• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.11a
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• pp.65-73
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• 1996

LPS/LOS, the compound found only in gram-negative bacterial outer membrane, plays important roles in bacterial maintenance as well as its pathogenesis. We isolated and characterized several genes required for NTHi 2019 LOS biosynthesis, which encode enzymes required for sugar substrate synthesis or the transfer of substrates to receptor molecules. The htrB gene, however, appears to have more complex role. It has acryltransferase activity as well as various other activity, which may control regulation of LOS biosynthesis as well as its pathogenicity. Evidences supporting the latter come from the observations that the lipid A of the B29 induced significantly less TNF ${\alpha}$ from macrophages than that of the wild type LOS (unpublished data). H. influenzae A2-htrB mutant strain was also significantly less invasive than the wild type strain. The structural similarities of the enterobacterial LPS and the Haemophilus LOS enabled us to isolate the NTHi 2019 genes involved in LOS biosynthesis genes by using the S. typhimurium LPS deep core mutants. While a similar approach has been used for E. coli, this technique for selection of an LPS phenotype has not been applied to nonenterobacterial species. The difficulties inherent in the molecular manipulation of organism such as Neisseria and Haemophilus species make this approach particularly attractive in the identification and cloning LOS genes. Studies on genetic features of LPS/LOS biosynthesis would be useful for understanding bacterial pathogenesis as well as for developing vaccines for these gram-negative pathogenic bacteria.

• Korean Journal of Veterinary Service
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• v.39 no.3
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• pp.159-166
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• 2016

Clostridium perfringens, Salmonella thyphimurium and S. Montevideo isolated from the intestines of dead broiler chickens in Korea were tested for antibacterial effects to purifed bee venom. Purified bee venom from Apis mellifera L. has been used as natural antimicrobial compounds in pigs, cows, dairy cattle and chicken farms in Korea. To investigate antibacterial effect of purified bee venom was evaluated by agar well diffusion method, minimum inhibitory concentraion (MIC), minimum bactericidal concentration (MBC), and postantibiotic effect (PAE). Purified bee venom exhibited significant inhibition of bacterial growth of C. perfringens, S. thyphimurium and S. Montevideo with MIC value of 0.85, 0.68 and $0.69{\mu}g/mL$, respectively. The MBC value of purified bee venom against C. perfringens, S. thyphimurium and S. Montevideo were 3.33, 2.66 and $2.86{\mu}g/mL$. Furthermore, the results of PAE values against C. perfringens, S. thyphimurium and S. Montevideo showed the bacterial effect with 3.5, 4.0 and 3.5 hr. Stability of pufifed bee venom at acidity from pH 1 to pH 8 for 24 hr was the antibacterial activity for C. perfringens, S. thyphimurium and S. Montevideo and melittin contents. Also purified bee venom processed through the heating for 15 min, there was no signification loss of the antibacterial activity and melittin at below $100^{\circ}C$. These results obtained in this study suggest that purified bee venom might be utilized as a feed additive in poultry diets.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.763-768
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• 2012

This study has been performed to measure the prevalence and microbial flora on chicken slaughtering as well as the processing process from the months of October to November. Whole-chicken rinsing technique was used in order to analyze the incidence of microorganisms on chicken carcass at the stage before chilling (after evisceration), after chilling and after cutting. The swab technique was used on processing the processed samples, such as working plates and cutting knives. Brine and cooling water from four cooling tubs were taken from each processing processes and were used as samples. Furthermore, the matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for whole cell fingerprinting in combination with a dedicated bioinformatic software tool was used to identify the isolated microorganisms. Of the tested samples and processes, brine ($4.50{\pm}0.64$ Log CFU/mL) and chicken carcass before chilling ($4.15{\pm}0.46$ Log CFU/mL) showed the highest population of microorganisms; the predominant microbial flora of them were Moellerella wisconsensis (54.84%), a member of the Enterobacteriaceae family, and Escherichia coli (60.36%), respectively. However, the predominant microbial flora of cut carcass was changed to Staphylococcus aureus (27.32%), which is a kind of pathogenic microorganism that can cause a food-borne illness. Therefore, the slaughtering and processing procedure of chicken are needed to be controlled more hygienically.

• Korean Journal of Veterinary Service
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• v.32 no.2
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• pp.147-153
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• 2009

Salmonella species are the most important etiologic agents of food-borne acute gastroenteritis. The most common serotypes isolated from humans are Salmonella enterica serotype Typhimurium (S. Typhimurium) and S. Enteritidis. Traditional detection methods for Salmonella are based on cultures using selective media and characterization of suspicious colonies by biochemical and serological tests. These methods are generally time-consuming and not so highly sensitive. Recently, the polymerase chain reaction (PCR) has been used as a highly sensitive, specific, and rapid test for the presence of pathogenic bacteria. In this study, a multiplex PCR (m-PCR) was used to detect S. Typhimurium and S. Enteritidis. We selected m-PCR target genes, which were the spv (virulence plasmid specific for S. Enteritidis) and sefA (S. Enteritidis fimbrial antigen) genes, fliC (H1-i antigen specific for S. Typhimurium) and a randomly cloned sequence specific for the genus Salmonella. With m-PCR, random sequence was detected from all strains of Salmonella spp, spv and sefA were detected from all strains of S. Enteritidis (100%), and fliC was detected from all strains of S. Typhimurium (100%). This assay indicate that the specificity of the m-PCR make them potentially valuable tools for detection of S. Typhimurium and S. Enteritidis.

• Korean Journal of Veterinary Research
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• v.33 no.3
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• pp.393-406
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• 1993

Antibacterial substances produced by Lactobacillus casei subsp. and Streptococcus faecium were examined for its antibacterial effects against some pathogenic bacteria. They were partially purified with ammonium sulfate precipitation, methanol-acetone extraction, G-50 gel filtration and examined its characteristics. When L casei subsp. and Str faecium were cultivated in MRS broth, stationary phase of L casei is until 24 hours and Str faecium is 20 hours. pH change of the cultured medium was both decreased after 12 hours and then constant at pH 4.5~4.6 after 28 hours. MRS broth culture fluids of L casei subsp. and Str faecium appeared the antibacterial effects by the spot-on-the-lawn method against ETEC, Sal pullorum and Sta aureus. Culture filtrates of L casei subsp. and Str faecium also appeared the antibacterial effects by the disc diffusion method. Culture filtrates of L casei sub. rhamnosus 7469 produced 0.032M of lactic acid and 0.01M of acetic acid. Str faecium 27273 also produced 0.027M of lactic acid and 0.01M of acetic acid. Protein concentrations of culture filtrates produced by L casei sub rhamnosus 7469 and Str faecium 27273 was $495{\mu}g/m{\ell}$ and $594{\mu}g/m{\ell}$, respectively. Antibacterial substances which are partially purified by ammonum sulfate precipitation, methanol-acetone extraction and G-50 gel filtration inhibit the growth of ETEC, Sal pullorum and Sta aureus. Characteristics of purified antibacterial substances was examined. Its molecular weight was about 31Kd, stabilized at $100^{\circ}C/20min.$ and some of proteolytic enzyme treatment.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.258-263
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• 2008

Late blight, one of the most important disease in many agricultural crops, is caused by Phytophthora infestans. Fusarium wilt is a vascular disease of many plants caused by Fusarium oxysporum. Some bacteria isolated from rhizosphere were screened for their ability to inhibit the growth of F. oxysporum and P. infestans. Productions of siderophore, $\beta-1$,3-glucanase, hydrogen cyanide and chitinase by 4 isolated strains were examined. Among them, Bacillus sp. RFO41 most effectively inhibited the growth of F. oxysporum. The highest productions of siderophore and $\beta-l$,3-glucanase were shown in the culture of Bacillus sp. RFO41. Bacillus strain PS2 was most effective against P. infestans. PS2 showed the highest production of chitinase and hydrogen cyanide. A significant relationship was shown between the antagonistic effects of isolates against F. oxysporum and P. infestans and their production level of siderophore, $\beta-1$,3-glucanase, hydrogen cyanide, and chitinase.

• Food Quality and Culture
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• v.3 no.1
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• pp.34-39
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• 2009

This study assessed the efficacy of aqueous chlorine dioxide ($ClO_2$) and commercial chlorine sanitizer in terms of its ability to eliminate Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7 on radish sprouts (Raphanus sativus L.). Radish sprouts were inoculated with a cocktail containing one each of three strains of three different foodborne pathogens, then treated with distilled water (control) or chemical sanitizers (100 ppm commercial chlorine, and 50, 100, 200 ppm $C1O_2$) for 1, 5, and 10 min at room temperature ($22{\pm}2^{\circ}C$). Populations of S. Typhimurium, E. coli O157:H7 and L. monocytogenes were counted at 4.64, 6.05, and 4.29 log CFU/g, respectively, after inoculation. Treatment with water did not significantly reduce the levels of any of the three foodborne pathogens. The levels of all three pathogens were reduced by treatment with chemical sanitizers; however, the observed levels of reduction of E. coli O157:H7 and L. monocytogenes were not significant as compared with the controls. The levels of the three pathogens were reduced most profoundly when treated for 10 min with 200 ppm of $C1O_2$, and the reduction levels of S. Typhimurium, E. coli O157:H7, and L. monocytogenes were 1.17, 1.63, and 0.96 log CFU/g, respectively. When chemically injured cells were investigated using SPRAB for E. coli O157 :H7 and by selective overlay methods for S. Typhimurium and L. monocytogenes, respectively, it was noted that commercial chlorine sanitizer generated more numbers of injured pathogens than did $C1O_2$. These data indicate that $C1O_2$ treatment may prove useful in reducing the numbers of pathogenic bacteria in radish sprouts.

• Korean Journal of Food Science and Technology
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• v.36 no.3
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• pp.514-517
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• 2004

O157:H7, Salmonella typhimurium, Listeria monocytogenes were treated with aqueous chlorine dioxide to elucidate effect of aqueous chlorine dioxide treatment on major food-borne pathogenic bacteria. Survival plot of E.coli O157:H7 at 5 ppm chlorine dioxide showed typical first-order rate. After 5 min of treatment, cell number decreased by 1.5 log cycle. Survival plot slope gave D value of 3.37 min. S. typhimurium and L. monocytogenes showed biphasic curve. Aqueous chlorine dioxide treatment on S. typhimurium and L. monocytogenes resulted in bactericidal effect for 5 min, and thereafter no effect was observed under experimental conditions of this study. These results suggest concentration of chlorine dioxide is more important than treatment time, and 5 ppm chlorine dioxide treatment is not sufficient for sanitizing fresh vegetables.

• Journal of fish pathology
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• v.21 no.3
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• pp.247-257
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• 2008

To understand the activity of non-specific defence factors in cultured Sebastes, the antibacterial effect of the serum, skin mucus and homogenate of various organs from cultured oblong rockfish (Sebastes oblongus) and rockfish(Sebastes schlegeli) against pathogenic bacteria, Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum, and Streptococcus sp. was compared with that of flounder(Paralichthys olivaceus) and seabass(Leteolabrax japonicus). And the activities of proteolytic enzyme, chitinolytic enzyme and haemolycin as non-specific defence factor were investigated on the oblong rockfish and rockfish. Samples from oblong rockfish showed the highest antibacterial activity by lysoplate assay on agar plate mixed with pathogens, followed in descending order by rockfish, seabass, and flounder. Turbidimetric assay was carried to evaluate the lysozyme activity of fish samples against lyophilized cells of Micrococcus lysodeiktikus. The serum, kidney, liver, stomach, intestine and eyeball of oblong rockfish and the mucus and gill of rockfish appeared to have the highest lysozyme activity among the fish strains investigated. All samples except skin mucus, liver, and eyeball of oblong rockfish and rockfish showed proteolytic enzyme activity. Chitinolytic enzyme activity was showed in random sampling and haemolytic activity was remarkable in oblong rockfish. Therefore, Sebastes strain was proved to have effective defense mechanisms based on the antibacterial activities, and lysozyme, proteolytic enzyme, chitinolytic enzyme, and haemolycin were considered to act as the non-specific defence factor of Sebastes.