• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.6
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• pp.644-649
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• 2019

Killed vaccines, developed by inactivation with formalin, have been investigated for many fish viruses. In this study, the inactivation of viral hemorrhagic septicemia virus (VHSV) by formalin was investigated based on the infectivity titer. When viral cell culture supernatants were used, the infectivity titer decreased 1,000-fold at 1 d after treatment with 0.1% (v/v) formalin, but was below the detection limit at 7 and 14 d. Moreover, neither the N nor G gene were detectable by RT-PCR immediately after formalin treatment. In western blot analysis, N protein was not detected by rabbit antiserum against VHSV KR-9225 from 2 d after formalin treatment. On the other hand, when we used a virus that was purified and concentrated ~100 times, the infectivity titer was maintained at 10^6.05 TCID₅₀/mL, even at 14 d after formalin treatment, and no change in the viral structural proteins was observed. This study provides important data on the production and use of formalin-inactivated vaccines.

• Journal of the Korean Society of Tobacco Science
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• v.18 no.2
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• pp.120-125
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• 1996

Dried tobacco leaf debris infected with tobacco mosaic virus (TMV) was subjected to heat treatment (6$0^{\circ}C$~10$0^{\circ}C$) with or without addition of moisture and to room temperature for natural decay to examine the periods of time required for the inactivation of PMV in the inoculum source. Wet conditions (60% moisture content of the debris) for heat treatment were more efficient than dry conditions to inactivate the virus at 7$0^{\circ}C$~10$0^{\circ}C$, and which decrease of temperature, the time needed for the viral inactivation increased greatly. At 6$0^{\circ}C$ and 7$0^{\circ}C$, the temperaturein a compost heap during the actively decomposing period, it takes about 15 days or more for the complete inactivation of the virus. However, considering the decrease of the viral infectivity during the decomposition, a shorter period of time will be required to inactivate TMV in the conditions mentioned above, suggesting that a well decomposed organic manure containing tobacco leaf debris may not have infective TMV and may not provide a potential inoculum source.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.186-192
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• 2000

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.377-382
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• 2009

Viral safety is an important prerequisite for clinical preparations of all biopharmaceuticals derived from plasma, cell lines, or tissues of human or animal origin. To ensure the safety, implementation of multiple viral clearance (inactivation and/or removal) steps has been highly recommended for manufacturing of biopharmaceuticals. Of the possible viral clearance strategies, Ultraviolet-C (UVC) irradiation has been known as an effective viral inactivating method. However it has been dismissed by biopharmaceutical industry as a result of the potential for protein damage and the difficulty in delivering uniform doses. Recently a continuous flow UVC reactor (UVivatec) was developed to provide highly efficient mixing and maximize virus exposure to the UV light. In order to investigate the effectiveness of UVivatec to inactivate viruses without causing significant protein damage, the feasibility of the UVC irradiation process was studied with a commercial therapeutic protein. Recovery yield in the optimized condition of $3,000\;J/m^2$ irradiation was more than 98%. The efficacy and robustness of the UVC reactor was evaluated with regard to the inactivation of human immunodeficiency virus (HIV), hepatitis A virus (HAV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), porcine parvovirus (PPV), bovine parvovirus (BPV), minute virus of mice (MVM), reovirus type 3 (REO), and bovine parainfluenza virus type 3 (BPIV). Non enveloped viruses (HAV, PPV, BPV, MVM, and REO) were completely inactivated to undetectable levels by $3,000\;J/m^2$ irradiation. Enveloped viruses such as HIV, BVDV, and BPIV were completely inactivated to undetectable levels. However BHV was incompletely inactivated with slight residual infectivity remaining even after $3,000\;J/m^2$ irradiation. The log reduction factors achieved by UVC irradiation were ${\geq}3.89$ for HIV, ${\geq}5.27$ for HAV, 5.29 for BHV, ${\geq}5.96$ for BVDV, ${\geq}4.37$ for PPV, ${\geq}3.55$ for BPV, ${\geq}3.51$ for MVM, ${\geq}4.20$ for REO, and ${\geq}4.15$ for BPIV. These results indicate that UVC irradiation using UVivatec was very effective and robust in inactivating all the viruses tested.

• Korean journal of applied entomology
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• v.29 no.4
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• pp.244-251
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• 1990

Three viral insecticides were differently formulated with a nuclear polyhedrosis virus isolated from Spdodoptera litura by addition of feeding attractant, anti-precipitate of polyhedra, spreading agent, and UV-protectants. Sucrose was effective for attraction of larval feeding to increase the mortality and for protection of polyhedra from inactivation by sunlight when added 1% to 5% of sucrose solution to the formulations. Contents of additives to the formulations were 0.5% in polyvinyl alcohol to prohibit the precipitation of polyhedra and 0.1% in Triton X-100 to spread and wet the formulations to the plant. Inactivation of the virus under sunlight was decreased when added 800g of white carbon to 100 L of water in the white carbon formulation and 30% of molasses to the molasses's. In the formulation of white carbon and molasses mixtures, activation of the virus was increased when mixtured 500g of the former with 10% of the latter. Three formulations were persisted their pathogenicity more than 95% of mortality at 3 days p.i. Encapsulation of the polyhedral surface was more distinctively coated with the carbon and showed more effective in the residual effects of the white carbon than others, but the molasses more attractive for larval feeding.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.997-1003
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• 2008

Viral safety is a prerequisite for manufacturing clinical antihemophilic factor VIII concentrates from human plasma. With particular regard to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor VIII concentrate. The loss of factor VIII activity during dry-heat treatment was of about 5%. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor VIII compared with those of the factor VIII before dry-heat treatment. The dry-heat-treated factor VIII was stable for up to 24 months at $4^{\circ}C$. The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV, murine encephalomyocarditis virus (EMCV), and human immunodeficiency virus (HIV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Bovine herpes virus (BHV) and bovine viral diarrhea virus (BVDV) were potentially sensitive to the treatment. However porcine parvovirus (PPV) was slightly resistant to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\geq}5.55$ for HAV, ${\geq}5.87$ for EMCV, ${\geq}5.15$ for HIV, 6.13 for BHV, 4.46 for BVDV, and 1.90 for PPV. These results indicate that dry-heat treatment improves the virus safety of factor VIII concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid-enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.155-161
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• 1983

The effects of ultraviolet light on bacteriophage f2 were investigated to determine the inactivation kinetics and its mechanism. The 260nm light showed a little higher inactivation rate than the one of 300 nm. In this work, our main concern was whether structural and/or conformational changes in the protein capsid could occur by UV irradiation. The inactivation for the first 20 minutes irradiation was rapid with a loss of about 4 logs and followed by a slower rate during the next 40 minutes with no survival noted in the samples irradiated for 90 minutes or longer. The structural change of the protein capsid was examined by optical spectroscopic techniques and electron microscopy. The absorption spectra of the UV irradiated phages showed no detectable differences in terms of the spectral shape and intensity from the control phage. However, the fluorescence emission spectroscopic data, i.e. 1) fluorescence quenching of tryptophan residues upon irradiation of 300 nm light, 2) enhancement of fluorescence emission of ANS (8-aniline-1-naphthalene sulfonate) bound to the intact phages compared to the one in the UV-treated phages, and 3) decrease of energy transfer efficiency from tryptophan to ANS in the UV-treated samples, presented remarkable differences between the intact and UV-treated phages. Such a structural alteration was also observed by electron microscopy The UV-treated phages appeared to be broken and empty capsids. Therefore, the inactivation of the bacteriophage f2 by UV irradiation is thought to be attributed to the structural change in the protein capsid as well as damage in the viral RNA by UV irradiation.

• Korean Journal of Microbiology
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• v.18 no.3
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• pp.123-132
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• 1980

Bacterial virus f2 and its RNA were examined to elucidate the mode of ozone utilizing sucrose density gradient analysis and electtron microscopic techniques. the inactivation kinetics of the virus f2 by ozonation showed that the viruses were inactivated during the first 5 sec of the reaction and were further inactivated at a slower rate during the next 10 min at 0.09 and 0.8mg/l ozone concentrations. The virus coat was broken by ozonation into many pieces of protein subunits and the adsorption of the viruses to the host pili was inversely related to the extent of the breakage of the virus. The viral RNA was released from the virus particles during ozone, but ozone inactivation of the RNA enclosed in the protein coat could not ruled out the possibility that the RNA was secondarily sheared by a reaction with the broken coat protein.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.858-864
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• 2000

The purpose of the present study was to examine the efficacy and mechanism of the fraction IV cold ethanol fractionation and pasteurization ($60^{\circ}C$ heat treatment for 10h) steps, involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of blood-born viruses. A variety of experimental model viruses for human pathogenic viruses, including the Bovine viral diarrhoea virus (BVDV), Bovine herpes virus (BHV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses, and the amount of virus in each fraction was then quantified using a 50% tissue culture infectious dose ($TCID_{50}$). The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction IV fractionation was inactivation rather than partitioning, however, it was partitioning in the case of the non-enveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction IV fractionation were ${\geq}6.9$ BHV, $\geq5.2$ for BBDV, 4.9 for EMC, and 4.0 for PPV. Pasteurization was found to be a robust and effective step in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during pasteurization were $\geq7.0$ for BHV, $\geq6.1$ for BVDV, $\geq6.3$ for EMCV, and 1.7 for PPV. These results indicate that the production process for albumin has sufficient virus-reducing capacity to achieve a high margin for virus safety.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.619-627
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• 2001

n order to increase the virus safety of a human intravenous immunoglobulin (IVIg) that was manufactured by a successive process of cold ethanol fractionation, polyethylene glycol precipitation, and pasteurization ($60^{\circ}C$ heat treatment for 10h), a low pH incubation process (pH 3.9 at $25{\circ}C$ for 14 days) was employed as the final step. The efficacy and mechanism of the fraction III cold ethanol fractionation, pasteurization, and low pH treatment steps in the removal and/or inactivation of blood-borne viruses were closely examined. A variety of experimental model viruses for human pathogenic viruses, including the Bovine herpes virus (BHV), Bovine viral diarrhoea virus (BVDV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), were selected for this study. The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction III fractionation was both inactivation and partitioning, however, it was partitioning in the case of the nonenveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction III fractionation were ${\geqq}$6.7 for BHV, ${\geqq}4.7$ for BVDV, 4.5 for EMCV, and 4.4 for PPV. Pasteurization was found to be a robust and effective step in inactivating all the viruses tested. The log reduction factors achieved during the pasteurization process were ${\geqq}7.5$ for BHV, ${\geqq}4.8$ for BVDV, 3.0 for EMCV, and 3.3 for PPV. A low pH incubation was very effective in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during low pH incubation were ${\geqq}7.4$ for BHV, ${\geqq}3.9$ for BVDV, 5.2 for EMCV, and 2.0 for PPV. These results indicate that the low pH treatment successfully improved the viral safety of the final products.