• Journal of Food Hygiene and Safety
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• v.16 no.4
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• pp.251-257
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• 2001

Listeria monocytogenes and L. ivanovii are important food-pathogens for human and animal. The diagnostic of Listeria in food using culture medium requires time and laborwork, because there are many other non-pathogenic species like L. innocua, L welshimeri, L. seeligeri and L. grayi in Genus Listeria. For these reasons, Lismix multiplex PCR method was developed as a rapid method for the detection and identification of Listeria. In this study we developed a practical system of Lis-mix PCR detection for the application to food samples and new developed Siw-mix III PCR system overall 69 listerial strains were successful species-identified and confirmed. Also, the Siw-mix III PCR system allows the species-specific identification among L. ivanovii, L. welshimeri and L seeligeri in a single PCR.

• Journal of Life Science
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• v.13 no.4
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• pp.390-399
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• 2003

Four species of the genus of Listeria were isolated from seawater and sea food in Kyungnam province, South Korea. These isolated strains were classified into Listeria sp. from different samples by appropriate cultivation conditions and biochemical tests including serological test. In a day enrichment cultivation, the following strains were found out of 100 samples: L. innocua (35%), L. ivanovii (4%), L. monocytogenes (4%), and L. welshimeri (1%). For seven days enrichment culture, L. innocua (38%), L. ivanoii (5%), L. monocytogenes (7%), and L. welshimeri (1%) were isolated. From these results, Listeria species were more efficiently isolated in seven day enrichment broth than in one day enrichment. However, these isolated Listeria species were less grown in the selective medium than in the enrichment medium. Isolation rates of Listeria species showed differency for each sample and Listeria species were more abundantly isolated in shrimps (80%) and crayfishes (80%) than little neck clams (50%), seawater (25%) and mussels (20%). From the results of serological classes for the seven L. monocytogenes, two strains were defined as type I and the other five strains as type IV.

• Journal of Life Science
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• v.25 no.2
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• pp.180-188
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• 2015

The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

• 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.

• 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 Food Hygiene and Safety
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• v.18 no.1
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• pp.25-29
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• 2003

The p60 protein of Listeria spp. is a Listeria-Genus-specific, major extra-cellular protein, which is used as an indicator protein for the detection of these bacteria from contaminated foods. In this study, p60 protein were recombinantly produced in E. coli and were purified using amylose resin based column chromatography. Purified recombinant-p6O was used to generate monoclonal antibody against native p60. Antibody from hybridoma cell line, 1H4, specificically reacted with native p60 protein isolated from pathogenic Listeria spp. such as L. monocytogenes, L. ivanovii, L. welshimeri II, but did not or relatively weakly reacted with non-pathogenic Listeia species, L. innocua or other bacterial proteins. Antibody from 1H4 was produced using ascites fluid method and it may be useful to develop the Listeria-detection kits based on immunological method.

• Food Science of Animal Resources
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• v.25 no.1
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• pp.13-19
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• 2005

The radio-sensitivity of pathogens and the effect of irradiation on microbiologican safety and mutagenicity of meat products such as seasoned and cooked beef and ham were investigated. Samples were radiation-sterilized and inoculated at 10/sup 7/ cfu/g with each of the four pathogens including Salmonella typhimurium, Escherichia coli, Staphylcoccus aureus, and Listeria ivanovii. No viable cells of pathogens were observed in the sample irradiated with 3 kGy. The D/sub 10/ value of inoculated pathogens in seasoned and cooked beef and ham were 0.24∼0.48 and 0.39∼0.45, respectively. Results of Ames test performed with non-irradiated and irradiated seasoned and cooked beef and ham were both negative at the level of 625, 1,250, 2,500, 50,000, and! 10,000 ㎍ sample/plate, respectively. Results indicate that low dose (2∼3 kGy) irradiation is effective to ensure safety for seasoned and cooked beef and ham with toxicological wholesomeness.

• Journal of Food Hygiene and Safety
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• v.10 no.2
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• pp.89-95
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• 1995

Highly lethal Listeria monocytogenes, causing bromatoxism through vegetables, dairy products, meat products and shellfish etc, was examined for possible contamination in market beef. USDA, FDA, Malthus and Modified Cold Enrichment methods were used for the detection of Listeria spp.. Samples of domestic and imported market beef were collected from local meat shopsat Seoul, Korea. Total two hundreds and six of Listeria spp. were isolated and identified from beef. Among 206 isolates, the number of L. welshimeri was one hundred and twenty-one(44.8%). The numbers of isolated L. innocua, L. murrayi, L. monocytogenes, L. grayi, L. seeligeri, and L. ivanovii were 49(18.1%), 14(5.2%), 12(4.4%), 6(2.2%), 2(0.7%), and 2(0.7%), respectively. Detection rates of Listeria spp. varied among four methods. The highest detection rate of Listeria spp. in market beef was found at USDA method and that of L. monocytogenes was found at Malthus method.

• Proceedings of the Zoological Society Korea Conference
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• 2000.10a
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• pp.286.1-286
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• 2000

No Abstract, See Full Text

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.254-259
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• 2004

Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.