• Title/Summary/Keyword: virus isolation

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Isolation and Characterization of White Spot Syndrome Baculovirus in Cultured Penaeid Shrimp (Penaeus chinensis) (양식새우(Penaeus chinensis)에서의 White Spot Baculovirus의 분리 및 특성)

  • Heo, M.S.;Sohn, S.G.;Sim, D.S.;Kim, J.W.;Park, M.A.;Lee, J.S.;Choi, D.L.;Jung, S.H.;Kim, Y.J.;Oh, M.J.
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.7-13
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    • 2000
  • Beginning in the summer of 1993, a serious mortality among cultured penaeid shrimp occurred in the western sea of Korea. The typical sign of this disease was white spots inside the surface of the carapace. Cytopathic effect (CPE) were not observed by virus in CHSE-214, RTG-2, but not by pH 11. A nonoccluded rod-shaped form virus was observed by electron microscopy in the lymphoid organ. The virion was bacilliform virus and sourrounded by a virion envelope. Its virion protein was found to be similar to hypodermal and hematopoietic necrosis virus (HHNBV) by analysis of virion proteins in SDS-PAGE. The genome of virus is double stranded DNA molecule whose full length was about 114kb. It was similar to penaeus acute viremia (PAV) of Japan.

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Molecular characterization of Malaysian fowl adenovirus (FAdV) serotype 8b species E and pathogenicity of the virus in specific-pathogen-free chicken

  • Sabarudin, Nur Syazana;Tan, Sheau Wei;Phang, Yuen Fun;Omar, Abdul Rahman
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.42.1-42.16
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    • 2021
  • Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions: FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

Isolation and identification of canine adenovirus type 2 from a naturally infected dog in Korea

  • Yang, Dong-Kun;Kim, Ha-Hyun;Yoon, Soon-Seek;Lee, Hyunkyoung;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.177-182
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    • 2018
  • Canine adenovirus type 2 (CAV-2) infection results in significant respiratory illness in dogs. Isolating and culturing CAV-2 allows for investigations into its pathogenesis and the development of vaccines and diagnostic assays. In this study, we successfully isolated a virus from a naturally infected dog in Gyeonggi-do, Korea. The virus was propagated in Madin-Darby canine kidney (MDCK) and Vero cells and showed a specific cytopathic morphology that appeared similar to a bunch of grapes. The virus was first confirmed as CAV-2 based on these cytopathic effects, an immunofluorescence assay, hemagglutination assay, and electron microscopy. The viral titer of the isolate designated APQA1601 reached $10^{6.5}$ 50% tissue culture infections dose per mL in MDCK cells and exhibited no hemagglutination units with erythrocytes from guinea pig. The virus was also confirmed by polymerase chain reaction and next-generation sequencing. The APQA1601 strain had the highest similarity (~99.9%) with the Toronto A26/61 strain, which was isolated in Canada in 1976 when the nucleotide sequences of the full genome of the APQA1601 strain were compared with those of other CAV strains. Isolating CAV-2 will help elucidate the biological properties of CAV-2 circulating in Korean dogs.

SARS-CoV-2 Antibody Neutralization Assay Platforms Based on Epitopes Sources: Live Virus, Pseudovirus, and Recombinant S Glycoprotein RBD

  • Endah Puji Septisetyani;Pekik Wiji Prasetyaningrum;Khairul Anam;Adi Santoso
    • IMMUNE NETWORK
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    • v.21 no.6
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    • pp.39.1-39.18
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    • 2021
  • The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.

Isolation of atypical Aeromonas salmonicida in cultured starry flounder (Platichthys stellatus) (양식산 강도다리(Platichthys stellatus)에서의 비정형 Aeromonas salmonicida 분리)

  • Wi-Sik, Kim;Min-Su, Kwon;Hwi-Jin, Kim;Myung-Joo, Oh;Kyoung-Hui, Kong
    • Journal of fish pathology
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    • v.35 no.2
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    • pp.247-250
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    • 2022
  • About 6.7% mortality was reported in a starry flounder (Platichthys stellatus) aquaculture farm in 2022. Most of the diseased fish showed a loss of pectoral fin, hemorrhages on muscle and gills, pale gills, enlarged spleen, and nodules on kidney. Parasites, fungi or viruses (viral hemorrhagic septicemia virus and hirame novirhabdovirus) were not detected from diseased fish. However, numerous bacteria were isolated from liver, spleen and kidney. Nucleotide sequences of the A-protein-encoding virulence array protein gene (vapA) of the bacteria showed 99.93% identity with Aeromonas salmonicida subsp. masoucida. This study is the first report of isolation of atypical A. salmonicida in cultured starry flounder in Korea.

Monitoring of viruses in cultured walleye pollock Gadus chalcogrammmus (양성 중인 명태(Gadus chalcogrammus)의 바이러스 모니터링)

  • Nam, U-Hwa;Jeon, Chan-Hyeok;Seo, Hyun-Joon;Choi, Da-Young;Seo, Joo-young;Kwon, O-Nam;Kim, Wi-Sik;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.30 no.1
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    • pp.1-9
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    • 2017
  • This study was conducted to monitor the prevalence of viral hemorrhagic septicemia virus (VHSV), nervous necrosis virus (NNV) and marine birnavirus (MABV) in cultured walleye pollock Gadus chalcogrammus by RT-PCR. All of the viruses tested were not detected by one-step PCR in 62 spleen sample sets, except for NNV in one brain sample set (1/55). By two-step PCR, VHSV was detected in 51.6%(32/62) and NNV was detected in 1.6%(1/62) spleen sample set, but MABV was not detected. In the brain sample sets, the detection rate of NNV was 3.6%(2/55). VHSV and NNV were detected for the first time in cultured walleye pollock in this study. However, the titers of viruses in these sample sets are thought to be very low, because most of the positive sample sets were detected by two-step PCR and none of the fish showed any clinical symptoms of each virus. Continuous monitoring, subsequent virus isolation and validation of carrier fish will be necessary.

Pseudomonas oleovorans Strain KBPF-004 Culture Supernatants Reduced Seed Transmission of Cucumber green mottle mosaic virus and Pepper mild mottle virus, and Remodeled Aggregation of 126 kDa and Subcellular Localization of Movement Protein of Pepper mild mottle virus

  • Kim, Nam-Gyu;Seo, Eun-Young;Han, Sang-Hyuk;Gong, Jun-Su;Park, Cheol-Nam;Park, Ho-Seop;Domier, Leslie L;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.393-401
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    • 2017
  • Efforts to control viral diseases in crop production include several types of physical or chemical treatments; antiviral extracts of a number of plants have also been examined to inhibit plant viral infection. However, treatments utilizing naturally selected microorganisms with activity against plant viruses are poorly documented. Here we report isolation of a soil inhabiting bacterium, Pseudomonas oleovorans strain KBPF-004 (developmental code KNF2016) which showed antiviral activity against mechanical transmission of tobamoviruses. Antiviral activity was also evaluated in seed transmission of two tobamoviruses, Pepper mild mottle virus (PMMoV) and Cucumber green mottle mosaic virus (CGMMV), by treatment of seed collected from infected pepper and watermelon, respectively. Pepper and watermelon seeds were treated with culture supernatant of P. oleovorans strain KBPF-004 or control strain ATCC 8062 before planting. Seeds germinated after treatment with water or ATCC 8062 yielded about 60% CGMMV or PMMoV positive plants, whereas < 20% of KBPF-004-treated seeds were virus-infected, a significantly reduced seed transmission rate. Furthermore, supernatant of P. oleovorans strain KBPF-004 remodeled aggregation of PMMoV 126 kDa protein and subcellular localization of movement protein in Nicotiana benthamiana, diminishing aggregation of the 126 kDa protein and essentially abolishing association of the movement protein with the microtubule network. In leaves agroinfiltrated with constructs expressing the coat protein (CP) of either PMMoV or CGMMV, less full-size CP was detected in the presence of supernatant of P. oleovorans strain KBPF-004. These changes may contribute to the antiviral effects of P. oleovorans strain KBPF-004.