• Korean Journal of Veterinary Service
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• v.43 no.4
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• pp.237-244
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• 2020

Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.

• KSBB Journal
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• v.23 no.6
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• pp.513-518
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• 2008

This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

• Journal of the East Asian Society of Dietary Life
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• v.24 no.1
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• pp.70-79
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• 2014

This study was conducted to develop natural additive for the improvement of lipid stability and the suppression of pathogenic contamination during the storage periods of fermented sausage. Fermented sausage prepared with grapefruit extract showed good suppression of lipid oxidation and pathogenic bacterial growth, and it was higher than that of fermented sausage prepared with nitrate until 7 days of storage. Fermented sausage prepared with treatment A (sodium nitrate and L. acidophilus), B (sodium nitrate and L. brevis), C (ascorbic acid and L. brevis) and D (grapefruit extract and L. brevis) were compared. As results, treatment C and D showed the lowest pH changes among treatments, and the highest growth of lactic acid bacteria was represented in treatment D. In the suppression effect of E. coli, the bacterial count was below 100 in all treatment except the treatment A. There were no differences among treatment in the brightness of meat color (p<0.05). Treatment D showed significantly high values in the redness, but showed the lowest values in the yellowness. In sensory and texture test, treatment C and D were superior compared to others. Therefore, grapefruit was represented as a good source of antioxidant for fermented sausage, and L. brevis isolated from kimchi was effect in lowering pH value of sausage during fermentation and it was also effective to persist fermentation during storage period and to reduce the occurrence of pathogenic bacteria by the suppression of their growth.

• Journal of Korean Neurosurgical Society
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• v.56 no.1
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• pp.21-27
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• 2014

Objective : Infectious spinal disease is regarded as an infection by a specific organism that affects the vertebral body, intervertebral disc and adjacent perivertebral soft tissue. Its incidence seems to be increasing as a result of larger proportion of the older patients with chronic debilitating disease, the rise of intravenous drug abuser, and the increase in spinal procedure and surgery. In Korea, studies assessing infectious spinal disease are rare and have not been addressed in recent times. The objectives of this study are to describe the epidemiology of all kind of spinal infectious disease and their clinical and microbiological characteristics as well as to assess the diagnostic methodology and the parameters related to the outcomes. Methods : A retrospective study was performed in all infectious spinal disease cases presenting from January 2005 to April 2010 to three tertiary teaching hospitals within a city of 1.5 million in Korea. Patient demographics, risk factors, clinical features, and outcomes were assessed. Risk factors entailed the presence of diabetes, chronic renal failure, liver cirrhosis, immunosuppressants, remote infection, underlying malignancy and previous spinal surgery or procedure. We comparatively analyzed the results between the groups of pyogenic and tuberculous spinal infection. SPSS version 14 statistical software was used to perform the analyses of the data. The threshold for statistical significance was established at p<0.05. Results : Ninety-two cases fulfilled the inclusion criteria and were reviewed. Overall, patients of tuberculous spinal infection (TSI) and pyogenic spinal infection (PSI) entailed 20 (21.7%) and 72 (78.3%) cases, respectively. A previous spinal surgery or procedure was the most commonly noted risk factor (39.1%), followed by diabetes (15.2%). The occurrence of both pyogenic and tuberculous spondylitis was predominant in the lumbar spine. Discs are more easily invaded in PSI. At initial presentation, white cell blood count and C-reactive protein levels were higher in PSI compared to TSI (p<0.05). Etiological agents were identified in 53.3%, and the most effective method for identification of etiological agents was tissue culture (50.0%). Staphyococcus aureus was the most commonly isolated infective agent associated with pyogenic spondylitis, followed by E. coli. Surgical treatment was performed in 31.5% of pyogenic spondylitis and in 35.0% of tuberculous spondylitis cases. Conclusion : Many previous studies in Korea usually reported that tuberculous spondylitis is the predominant infection. However, in our study, the number of pyogenic infection was 3 times greater than that of tuberculous spinal disease. Etiological agents were identified in a half of all infectious spinal disease. For better outcomes, we should try to identify the causative microorganism before antibiotic therapy and make every effort to improve the result of culture and biopsy.

• Korean Journal of Veterinary Research
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• v.24 no.2
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• pp.149-162
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• 1984

The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.165-171
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• 2006

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

• Journal of the Korean Society of Food Culture
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• v.34 no.2
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• pp.224-232
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• 2019

This study evaluated the physicochemical and microbial quality characteristic of seasonal commercial kimchi for hygienic safety levels. The pH of seasonal commercial kimchi was 3.84-6.36 and the titratable acidity and salinity of the samples were 0.21-1.16 and 1.19-1.54%, respectively. The content of nitrate and nitrite in the commercial kimchi were lower in the spring and summer, which was affected by acidic condition of the kimchi depending on fermentation. Heavy metal contents in commercial kimchi are not an issue because they were detected only at very low levels. The total aerobic bacteria and coliforms counts ranged from 5.25 to 8.44 Log CFU/g and 0.00 to 5.08 Log CFU/g, respectively. The total aerobic bacteria and coliforms were detected more in summer than in the other seasons. E. coli was detected in three of the samples tested. Food-borne pathogens were not detected in any of the samples except for B. cereus. B. cereus was detected in the fall in more than 70% of samples. These results suggest that commercial kimchi distributed in the fall maintain the quality properties and the microbiological safety of kimchi compared to the other seasons. Therefore, further studies as an effective distribution system for the particular seasons will be needed to guarantee the hygienic safety levels of commercial kimchi required by the consumers.

• The Journal of the Convergence on Culture Technology
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• v.8 no.6
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• pp.943-951
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• 2022

In this study, 8 kinds of fruits and vegetables such as apples, pears and radishes were cut and hot water extracts and Steamed hot water extract from fruits and vegetables were prepared and used as experimental substrates. As a result of fermenting with 1% (W/V) red ginseng extract (W/V) and 8 types of lactic acid bacteria mixed starter added to the lactic acid bacteria fermented extract, the pattern and content of ginsenosides were almost unchanged in the fruit and vegetable extract group and the steam treatment group. However, in the lactic acid bacteria fermented group, the TLC pattern was changed according to the fermentation process and treatment, and the content of ginsenosides converted into Rg₃(S) and Rg₅ increased. No change in the number of lactic acid bacteria (cfu) was observed in all four types of fruit and vegetable extracts. The number of lactic acid bacteria CFU was slightly decreased in the four fermented groups of fruit and vegetable extracts, but the growth inhibitory effect of beneficial bacteria was not significant. The growth inhibitory effect of the three harmful bacteria was not affected by the growth of E. coli and Pseudomonas in the four fruit and vegetable extracts. However, the proliferation of Salmonella was inhibited, which was confirmed as the growth inhibitory effect of the fruit and vegetable extract regardless of whether the steamed hot water extract or red ginseng extract was added.

• Journal of Life Science
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• v.22 no.6
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• pp.788-793
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• 2012

High concentration of antibiotics has been used to treat the outbreak of edwardsiellosis caused by Edwardsiella tarda in aquaculture. However, not all of the bacteria have been killed with high concentrations of antibiotics treatment by the formation of persister cells with a dormant state. The main objective of this study was to kill persister cell using antibiotics with the addition of natural plant compounds. Antibiotics used in this study consist of 100 mg/ml florfenicol and 100 mg/ml amoxicillin. Ten natural plant compounds with persister cell inhibitor activity to E. coli were obtained from Protein Engineering and Systems Biology Lab. of Sungkyunkwan University. The persister cell inhibition activities of those natural plant compounds were evaluated in test tube. Concentrations of the antibiotics were in the ranges of 25~200 ${\mu}g/ml$. The persister cell formation was observed after 16 hours of culture. Persister cells were killed by antibiotics with natural plant compounds. Among ten natural plant compounds, Gynostemma pentaphyllum, Mallotus japonicus, and Orixa japonica showed persister cell formation inhibition activities. The optimal concentrations of G. pentaphyllum, M. japonicus, and O. japonica for the inhibitor of persister cell formation were 100 ${\mu}g/ml$, 100 ${\mu}g/ml$, and 200 ${\mu}g/ml$, respectively. In vivo study was carried out to evaluate the effect of the antibiotics with natural plant compounds using aquacultural fish, olive flounder, as test animals. G. pentaphyllum, M. japonicus, and O. japonica of 30 ${\mu}g/ml$, 10 ${\mu}g/ml$, and 10 ${\mu}g/ml$ with antibiotics reduced cumulative mortalities, showing the effectiveness of persister cell inhibition.

• KSBB Journal
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• v.24 no.3
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• pp.273-278
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• 2009

The purpose of this work was to investigate the antibacterial activity of Lactobacillus plantarum YK-9 isolated from fermented Kimchi. Morphological and biochemical characteristics of L. plantarum YK-9 were examined. Phylogenetic analysis using 16S rRNA sequencing was performed to identify the strain, and the strain could be assigned to Lactobacillus plantarum, designated as L. plantarum YK-9. The strain was registered in GenBank as [FJ669130]. During the incubation period of L. plantarum YK-9, the changes of bacterial growth and residual organic acids were monitored. HPLC was used to confirm the organic acids produced in the cultures as metabolites. L. plantarum YK-9 produced both lactic acid and acetic acid, which were responsible for the pH decrease during growth. Initial pH 7.0 of the cultures decreased to 3.6 at the incubation after 72 hours, and concentrations of lactic acid and acetic acid increased to approximately 588.7 mM and 255.5 mM, respectively. The antibacterial activities against food-poisoning causing bacteria were examined with 20-fold concentrated culture supernatants from L. plantarum YK-9, and the antibacterial effects were clearly observed against all the bacteria tested in this work.