• Journal of Veterinary Clinics
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• v.20 no.2
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• pp.150-154
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• 2003

A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the intestines after two blind-passages in Vero cells supplemented with trypsin (10 ug/ml). and the biological and physicochemical properties of the isolate were characterized. The virion was roughly spherical in shape and had spike peplomers on its outer surface. The virus exhibited cytopathic effects such as rounding degeneration at initiation of infection and syncytia formation later in Vero cells. The virus was labile to 20% ether and 5% chloroform but stable in acid with pH 4-7 at $4^{\circ}C$. The infectivity of the virus was maintained at $50^{\circ}C$ for 180 min, and the buoyant density of the virus in sucrose was 1.180 g/ml. All biological and physicochemical properties of the virus were typical features of coronaviruses.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.284-288
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• 2001

Respiratory viruses are one of the most infectious agent in human. Six different respiratory tract viruses were detected from Busan while working on the preventive surveillance in 1997-2000. The isolation rate from suspected specimens were 8.4%. Influenza virus A, B type, parainfluenza virus, adenovirus, mumps virus, and measles virus were examined from throat swabs, serum, and secretions of patients. Influenza A/Sydney/05/97(H3N2)-like, A/Johanesburg/33/94(H3N2)-like, A/Beijing/262/95(H1N1)-like and Influenza B/Beijing/262/95-like, B/Harbin/07/94-like, B/Guangdong/08/93-like were found. Adenovirus serotype 1, 2, 3 and 5 were detected, antibody of mumps both IgM and IgG were shown and outbreaks of measles were confirmed. Different antigenic types of influenza virus were detected every year, one outbreak of parainfluenza in 1999, mumps outbreak in 1999 and 2000, and incidence of measles in 2000 were noticeable. Monthly outbreaks were November through following March with influenza virus, January through June with adenovirus, February through May and December with mumps, April through August and November, December with measles, respectively. The size of isolated viruses were 130 nm with influenza virus B type, non-enveloped, icosahedron with 70 nm with adenovirus, 170 nm with mumps virus and 180 nm with parainfluenza virus in diameter, respectively.

• Journal of fish pathology
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• v.11 no.2
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• pp.129-136
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• 1998

In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

• Journal of Veterinary Science
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• v.25 no.2
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• pp.21.1-21.15
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• 2024

Background: Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial. Objectives: To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication. Methods: The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway. Results: The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly downregulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010. Conclusions: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.

• Korean Journal of Veterinary Research
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• v.52 no.4
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• pp.223-230
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• 2012

The worldwide distribution and continuing genetic mutation of avian influenza virus (AIV) has been posed a great threat to human and animal health. A comparison of 3 isolates of AIV H9N2, A/chicken/Korea/KBNP-0028/00 (H9N2) (KBNP-0028), A/chicken/Korea/SNU8011/08 (H9N2) (SNU 8011) and an inactivated oil vaccine strain A/chicken/Korea/01310/01 (H9N2) (01310), was performed. The former 2 AIVs were isolated from field cases before and after the application of an inactivated H9N2 vaccine in 2007, respectively. The antigenic relationship, viral shedding, tissue tropism and genetic analysis were examined. The comparison of virus shedding from the cloaca and the oropharynx revealed that both isolates were more frequently isolated from the upper respiratory tract (90~100%) 1 day post inoculation (DPI) compared with isolation 5 DPI from gastrointestinal tracts (10~60%). Moreover, the isolate KBNP-0028 were recovered from all organs including bone marrow, brain and kidneys, indicating higher ability for broad tissue dissemination than that of SNU 8011. KBNP-0028 replicated earlier than other strains and with a higher titer than SNU 8011. In full-length nucleotide sequences of the NA gene and a partial sequence of the HA gene of SNU 8011, we found that there might be significant changes in tissue tropism, virus replication and genetic mutation in AIV H9N2 isolates.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.150.3-151
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• 2003

To analyze the genome of Cucumber mosaic virus(CMV) in pepper, we developed a new extraction method for double-stranded RNA(dsRNA). To isolate the dsRNA, 0.1g of pepper leaves homogenized with 1ml of 5${\times}$EXB extraction buffer[0.5M glycin, 0.5M NaCl, 5mM EDTA(pH9.0/NaOH), 10％ Sodium N-lauryl salcosinate(NLS), 10％ Sodium dodecylsulfate(SDS)] and purified with the 1/4 volume of phenol： chloroform： isoamylalcohol(25：24：1). dsRNAs from the aqueous phase was precipitated with isopropanol. This procedure was able to detect a minimal amount of dsRNA from CMV infected plant tissue and to distinguish different CMV satellite RNAs by polyacrylamide gel electrophoresis(PAGE). Moreover, this method can be applied CMV infected in pepper or Rice dwarf virus (RDV) infected rice.

• Korean Journal of Poultry Science
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• v.46 no.3
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• pp.173-183
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• 2019

Based on their virulence, the avian influenza viruses (AIVs) are classified into two pathotypes: low pathogenic avian influenza (LPAI) virus and highly pathogenic avian influenza (HPAI) virus. Among the 16 HA subtypes of AIV, only the H5 and H7 subtypes are classified as HPAI. Some AIVs, including H5 and H7 viruses, can infect humans directly. Six H7 subtype isolates from wild birds of the H7N7 (n=4) and H7N1 (n=2) subtypes were characterized in this study. Phylogenetic analysis showed that eight viral genes (HA, NA, PB2, PB1, PA, NP, M, and NS) of the H7 isolates clustered in the Eurasian lineage, the genetic diversity of which is indicated by its division into several sublineages. The Korean H7 isolates had two motifs, PEIPKGR and PELPKGR, at the HA cleavage site, which have been associated with LPAI viruses. Six H7 isolates encoded glutamine (Q) and glycine (G) at positions 226 (H3 numbering) and 228 of HA, suggesting avian-type receptor-binding specificity. None of the Korean H7 isolates had the amino acid substitutions E627K in PB2 and I368V in PB1, which are critical for efficient replication in human cells. The Korean H7 isolates showed no deletions in the NA stalk region and in NS. These results suggest that the Korean H7 isolates from wild birds are different from the H7N9 influenza viruses isolated in China in 2013, which are capable of infecting humans.

• Journal of Microbiology and Biotechnology
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• v.21 no.5
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• pp.449-454
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• 2011

The infection of pandemic influenza viruses such as swine flu (H1N1) and avian flu viruses to the host cells is related to the following two factors: First, the surface protein such as HA (hemagglutinin) and NA (neuraminidase) of the influenza virus. Second, the specific structure of the oligosaccharide [sialic acid(${\alpha}2$-6) galactose(${\beta}1$-4)glucose or sialic acid(${\alpha}2$-3)galactose(${\beta}1$-4)glucose] on the host cell. After recognizing the specific structure of the oligosaccharide on the surface of host cells by the surface protein of the influenza virus, the influenza virus can secrete sialidase and cleave the sialic acid attached on the final position of the specific structure of the oligosaccharide on the surface of host cells. Tamiflu (oseltamivir), known as a remedy of swine flu, has a saccharide analog structure, especially the sialic acid analog. Tamiflu can inhibit the invasion of influenza viruses (swine flu and avian flu viruses) into the host cells by competition with sialic acid on the terminal position of the specific oligosaccharide on the surface of the host cell. Because of the emergence of Tamiflu resistance, the development of new potent anti-influenza inhibitors is needed. The inhibitors with positive-charge groups have potential as antiviral therapeutics, and the strain specificity must also be resolved.

• Korean Journal of Microbiology
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• v.43 no.1
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• pp.23-30
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• 2007

The most rapid Real-time PCR based detection method for Avian influenza A virus (AIV) subtype H5N1 was developed. The target DNA sequence in this study was deduced from H5N1 subtype-specific 387 bp partial gene of hemagglutinin, and was synthesized by using PCR-based gene synthesis on the ground of safety. Real-Time PCR was performed by $GenSpector^{TM}$ using microchip-based, total $1{\mu}l$ of reaction mixture with extremely short time in each steps in PCR. The detection including PCR-amplication and analysis of melting temperature was totally completed within 13 min. The H5N1-specific 189 bp PCR product was correctly amplified until 2.4 molecules of hemagglutinin gene as minimum of templates. This kind of PCR was designated as Quick Real-Time PCR in this study and it could be applied to detect not only AIV H5N1, but also other pathogens using PCR-based detection.

• Journal of Veterinary Science
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• v.24 no.5
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• pp.73.1-73.16
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• 2023

Background: Highly pathogenic avian influenza virus (HPAIV) is considered a global threat to both human health and the poultry industry. MicroRNAs (miRNA) can modulate the immune system by affecting gene expression patterns in HPAIV-infected chickens. Objectives: To gain further insights into the role of miRNAs in immune responses against H5N1 infection, as well as the development of strategies for breeding disease-resistant chickens, we characterized miRNA expression patterns in tracheal tissues from H5N1-infected Ri chickens. Methods: miRNAs expression was analyzed from two H5N1-infected Ri chicken lines using small RNA sequencing. The target genes of differentially expressed (DE) miRNAs were predicted using miRDB. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were then conducted. Furthermore, using quantitative real-time polymerase chain reaction, we validated the expression levels of DE miRNAs (miR-22-3p, miR-146b-3p, miR27b-3p, miR-128-3p, miR-2188-5p, miR-451, miR-205a, miR-203a, miR-21-3p, and miR-200a3p) from all comparisons and their immune-related target genes. Results: A total of 53 miRNAs were significantly expressed in the infection samples of the resistant compared to the susceptible line. Network analyses between the DE miRNAs and target genes revealed that DE miRNAs may regulate the expression of target genes involved in the transforming growth factor-beta, mitogen-activated protein kinase, and Toll-like receptor signaling pathways, all of which are related to influenza A virus progression. Conclusions: Collectively, our results provided novel insights into the miRNA expression patterns of tracheal tissues from H5N1-infected Ri chickens. More importantly, our findings offer insights into the relationship between miRNA and immune-related target genes and the role of miRNA in HPAIV infections in chickens.