• The Plant Pathology Journal
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• v.39 no.1
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

• Journal of Dairy Science and Biotechnology
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• v.41 no.3
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• pp.103-112
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• 2023

This study aimed to assess the effectiveness of 10 different pre-enrichment methods using Real-Time polymerase chain reaction (PCR) in support of the FDA method. When the initial Cronobacter spp. (Enterobacter sakazakii) inoculation was 7.2 CFU/g, the Ct values were observed in the following order: 21.37 (Enterobacteriaceae enrichment [EE] broth), 21.95 (brain heart infusion [BHI]), 22.72 (tryptic soy broth [TSB]), 23.02 (violet red bile lactose [VRBL]), 22.31 (TSB-0.1% sodium pyruvate [SP]), 23.43 (distilled water [DW]), 24.34 (phosphate buffered saline [PBS]), 24.95 (nutrient broth [NB]), 25.82 (TSB-0.6% yeast extract [YE]), and 28.27 (violet red bile glucose [VRBG]). For an inoculation of 1.82% CFU/g of Cronobacter spp. (E. sakazakii), the Ct values were recorded in this sequence: 20.34 (EE broth), 22.16 (TSB-0.6% YE), 22.37 (BHI), 22.71 (VRBL), 22.88 (TSB), 23.01 (DW), 23.19 (NB), 23.79 (TSB-0.1% SP), 24.66 (VRBG), and 24.70 (PBS). Finally, when the inoculum of Cronobacter spp. (E. sakazakii) was 0.182 CFU/g, the Ct values followed this order: 21.93 (VRBL), 23.07 (TSB-0.6% YE), 23.31 (DW), 23.47 (PBS), 23.70 (BHI), 24.14 (TSB-0.1% SP), 25.14 (TSB), 29.00 (VRBG), 31.55 (EE broth), and were undetected in the case of NB. Consequently, these results indicate that there were no significant differences among the 10 different pre-enrichment broths. Future studies should focus on exploring pre-enrichment broths that can improve the limit of detection at very low Cronobacter spp. (E. sakazakii) concentrations and enhance the selective recovery of Cronobacter spp. (E. sakazakii) under acid, antibiotic, cold, and heat damage conditions.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.2
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• pp.158-167
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• 2015

A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1377-1383
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• 2006

Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.176-182
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• 2006

Sensitive and specific monoclonal antibody (MAb) was produced from hybridoma (1H11-5) obtained by fusion of myeloma cell (V653) and spleen cell isolated from mouse immunized sulfamthazine (SMZ)-HG-KLH. Direct competitive ELISA was developed for rapid detection of SMZ in milk samples using MAb against SMZ with optimized conditions between MAb and SMZ-HG-HRP conjugate, and applicable conditions for analysis of milk samples were established. Detection range of immunoassay was 0.1 to 100 ppb. Recoveries from spiked raw milk and processed milk samples averaged 82.1-120.7 and 82.1-97.1%, respectively.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.10
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• pp.845-854
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• 2009

There has been increased concern regarding the release of antibiotics to different environmental compartments due to the possibility of the development of antibiotic resistant bacteria. However, limited information is available regarding the occurrence, fate, and transport of antibiotics in Korea in both the aqueous phase and in solid phases such as sediment and soil. Therefore, this study was conducted to monitor the concentration of released antibiotics in surface water, sediment, and soil adjacent to a cattle manure composting facility in Korea. Specifically, the following six antibiotics were monitored: tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC), sulfamethazine (SMT), sulfamethoxazole (SMX), and sulfathiazole (STZ). To extract and quantify the antibiotics from different environmental compartments, solid phase extraction (SPE) and high performance liquid chromatography mass spectrometry (HPLC/MS) techniques were adopted. The concentration of the six antibiotics ranged from below the detection limit (BDL) to 0.71 ${\mu}g$/L in surface water, from BDL to 27.61 ${\mu}g$/L in sediment, and from 0.12 to 157.33 ${\mu}g$/L in soil. In addition, higher concentrations of antibiotics were observed in surface water and sediment at locations closer to the composting facility indicating that composting is the source of the antibiotics found in the environment. Furthermore, higher concentrations of antibiotics were observed in the solid phase (sediment and soil) than the aqueous phase. These findings indicate that the possibility of antibiotic resistant bacteria is increased because such bacteria are more stable in the solid phase. Overall, longterm monitoring of the aqueous phase and solid phase is necessary to gain a better understanding of the impact of antibiotics from source on the environment in Korea.

• Journal of Food Hygiene and Safety
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• v.34 no.3
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• pp.235-241
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• 2019

An analytical method was developed for the determination of an antibiotic fungicide, kasugamycin, in agricultural products (hulled rice, potato, soybean, mandarin and green pepper) using liquid chromatographytandem mass spectrometry (LC-MS/MS). Samples were extracted with methanol adjusted to pH 13 using 1 N sodium hydroxide, and purified with a HLB (hydrophilic lipophilic balance) cartridge. Linearity of a matrix-matched calibration curve using seven concentration levels, from 0.001 to 0.1 mg/kg, was excellent with a correlation coefficient ($R^2$) of more than 0.9998. The limits of detection (LOD) and quantification (LOQ) of instrument were 0.0005 and $0.001{\mu}g/mL$, respectively, and the LOQ of analytical method calculated as 0.01 mg/kg. The average recoveries at three spiking levels (LOQ, $LOQ{\times}10$, $LOQ{\times}50$, n=5) were in the range of 71.2~95.4% with relative standard deviation of less than 12.1%. The developed method was simple and all optimized results was satisfied with the criteria ranges requested in the Codex guidelines and Food Safety Evaluation Department guidelines. The present study could be served as a reference for the establishment of maximum residue limits (MRL) of kasugamycin and be used as basic data for safety management relative to kasugamycin residues in imported and domestic agricultural products.

• Korean Journal of Microbiology
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• v.1 no.1
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• pp.38-44
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• 1963

According to the experiment on pure-isolation and the related contaminants of Chlorella, the phenomena of the ecological distributions of Chlorella in Korea have been manifested in several areas and also the aim that in going to do culture, biological and physiological study of Chlorella is carried out. Contaminants very oftenly occupied on the colony of the strains taken in order to fulfil pure-isolation of Chlorella, but in accordance with being piled up the minute research on this subject, I can obtain the desirable results as follows: 1. For the pure-isolation, the duration chose the time from May to September 1957 so that may easily isolate from contaminant water with utilizing the antibiotic substances. 2. To take long time, 36-48 hours until growth of nascent through the non-sporulated, it originates from the difference of the cultured media. In addition to the above mention, the mechanism of growth until nascent through the sporulated must not always require the ligh. However the supply of metabolic energy depend upon its nutritional conditions per phase. 3. The culture of Chlorella should be based on the lower culturing except adding especial conditions such as reagent concentration of media, artifical shake of media and other facts due to the natural conditions. And also these strains grew not only in distilled water but 2% NaCl solution without any abnormality in cell it self. I, therefore, guess it is possible to culture in sea-water under phasic environment. 4. In the experiment of ammonia detection, it is caused by the sampling surroundings to contain the minute quantity of ammonia in strain No. M 918; that is the place to be plenty of Carbohydrate on behalf of protein. 5. To compare the absorption curve of chlorophyll of higher plant with that of Chlorella, the absorption zone made mostly the Same ones each other but a little absorption grade dose not clearly appear. The colony which formed giant type grows with intensive colour and green band on surrounding of the colony and after that it was changed into all the green colour and developed up to end. 6. At first phase for a week, the development of Chlorella suspends the normal condition as in vivo but after a few days, the colour of chlorophyll gradually changed into blue-yellow which secrete the mucous substances on the agar media. The cell was flew out the contained substances itself on leaving the cell wall only, or the various micro-organism diffused on the outer-region of the cell.

• The Korean Journal of Nuclear Medicine
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• v.26 no.1
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• pp.14-25
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• 1992

Thallium-201 $(^{201}T1)$ SPECT studies were performed on a normal volunteer and 12 patients with intracerebral lesions: 3 patients with gliomas, 3 patients with meningiomas, 1 patient each with metastatic tumor, brain abscess, and cerebral infarction, and 3 postirradiation patients. (2 with metastatic tumors, 1 with lymphoma). A $^{201}T1$ index, based on the ratio of $^{201}T1$ uptake in the brain lesion versus the homologous contralateral brain, was calculated and compared with tumor histology and CT/MRI findings. The SPECT $^{201}T1$ scan showed minimal uptake of tracer in a normal brain. There was substantial uptake of $^{201}T1$ in high-grade gliomas (index>1.5) with little uptake in low-grade lesions. A previously irradiated patient with recurrent astrocytoma, in whom MRI study was unable to distinguish tumor recurrence from necrosis, showed the lesions with high $^{201}T1$ indices in both hemispheric regions (2.50/1.93), suggesting tumor recurrence. Two meningiomas and a metastatic tumor showed varying degrees of $^{201}T1$ uptake (index $1.71\sim8.15$), revealing that $^{201}T1$ uptake is not exclusive to high-grade gliomas. In 2 postirradiation patients with metastatic tumors, no abnormal $^{201}T1$ uptake was found in the cerebral lesions, shortly after the initiation of radiation therapy or despite the persistence of enhancing lesions-though improved-on MR images, suggesting that $^{201}T1$ uptake may reflect the metabolic and possibly clonogenic activities of tumors and the brain $^{201}T1$ SPECT imaging might be valuable for the evaluation of tumor responsiveness to the therapy and for early detection of tumor recurrence. A patient with brain abscess on antibiotic treatment, showig increased uptake of $^{201}T1$ in the resolving lesions (index 2.87/1.52) is discussed. In a patient with cerebral infarction, there was no abnormal uptake of $^{201}T1$ in infarcted tissue. When using a threshold index of 1.5, correlation rate between $^{201}T1$ uptake and contrast enhancement of the cerebral lesions on CT/MRI was 73% (8/11). In conclusion, the brain $^{201}T1$ SPECT imaging may be useful for assessment of tumor response to the therapy and to predict low-or high-grade lesions.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.309-313
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• 2012

The purpose of this study is to simultaneously detect and identify specific genus of Lactobacillus distributed in vagina of healthy women in their twenties by using multiplex primer. Vaginal fluid samples were taken from 166 women who were healthy and did not have vaginosis symptoms caused by bacterial infection. Six species of lactic acid bacteria belong to Lactobacillus genus that are known to inhabit in healthy vagina were selected to make a species-specific multiplex primer. Multiplex primer I was specified to selectively detect L. iners, L. crispatus, L. gasseri and multiplex primer II was specified to selectively detect L. acidophilus, L. jensenii, L. fermentum. L. crispatus (77%) was most frequently detected, and L. acidophilus (57%) and L. jensenii (57%) were relatively higher than others. Although the proportion of L. iners (59%) was a little higher than those of L. acidophilus and L. jensenii, this should be further validated in healthy women's vagina to be sure since they often appear in women having bacterial vaginosis and/or during the antibiotic therapy. Conclusively, the multiplex PCR technique using the species-specific primer could a useful tool to predict variation of vaginal health condition and process of recovery from vaginal disease.