• Journal of Veterinary Science
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• 제22권4호
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• pp.56.1-56.10
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• 2021

Background: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. Objectives: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). Methods: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. Results: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (10^8.0 TCID₅₀/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). Conclusions: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

• 대한수의학회지
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• 제58권4호
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• pp.211-217
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• 2018

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.

• 대한수의학회지
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• 제61권2호
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• pp.19.1-19.7
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• 2021

Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:2¹⁵. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.

• 대한수의학회지
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• 제62권1호
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• pp.10.1-10.9
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• 2022

Feline panleukopenia virus (FPV) causes fatal leukopenia and severe hemorrhagic diarrhea in cats. Although FPV isolates have been reported worldwide from several animals, the biological and genetic features of South Korean FPVs remain unclear. We characterized molecularly South Korean FPV isolates. Crandell-Rees feline kidney (CRFK) cells were used to isolate FPV from 60 organ homogenates. The isolates were confirmed to be FPVs via analyses of cytopathic effects, immunofluorescence studies, electron microscopy, and polymerase chain reaction. Viral genetic analyses used the full VP2 sequences. Eight isolates propagated in CRFK cells were confirmed to be FPVs. All isolates yielded viral titers ranging from 10^4.5 to 10^6.0 TCID₅₀/mL 5 days after inoculation into CRFK cells and exhibited hemagglutination titers ranging from 2⁷ to 2¹² (using pig erythrocytes). The Korean FPV isolates grew well in cat cells such as CRFK and Fcwf-4 cells. The FPV isolates were most similar to the KS42 strain isolated from a Korean cat in 2008. The FPV isolates will serve as useful antigens in future sero-epidemiological studies and will aid in the development of diagnostic tools.

• 한국동물위생학회지
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• 제45권1호
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• pp.39-45
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• 2022

The total bacteria count is significantly important factor for hygienic quality in raw milk. BactoScan FM^TM and BactoCount IBC^TM are the automated instruments for the determination of the total bacterial count in raw milk. They have been used after calibration by standard plate count (SPC) method in South Korea since 2000. It is necessary to re-evaluate for total bacterial counter according to the improvement of milk quality and the change of milk quality grade. Therefore, this study was evaluated the conversion mode of the two machines by SPC method. We collected 921 bulk-tank milk samples throughout the concentration range of 1,000~1,000,000 CFU/mL from June 2020 to October 2021. As a result, when compared by the SPC value, there was a slight difference in total bacterial count in BactoScan below 10,000 CFU/mL and above 200,000 CFU/mL and in BactoCount above 100,000 CFU/mL, respectively. Therefore, the conversion factor for BactoScan and the conversion equation for BactoCount were newly adjusted based on SPC value, and then the correlation coefficients (R²) was 0.85 or higher. In addition, the correlation (R²) between BactoScan and BactoCount was 0.91, which means the results were high positive correlation. These results are expected to contribute to improving the accuracy of the automated instruments for determining of total bacterial count in raw milk.

• 대한수의학회지
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• 제57권4호
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• pp.249-252
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• 2017

Rabies virus (RABV) causes a neurological disease in warm-blooded animals that is nearly always fatal. In this study, we analyzed the matrix (M) genes in 10 Korean street RABV strains isolated from two Provinces during 2011-2013. The M genes in these 10 Korean strains were highly conserved during 1999-2013. Phylogenetic analysis revealed they were closely related to the M genes of RABVs isolated in northeastern China. Specific amino acid substitutions were identified in the KRVB1206, KRVF1301, and BV9901PJ strains. However, functional domains, including those involved in virus production and pathogenicity, were conserved in all 10 strains.

• The Plant Pathology Journal
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• 제21권2호
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• pp.111-118
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• 2005

Weather data for disease forecasts are usually derived from automated weather stations (AWS) that may be dispersed across a region in an irregular pattern. We have developed an alternative method to simulate local scale, high-resolution weather and plant disease in a grid pattern. The system incorporates a simplified mesoscale boundary layer model, LAWSS, for estimating local conditions such as air temperature and relative humidity. It also integrates special models for estimating of surface wetness duration and disease forecasts, such as the grapevine downy mildew forecast model, DMCast. The system can recreate weather forecasts utilizing the NCEP/NCAR reanalysis database, which contains over 57 years of archived and corrected global upper air conditions. The highest horizontal resolution of 0.150 km was achieved by running 5-step nested child grids inside coarse mother grids. Over the Finger Lakes and Chautauqua Lake regions of New York State, the system simulated three growing seasons for estimating the risk of grape downy mildew with 1 km resolution. Outputs were represented as regional maps or as site-specific graphs. The highest resolutions were achieved over North America, but the system is functional for any global location. The system is expected to be a powerful tool for site selection and reanalysis of historical plant disease epidemics.

• The Plant Pathology Journal
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• 제18권6호
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• pp.301-307
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• 2002

Graft transmission and cytopathological studies of a severe pear disease, pear black necrotic leafspot（PBNLS), were carried out to determine the causal agent of the disease. No evidence was found that a fungal or bacterial pathogen could be the causal agent of the disease. Attempts to transmit the agent by sap-inoculation to other plants including herbaceous hosts failed. How-ever, the pathogen was readily graft-transmitted from symptomatic diseased pears to healthy pears. Graft transmission of the pathogen was also demonstrated by using an indicator plant, PS-95, developed in the laboratory through various grafting methods. Ultrastructural study of the disease revealed the consistent presence of flexuous rod-shaped virus-like particles (VLP) in the symptomatic leaves of both Niitaka cultivar and indicator pear, PS-95. The particles, approximately 12 nm in diameter with undetermined length, occurred in the cytoplasm of mesophyll parenchyma cells. Cells with VLPs also contained fibril-containing vesicles, which are common in cells infected with plant viruses with ssRNA genome. The vesicles were formed at the tonoplast. Based on the symptomatology, the presence of fibril-containing vesicles, and graft-transmissibility, it is believed that the VLPs that occurred on symptomatic leaves of black necrotic leafspot of pear are viral in nature, possibly those of a capillovirus.

• The Plant Pathology Journal
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• 제32권3호
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• pp.216-227
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• 2016

Biological control agents (BCAs) from different microbial taxa are increasingly used to control bacterial wilt caused by Ralstonia solanacearum. However, a quantitative research synthesis has not been conducted on the role of BCAs in disease suppression. Therefore, the present study aimed to meta-analyze the impacts of BCAs on both Ralstonia wilt disease suppression and plant (host) growth promotion. The analysis showed that the extent of disease suppression by BCAs varied widely among studies, with effect size (log response ratio) ranging from -2.84 to 2.13. The disease incidence and severity were significantly decreased on average by 53.7% and 49.3%, respectively. BCAs inoculation also significantly increased fresh and dry weight by 34.4% and 36.1%, respectively on average. Also, BCAs inoculation significantly increased plant yield by 66%. Mean effect sizes for genus Pseudomonas sp. as BCAs were higher than for genus Bacillus spp. Among antagonists tested, P. fluorescens, P. putida, B. cereus, B. subtilis and B. amyloliquefaciens were found to be more effective in general for disease reduction. Across studies, highest disease control was found for P. fluorescens, annual plants, co-inoculation with more than one BCA, soil drench and greenhouse condition were found to be essential in understanding plant responses to R. solanacearum. Our results suggest that more efforts should be devoted to harnessing the potential beneficial effects of these antagonists, not just for plant growth promoting traits but also in mode of applications, BCAs formulations and their field studies should be considered in the future for R. solanacearum wilt disease suppression.

• The Plant Pathology Journal
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• 제35권3호
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• pp.189-199
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• 2019

In this research, the relationships among the 31 microsatellite markers with charcoal rot disease resistance related indices in 130 different soybean cultivars and lines were evaluated using association analysis based on the general linear model (GLM) and the mixed linear model (MLM) by the Structure and Tassel software. The results of microsatellite markers showed that the genetic structure of the studied population has three subpopulations (K=3) which the results of bar plat also confirmed it. In association analysis based on GLM and MLM models, 31 and 35 loci showed significant relationships with the evaluated traits, respectively, and confirmed considerable variation of the studied traits. The identified markers related to some of the studied traits were the same which can probably be due to pleiotropic effects or tight linkage among the genomic regions controlling these traits. Some of these relationships were including, the relationship between Sat_252 marker with amount of charcoal rot disease, Satt359, Satt190 and Sat_169 markers with number of microsclerota in stem, amount of charcoal rot disease and severity of charcoal rot disease, Sat_416 marker with number of microsclerota in stem and amount of charcoal rot disease and the Satt460 marker with number of microsclerota in stem and severity of charcoal rot disease. The results of this research and the linked microsatellite markers with the charcoal rot disease-related characteristics can be used to identify the suitable parents and to improve the soybean population in future breeding programs.