• Title/Summary/Keyword: cytopathic effect

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Isolation of porcine reproductive and respiratory syndrome virus(PRRSV) in Korea (돼지생식기 및 호흡기증후군(Porcine Reproductive and Respiratory Syndrome ; PRRSV) 바이러스의 국내분리주 작성에 관한 연구)

  • Kweon, Chang-hae;Kwon, Byung-joon;Lee, Han-jung;Cho, Jae-jin;Hwang, Eui-kyung;Shin, Jin-ho;Yoon, Yong-dhuk;Kang, Yung-bai;An, Soo-hwan;Kim, Yong-hee;Huh, Won;Jun, Moo-hyung;Wensvoort, G.
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.77-83
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    • 1994
  • Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.

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Antiviral activity of ginsenosides against coxsackievirus B3, enterovirus 71, and human rhinovirus 3

  • Song, Jae-Hyoung;Choi, Hwa-Jung;Song, Hyuk-Hwan;Hong, Eun-Hye;Lee, Bo-Ra;Oh, Sei-Ryang;Choi, Kwangman;Yeo, Sang-Gu;Lee, Yong-Pyo;Cho, Sungchan;Ko, Hyun-Jeong
    • Journal of Ginseng Research
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    • v.38 no.3
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    • pp.173-179
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    • 2014
  • Background: Ginsenosides are the major components responsible for the biochemical and pharmacological actions of ginseng, and have been shown to have various biological activities. In this study, we investigated the antiviral activities of seven ginsenosides [protopanaxatriol (PT) type: Re, Rf, and Rg2; protopanaxadiol (PD) type: Rb1, Rb2, Rc, and Rd)] against coxsackievirus B3 (CVB3), enterovirus 71 (EV71), and human rhinovirus 3 (HRV3). Methods: Assays of antiviral activity and cytotoxicity were evaluated by the sulforhodamine B method using the cytopathic effect (CPE) reduction assay. Results: The antiviral assays demonstrated that, of the seven ginsenosides, the PT-type ginsenosides (Re, Rf, and Rg2) possess significant antiviral activities against CVB3 and HRV3 at a concentration of $100{\mu}g/mL$. Among the PT-type ginsenosides, only ginsenoside Rg2 showed significant anti-EV71 activity with no cytotoxicity to cells at $100{\mu}g/mL$. The PD-type ginsenosides (Rb1, Rb2, Rc, and Rd), by contrast, did not show any significant antiviral activity against CVB3, EV71, and HRV3, and exhibited cytotoxic effects to virus-infected cells. Notably, the antiviral efficacies of PT-type ginsenosides were comparable to those of ribavirin, a commonly used antiviral drug. Conclusion: Collectively, our findings suggest that the ginsenosides Re, Rf, and Rg2 have the potential to be effective in the treatment of CVB3, EV71, and HRV3 infection.

Mass Mortalities of Cultured Striped Beakperch, Oplegnathus fasciatus by Iridoviral Infection (이리도 바이러스 감염에 의한 양식 돌돔, Oplegnathus fasciatus의 대량폐사)

  • Sohn, Sang-Gyu;Choi, Dong-Lim;Do, Jeung-Wan;Hwang, Jee-Youn;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.13 no.2
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    • pp.121-127
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    • 2000
  • From August to October 1998, over 60% mortality of cultured striped beakperch Oplegnathus fasciatus was occurred in net cages along the southern coast of Korea. Moribund fish showed some clinical signs of lethargic behavior, dark coloration or decoloration, severe gill anemia and enlargement of spleen. Also enlarged basophilic cells showing Feulgen -positive reaction were observed in the tissue section of spleen, kidney, liver and heart of the diseased fish. GF cells inoculated with spleen homogenate of diseased fish produced cytopathic effect of enlarged and rounded cells, therefore the causative virus was isolated from diseased fish. Striped beakperch fingerlings intraperitoneally inoculated with the causative virus ($10^4TCID_{50}$/0.1 ml) revealed symptoms similar to those of naturally infected fish and died from 7 to 14 days post injection. Transmission electron microscopy revealed that the causative virus was enveloped icosahedral particle with 120~130 nm in diameter. PCR products of the expected size (500 bp) were amplified with a primer set based on the ATPase gene of RSIV(red sea bream iridovirus) using template DNAs which were extracted from the spleen of diseased fish and GF cells inoculated with the causative virus. According to the analysis of nucleotide sequence of these PCR products, the sequence from ATPase cDNA gene of the causative virus showed 95% homology with that of RSIV. These results indicate that the mass mortality in the cultured striped beakperch was caused by the infection of iridovirus similar to RSIV.

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Characterization of an isolated reovirus from the paradise fish Macropodus opercularis imported from Southeast Asia (동남아시아로부터 수입된 paradise fish Macropodus opercularis로부터 분리한 reovirus의 특성)

  • Kim, Wi-Sik;Kim, Soo-Jin;Kim, Jeong-Ho;Jung, Sung-Ju;Kim, Do-Hyung;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.335-342
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    • 2010
  • In 2008, mass mortality was observed in paradise fish Macropodus opercularis which was imported from Indonesia. PCR of these fish found positive for megalocytivirus and Mycobacterium sp., while an unidentified virus was culture-isolated using CHSE-214 cells. In the present study, we investigated characterization of the unidentified virus and its pathogenicity to determine whether the virus was the causative agent of the mass mortality of paradise fish. The unidentified virus induced cytopathic effect (CPE) with syncytia in CHSE-214 and other fish cells, BF-2, GF, SSN-1, FSP and FFN. The virus was resistant against treatments with IUdR, chloroform, acidity at pH 3, basicity at pH 11 and high temperature at $56^{\circ}C$ for 3h. By electron microscopy, the viral particles were spherical having a double capsid structure with approximately 65 nm in external diameter. Viral genome was composed of at least 10-segmented RNA with sizes ranging from 0.7 kb to 3.6 kb. Based on these characters, this virus can be classified into family Reoviridae. This reovirus did not cause any mortality in an artificial experiment conducted by injecting the virus to paradise fish. This indicates that the reovirus is not only responsible for the mass mortality of paradise fish in 2008.

Comparison of Microbiological Safety of Porcine Grafts on Gamma Irradiation for Use of Xenografts (돼지 유래 생체 조직의 이식재 활용을 위한 방사선 조사 미생물 제어 평가)

  • Jo, Eu-Ri;Kim, Jeongsoo;Choi, Jong-il;Kim, Jae-Hun;Sung, Nak-Yun;Song, Beom-Seok;Kim, JaeKyung;Park, Jong-Heum;Lee, Ju-Woon
    • Journal of Radiation Industry
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    • v.5 no.3
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    • pp.279-283
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    • 2011
  • This study was compared microbiological safety with gamma-irradiated porcine tendon and skin, as materials for the development of xenografts to regenerate damaged tissues and protect secondary contamination. The porcine tendon and skin were gamma-irradiated after inoculation of bacteria and virus to evaluate irradiation sensitivity of microorganisms. The result showed that the porcine tendon and skin were not different on the sensitivity of microorganisms by gamma irradiation. Bacteria inoculated in the porcine tendon and skin were confirmed that E. coli was the $D_{10}$ values of $0.32{\pm}0.082$ and $0.25{\pm}0.1kGy$ on tendon and skin, and B. subtilis was $4.00{\pm}0.312$ and $3.88{\pm}0.3kGy$ on gamma irradiation, respectively. Moreover, Virus inoculated in the porcine tendon and skin was observed that poliovirus (PV) was $6.26{\pm}0.332$ and $6.88{\pm}0.3kGy$, and porcine parvovirus (PPV) was $1.75{\pm}0.131$ and $1.73{\pm}0.2kGy$ and bovine viral diarrhoea virus (BVDV) was $3.70{\pm}0.212$ and $3.81{\pm}0.2kGy$ on gamma irradiation, respectively. Virus showed higher resistance compared to bacteria on gamma irradiation, but was not detected CPE (cytopathic effect) by virus both tendon and skin at 25 kGy, a standard dose recommended from IAEA for sterilization of medical products. Therefore, These results were considered that gamma irradiation could control effectively bacteria and virus to develop safe porcine xenograft, and apply same irradiation doses to all tissues including tendon and skin of porcine.

CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))

  • Kim, Uh-Ho
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.23-62
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    • 1969
  • Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

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Antioxidant and Antiviral Activities of Polyphenolics in Plum Wine (자두와인 내 폴리페놀 화합물의 항산화 및 항바이러스 활성)

  • Kang, Byung-Tae;Kwon, Dur-Han;Choi, Wha-Jung;Kim, Soon-Hee;Park, Dong-Cheol
    • Food Science and Preservation
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    • v.15 no.6
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    • pp.891-896
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    • 2008
  • Total phenolics and flavonoids, and the antioxidant capacity of plum cultivar wines (Prunus salicina L. cv. Soldam and P. salicina L. cv. Formosa) were determined using spectrophotometric methods. The total phenolic and flavanoid contents of Soldam wine were $478.4\;{\pm}\;5.6\;mg$ GAE and $202.4\;{\pm}\;7.5\;mg$ CE per L,respectively, and in Formosa wine were $200.6\;{\pm}\;7.5\;mg$ GAE and $64.4\;{\pm}\;6.8\;mg$ CE per L, respectively. Neutral and acidic phenolics in Soldam wine were extracted with ethyl acetate and 0.01 N HCl, respectively. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay, neutral phenolics (64.5 EDA%) had $3{\sim}4$ times higher antioxidant activity than acidic phenolics (21.5 EDA%) and other related phenolic compounds such as chlorogenic acid (15.5 EDA%) and quercetin (24.6 EDA%) at a concentration of $100\;{\mu}g/mL$. The antiviral activities of neutral and acidic phenolics in Soldam wine were investigated in vitro using a virus-induced cytopathic effect (CPE) inhibition assay. Results showed that neutral and acidic phenolics at concentrations of $100\;{\mu}g/mL$ inhibited porcine epidemic diarrhea virus (PEDV) replication at rates of 78.12% and 58.37%, respectively. The inhibition rate of 10 g/mL neutral phenolics (69.42%) was higher than that of ribavirin as an antiviral reagent (57.86%). At concentrations of $100\;{\mu}g/mL$ or less, neutral and acidic phenolics of Soldam wine had no cytotoxic effect against vero cells.

Studies on Host-Virus Interaction of Poxviruses 1. Cytochemical, Autoradiographic and Immunocytological Analysis in Cowpox Virus-FL Cell System (Poxvirus 감염(感染)에 있어서의 Virus-숙주세포(宿主細胞)의 상호관계(相互關係) 1. Cowpox Virus-FL 세포계(細胞系)의 세포화학적(細胞化學的) Autoradiography 및 세포면역학적해석(細胞免疫學的解析))

  • Kim, Uh Ho
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.57-67
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    • 1975
  • The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

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An investigation of the congestion on the gills of eel (뱀장어 아기미 울혈증에 관한 연구)

  • Kim, Young-Gill;Choi, Min-Soon;Park, Sung-Woo;Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.7 no.2
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    • pp.161-171
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    • 1994
  • In order to elucidate the outbreak mechanisms of a new disease which is characterized by an intense congestion in central venus sinuses(CVS) of gill filaments in cultured eel. these experiments were carried out; epidemically surveyed on the cultured eel farms in the vicinity of Kunsan city and experimentaliy outbreaked the disease in the stressful condition such as thermal and handling shock and innoculated the supernatant from the homogenate of naturally severe congested gill into eels and onto the monolayer of the CHSE-214. Although the frequency of congestion in eels of B, C, D and E farms were higher than in eels of A farms, the water qualities(stocked and cultured water) among farms were not a great difference. In eels of B, C, D and E farms, the values of haematocrit(Ht), haemoglobin(Hb), total protein(Tp), albumin(Alb), glucose(Glu), magnesium(Mg) were lower and the values of calcium(Ca), methemoglobin(Met-Hb), glutamic pyruvic transminase(GPT), glutamic oxalacetic transminase(GOT) higher than in eels of A farms. These valules have not related to the frequency of congestion. An intensive congestion and dilataton in CVS of gill filaments in experimentally handling-stressed eels produced similar histopathological changes to those observed in the spontaneously diseased eel, but not in eels experimentally injected with filtering contents. The cytopathic effect on the CHSE-214 was not observed. In stressed eels the congestion of gill was increased in relation to either the decrease ranges of water temperature or the incerase in accllimated times. And increase in Ht, Met-Hb, Alb, Glu, GOT and GPT and decrease in Mg, Hb and Tp were found, which had a close relationship to congestion of gill. Cortisol were increased according to the decrease ranges in acclimated water temperature. From these results, decrease in water temperature during selection placed eels upon the stressed condition, made increase in ionic strength in blood stream, and CVS was dilatated owing to the increased blood inflow.

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