• Journal of Korean Society of Water and Wastewater
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• v.29 no.6
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• pp.633-641
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• 2015

Waterborne infectious disease is induced by several pathogenic microbes such as bacteria, viruses and protozoans, and the cases caused by viral infection is currently increasing. Water treatment process could reduce the number of virus in the water, but there were many difficulties to completely remove the virus particles from water. Therefore, the membrane separation technology which was reported to effectively remove pollutants from raw water has attracted increasing attention and demand. Since its efficiency has been introduced, demands for evaluation method toward the membrane filtration process are increasing. However, progression of the method development is slow due to the difficulties in cultivation of several waterborne viruses from animal models or cell culture system. To overcome the difficulties, we used adenovirus, one of the commonly isolated pathogenic waterborne viruses which can grow in cell culture system in vitro. The adenovirus used in this study was identified as human adenovirus C strain. The adenovirus was spiked in the raw water and passed through the microfiltration membrane produced by Econity, a Korean membrane company, and then the viral removal rate was evaluated by real-time PCR. In the results, the amount of virus in the filtered water was decreased approximately by 5 log scale. Because coagulant treatment has been known to reduce filtering function of the membrane by inducing fouling, we also investigated whether there was any interference of coagulant. In the results, we confirmed that coagulant treatment did not show significant interference on microfiltration membrane. In this study, we found that waterborne virus can be effectively removed by membrane filtration system. In particular, here we also suggest that real-time PCR method can rapidly, sensitively and quantitatively evaluate the removal rate of virus. These results may provide a standard method to qualifying membrane filtration processes.

• Journal of fish pathology
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• v.6 no.2
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• pp.205-218
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• 1993

On the development of hirame(Paratichtys olivaceus) culture, outbreak of scuticociliata infection was reported to cause severe damage in Japan. To establish effective measures for isolation and cultivation of this ciliate, we tried to culture this pathogenic ciliate using medium for bacteria and fish cell lines in vitro. Scuticociliata from the brain tissues of infected fish was aseptically inoculated to CHSE-214 cells cultured in MEM-10 without antibiotic. Scuticociliata grew well and the number of ciliate reached $10^6\;cells/ml$ at temperatures of $15^{\circ}C$ to $20^{\circ}C$ for 10d. The number of ciliate cultured in the cell lines is 10 times higher than the numbers cultured in the liquid medium alone. This ciliata could be cloned by dilution method. Scuticociliata isolated could grow well on 42 different cell lines that were established from marine fish, warm freshwater fish, and salmonids. This ciliate could be preserved in liquid nitrogen for more than 6 months. Subsequently, we observed the optimal temperature and salinity for growth, and tested the sensitivities of this organism to formaldehyde, flagyl(Metronidazole), Ekuteshin(Combination compound of sulfamonometoxin and ormethoprim), and ozonixation. Optimal temperature for growth was $25^{\circ}C$ and salinity was 1.0 to 1.5%. Washed scuticociliata was killed by formaldehyde at the concentration of 50ppm for 10min, but was not completely killed even at a high concentration of 400ppm for 20min in MEM-5. Flagyl and Ekuteshin can inhibit the growth of scuticociliata at the concentration of 1,000 and $100{\mu}g/ml$ in MEM-10, respectively. More than 99% of this scuticociliata could be killed by ozonization at a dose equivalent to $1.0mg/\ell$ oxidant for 30sec in sea water. Isolated scuticociliata showed the pathogenicity to the cultured hirame by artificial infection(I. P. injection, $10^5\;cells$/fish). The number of scuticociliata in the water could be counted by most probable number(MPN) method using tissue culture, and the minimum detectable number was $1.8\;cells/\ell$. The number in the reservoir tank for water supply to the culture tank was 110 cells/l. After cleaning by elimination of the sediments from of the reservoir tank and disinfected with formaldehyde, number of scuticociliata decreased and was counted less than $1.8\;cells/\ell$ and infection rate of cultured hirame was decreased.

• FLOWER RESEARCH JOURNAL
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• v.18 no.3
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• pp.216-219
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• 2010

An Asiatic lily cultivar 'Yeri' was developed in 2005 at National Institute of Horticultural and Herbal Science (NIHHS), Rural Development Administration (RDA), Korea. The cross was made in 1993 between Asiatic lily 'Geneve', a light pink colored cultivar, and 'Montreux', deep purple colored cultivar. The first selection was done and was tentatively named as 'A95-68' in 1995. After in vitro multiplication and bulbing production, growth and flowering characteristic tests were conducted from 1996 to 2003. The evaluation of characteristics was performed and named as 'Wongyo C1-19' in 2005 that was registered as 'Yeri' to the registration office of Korea Seed & Variety Service. 'Yeri' flowered at the first of July and grew average 34.6 cm stem in length. Flowers bloomed facing upward, unspotted in petals and deep purple (RHS, RP58A). The size of flower was 13.3 cm. Mean petal length and width was 7.7 cm and 2.7 cm, respectively. Leaves were 5.1 cm long and 0.5 cm wide, respectively. The weight and size of bulb were 9.6 g and 11.7 cm, respectively. Year-round flowering can be done by storing the bulb under $-1.5^{\circ}C$ conditions. For forced cultivation, it was necessary to add calcium to the fertilizer or remove side scales to prevent leaf scorch. It was needed to control Botrytis disease in wet season.

• Research in Plant Disease
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• v.22 no.1
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• pp.32-37
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• 2016

In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice black-streaked dwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted from RBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBank JX994211), a pair of S10 specific primers were designed and used to amplify the fragment encoding the N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR. Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). Recombinant RBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified protein was used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV from infected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, we provide purified RBSDV P10 (1-278 aa), which would be good material for the serological study of RBSDV-miryang isolates.

• Horticultural Science & Technology
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• v.30 no.4
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• pp.432-436
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• 2012

This study was conducted to determine the optimal aeration rate for mass propagation of ever-bearing strawberry by bioreactor culture. The aeration rate was treated in four levels: 0.1 vvm (air volume/medium volume/min), 0.2 vvm, 0.3 vvm, and 0.4 vvm. In 0.2 vvm conditions, shoot length was the longest at 9.03 cm in bioreactor culture, leave numbers were 40.4 ea and fresh weight was 6,106 mg. Plant growth rate at 0.2 vvm condition was faster than other treatments. In the aeration condition, 0.2 vvm was most effective to increase aerial part growth and to decrease medium consumption. As the culture periods increased, the fresh weight also increased rapidly. After six weeks of cultivation, shoots were emerged with 10.4 ea per plantlet, resulting in developing a complete plant. As a result, the bioreactor culture system for mass propagation of strawberry is required to continuously supply the air by 0.2 vvm speed and cultivate at least for six weeks.

• The Korean Journal of Mycology
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• v.46 no.3
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• pp.295-306
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• 2018

In the present study, we compared the effects of 50% ethanolic extracts of Chinese and Korean Wolfiporia cocos (CPE and KPE) on in vitro lipid accumulation in 3T3-L1 cells and their anticancer activities in Sarcoma 180 cells. We further compared the anticancer activities and the 50% inhibitory concentrations ($IC_{50}$) of CPE with KPE with cultivated for one and two years in a landfill and a facility (LPE and FPE), respectively. In addition, the single oral dose toxicities of CPE and KPE were evaluated in mice. Lipid accumulation was inhibited after 48 hours, in CPE and KPE treated 3T3-L1 cells; however, no significant difference was observed between CPE and KPE in their lipid accumulation inhibitory activities. The anticancer activity of KPE was higher than that of CPE at $300{\mu}g/mL$ (p<0.05), revealing the possibility of an auxiliary biological means for origin identification. The anticancer activities of LPE and FPE were significantly stronger than that of CPE (p<0.05) but there was no difference between extracts from one- and two-year-old W. cocos, irrespective of the cultivation method. In single oral dose toxicity tests, CPE and KPE did not induce mortality during the 14-day observation. Thus, the 50% of lethal dose ($LD_{50}$) of CPE and KPE were estimated to be higher than 2,000 mg/kg. Taken together, our results indicate that the anticancer assay could be an auxiliary means of identifying the origin of W. cocos. In addition, artificial cultivation could be an alternative way to reduce the import of W. cocos. Lastly, 50% ethanolic W. cocos extracts could be potential candidates for obesity and cancer managements.

• Journal of The Korean Society of Grassland and Forage Science
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• v.30 no.2
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• pp.115-120
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• 2010

This study was carried out to determine the forage production and quality of Italian ryegrass (IRG) and forage barley sown in early spring in Suwon, 2009. The five treatments used in this experiment were IRG Kowinearly (early maturity), IRG Kowinmaster (medium maturity), Yuyeon barley, Kowinearly + Yuyeon mixture, and Kowinmaster + Yuyeon mixture. The lodging was observed in IRG, but no lodging was found IRG + barley mixtures. The heading date of Kowinearly and Kowinmaster were 16 May and 22 May, respectively, and that of barley was 13 May. The dry matter (DM) percentage at harvest was 22.2~27.6%. The forage quality among treatments were similar, but the crude protein (CP) content of IRG was higher than that of barley, and in vitro DM digestibility was a little low in Kowinmaster. The yields of DM, CP and digestible DM were high in Kowinearly + Yuyeon barley mixtures as a 13,816 kg, 1,384 kg and 10,387 kg per ha, respectively (p<0.05). In conclusion, the mixture cultivation of IRG and forage barley was very effective, because of preventing of IRG lodging, increasing of forage yield, and stable production of forages. The optimum harvest date for silage manufacture of IRG and barley sown in early spring was recommended early June instead of May.

• Journal of The Korean Society of Grassland and Forage Science
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• v.22 no.4
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• pp.241-246
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• 2002

Rye-hairy vetch mixture would be recommended fur produce higher yield and conserving soil environment. This experiment was conducted to investigate the effect of planting and harvest dates on quality and productivity of rye-hairy vetch mixture. Plant height of rye and hairy vetch was increased with delayed harvest dates, but it found that there was on significant difference among seeding dates. The dry matter(DM) content was increased with delayed harvest dates, and it showed 30% DM in mid-May. Crude protein(CP) content was decreased sharply from 17~18% in heading stage to 9~10% in flowering stage. The content of ADF(Acid detergent fiber) and NDF(Neutral detergent fiber) were increased with delayed harvest date, but IVDMD(In vitro dry matter digestibility) and TDN(Total digestible nutrient) were decreased. The change of dry matter yield was affected significantly by harvest date but was not by seeding time. The results of this experiments indicated that harvest in late-April would be recommended to produce the highest yield and quality if it is considered to cultivate com fur silage. Harvest in mid-May would be recommendable with the cultivation of early maturity silage corn or sorghum $\times$ sudangrass hybrids.

• Journal of Forest and Environmental Science
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• v.30 no.2
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• pp.243-252
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• 2014

Forest tree species usually takes for long periods to be harvested and cultivated but spearmints are a good model system for woody plant because of reducing and shortening cultivation time. Spearmints are good model plants (Mentha species) for research about terpenoids production and industrial essential oil manufacture. Isopentenyl pyrophosphate isomerase (Iso) and limonene synthase (Limo) are the key enzymes of terpenoid biosynthesis pathway. Transgenic and wild spearmints (Mentha spicata, MS) were cultured in vitro and assessed for the essential oil contents. The content of essential oil of transgenic spearmint also was enhanced slightly depending on the target terpenoid genes. In an attempt to increase productivity of terpenoids further, salt and fungal elicitation strategy was adopted on transgenic Mentha spicata. The salt (800 mM NaCl) as abiotic and two fungi (Botrytis cinerea and Glomerella cingulata) as biotic were used for elicitors. In the absence of salt stress four terpenoids were detected from the spearmint extracts, all of them being monoterpenes. On the other hand, the transgenic (MSIso) extracts contained eleven terpenoids (10 monoterpenes and 1 phenylpropene) while transgenic (MSLimo) extracts contained seven monoterpenes. After 3 days of fungal infection, the resistance indices further increased to 4.38, 3.89 and 2.04 for wild type, MSIso and MSLimo, respectively. The salt and fungal elicitators proved beneficial towards modifying both the terpenoids profile and improvement in the composition of essential oil. These results have important applications for the large-scale production of essential oils and forest biotechnology with respect to spearmint.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.75-80
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• 2008

In vitro culture of shoot tip of garlic (Allium sativium L. cv. Seosan) was carried out to find medium condition of the induction of multiple shoots and bulbing for muliproduction of virus-free seed bulbs. For this work, tank culture was introduced. In shoot tip culture on MS solid medium the induction of multiple shoots and bulbing were better by adding 3% sucrose than 8%. Supplementation with 2mg/L 2ip and 0.2 mg/L IAA in this medium was effective. Three point three shoots including 2.7 bulbs were formed from a shoot tip after cultivation for 30 days on this medium. Bulbing of garlic in liquid culture with plastic water tank of 20L supplied air at the side of the lower part was better by adding 3% sucrose than 8% by subculture for 45 days with shoots obtained from shoot tip culture for 30 days on soid MS medium. Shoot growth was vigorous at 3% sucrose however bulb growth was more effective on the medium of 8% sucrose. Because of the effectiveness on solid medium added 3% sucrose, 2 mg/L 2ip and 0.2 mg/L IAA for initial production of multi-shoot in stem tip culture and the effectiveness in liquid culture with water tank for growth of bulbs, the method of two-step culture could be introduced for the multiple production of seed bulb of high quality. It was more desirable by supply of 0.2 mg/L BA and 0.02 mg/L NAA at tank culture time. But growth of the bulbs became poor by increasing concentration of NAA of the medium.