• Title/Summary/Keyword: virus isolation

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Isolation and identification of infectious bursal disease virus from broiler and layer chickens during the outbreak year 2007 in Bangladesh

  • Islam, Md. Taohidul;Mohiuddin, Mohammad;Hossain, Muhammad Tofazzal;Rahman, Md. Bahanur;Rahman, Md. Mostafizur;Rahman, Md. Siddiqur;Song, Hee-Jong;Islam, Md. Alimul
    • Korean Journal of Veterinary Service
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    • v.35 no.1
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    • pp.9-17
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    • 2012
  • The objective of the present study was to isolate and identify infectious bursal disease viruses (IBDVs) from broiler and layer chickens of outbreaks of infectious bursal disease (IBD) in three districts of Bangladesh. A total of 70 bursal samples were collected from dead broiler (n=40) and layer (n=30) chickens showing specific lesions of IBD from seven commercial poultry farms of three different districts (Mymensingh, Chittagong and Tangail) of Bangladesh during the year 2007. Five representative bursal samples from each farm were used for the isolation of IBDVs using 9-day-old embryonated eggs of seronegative flock of layer birds and for identification the samples were subjected to agar gel immunodiffusion test (AGIDT), immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR). Out of 35 bursal samples, IBDVs were successfully isolated from 28 (80%) samples. By AGIDT, 32 (91.4%) samples were found positive for IBDV antigen. Results of AGIDT clearly indicated that IBDVs detected in 29 bursal samples of six affected farms were identical to each other but not to IBDVs present in the remaining three samples of another farm. Indirect immunoperoxidase staining of the bursal sections revealed the presence of IBDV antigen in 32 (91.4%) samples and the IBDV antigen was detected mainly in the cortex of the lymphoid follicles of the bursal tissues. In histopathology, cell depletion, atrophy and necrosis were observed in many bursal follicles with severe edema of interfollicular septa. Of the 35 bursal samples, 34 (97.1%) samples generated 254 bp product by RT-PCR. In conclusion, the results of virus isolation and identification by AGIDT, IHC and the analysis of viral genome by RT-PCR confirmed the outbreaks of acute IBD in commercial poultry of Bangladesh. Moreover, histopathological findings and results of AGIDT gave a clear indication that the isolates from six outbreaks were different from classical strain and it seems to be of very virulent strain. On the other hand, the isolates from the other outbreak were similar to the classical strain.

Studies on Garlic Mosaic Virus -lts isolation, symptom expression in test plants, physical properties, purification, serology and electron microscopy- (마늘 모자이크 바이러스에 관한 연구 -마늘 모자이크 바이러스의 분리, 검정식물상의 반응, 물리적성질, 순화, 혈청반응 및 전자현미경적관찰-)

  • La Yong-Joon
    • Korean journal of applied entomology
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    • v.12 no.3
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    • pp.93-107
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    • 1973
  • Garlic (Allium sativum L.) is an important vegetable crop for the Korean people and has long been cultivated extensively in Korea. More recently it has gained importance as a source of certain pharmaceuticals. This additional use has also contributed to the increasing demand for Korean garlic. Garlic has been propagated vegetatively for a long time without control measures against virus diseases. As a result it is presumed that most of the garlic varieties in Korea may have degenerated. The production of virus-free plants offers the most feasible way to control the virus diseases of garlic. However, little is known about garlic viruses both domestically and in foreign countries. More basic information regarding garlic viruses is needed before a sound approach to the control of these diseases can be developed. Currently garlic mosaic disease is most prevalent in plantings throughout Korea and is considered to be the most important disease of garlic in Korea. Because of this importance, studies were initiated to isolate and characterize the garlic mosaic virus. Symptom expression in test plants, physical properties, purification, serological reaction and morphological characteristics of the garlic mosaic virus were determined. Results of these studies are summarized as follows. 1. Surveys made throughout the important garlic growing areas in Korea during 1970-1972 revealed that most of the garlic plants were heavily infected with mosaic disease. 2. A strain of garlic mosaic virus was obtained from infected garlic leaves and transmitted mechanically to Chenopodium amaranticolor by single lesion isolation technique. 3. The symptom expression of this garlic mosaic virus isolate was examined on 26 species of test plants. Among these, Chenopodium amaranticolor, C. quince, C. album and C. koreanse expressed chlorotic local lesions on inoculated leaves 11-12 days after mechanical inoculation with infective sap. The remaining 22 species showed no symptoms and no virus was recovered from them whet back-inoculated to C. amaranticolor. 4. Among the four species of Chtnopodium mentioned above, C. amaranticolor and C. quinoa appear to be the most suitable local lesion test plants for garlic mosaic virus. 5. Cloves and top·sets originating from mosaic infected garlic plants were $100\%$ infected with the same virus. Consequently the garlic mosaic virus is successively transmitted through infected cloves and top-sets. 6. Garlic mosaic virus was mechanically transmitted to C, amaranticolor when inoculations were made with infective sap of cloves and top-sets. 7. Physical properties of the garlic mosaic virus as determined by inoculation onto C. amaranticolor were as follows. Thermal inactivation point: $65-70^{\circ}C$, Dilution end poiut: $10^-2-10^-3$, Aging in vitro: 2 days. 8. Electron microscopic examination of the garlic mosaic virus revealed long rod shaped particles measuring 1200-1250mu. 9. Garlic mosaic virus was purified from leaf materials of C. amaranticolor by using two cycles of differential centrifugation followed by Sephadex gel filtration. 10. Garlic mosaic virus was successfully detected from infected garlic cloves and top-sets by a serological microprecipitin test. 11 Serological tests of 150 garlic cloves and 30 top-sets collected randomly from seperated plants throughout five different garlic growing regions in Korea revealed $100\%$ infection with garlic mosaic virus. Accordingly it is concluded that most of the garlic cloves and top-sets now being used for propagation in Korea are carriers of the garlic mosaic virus. 12. Serological studies revealed that the garlic mosaic virus is not related with potato viruses X, Y, S and M. 13. Because of the difficulty in securing mosaic virus-free garlic plants, direct inoculation with isolated virus to the garlic plants was not accomplished. Results of the present study, however, indicate that the virus isolate used here is the causal virus of the garlic mosaic disease in Korea.

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Detection, isolation, and characterization of the cucumber mosaic virus in Pseudostellaria heterophylla from Korea

  • Lee, Da Hyun;Kim, Jinki;Han, Jun Soo;Lee, Jae-Hyeon;Lee, ByulHaNa;Park, Chung Youl
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.150-156
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    • 2020
  • Weeds play an important role in the survival of viruses and are potential inoculum sources of viral diseases for crop plants. In this study, specimens of Pseudostellaria heterophylla exhibiting symptoms of the cucumber mosaic virus (CMV) were collected in Bonghwa, Korea. The characteristics of the disease were described and leaf RNA was extracted and sequenced to identify the virus. Three CMV contigs were obtained and PCR was performed using specific primer pairs. RNA from positive samples exhibiting CMV leaf symptoms was amplified to determine the coat protein. A sequence comparison of the coat protein gene from the CMV BH isolate shared the highest nucleotide identity (99.2%) with the CMV ZM isolate. Phylogenetic analysis showed that CMV-BH belonged to subgroup IA and that the most closely-related isolate was CMV-ZM. All test plants used for the biological assay were successfully infected with CMV and exhibited CMV disease symptoms such as blistering, mosaic, and vein yellowing. To our knowledge, this is the first report of CMV infection in P. heterophylla from Korea.

Isolation and Identification of Barley Yellow Mosaic Virus in Korea (보리누른모자이크바이러스(BaYMV)의 분리 및 동정)

  • 이귀재;소인영;백기철
    • Korean Journal Plant Pathology
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    • v.14 no.1
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    • pp.62-67
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    • 1998
  • Barley yellow mosaic virus (BaYMV-HN) occurring Haenam area was isolated by mechanical inoculation onto barley cultivars, purification and production of antibody. BaYV-HN were purified from infected plants a filamentous virus with 13 nm in diameter and 250∼300 nm and 500∼650 nm in length. Specific antibody made by injecting the purified virus to the muscle of a rabbit. In gel-diffusion tests antibody to BaYMV-HN did not make spur with tow Japanese BaYMV isolates BaYMV-II-1 or BaYMV-III. BaYMV-HN showed the symptom of yellowing and necrosis in host plants. Mechanical inoculation tests with Japanese barley cultivars showed that BaYMV-HN infected New Golden, Akagi Nijo and Tosan Kawa 73, but did not infect Amagi Nijo, Haruna Nijo, Ishukushirazu (ym3), Misato Golden (Ym1), Kashimamugi, Joshushiro Hadaka and Mokusekko 3 (ym1). In Korean barley cultivars, some of the naked barleys which are Hinssalbori, Kinssalbori, Saessalbori and Saechalssalbori were not infected by BaYMV-HN. However, it infected all the covered barley cultivars and the beer barley cultivars. BaYMV-HN had two RNAs, RNA 1 (7.6 Kb) and RNA 2 (3.5 Kb), and one coat protein (33 KDa).

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Preparedness of Siddha system of medicine in practitioner perspective during a pandemic outbreak with special reference to COVID-19

  • Rajalakshmi, S.;Samraj, K.;Sathiyarajeswaran, P.;Kanagavalli, K.
    • CELLMED
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    • v.10 no.4
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    • pp.29.1-29.6
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    • 2020
  • COVID-19 (Corona Virus Disease-2019) is an infectious respiratory disease caused by the most recently discovered coronavirus, SARS-CoV-2 (Severe Acute Respiratory Syndrome Corona virus-2). This new viral disease was unknown before the outbreak began in Wuhan, China, in December 2019. As of November 16th 2020, it affects about 54.3 million populations, death troll increased to 1.32 million cases in worldwide. Whereas in India 8.85 cases are infected with COVID-19, of which 1, 30, 112 cases were died. Till now there has been no specific anti-virus drug or vaccines are available for the treatment of this disease, the supportive care and non-specific treatment to the symptoms of the patient are the only options in Biomedicine, the entire world turns its attention towards alternative medicine or Traditional medicine. Siddha medicine is one of the primordial systems of medicine practiced in the southern part of India, it dealt a lot about pandemic, and its management. This review provides an insight into Pandemic in Siddha system and its management in both ancient history and modern history, National and state level Government policies related to current pandemic, World Health Organization (WHO) guidelines on usage of unproven drug during infectious disease outbreak, Preparedness of Siddha system during a pandemic outbreak Challenges and Recommendations.

A Reliable Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Apple stem grooving virus in Pear

  • Lee, Hyo-Jeong;Jeong, Rae-Dong
    • Research in Plant Disease
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    • v.28 no.2
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    • pp.92-97
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    • 2022
  • Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63℃ for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.

Studies with the tobacco mosaic viruses (한국산 연초 "바이러스"에 관한 연구)

  • 김은수;소인영
    • Korean Journal of Microbiology
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    • v.1 no.1
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    • pp.10-18
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    • 1963
  • Studies with the Tobacco Mosaic Viruses; W. S Kim, and So, I Y., (Dept. of biology Sung Kyun Kwan Univer. Seoul, Korea.). Using the common strain of tobacco mosaic virus (TMV) which was sent from the Dept. of Plant Pathology, University of Wisconsin, U.S.A. as control virus, a possible new strain of tobacco mosaic virus (SMV) was isolated from tobacco leaves collected from Tobacco Experiment Station farms as well as from various blends of manufactured Korean cigaretts. SMV was isolated by single lesion isolation method and by inoculating the virus through various species of host plants. The two viruses, TMV and SMV were indentified by the difference in symptoms, host range, serological reaction, and electron micrograpy. As the results of the above experiment the author believes the virus isolate SMV is a different strain of TMV. The experimental evidences that SMV belongs to the TMV group are as follows; 1. Both viruses produced local necrotic lesions on Nicotiana glutimosa L. 2. Both showed a dilution end point of $10^8$. 3. Aphid transmission was failed with the viruses. 4. Both had an isoelectric point around pH 3.3. 5. Two viruses were serological reactive. 6. The size of the virus particles was around 270-300mu as they were observed under the electron microscope. The virus SMV, however, is different from the common strain of TMV and the experimental evidences are as follows; 1. SMV produced quite different symptoms from TMV on various host plants like tobacoo(Nicotiana tabacum L., White Burley), Nicotiana rustica L., Chenopodium Koreanse Nakai. Bata vulgaris L., and Datura tatula L., SMV produced distinct local lesions on these host plants whereas TMV incited largely mosaic diseases. 2. The serological titers obtained from the heterologous combinations were lower than those from homologous combinations of antigens and antiser.

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Isolation of novel bovine parainfluenza virus type 5 (bPIV5) and its incidence in Korean cattle

  • Yang, Dong-Kun;Nah, Jin-Ju;Kim, Ha-Hyun;Choi, Sung-Suk;Bae, You-Chan;Park, Jung-Won;Song, Jae-Young
    • Korean Journal of Veterinary Research
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    • v.54 no.2
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    • pp.107-112
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    • 2014
  • Four viruses showing cytopathic effects in MDBK cells were isolated from brains of cattle showing downer cattle syndrome in 2012. The isolates were confirmed to belong to the genus Rubulavirus of the subfamily Paramyxovirinae. Isolate QIA-B1201 had the ability to hemagglutinate red blood cells from several species of animals and was capable of adsorbing guinea pig erythrocytes on the surface of infected Vero cells. Nucleotide sequence analysis showed that two isolates (QIA-B1201 and QIA-B1204) had high similarity with other human and animal PIV5 isolates ranging from 98.1 to 99.8%. The highest sequence similarity of the two isolates corresponded to strain KNU-11 (99.8% at the nucleotide and amino acid level) isolated from suckling piglets in Korea in 2012. To evaluate the virulence of strain QIA-B1201, we inoculated bPIV5 into 5 week-old mice via both the intraperitoneal and intracranial route. Body weight was not significantly altered in mice inoculated with QIA-B1201. In this study, we isolated and characterized novel bPIV5s from brain samples showing downer cattle syndrome, but were not able to elucidate the pathogenicity of the bPIV5s in mice.

Recommendation for conducting process of an epidemiological survey in respiratory syncytial virus infection (호흡기세포융합바이러스감염증 역학조사 수행절차 제안)

  • Kim, Dae Soon;Bae, Jong-Myon
    • Journal of Medicine and Life Science
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    • v.17 no.1
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    • pp.29-32
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    • 2020
  • As respiratory syncytial virus(RSV) is transmitted either via directly contact with an infected case or via indirectly contaminated fomites or skin, the major preventive measures are strict hand hygiene, early detection of transmitted sources, and rapid isolation of RSV patients. Especially early detection of hidden cases is the most critical control measure when an index case was notified in a postpartum center. The Guideline of Korea Centers for Diseases Control and Prevention defines potential contacts in an epidemiologic survey as admitted newborns, parents of index cases, center's workers, and visitors for 10 days before the first diagnosis day of index case. However, it needs to classify potential contacts in more detail in order to conduct a successful survey. Authors conducted to search related literatures and appraise the evidences. Firstly, potential contacts would be classified into RSV-related symptomatic contacts(SxC) and asymptomatic contacts. And then, mother, caring workers, and visitors of the index cases among asymptomatic contacts would be defined as the asymptomatic close contacts(ASCC). Finally, the rest would be defined as the asymptomatic regular contacts(ASRC). The defined test using reverse transcription-PCR is applied to SxC and ASCC, and decision of isolation or regular activities are made according to the results. The rapid antigen detection test kits are applied to ASRC. These suggestions might be helpful to detect hidden cases earlier and prevent a further infection.

Identification of Reassortant Pandemic H1N1 Influenza Virus in Korean Pigs

  • Han, Jae-Yeon;Park, Sung-Jun;Kim, Hye-Kwon;Rho, Se-Mi;Nguyen, Giap Van;Song, Dae-Sub;Kang, Bo-Kyu;Moon, Hyung-Jun;Yeom, Min-Joo;Park, Bong-Kyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.699-707
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    • 2012
  • Since the 2009 pandemic human H1N1 influenza A virus emerged in April 2009, novel reassortant strains have been identified throughout the world. This paper describes the detection and isolation of reassortant strains associated with human pandemic influenza H1N1 and swine influenza H1N2 (SIV) viruses in swine populations in South Korea. Two influenza H1N2 reassortants were detected, and subtyped by PCR. The strains were isolated using Madin-Darby canine kidney (MDCK) cells, and genetically characterized by phylogenetic analysis for genetic diversity. They consisted of human, avian, and swine virus genes that were originated from the 2009 pandemic H1N1 virus and a neuraminidase (NA) gene from H1N2 SIV previously isolated in North America. This identification of reassortment events in swine farms raises concern that reassortant strains may continuously circulate within swine populations, calling for the further study and surveillance of pandemic H1N1 among swine.