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Use of Serological-Based Assay for the Detection of Pepper yellow leaf curl Indonesia virus

  • Hidayat, Sri Hendrastuti;Haryadi, Dedek;Nurhayati, Endang
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.328-332
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    • 2009
  • Diseases caused by Pepper yellow leaf curl virus infection is considered to be emerging plant diseases in Indonesia in the last five years. One key factor for disease management is the availability of accurate detection of the virus in plants. Polyclonal antibody for Pepper yellow leaf curl Indonesia virus-Bogor (PYLCIV-Bgr) was produced for detection of the virus using I-ELISA and DIBA methods. The antibody was able to detect PYLCIV-Bgr from infected plants up to dilution 1/16,384 and cross reaction was not observed with Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), and Chilli veinal mottle virus (ChiVMV). Positive reaction was readily detected in membrane containing Begomovirus samples from Yogyakarta (Kaliurang and Kulonprogo) and West Java (Bogor and Segunung). Infection of PYLCIV-Bgr in chillipepper, tomato, and Ageratum conyzoides was also confirmed using polyclonal antibody for PYLCIV-Bgr in DIBA. Polyclonal antibody for PYLCIV-Bgr is suggested to be included in disease management approach due to its good detection level.

Monitoring of viruses (IHHNV, TSV, IMNV, YHV, and CMNV) in cultured whiteleg shrimp (Litopenaeus vannamei) between 2018 and 2019 (2018-2019년 양식산 흰다리새우의 바이러스 (IHHNV, TSV, IMNV, YHV, CMNV) 모니터링)

  • Kokkattunivarthil, Shyam;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.33 no.1
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    • pp.71-75
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    • 2020
  • A survey was conducted to investigate viral infections in 184 whiteleg shrimp (Litopenaeus vannamei) collected from nine farms and one wholesale fish vendor during 2018 and 2019. Gill and abdominal muscle of shrimp were tested for the presence of five viruses, viz. infectious hypodermal and haematopoietic necrosis virus, taura syndrome virus, infectious myonecrosis virus, yellow head virus genotype 1, and covert mortality nodavirus by reverse transcription-polymerase chain reaction (RT-PCR) and PCR. These viruses were not detected in any of 184 samples, screened under the study.

The Viruses in Gladiolus hybridus cultivated in Korea 2. Broad Bean Wilt Virus, Cucumber Mosaic Virus and Tobacco Rattle Virus (한국산 글라디올러스에 발생하는 바이러스)

  • 박인숙;김규원;권현정;장무웅
    • Korean Journal Plant Pathology
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    • v.14 no.1
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    • pp.83-91
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    • 1998
  • Gladioli (Gladiolus hybridus) showing flower colour breaking leaf mosaics, notched leaf, and dwarfing or lack of visible symptoms were collected from gladioli growing areas in Taegu and Kyungpook province, Korea. The three viruses isolated from the naturally infected gladioli were identified as broad bean wilt virus (BBWV), cucumber mosaic virus (CMV), and tobacco rattle virus (TRV) by their host range, immunosorbent electron microscopy (ISEM), enzyme-linked immunosorbent assay (ELISA), direct tissue blotting immunoassay (DTBIA), and intracellural symptoms. By DTBIA and ISEM, TRV was detected in gladiolus showing notched leaf, while CMV was frequently detected in gladioli with dwarfing, color breaking and malformation of flowers. BBWV was also often detected in many symptomless gladiolus plants, but TRV was detected in notched-leaf of gladiolus. Electron microcopic examination of negatively stained preparations showed that BBWV and CMV are spherical particles of 28 nm and 30 nm in diameter, and TRV is rigid rod-shaped particles of 40∼200 nm in length. The rigid rodshaped virus particles reacted positively with TRV antiserum in ISEM and DTBIA, respectively.

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Mural folliculitis and alopecia caused by infection with malignant catarrhal fever virus in goat (Capra hircus) (Malignant catarrhal fever virus 감염과 관련된 goat (Capra hircus)의 mural folliculitis와 alopecia)

  • Kim, Ok-Jin;Crawford, Timothy B.
    • Korean Journal of Veterinary Pathology
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    • v.7 no.1
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    • pp.5-9
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    • 2003
  • Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Four 1-year old goats (Capra hircus), which were infected with MCF virus, OvHV-2, by being housed together with MCF virus-infected seep, were referred with a I-month history of chronic dermatitis. On the other hand, MCF virus-negative goats, which were isolated for negative control, had not those kinds of skin problems. Examination of the affected goats revealed generalized alopecia, patchy erythema, and superficial erosions with histologic evidence of mural folliculitis. Fungal culture tests and external parasite tests with the scraping skin samples were negative. However, polymerase chain reaction revealed the existence of MCF virus DNAs in the lesion. These results suggested that MCF virus may induce mural folliculitis and alopecia in goat.

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Characterizations of Disease Symptoms and Virus Replication Shown in the Interactions Between Arabidopsis (Arabidopsis ecotype에서 3종의 BCTV 분리주의 병증 및 복제 특성)

  • 박을용;박종범;이석찬
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.507-512
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    • 1998
  • Molecular analysis has been done for characterization of the interactions between three beet curly top virus (BCTV) strains and two Arabidopsis ecotypes in terms of virus inducible disease symptoms and infectivities. The total DNA was isolated from three tissues (shoot tips, infection origins and roots) of virus infected plants and this DNA was analyzed by quantitatively and qualitatively to elucidate virus movement and symptom development. CTV-Worland infected Col-O and Sei-O showed only symptom shown in hypersusceptible ecotype Sei-O by BCTV-worland was shoot tip stunting. Kinetics of virus DNA accumulation of three different viruses indicated that roots contained more virus DNA than shoot tips or infection origins, and that disease symptom severity was strongly correlated with virus DNA accumulation. These results suggest that the mild and Worland-specific symptoms shown in Sei-O by BCTV-worland are caused by the interactions of host factors provided by hypersusceptible ecotype and viral factors of mild strain.

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Immunohistochemical diagnosis on rabies virus using its monoclonal antibody in mice (단크론항체를 이용한 광견병바이러스의 면역병리조직학적 진단)

  • Kang, Mun-il;Park, Nam-yong;Song, Jae-yeong
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.255-261
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    • 1993
  • For a immunohistochemical diagnosis of the frozen and paraffin-embedded tissues against rabies virus, mice were intracerebrally inoculated with challege virus standard(CVS) rabies virus and then were used to detect the rabies viral antigen by the immunoperoxidase(IP) and the avidin-biotin complex(ABC) method. In this study, the results confirmed that ABC and IP methods, although the former showed more specific and sensitive than the latter, were reliable and effective for the demonstration of rabies virus in both frozen and paraffin-embedded brain tissues prepared from rabies-infected mice. Additionally, IP technique using the monoclonal antibody against rabies virus could be recommended as a standard diagnostic tool instead of the present immunofluorescent method for the local veterinary services in Korea.

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First report of Cycas necrotic stunt virus from cultivated Daphne plants

  • Lee, B.Y.;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.148.1-148
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    • 2003
  • Natural virus infection of cultivated Daphe odora plants showing chlorosis and stunting was observed and their causal agent was investigated. An isolate of isometic virus was purified from infected leaf tissues, and it could infect systemic severe mosaic on Chenopodium quinoa and C. amaranticolor. cDNA library was generated from partially purified viral RNAs and oligo dT primer-pSPORTl system, and recombinant clones were selected and their inserts were sequenced randomly. Nucleotide sequences of the virus were analyzed by BLAST, and it was closely related to members of subgroup B in the genus Nepovirus. The sequence analysis suggest that the virus was identified as an isolate of Cycas necrotic stunt virus (CNSV) because it was 89.7 % and 94.7 % identical to known CNSV for the CP and 3' noncoding region, respecitively. RT-PCR was performed to screen disease incidence of CNSV in Daphe plants, and five out of 10 plants (50 %) were infected by CNSV This is the first sequence information of CNSV from Daphe plants.

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Combining ex vitro thermotherapy with shoot-tip grafting for elimination of virus from potted apple plants (기외 열처리와 경정접목을 이용한 사과 폿트묘에서의 바이러스 제거)

  • Chun, Jae An;Gwon, Jiyeong;Lee, Seon Gi
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.222-229
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    • 2022
  • Apples are the most grown fruit crops in the fruit industry of Korea. However, virus or viroid infection such as apple mosaic virus (ApMV), apple stem grooving capillovirus (ASGV), apple stem pitting virus (ASPV), apple chlorotic leaf spot virus (ACLSV), apple scar skin viroid (ASSVd) causes fruit yield reduction and poor fruit quality. Therefore, in this study, we examined to established an efficient virus-free system to eliminate the most infected ASGV virus in domestic apple orchard. We investigated that the shoot growth rate and the virus removal rate in ASGV infected potted apples that were treated with heat treatment in a growth chamber (constant temperature/humidity device) maintained at 36℃, 38℃ and 40℃ for 4 weeks. Here we found that the shoot growth rate was the highest in the heat treatment group (36℃) and the virus was removed in the middle and top of the shoot but not in the bottom. The virus was did not removed in the 38℃ and 40℃ heat treatment group in all section of shoots, and the heat treatment group (40℃) died after 4 weeks of heat treatment without growth of shoots. We performed in vivo shoot-tip grafting using the shoot-tip of potted apple heat-treated at 36 ℃, and we also investigated the viability and virus removal rate, which showed 94% viability and 20% virus removal rate. Collectively, our results suggest that it would be possible to produce the virus-free apple plants through heat treatment and shoot-tip grafting.

Effect of virus infectivity titer following centrifugation and filtration during virus extraction from fish samples

  • Kim, Wi-Sik;Kim, Jong-Oh;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.28 no.2
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    • pp.113-116
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    • 2015
  • A $0.45-{\mu}m$ membrane filter is generally used to remove bacterial contamination during virus extraction from fish samples. However, the number of fish viruses is drastically reduced after filtration with a $0.45{\mu}m$ filter. In this study, we investigated the effect of filters on virus infectivity titer and the change in virus titer and bacterial number following different centrifugation conditions to determine a suitable procedure for virus extraction from fish samples. $10^{4.05}$ and $10^{5.05}TCID_{50}/ml$ of infectious hematopoietic necrosis virus (IHNV) and $10^{4.05}$ and $10^{4.55}TCID_{50}/ml$ of Oncorhynchus masou virus (OMV) were not detectable after filtration with two types of $0.45-{\mu}m$ filters, except the IHNV titer was reduced by about 10 fold after filter use (company A). No significant difference was found in the virus titer following centrifugation at $880{\times}g$ (30 min) or $3,500{\times}g$ (30 min), whereas IHNV and OMV titers were reduced by about 10 and 10-1000 fold by centrifugation at $14,000{\times}g$ (30 min) and $14,000{\times}g$ (10 and 30 min), respectively. A total of 97.7-99.9% Escherichia coli were eliminated by centrifugation at $880 {\times}g$ (30 min) and $3,500{\times}g$ (30 min). These results show that fish viruses were affected by filtering, even though the effect differed by virus species and filter type. Therefore, centrifugation at $3,500{\times}g$ (30 min) and use of medium with antibiotics may be useful for virus extraction along with a reduction in bacteria.

Titer Amplification of GALV (Gibbon Ape Leukemia Virus) Pseudotyped Retrovirus Vectors Produced from PG13 Cells (PG13 Cell로부터 생산된 GALV (Gibbon Ape Leukemia Virus)-pseudotyped Retrovirus Vector의 증폭)

  • 김태완;박윤엽;권모선;염행철;김경화;박영식;박세필
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.397-403
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    • 1997
  • For the ultimate goal of efficient retrovirus vector-mediated transgenic animal production, we tried to increase virus titer by employing three methods: boosting virus production by treating virus-producing cells with sodium butyrate, concentration of virus stock by either filtration or ultracentrifugation. Compared to the control, applications of sodium butyrate (5 mM) treatment and filtration resulted in only 3 and 3. 6 folds of titer increases on bovine EBTr target cells, respectively. However, concentration of virus-containing medium by ultracentrifugation showed 12.5 folds of titer increase compared to the control (10${\times}$10$^5$ LacZ$^+$ TU Im), indicating the best method which can enhance retrovirus vector-mediated transgenic animal production.

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