• Journal of the Korean Chemical Society
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• v.57 no.1
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• pp.81-87
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• 2013

Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it's a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2' for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.

• Journal of Life Science
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• v.22 no.9
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• pp.1137-1144
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• 2012

Inflammation is a host defense mechanism that is activated in response to harmful substances or pathogens. However, an excessive inflammatory response is a problem in itself. Macrophages secrete inflammatory mediators such as nitric oxide (NO) or cytokines through various pathways such as the nuclear factor kappa B (NF-${\kappa}B$)-activated pathway after recognizing pathogen-like lipopolysaccharides (LPSs). In this study, anti-inflammatory effects of Eupatorium chinensis var. simplicifolium (EUC) extracts were investigated using LPS-stimulated RAW 264.7 macrophages. The EUC root extract significantly reduced NO production, inducible nitric oxide synthase (iNOS) expression, and cyclooxygenase-2 expression in a concentration-dependent manner. In addition, the EUC root extract reduced phosphorylation of mitogen-activated protein kinases and protein kinase B, which is upstream of NF-${\kappa}B$. The EUC root extract also reduced the degradation of inhibitory kappa B. These results indicate that EUC root extract exerts anti-inflammatory effects, which are mediated by inhibition of iNOS expression and the NF-${\kappa}B$ pathway.

• Research in Plant Disease
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• v.18 no.2
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• pp.86-92
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• 2012

Clubroot caused by Plasmodiophora brassicae Woron. is one of the most important diseases in Brassica crops worldwide. To establish more simple and reliable screening method for resistant cabbage and broccoli to P. brassicae, the development of clubroot on the plants according to inoculum concentration and incubation period after inoculating with the pathogen was investigated using P. brassicae GN1 isolate (race 9). To facilitate and acquire precise result of resistance screening of cabbage and broccoli to clubroot, 14-day-old seedlings were inoculated by drenching roots with the spore suspension of P. brassicae to give inoculum density of $2.5{\times}10^9$ spores/pot. To develop the disease, the inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days, and then cultivated in a greenhouse ($20{\pm}5^{\circ}C$) for five weeks. Under the optimum conditions, 16 cabbage and 17 broccoli cultivars were tested for resistance to four field isolates (GN1, GN2, GS and YC) of P. brassicae collected from four regions in Korea. Among them, some cabbage and broccoli cultivars showed different resistance response to three isolates (GN1, GN2 and GS) determined as race 9 by using the differential varieties of Williams. On the other hand, all the tested cultivars were highly susceptible to YC isolate (race 2). The results suggest that this method is efficient screening method of cabbage and broccoli for resistance to P. brassicae.

• Horticultural Science & Technology
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• v.31 no.5
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• pp.626-632
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• 2013

In the course of a developing screening method for resistant radish to Fusarium oxysporum f. sp. raphani, we found that the fungus produces phytotoxic compound against Raphanus sativus. The culture filtrate of F. oxysporum f. sp. raphani KR1 represented the strongest phytotoxicity when the fungus was incubated in the malt extract broth with 150 rpm at $25^{\circ}C$ for 14 days. Under bioassay-guided purification, we isolated a substance from liquid culture of F. oxysporum f. sp. raphani KR1, with phytotoxic effect against R. sativus. The compound was identified as fusaric acid by mass and nuclear magnetic resonance spectral analyses. Phytotoxicity of the compound against cruciferous vegetable crops, including radish, cabbage, and broccoli, was investigated. Fusaric acid represented phytotoxicity on radish seedlings by concentration dependant manner. And the phytotoxin demonstrated strong phytotoxicity on the resistant cultivars as well as susceptible cultivars of radish to F. oxysporum f. sp. raphani. In addition, fusaric acid isolated from the fungus also showed a potent phytotoxic efficacy against non-host Brassicaceae crops of the fungus such as cabbage and broccoli. The results demonstrate that fusaric acid produced by F. oxysporum f. sp. raphani is non-host-specific toxin and for screening of resistant radish to the fungal pathogen, spore suspension of the fungus without the phytotoxin has to be used.

• Korean Journal of Environmental Agriculture
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• v.40 no.4
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• pp.248-259
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• 2021

BACKGROUND: Contaminated water was a major source of food-borne pathogens in various recent fresh produce-related outbreaks. This study was conducted to investigate the microbial contamination level and correlations between the level of sanitary indicator bacteria and the detection ratio of pathogens in agricultural water by logistic regression analysis. METHODS AND RESULTS: Agricultural water was collected from 457 sites including surface water (n=300 sites) and groundwater (n=157 sites) in South Korea from 2018 to 2020. Sanitary indicator bacteria (total coliform, fecal coliform, and Escherichia coli) and food-borne pathogens (pathogenic E. coli, E. coli O157:H7, Salmonella spp., and Listeria monocytogenes) were analyzed. In surface water, the coliform, fecal coliform, and E. coli were 3.27±0.89 log CFU/100 mL, 1.90±1.19 log CFU/100 mL, and 1.39±1.26 log CFU/100 mL, respectively. For groundwater, three kinds of sanitary indicators ranged in the level from 0.09 - 0.57 log CFU/100 mL. Pathogenic E. coli, Salmonella and Listeria monocytogenes were detected from 3%-site, 1.5%- site, and 0.6%-site water samples, respectively. According to the results of correlations between the level of sanitary indicator bacteria and the detection ratio of pathogens by logistic regression analysis, the probability of pathogen detection increased individually by 1.45 and 1.34 times as each total coliform and E. coli concentration increased by 1 log CFU/100mL. The accuracy of the model was 70.4%, and sensitivity and specificity were 81.5% and 51.7%, respectively. CONCLUSION(S): The results indicate the need to manage the microbial risk of agricultural water to enhance the safety of fresh produce. In addition, logistic regression analysis is useful to analyze the correlation between the level of sanitary indicator bacteria and the detection ratio of pathogens in agricultural water.

• Journal of Environmental Science International
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• v.28 no.2
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• pp.225-233
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• 2019

In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

• Journal of fish pathology
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• v.35 no.1
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• pp.57-63
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• 2022

Vibrio parahaemolyticus and V. vulnificus are known to be infected to human via fisheries products. Therefore, food safety of fisheries products is important for public health and fish industry. This paper was conducted to know how well these human isolates can survive in olive flounder (Paralichthys olivaceus). The growth of V. parahaemolyticus and V. vulnificus showed about 50~60% reduced rates at 25℃ than at 37℃ and did not show any differences according to NaCl concentration of media except the increasing in the growth of V. vulnificus in medium containing 3% NaCl. Artificial infection of 1×10⁶ CFU/fish was carried out to confirm the sensitivity of olive flounder against V. parahaemolyticus and V. vulnificus. After 1 week from injection, no fish was dead. To evaluate nonspecific defense of olive flounder against V. parahaemolyticus and V. vulnificus, the antibacterial potency of serum and epidermal mucus were tested. The number of the vibrios exposed to serum obtained from olive flounder significantly decreased after 3 hours, and epidermal mucus showed decrease of the bacteria over than 90% until 12 hours from exposure. Phagocytosis of head kidney leucocytes of healthy olive flounder against V. parahaemolyticus and V. vulnificus showed in over 70% of leucocytes at the 2 hours. Therefore, cultured olive flounder only as vehicle for human pathogen in environmental water is well developed its antibacterial potency against human pathogens, so the viability of V. parahaemolyticus and V. vulnificus in cultured olive flounder was considered very low.

• Research in Plant Disease
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• v.30 no.2
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• pp.199-205
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• 2024

Erwinia amylovora, a causal pathogen of fire blight, has been continuously inducing damage to the apple and pear trees in South Korea since 2015. Farmers apply antibiotics during blooming season to prevent the fire blight. However, continuous use of antibiotics can induce the emergence of resistant bacteria, which consequently reduces control efficacy. In this study, we assessed the minimal inhibitory concentration (MIC) of streptomycin, using a total of 361 E. amylovora isolates that were collected from the six provinces of South Korea from 2019 to 2023. As a result, the MIC of streptomycin ranged from 0.5 to 4 ㎍/ml and the strA-strB genes were not identified from the isolates. The MIC was higher in the isolates from Gyeonggi-do, Gangwondo, and Chungcheongbuk-do compared to those from other three provinces. These results may bring broad attention to the use of streptomycin and aid in developing a management protocol for the occurrence of fire blight in South Korea.

• Research in Plant Disease
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• v.30 no.3
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• pp.288-293
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• 2024

Fusarium basal rot (FBR), caused by the ascomycete fungus Fusarium oxysporum, is an economically important disease of onion worldwide. The most economical and effective way to manage FBR would be the use of FBR resistant onion cultivars. This study was carried out to develop a rapid screening method for resistant onion cultivars in seedling stage. We used the F. oxysporum 19-385 isolate, which causes damping-off in onion seedlings and basal rot in onion bulbs. We optimized broth incubation and medium composition for the production of inoculum, and determined conidial concentration for the preparation of F. oxysporum infected soil. Ten commercial cultivars of onion were evaluated the seedling survival rates and heights by infected soil inoculation methods. As a result, 'K-force' was the most resistant cultivar with 97.4% of relative seedling survival rate against the pathogen, whereas 'Sunpower' was the most susceptible cultivar with 20.0% of relative seedling survival rate.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.29-36
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• 2011

Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.