• Title/Summary/Keyword: virus inactivation

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Identication of Bean Yellow Mosaic Virus from Trifolium repens (흰 토끼풀에서 분리한 강낭콩황반모자이크 바이러스의 동정)

  • Ryu Gab Hee;Lee Soon Hyung;La Yong Joon
    • Korean Journal Plant Pathology
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    • v.2 no.1
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    • pp.48-52
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    • 1986
  • The virus isolated from white clover, Trifolium repens showing mosaic symptom was identified as bean yellow mosaic virus (BYMV) based on the host range, physical properties, aphid transmission, serology and morphology of the virus particles. Chenopodium amaranticolor and C. quinoa produced local lesions on the inoculated leaves and chlorotic spot on the upper leaves. Broad bean and cowpea produced local lesions on the inoculated leaves and mosaic with vein necrotic symptoms on the upper leaves. French bean showed vein necrosis on the inoculated leaves, yellow mosaic on the upper leaves and bud blight. The average size of virus particles was 740nm in length. The virus was also transmitted by Myzus persicae. The thermal inactivation point of the virus isolate was $60\;to\;65^{\circ}C$, the dilution end point $10^{-3}\;-\;10^{-4}$ and the longevity in vitro was 3 days Serological tests with the virus purified from Trifolium repens were positive to BYMV antiserum.

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Process development of a virally-safe dental xenograft material from porcine bones (바이러스 안전성이 보증된 돼지유래 골 이식재 제조 공정 개발)

  • Kim, Dong-Myong;Kang, Ho-Chang;Cha, Hyung-Joon;Bae, Jung Eun;Kim, In Seop
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.140-147
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    • 2016
  • A process for manufacturing virally-safe porcine bone hydroxyapatite (HA) has been developed to serve as advanced xenograft material for dental applications. Porcine bone pieces were defatted with successive treatments of 30% hydrogen peroxide and 80% ethyl alcohol. The defatted porcine bone pieces were heat-treated in an oxygen atmosphere box furnace at $1,300^{\circ}C$ to remove collagen and organic compounds. The bone pieces were ground with a grinder and then the bone powder was sterilized by gamma irradiation. Morphological characteristics such as SEM (Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images of the resulting porcine bone HA (THE Graft$^{(R)}$) were similar to those of a commercial bovine bone HA (Bio-Oss$^{(R)}$). In order to evaluate the efficacy of $1,300^{\circ}C$ heat treatment and gamma irradiation at a dose of 25 kGy for the inactivation of porcine viruses during the manufacture of porcine bone HA, a variety of experimental porcine viruses including transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), and porcine parvovirus (PPV) were chosen. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the $1,300^{\circ}C$ heat treatment. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.81$$ for PRV, $${\geq_-}6.28$$ for PRoV, and $${\geq_-}5.21$$ for PPV. Gamma irradiation was also very effective at inactivating the viruses. TGEV, PRV, PRoV, and PPV were completely inactivated to undetectable levels during the gamma irradiation. The mean log reduction factors achieved were $${\geq_-}4.65$$ for TGEV, $${\geq_-}5.87$$ for PRV, $${\geq_-}6.05$$ for PRoV, and $${\geq_-}4.89$$ for PPV. The cumulative log reduction factors achieved using the two different virus inactivation processes were $${\geq_-}9.30$$ for TGEV, $${\geq_-}11.68$$ for PRV, $${\geq_-}12.33$$ for PRoV, and $${\geq_-}10.10$$ for PPV. These results indicate that the manufacturing process for porcine bone HA from porcine-bone material has sufficient virus-reducing capacity to achieve a high margin of virus safety.

Quantitative Analysis of Feline Calicivirus Inactivation using Real-time RT-PCR (Real-time RT-PCR을 이용한 Feline Calicivirus 불활성화의 정량적 분석)

  • Jeong, Hye Mi;Kim, Kwang Yup
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.31-39
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    • 2014
  • Norovirus causes acute gastroenteritis in all age groups and its food poisoning outbreaks are rapidly increasing in Korea. Reverse transcription-polymerase chain reaction (RT-PCR) is most widely used for the rapid detection of foodborne viruses due to high sensitivity. However, the false positive results of RT-PCR obtained against already inactivated viruses could be a serious drawbacks in food safety area. In this study, we investigated a method to yield true positive RT-PCR results only with alive viruses. To decompose the RNA genes from dead viruses, the enzymatic treatments composed of proteinse K and Ribonuclease A were applied to the sanitized and inactivated virus particles. Another aim of this study was to quantify the efficiencies of several major sanitizing treatments using real-time RT-PCR. Feline calicivirus (FCV) that belongs to the same Caliciviridae family with norovirus was used as a surrogate model for norovirus. The initial level of virus in control suspension was approximately $10^4$ PFU/mL. Most of inactivated viruses treated with the enzymatic treatment for 30 min at $37^{\circ}C$ were not detected in RT-PCR, Quantification results to verify the inactivation efficiencies of sanitizing treatments using real-time RT-PCR showed no false positive in most cases. We could successfully develope a numerical quantification process for the inactivated viruses after major sanitizing treatments using real-time RT-PCR. The results obtained in this study could provide a novel basis of rapid virus quantification in food safety area.

Recent (2010-2019) foodborne outbreaks caused by viruses in the Republic of Korea along with their detection and inactivation methods (바이러스에 의한 최근(2010-2019) 국내 식중독 사고와 검출법 및 제어법에 대한 동향 조사)

  • Kwon, Seung-Wook;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.1
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    • pp.1-11
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    • 2021
  • In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.

Occurrence of Three Strains of Cucumber mosaic virus Affecting Tomato in Kuwait

  • Montasser Magdy Shaban;Dashti Narjes Haji;Ali Neda Yousef;Bhardwaj Radhika Guleri;Al-Hamar Bader
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.51-62
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    • 2006
  • Three strains of Cucumber mosaic virus (CMV) have been found to cause a lethal disease, referred to as fern leaf syndromes and mild mosaic symptoms in tomato (Lycopersicon esculentum Mill.) crops grown in Kuwait. CMV strains were detected and identified based on host range, symptomatology, serology, electron microscopy, and ribonucleic acid (RNA) electrophoresis in polyacrylamide gels. A high degree of viral genomic heterogeneity was detected among CMV strains isolated in Kuwait, with no apparent correlation to symptomatology in tomato host plants. Two different virus satellites of 'CMV associated RNA 5', designated CARNA 5, were detected in two virus strains that caused both lethal disease and mild symptoms, designated CMV-D1 and CMV-S1 respectively. CARNA5 was not detected in the third CMV strain that caused fern leaf syndromes designated CMV-F. All the three isolated strains were serologically indistinguishable from each other and may belong to one serotype according to Ouchterlony gel diffusion tests. These strains transmitted via aphids (Myzus persicae Sulz) in a non-persistent manner. Physical properties of the virus strains were very similar where thermal inactivation test showed that virus withstood heating for 10 min at $70^{/circ}$, dilution end point was $10^{-4}$, and the longevity in vitro at room temperature was less than 5 days for all virus strains. CMV-D1 and CMV-F were the most devastating diseases spreading in both greenhouse and field-grown tomato where aborted flower buds failed on fruit setting due to the viral infection. This is the first report to isolate three different strains of CMV in Kuwait.

Studies on Ginger Mosaic Virus (생강모자이크바이러스병에 관한 연구)

  • So In Young
    • Korean journal of applied entomology
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    • v.19 no.2 s.43
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    • pp.67-72
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    • 1980
  • A mosaic virus disease of ginger plant was investigated to determine its virus group on the basis of host range, physical and chemical properties, serological behavior and electron-microscopic morphology. The disease gave rise to yellowsih and dark-green mosaic on the leaves in the early stage and stunted all the leaves as well as rhizomes in the late stage. In the field about 43\% of the plants were observed to be diseased The disease was able to be artificially infected to the ginger plants by the sap and transmission as well as to 23 other species of plants which were known to be the CMV susceptible plants by the sap transmission; Chenopodium amaranticolar, Nicotiana tabaccum var. Havana, cow pea, cucumber, tomato,... etc. The dilution end point of the virus ranged $10^{-4}-10^{-5}$ and the thermal inactivation point $65-70^{\circ}C$. Serological test showed a positive reaction by a CMV antiserum. An electron microscopy of the purified virus showed that the virus particles were spherical with a diameter of $28-32m\mu$. Virus particles from the infected tissue were observed to be free or aggregated in the mesophyll tissue of artificially infected tobacco plant. The mosaic disease of ginger plants were conclusively suggested to the CMV group.

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Inactivation of various bacteriophages in wastewater by chlorination; Development of more reliable bacteriophage indicator systems for water reuse (하수 처리 과정의 염소 소독에 대한 여러 박테리오파지들의 저항성 평가; 물 재이용 과정의 안전성 관리를 위한 바이러스 지표미생물의 개발)

  • Bae, Kyung-Seon;Shin, Gwy-Am
    • Journal of Korean Society of Water and Wastewater
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    • v.30 no.3
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    • pp.285-291
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    • 2016
  • There has been an accelerating increase in water reuse due to growing world population, rapid urbanization, and increasing scarcity of water resources. However, it is well recognized that water reuse practice is associated with many human health and ecological risks due to numerous chemicals and pathogenic microorganisms. Especially, the potential transmission of infectious disease by hundreds of pathogenic viruses in wastewater is one of the most serious human health risks associated with water reuse. In this study, we determined the response of different bacteriophages representing various bacteriophage groups to chlorination in real wastewater in order to identify a more reliable bacteriophage indicator system for chlorination in wastewater. Different bacteriophages were spiked into secondary effluents from wastewater plants from three different geographic areas, and then subjected to various doses of free chlorine and contact time at $5^{\circ}C$ in a bench-scale batch disinfection system. The inactivation of ${\phi}X174$ was relatively rapid and reached ~4 log10 with a CT value of 5 mg/L*min. On the other hand, the inactivation of bacteriophage PRD1 and MS2 were much slower than the one for ${\phi}X174$ and only ~1 log10 inactivation was achieved by a CT value of 10 mg/L*min. Overall, the results of this study suggest that bacteriophage both MS2 and PRD1 could be a reliable indicator for human pathogenic viruses for chlorination in wastewater treatment processes and water reuse practice.

A Novel Role of Classical Swine Fever Virus Erns Glycoprotein in Counteracting the Newcastle Disease Virus (NDV)-mediated IFN-β Induction

  • Xia, Yan-Hua;Chen, Liu;Pan, Zi-Shu;Zhang, Chu-Yu
    • BMB Reports
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    • v.40 no.5
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    • pp.611-616
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    • 2007
  • $E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

Biological, Physical and Cytological Properties of Pepper mottle virus-SNU1 and Its RT-PCR Detection

  • Han, Jung-Heon;Choi, Hong-Soo;Kim, Dong-Hwan;Lee, Hung-Rul;Kim, Byung-Dong
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.155-160
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    • 2006
  • A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

The Change of Cytosolic Free Calcium Concentration Following Herpes Simplex Virus Type-1 (HSV-1) Infection (Herpes Simplex Virus Type-1 (HSV-1) 감염에 따른 세포내 유리 $Ca^{2+}$농도의 변화)

  • 남윤정;이규철;이찬희
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.306-311
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    • 2000
  • Infection of Vero cells with herpes simplex virus type-1 (HSV-1) resulted in a series of changes in intra-cellular free calcium concentration $([Ca^{2+}]_i)$. A significant and maximal decrease $[Ca^{2+}]_i$ was observed at 4 hours postinfection (hr p.i.) in HSV-1-infected in Vero cells. Inactivation of HSV-1 with UV irradiation and heat treatment abolished HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells. And the degree of the decrease in $[Ca^{2+}]_i$ was dependent on the amount of input virus. Taxol, which stabilizes the polymerization of microtubule blocked HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i., suggesting that microtubule may mediate the transport of HSV-1 nucleocapsid to the nucleus of infected cell. Treatment of HSV-1-infected Vero cells with metabolic inhibitors such as cycloheximide, cordycepin, or acyclovir partially reversed the decrease in $[Ca^{2+}]_i$ at 4 hr p.i.. Thus, it is suggested that HSV-1 induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells may play an important role in the multiplication of HSV-1.

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