, 81 samples (29.6%) were positive for L. monocytogenes. Listeria innocua (50.1%), Listeria welshimeri (6.9%), and Listeria grayi (11.3%) were also detected. PFGE analysis, using restriction enzymes ApaI and AscI, showed 27 pulsotypes of L. monocytogenes. Antimicrobial resistance analysis confirmed the existence of antimicrobial resistance for penicillin G and tetracycline in isolated L. monocytogenes strains.

• Preventive Nutrition and Food Science
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• v.7 no.2
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• pp.168-173
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• 2002

Gastrointestinal infections kill over two million people each year, and pathogen contamination of livestock causes many cases of food poisoning. Two candidate intestinal probiotic strains, L. rhamnosus GR-1 and L. fermentum RC-14 were found to inhibit the growth of Salmonella typhi, Shigella dysenteriae, E. coli O157:H7, Listeria monocytogenes, L. innocua, Enterococcus faecalis, and Bacteroides fragilis. In a series of mouse experiments, L. rhamosus GR-1 and L rhamnosus GG protected against S. typhimurium infection and translocation to the liver and spleen, reduced mortality and induced intestinal phagocytic and immunoglobulin responses. In a second series of experiments, the combination of L. rhamnosus GR-1 and L. fermentum RC-14 was superior to L. rhamnosus GG and placebo in protecting the mice from the lethal effect of salmonella. In summary, the use of combinations of probiotic lactobacilli as dietary supplements or foods could be considered for people at high risk of salmonella intestinal infection. Given the post-infection complications that can arise, such natural methods warrant further exploration especially given the increasing problem of antibiotic resistance and the lack of alternative measures available to many developing countries.

• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.680-686
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• 2003

This study was carried out to characterize the antilisterial substances produced by Leuconostoc sp. W65 and to evaluate the effects of pH, temperature, and time on inhibitory activity using response surface methodology. Leucocin W65, an antilisterial substance produced by Leuconostoc sp. W65, markedly inhibited the growth of Listeria monocytogenes, L. innocua, and L. ivanovii, whereas other pathogens including Gram-negative bacteria were not susceptible. The pH was the most effective factor with regard to bacteriocin activity, while temperature and time of heat treatment had no significant effect. Fifty percent of inhibitory activity remained after 22.8 min at pH 4.2 and $121^{\circ}C$. Leucocin W65 was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and tricine-SDS-PAGE. Compositional analysis originally estimated the peptide to be 56 amino acids in length without asparagine, glutamine, and tryptophane. The sequence of partial N-terminal amino acid residues of purified bacteriocin was identified as follows: $NH_{2}-XGXAGVXPXGGQQPXVPLXYP$.

• Food Science of Animal Resources
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• v.39 no.1
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• pp.84-92
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• 2019

Listeria monocytogenes is a major cause of serious foodborne illness in the dairy foods. Although Caenorhabditis elegans model is well established as a virulence model of pathogenic bacteria, its application on L. monocytogenes is critically unclear. The objective of this study was to carry out an evaluation of L. monocytogenes toxicity using C. elegans nematode as a simple host model. We found that C. elegans nematodes have high susceptibility to L. monocytogenes infection, as a consequence of accumulation of bacteria in the worms' intestine. However, L. innocua, which is known to be non-toxic, is not accumulate in the intestine of worms and is not toxic similarly to Escherichia coli OP50 known as the normal feed source of C. elegans. Importantly, immune-associated genes of C. elegans were intensely upregulated more than 3.0-fold when they exposed to L. monocytogenes. In conclusion, we established that C. elegans is an effective model for studying the toxicity of L. monocytogenes and we anticipate that this system will result in the discovery of many potential anti-listeria agents for dairy foods.

• Journal of the Korean Dietetic Association
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• v.24 no.4
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• pp.330-343
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• 2018

Recently, there has been a growing demand for natural preservatives because of increased consumer interest in health. In this study, we produced Lactobacillus rhamnosus cell-free supernatant (LCFS) and evaluated and compared its antimicrobial activity with existing natural preservatives against pathogenic microorganisms and in chicken breast meat contaminated with Escherichia coli and Staphylococcus aureus. Lactobacillus rhamnosus cell-free supernatant possessed 30 units of lysozyme activity and contained 18,835 mg/L of lactic acid, 2,051 mg/L of citric acid and 5,060 mg/L of acetic acid. Additionally, LCFS inhibited the growth of fourteen pathogenic bacteria, S. aureus, Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Listeria innocua, S. epidermidis, L. ivanovii, E. coli, Pseudomonas aeruginosa, Shigella sonnei, Shi. flexneri, Proteus vulgaris, Pseudomonas fluorescens, and Klebsiella pneumoniae. The antibacterial activity of LCFS was stronger than that of egg white lysozyme (EWL), Durafresh (DF) and grapefruit seed extract (GSE). Additionally, LCFS maintained its antimicrobial activity after heat treatment at $50^{\circ}C{\sim}95^{\circ}C$ and at pH values of 3~9. Moreover, LCFS inhibited the growth of E. coli and S. aureus in chicken breast meat. In conclusion, it is expected that LCFS, which contains both lysozyme and three organic acids, will be useful as a good natural preservative in the food industry.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1026-1034
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• 2016

In the current study, a total of 135 enterococci strains from different sources were screened for the presence of the enterocin-encoding genes entA, entP, entB, entL50A, and entL50B. The enterocin genes were present at different frequencies, with entA occurring the most frequently, followed by entP and entB; entL50A and L50B were not detected. The occurrence of single enterocin genes was higher than the occurrence of multiple enterocin gene combinations. The 80 isolates that harbor at least one enterocin-encoding gene (denoted "Gene⁺ strains") were screened for antimicrobial activity. A total of 82.5% of the Gene⁺ strains inhibited at least one of the indicator strains, and the isolates harboring multiple enterocin-encoding genes inhibited a larger number of indicator strains than isolates harboring a single gene. The indicator strains that exhibited growth inhibition included Listeria innocua strain CLIP 12612 (ATCC BAA-680), Listeria monocytogenes strain CDC 4555, Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, S. aureus ATCC 29213, S. aureus ATCC 6538, Salmonella enteritidis ATCC 13076, Salmonella typhimurium strain UK-1 (ATCC 68169), and Escherichia coli BAC 49LT ETEC. Inhibition due to either bacteriophage lysis or cytolysin activity was excluded. The growth inhibition of antilisterial Gene⁺ strains was further tested under different culture conditions. Among the culture media formulations, the MRS agar medium supplemented with 2% (w/v) yeast extract was the best solidified medium for enterocin production. Our findings extend the current knowledge of enterocin-producing enterococci, which may have potential applications as biopreservatives in the food industry due to their capability of controlling food spoilage pathogens.

• Journal of Food Hygiene and Safety
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• v.19 no.3
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• pp.151-160
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• 2004

Enterococcus faecium MJ5-14 isolated from Meju produced a bacteriocin, which was antagonistic towards Listeria monocytogenes. Bacteriocin activity reached a maximum (640 BU/mL) after incubation for 12 hr, the early stationary phase, then dropped after the late stationary phase. Bacterocin of E. faecium MJ5-14 was extremely active against a wide range of Listeria species, including L. monocytogenes with sensitives up to about 640 BU/mL. In case of mixed culture with 105 CFU/mL L. monocytogenes and 105 CFU/mL E. faecium MJ5-14, the inhibitory effect against L. monocytogenes at $37^{\circ}C$ was higher than at $25^{\circ}C$. The mode of action was identified as bactericidal, because the addition of 100 BU/mL this bacteriocin to cell suspensions of L. monocytogenes KCTC 3569, led to a marked decrease in the number of viable cells. Further, when held in contact with bacteriocin of E. faecium MJ15-14 for 12 hr, L. monocytogenes KCTC 3569 displayed the disruption of the cells and an important efflux of the intracellular material.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.6
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• pp.642-647
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• 2010

This study examined the antimicrobial activity of Myagropsis yendoi (MY). The antimicrobial activity of ethanol and aqueous MY extracts were measured using a paper disc assay. The MY ethanol extract had the best antimicrobial activity. It inhibited the growth of Candida tropicalis and Gram-positive bacteria such as Bacillus subtilis, Listeria innocua, L. monocytogenes, and Staphylococcus aureus at 4 mg/mL. The minimum inhibitory concentration (MIC) of the MY ethanol extract ranged from 0.1 to 0.5%. The MY ethanol extract inhibited the growth of B. subtilis and L. innocuaat concentrations of 0.15% and 0.1%, respectively. The antimicrobial activity of the MY ethanol extract was not affected by heating at $121^{\circ}C$ for 15 min or pH 2.10. Therefore, the main substances responsible for the antimicrobial activity of the MY ethanol extract are believed to be stable with changing heat and pH.

• Journal of Food Hygiene and Safety
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• v.30 no.2
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• pp.178-188
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• 2015

Microbial quality of baked egg products was evaluated by counting the levels of sanitary indicative bacteria (aerobic plate counts, coliforms, and E. coli), L. monocytogenes and Salmonella spp. at the critical control points (CCPs) of manufacturing process. In addition, the survival and growth of foodborne pathogens in various egg products (cheese, tuna, tteokgalbi, pizza omelets, baked egg, and steamed egg) were investigated at 4, 10, and $15^{\circ}C$. The contamination level of aerobic plate counts decreased from 4.67 log CFU/g at CCP 1 to 0.56 log CFU/g at CCP 3 in baked egg products. No coliforms and E. coli were detected at all CCPs. Although L. innocua and Salmonella spp. were identified at CCP 1, no L. monocytogenes and Salmonella spp. were detected in the final products. The contamination levels of aerobic plate counts and coliforms in egg strips and number of aerobic plate counts in Tteokgalbi omelet are higher than the microbiological standard of processed egg products. At $10^{\circ}C$, the growth of all pathogens was not prevented in omelet and baked egg, but the populations of S. Typhimurium and E. coli were reduced in steamed egg at $10^{\circ}C$, regardless of the presence of other pathogens. The growth of L. monocytogenes was faster than that of S. Typhimurium and E. coli in omelet. More rapid growth of S. Enteritidis than S. Typhimurium was observed in egg products, indicating the greater risk of S. Enteritidis than S. Typhimurium in egg products.