• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.42 no.3
• /
• pp.238-242
• /
• 2009

Recently coastal seawater has been contaminated by enteric viruses such as the norovirus via untreated groundwater globally. Accordingly, the consumption of molluscan shellfish from seawater that has been contaminated with fecal material has become an important issues. The levels of enteric viruses in seawater are low and recovery and concentration of the virus from large volumes of water is difficult. We compared the effectiveness of two representative method of concentrating virus using negatively and positively charged filters. The mean retention of seeded FCV by HAMF and NCCF was 48% and 78%, respectively. Overall, the recovery of NCCF was 43.3$\pm$11% better than that of HAMF. However, the eluate obtained by using beef extract solution in the NCCF procedure caused an inhibitory effect on the RT-PCR; therefore, it was necessary to employ a PCR inhibitor removal procedure. The HAMF eluate contained no PCR inhibitors, but HAMF was not an effective method of concentrating the virus from large volumes of natural seawater due to clogging.

• Korean journal of applied entomology
• /
• v.19 no.2 s.43
• /
• pp.91-95
• /
• 1980

The experiment was conducted to culture callus tissue induced from foliage leaf of garlic bulb for the production of virus-free stocks and for the reduction of expenses for seeds, The following results were reached. 1. Linsmaier-Skoog basal medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) $10^{-5}M$ and benzyladenine $10^{-5}M$ showed the most effective for the induction for the induction of garlic callus. 2. The growth rate of callus was the highest in Linsmaier-Skoog basal medium containing kinetin $10^{-6}M\;and\;2,4-D\;10^{-6}M$ 3. The results of periodical assay of virus concentration in callus tissues showed that virus was almost eliminated by repeated transfer of translucent and soft tissue for eight generations. 4. When virus-free garlic callus tissues were transfered to Murashige-Skoog basal medium containing kinetin $10^{-5}M$ and naphthaleneacetic acid $5\times10^{-6}M$, the tissues were redifferentiated and formed plantlet.

• Korean Journal of Veterinary Research
• /
• v.45 no.4
• /
• pp.569-574
• /
• 2005

Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

• BMB Reports
• /
• v.35 no.2
• /
• pp.206-212
• /
• 2002

The NS2B-NS3(pro) polyprotein segment from the dengue virus serotype 2 strain 16681 was purified from overexpressing E. coli by metal chelate affinity chromatography and gel filtration. Enzymatic activity of the refolded NS2B-NS3(pro) protease complex was determined in vitro with dansyl-labeled peptide substrates, based upon native dengue virus type 2 cleavage sites. The 12mer substrate peptides and the cleavage products could be separated by reversed-phase HPLC, and were identified by UV and fluorescence detection. All of the peptide substrates (representing the DEN polyprotein junction sequences at the NS2A/NS2B, NS2B/NS3, NS3/NS4A and NS4B/NS5 sites) were cleaved by the recombinant protease NS2B-NS3(pro). No cleavage was observed with an enzymatically inactive S135A mutant of the NS3 protein, or with a modified substrate peptide of the NS3/NS4A polyprotein site that contained a K2093A substitution. Enzymatic activity was dependent on the salt concentration. A 50% decrease of activity was observed in the presence of 0.1M sodium chloride. Our results show that the NS3 protease activity of the refolded NS2B-NS3(pro) protein can be assayed in vitro with high specificity by using cleavage-junction derived peptide substrates.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2013.02a
• /
• pp.87-87
• /
• 2013

Sample concentration and purification processes are essential in the bio-analytical and pharmaceutical fields because most bio samples or media are extremely sophisticated. To concentrate and purify specific substances, passive membrane type filters have been utilized, which is driven by size or charge differences between target and others. The traditional and representative method to identify nucleic acid sequences in the complex biosample is gel electrophoresis, which has been worked by size and net charge of molecules. The adsorption phenomena have been also utilized to concentrate and purify biomolecules. This adsorption of biomolecule can be controlled under specific salts and surfaces as well as surface area. To utilize the differences of physical properties of molecules or bio-targets such as virus, bacteria, and cells, the nanotechnologies can be introduced in target concentration, purification, and isolation processes. In here, I'd like to briefly survey typical examples of nanobiotechnologies which are introduced in sample treatment. Also I specifically demonstrate two different simple techniques to concentrate and detect bacteria from the samples using multifunctional silica nanotube (SNT).

• Microbiology and Biotechnology Letters
• /
• v.40 no.4
• /
• pp.396-402
• /
• 2012

The chemical composition, anti-porcine epidemic diarrhea virus (PEDV) activity and antimicrobial activity of Artemisia dubia essential oil were evaluated in this study. Fifty eight compounds from A. dubia essential oil were identified through analysis by gas chromatography-mass spectrometry (GC-MS). The major constituents of the oil were camphor (17.18 %), germacrene-D (15.70%), trans (${\beta}-$) racaryophyllene (6.79%), ene thujones (6.57%), 1, 8-cineole (5.94%) and camphene (5.08%). The essential oil was evaluated for antiviral activity against PEDV in Vero cells using a cytopathic effect (CPE) reduction method. The oils actively inhibited PEDV replication with a 50% inhibitory concentration ($IC_{50}$) of 43.7 ${\mu}^3/mL$. The 50% cytotoxicity concentration ($CC_{50}$) of the oils was over 100 ${\mu}/mL$ and the derived therapeutic index was >2.3. Similar analysis of the ribavirin revealed that they have a relatively weaker efficacy when compared to the oils. The antimicrobial activity of the essential oil against 5 microorganisms was evaluated by the disc diffusion method. The essential oil exhibited antimicrobial activity against 5 tested microorganisms with a clear zone of 8-22 mm. Among the tested microorganisms, Streptococcus pyogenes was the most sensitive and Candida albicans the least. Therefore, in can be concluded that essential oils of A. dubia may have interesting applications for microbial control or the control of PEDV-derived diseases.

• Food Science and Preservation
• /
• v.21 no.2
• /
• pp.170-174
• /
• 2014

Hepatitis A virus (HAV) infection leads to acute liver failure and death through the intake of contaminated food. The polymerase chain reaction (PCR) has been used to detect HAV in food samples. HAV detection takes a long time, however, due to the virus concentration step required before PCR assay. In this study, a rapid method of detecting the HAVs present in lettuce using immunomagnetic separation combined with quantum dots (IMS-QDs) assay was developed. The detection limit of IMS-QDs for HAV was 10 $TCID_{50}/mL$, similar to the result that was obtained using RT-PCR combined with PEG or IMS. The application of IMS-QDs assay completed the viral detection within one hour, but this was not possible using PEG combined with RT-PCR. In conclusion, IMS-QDs assay is a rapid and efficient method for detecting HAV at a low concentration in agricultural products.

• Korean Journal of Plant Resources
• /
• v.26 no.2
• /
• pp.303-309
• /
• 2013

This study was performed to clarify the effect of medium compositions and plant growth regulators on the shoot, root formation and growth of 'Jarang' grape for mass propagation of virus-free plant. The formation and growth of shoot were considerably favorable in half-concentration of MS medium. However, the formation of adventitious root per explants (avg. 2.1) was effective in higher concentration (two times) of MS medium. For sucrose concentration, 1% for the shoot formation, 3% for the adventitious root formation and 1% for the growth were observed as yield significant results. With the addition of 0.05% of activated carbon, the shoot growth was improved, and it was effective for the adventitious root formation and growth as well. A pH of 6.8 in the medium was the most suitable for mass propagation; the results showed significant enhancement in the number of nodes and the length of the shoot, 3.9 and 1.3 cm, respectively. The shoot growth was the most vigorous in BA 1.0 mg/L due to the impact of the growth regulator on the mass propagation in it. Consequently, 16.9 shoots per explant were formed in NAA 1.0 mg/L so good results were obtained.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.3
• /
• pp.497-503
• /
• 2001

The purpose of this study was to examine the efficacy and mechanism of the cryo-precipitation, solvent/detergent (S/D) treatment, monoclonal anti-FVIIIc antibody (mAb) column chromatography, Q-Sepharose column chromatography, and lyophilization involved in the manufacture of antithemophilic factor VII(GreenMono) from human plasma, in the removal and/or inactivation of blood-borne viruses. A variety of experimental model viruses for human pathogenic viruses, including the bovine viral diarrhoea virus (BVDV), bovine herpes virus (BHV), murine encephalomyocarditis virus (EMCV), and porcine parvovirus (PPV), were all selected for this study. BHV and EMCV were effectively partitioned from a factor VII during the cryo-precipitation with a log reduction factor of 2.83 and 3.24, respectively. S/D treatment using the organic solvent, tri(n-butyl) phosphate (TNBP), and the detergent, Triton X-100, was a robust and effective step in inactivating enveloped viruses. The titers of BHV and BVDV were reduced from the initial titer of 8.85 and $7.89{log_10} {TCID_50}$, respectively, reaching undetectable levels within 1 min of the S/D treatment. The mAb chromatography was the most effective step for removing nonenveloped viruses, EMCV and PPV, with the log reduction factors of 4.86 and 3.72, respectively. Q-Sepharose chromatography showed a significant efficacy for partitioning BHV, BVDV, EMCV, and PPV with the log reduction the log reduction factors of 2.32, 2.49, 2.60, and 1.33 respectively. Lyophilization was an effective step in inactivating g nonenveloped viruses rather than enveloped viruses, where the log reduction factors of BHV, BVDV, DMCV, and PPV were 1.41, 1.79, 4.76, and 2.05, respectively. The cumulative log reduction factors of BHV, BVDV, EMCV, and PPV were ${\geqq}$11.12, ${\geqq}$7.88, 15.46, and 7.10, respectively. These results indicate that the production process for GreenMono has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

• Korean journal of applied entomology
• /
• v.18 no.1 s.38
• /
• pp.1-10
• /
• 1979

An inclusion forming virus isolated from a fan webworm, Hyphantria cunea, in 1975 was identified as a nuclear polyhedrosis virus. With the virus isolated in Korea, it was considered that the virus would be one of the valuable microorganism in microbial control. In this connection, 1) the shape and size of the virus for identification, 2) susceptibility of the various instar larvae to the virus, 3) the effects of storage condition on the pathogenicity and the cross infection of the virus to the larvae of Bombyx mori were examined. The results are summarized as follows; 1. The polyhedron was tetrahedron or hexahedron of $2\mu$ in size and the rod-shaped virus particles consisting of $2\~14$ rods in a bundle were $330m{\mu}\times35m{\mu}$ in size. 2. The hexagonal nuclear polyhedra were found only in the nucleus of the midgut cells but were variable in size. 3. The $LD_{50}$ values for the various instar larvae of H. cunea were $8.377\times10^4\;PIBs/ml$ for the second, $4.974\times10^5\;PIBs/ml$ for the fifth instar larvae. The $LT_{50}values$ for $10^6\;PIBs/ml$ were 9.6 days for the second, 11.5 days for the third, 12.0 days for the fourth and 17 days for the fifth instar larvae. 4. The susceptibility of H. cunea to the nuclear polyhedrosis virus was greater in the first generation than in the second generation. 5. The effect of the storage conditions on the pathogenicity of the nuclear polyhedra was less in refrigerator $(5^{\circ}C)$ and in freezing $(-80^{\circ}C)$ than in room temperature $(18.5^{\circ}C)$, especially as air-dried polyhedra than as suspension. The pathogenicity of the polyhedra seemed to decrease by sunlight during storage as cadavers, since rather greater decrease in pathogenicity was found in sunny condition than in shady condition. 6. The effective spray concentration was $6.4\times10^7\;PIBs/ml$ in the field and its $LT_50$ values for the third and the fifth instar larvae were 4.8 days and 14.2 days, respectively. 7. No cross infections were found in the nuclear polyhedrosis virus between H. cunea and B. mori. larvae.