• Korean Journal of Veterinary Service
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• v.44 no.3
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• pp.149-155
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• 2021

In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disinfectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under biological safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD₆₀₀ 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.

• Research in Plant Disease
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• v.21 no.4
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• pp.290-296
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• 2015

This study was conducted to establish an efficient screening system for resistant tomato to bacterial wilt caused by Ralstonia solanacearum. Under several conditions such as inoculation methods, growth stages of tomato seedlings, inoculum concentrations, and incubating temperatures after inoculation, development of bacterial wilt on nine resistant or susceptible cultivars of tomato was investigated. To inoculate by drenching the non-cut roots with the bacterial suspension was better to distinguish resistance and susceptibility of tomato cultivars than by drenching the cut roots using scalpel. And 'Hawaii7996' a resistant tomato to R. solanacearum showed high resistance at all the tested conditions including growth stages (3-, 6-, 8-, 10-leaf stages), inoculum concentrations ($OD_{600}=0.1-0.4$) and incubation temperatures (25, 30, $35^{\circ}C$). On the other hands, susceptible cultivars represented disease index of 3.7 and 3.9 at 6- and 8-leaf stages, respectively. At 3- and 10-leaf stages, the cultivars demonstrated lower disease severity of 2.1 and 0.5, respectively, than at 6- and 8-leaf stages. When the inoculated seedlings were incubated in growth chambers of 25, 30 and $35^{\circ}C$, disease severity of susceptible cultivars was significantly greater at 30 and $35^{\circ}C$ than at $25^{\circ}C$. In addition, the level of resistance of the tomato cultivars was not significantly affected by inoculum concentrations of $OD_{600}=0.1-0.4$. On the basis of the results, we suggest an efficient screening method to measure resistance level of tomato cultivars to bacterial wilt. The eight-leaf stage seedlings transplanted 7 days before inoculation, are inoculated with R. solanacearum by drenching the non-cut roots with a bacterial suspensions ($OD_{600}=0.4$) to give inoculum volume of 50 ml/soil l. The inoculated plants are incubated in a growth room at $30^{\circ}C$ for 12-13 days with 12-hour light a day.

• Geophysics and Geophysical Exploration
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• v.9 no.1
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• pp.19-29
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• 2006

The first Japanese pilot-scale $CO_2$ sequestration project has been undertaken in an onshore saline aquifer, near Nagaoka in Niigata prefecture, and time-lapse well logs were carried out in observation wells to detect the arrival of injected $CO_2$ and to evaluate $CO_2$ saturation in the reservoir. $CO_2$ was injected into a thin permeable zone at the depth of 1110m at a rate of 20-40 tonnes per day. The total amount of injected $CO_2$ was 10400 tonnes, during the injection period from July 2003 to January 2005. The pilot-scale demonstration allowed an improved understanding of the $CO_2$ movement in a porous sandstone reservoir, by conducting time-lapse geophysical well logs at three observation wells. Comparison between neutron well logging before and after the insertion of fibreglass casing in observation well OB-2 showed good agreement within the target formation, and the higher concentration of shale volume in the reservoir results in a bigger difference between the two well logging results. $CO_2$ breakthrough was identified by induction, sonic, and neutron logs. By sonic logging, we confirmed P-wave velocity reduction that agreed fairly well with a laboratory measurement on drilled core samples from the Nagaoka site. We successfully matched the history changes of sonic P-wave velocity and estimated $CO_2$ saturation a(ter breakthrough in two observation wells out of three. The sonic-velocity history matching result suggested that the sweep efficiency was about 40%. Small effects of $CO_2$ saturation on resistivity resulted in small changes in induction logs when the reservoir was partially saturated. We also found that $CO_2$ saturation in the $CO_2$-bearing zone responded to suspension of $CO_2$ injection.

• Tuberculosis and Respiratory Diseases
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• v.60 no.6
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• pp.663-672
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• 2006

Background: $PM_{10}$(Particulate matter with a diameter ($<10{\mu}m$), which is characterized by different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also produce unique particulate matter in affected areas. This study investigated the cytokine produced by A549 epithelial cells exposed to particles collected during both the Asian dust pfenomenon and ambient air particles in a non-dusty period. Method: Air samples were collected using a high volume air sampler(Sibata Model HV500F) with an air flow at $500{\ell}/min$ for at least 6 hours. The cytokine messenger RNA(mRNA) was measured using a reverse transcriptase polymerase chain reaction(RT-PCR). The A549 cells were exposed to 10 to $500{\mu}g/m{\ell}$ of a suspension containing $PM_{10}$ for 24 hours. Each was compared with those in the non-exposed control cells. Result: The mRNA levels of interleukin(IL)-$1{\alpha}$, $IL-I{\beta}$, IL-8, and the granulocyte macrophage colony stimulating factor(GM-CSF) increased after veing exposed to $PM_{10}$ in the ambient air particles, compared with those in the non-exposed control cells. The increase in $IL-1{\alpha}$ and IL-8 were dose dependent at a $PM_{10}$ concentration between $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$. The mRNA level of IL-8 in the A549 epithelial cells was higher during the in the Asian dust period($500{\mu}g/m{\ell}$) than during the non dust period. Conclusion: A549 cells exposed to the $PM_{10}$ collected during the Asian dust period produce more proinflammatory cytokine than during non-dusty period. This cytokine enhances the local inflammatory response in the airways and can also contribute to the systemic component of this inflammatory process.

• Korean Journal of Ecology and Environment
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• v.56 no.1
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• pp.94-103
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• 2023

Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria's early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.