• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.263-268
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• 2002

Bacillus polyfermenticus SCD, which is commonly called a 'Bisroot' strain, has been appropriately used for the treatment of long-term intestinal disorders, since the live strains, in the form of active endospores, can successfully reach the target intestine. Goal of this study was to develop an industrial medium for growth and sporulation of B. polyfermenticus SCD. From the results of effect of mixed carbon sources on growth and sporulation of B. polyfermenticus SCD, glucose 2% and starch 2% was particularly found to be the most effective for the maximum number of spore production, resulting in spore cells of $4.3{\times}10^9\;spores/mL$ with a sporulation yield of 91%. For the effect of nitrogen sources, the maximum spore cells of $5.7{\times}10^9\;spores/mL$ of B. polyfermenticus SCD with a sporulation yield of 97% was obtained when B. polyfermenticus SCD was cultivated in an optimum nitrogen source medium containing 5% soybean flour. A medium involving proper phosphate salt yielded the maximum number of a spore cells of $6.0{\times}10^9\;spores/mL$ with a sporulation yield of 95%. Finally, the efficacy of an industrial medium (KH5 medium) on growth and sporulation of B. polyfermenticus SCD was investigated in jar fermenter. The higher number of viable cells $(3.3{\times}10^{10}\;cells/mL)$ and spore cells $(3.0{\times}10^{10}\;spores/mL)$ were obtained in 5 L fermenter when compared with a 500 mL baffle flask cultivation. Thus, KH5 medium developed in this study shows promise as an industrial medium because of higher cells and sporulation yield.

• The Korean Journal of Mycology
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• v.24 no.1 s.76
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• pp.38-48
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• 1996

The survival or colonization of beneficial organsisms and suppression of root rot of ginseng (Panax ginseng) by two distinct bacteria, Pseudomonas cepacia, Bacillus cereus and three mycorrhiza in pot soil were investigated and compared with uninoculated root. In separate inoculation, colonization of roots by P. cepacia was maintained at 6.25 (log cfu/g root) during growth for 10 days under pot culture conditions comparing to $5.62{\sim}6.19$ by mixed treatment with other organisms. Colonizations of P. cepacia were gradually decreased from 6.25 (log cfu/g root) in 10 days growth to 3.01 (log cfu/g root) in 270 days incubation period. This reduction was also investgated in combination treatments by B. cereus or F. solani. The numbers of Fusarium spp. were colonized high number in rhizosphere soil from 3.33 to 3.67 (log cfu/g root) in control within $10{\sim}60$days after treatment of pathogen F. solani, but it's numbers were markedly decreased in 270 days cultivation of plant from 3.33 to 1.02 (log cfu/g root) after treatment. In treatment of beneficial strains of P. cepacia and B. cereus, P. cepacia significantly suppressed the development of root rot from 4.3 in control to 1.2 in treatment, whereas B. cereus alone had no effect on the rate of disease suppression. The disease index $(1.8{\sim}2.3)$ in combination of two bacteria was reduced in plants inoculated with both P. cepacia and B. cereus comparing to the index (4.3) of control. As an effect of inoculation with mycorrhiza on disease suppression, suppression of root rot by F. solani was reduced to $1.2{\sim}1.6$ in disease index in treatment of Glomus albidum and Acaulospora longular comparing to 4.3 of control. In the treatment of bacterial strain P. cepacia and mycorrhizal fungus Glomus albidum, the disease suppression was apparent to 1.2 and 1.2 comparing to 4.3 of control in disease index respectively.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.111-115
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• 2006

A feeding trial was conducted to test the use of marine yeasts isolated from seawaters and sediments as a dietary source in cultivating a Cladocera, Moina macrocopa which is available as an alternative live food for fish larvae. The marine yeast-fed M. macrocopa had similar essential amino acid profiles to the documented values for Rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. $S{\pi}-1$ lacked ${\omega}-3$ high unsaturated fatty acids (HUFAs), $20:5{\omega}-3$ (EPA) and $22:6{\omega}-3$ (DHA), which were also poor but detected in both the marine yeasts. An increase in the $20:5{\omega}-3$ and $22:6{\omega}-3$ levels, compared with the levels in marine yeast strains themselves, was more pronounced in the $22:6{\omega}-3$ level of Moina fed the Candida sp. Y-16, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their ${\delta}^{13}C$ values shifted gradually toward the values of the switched diets. Diet switch from Erythrobacter sp. $S{\pi}-1$to Candide sp. Y.16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the ${\delta}^{13}C$ values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the ${\delta}^{13}C$ values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly Candida sp. Y-16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietaty values than the commercial diet (Erythrobacter sp. $S{\pi}-1$).

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.340-346
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• 2014

In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.552-558
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• 2021

Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.