• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.660-661
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• 1990

A stable preservation method of extracellular non-occluded virion of Autographa californica nuclear polyhedrosis virus (AcNPV) was studied. AcNPVL-1 strain infected to Spodoptera frugiperda cell line and then the culture media were centrifuged. After centrifugation the supernatant containing extracellular nonoccluded virions of the AcNPV was harvested and incubated at $4^{\circ}C$ . Even after the extracellular nonoccluded virions were incubated at $4^{\circ}C$ for about 11 years, the infectivity and multiplication property of the nonoccluded virions in the S. frugiperda cell line were normal. However the titers of the nonoccluded virions in TC-100 medium measured about 11 years ago decreased from $8.9 \times 10^7\; to \;3.8 \times 10^5$ pfu per ml. The AcNPV genome DNA fragment patterns from digestion with Hind11 and EcoRI restriction endonucleases did not change. The AcNPV nonoccluded virions were stable at $4^{\circ}C$ in the cultured medium of more than 10 years and the preservation of AcNPV nonoccluded virions at $4^{\circ}C$ is easy and useful for handling.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.2
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• pp.220-227
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• 2007

Sweetpotato fields in Korea are highly infected with virus and virus like diseases that greatly diminish both yield and quality as indicated by field observations and laboratory tests. In order to solve this problem, there is an urgent need to produce and mass propagate virus-free planting materials for distribution to the farmers. These experiments were conducted, firstly, to determine the most appropriate culture media, nutrient solution, and cutting intervals to maintain growth and vigor of tissue cultured plantleta as mother plants for propagation in insect-proof greenhouse. And as a labor saving method, the production efficiency of plug trays for rapid propagation of stem cuttings as a source of planting materials was likewise evaluated. Results showed that plants grown in medium B supplied with 0.5 and 1.0 strength of MS nutrients had high growth rate, and 20-day cutting interval was the best. 72-plug tray was better than 128-plug. Secondly, it was to develop a technique for the production of first-generation seed roots using hydroponics cultivation system. The yield of virus-free plants propagated in the non-insect proof and open-field cultivation was 2,402 kg/10a, 6% higher than those in the insect-proof cultivation, and the rate of virus re-infection was 18% higher compared to 3.3% with insect-proof cultivation. Lastly, it was to investigate the growth performance of virus free plants in farmers' field. Differences were existed in the yield depending on the variety used, but virus free plants showed an increase of $6{\sim}24%$ over virus infected plants.

• Korean Journal of Veterinary Research
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• v.1 no.1
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• pp.36-53
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• 1961

This experiment was conducted on the fowl pox embryo vaccine for the production immunity, and stability, using an attenuated fowl pox virus (Nakano strin). Burnet's window method was applied, that is, 0.1 ml. of seed virus was inoculated on the chorioallantoic membrane of 12-day old chicken embryos, and incubated for 5 to 6 day, and then the result were read. Four kinds of suspensions of different embyo tissue were prepared and tested for the infectivity in chickens. Finally the suspension of chorioallantoic membrane was used as the vaccine throughout the experiment. Results obtained in this experiment are summarized as follows: (1) Of embryo tissue infected with the vaccine virus, chorioallantoic membrane had the highest virus titer of $10^{-5.4}$ $EID_{50}$, and albumen the lowest titer of $10^{-0.7}$ $EID_{50}$. (2) Suspensions of infected whole embryo with or without saline, and de-embryonated whole egg had about the same virus titer of $10^{-4.4}$ $EID_{50}$, whereas the chorioallantoic membrane had $10^{-5.7}$ EID 50 or higher. The virus titer droped one log from $EID_{50}$ when inoculated into chickens. Takes were observed 35.6% of 500 chickens by stick method and 89% of 500 chickens by brush method. (3) The chorioallantoic membrane conferred almost perfect immunity for chickens by 10 days after vaccination. (4) Satisfactory immunity was observed in the chickens when eruption in a single follicle. (5) Eight of 10 vaccinated chickens revealed durable immunity for 307 days following vaccination. (6) The vacuum-dried vaccine maintained its infectiviy for 899 days at $5^{\circ}C$ or below and maintained the vius titer of $10^{-3.6}$ $EID_{50}$. On the other hand, non-desiccated wet vaccine maintained the titer of $10^{-3.0}$ $EID_{50}$ for 50 days of preservation period at $5^{\circ}$. However, in 50% glycerin-saline the infectivtiy of the same wet vaccine dropped to $10^{-1.5}$ $EID_{50}$ (7) The vartation of virus titer of the vaccine before and after desiccation was $10^{-0.5}$ $EID_{50}$ on the average. (8) As suspending media, 0.85 per cent saline and distilled water showed nearly the same effect on the infectivity of the vaccine by retaining the titer $10^{-3.0}$ $EID_{50}$ after 50 days of preservation both at $5^{\circ}C$ and $20^{\circ}C$, while 50 percent cent glycerine-saline dropped the titer to $10^{-2.5}$ EID and $10^{-1.5}$ $EID_{50}$ respectively at $5^{\circ}C$ and $2^{\circ}C$ after the same period.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.4
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• pp.498-504
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• 2014

Potato virus Y (PVY) and potato leafroll virus (PLRV) are among the most damaging potato viruses and prevalent in most potato growing areas. In this study, cryopreservation was used to eradicate PVY and PLRV using two cryogenic methods. Potato shoot tips proliferated in vitro were cryopreserved through droplet-vitrification and encapsulation-vitrification using plant vitrification solution 2 (PVS2; 30% glycerol + 15% dimethyl sulfoxide + 15.0% ethylene glycol + 13.7% sucrose) and modified PVS2. Both cryogenic procedures produced similar rates of survival and regrowth, which were lower than those from shoot tip culture alone. The health status of plantlets regenerated from shoot tip culture alone and cryopreservation was checked by reverse transcription-polymerase chain reaction. The frequency of virus-free plants regenerated directly from highly proliferating shoot tips reached 42.3% and 48.6% for PVY and PLRV, respectively. In comparison, the frequency of PVY and PLRV eradication after cryopreservation was 91.3~99.7% following shoot-tip culture. The highest cryopreserved shoot tip regeneration rate was observed when shoot tips were 1.0~1.5 mm in length, but virus eradication rates were very similar (96.4~99.7%), regardless of shoot tip size. This efficient cryotherapy protocol developed to eliminate viruses can also be used to prepare potato material for safe long-term preservation and the production of virus-free plants.

• Food Science and Preservation
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• v.21 no.5
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• pp.735-739
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• 2014

Ephedra sinica Stapf, known as a medicinal plant, inhibited not only syncytium formation, but also trafficking of viral glycoprotein, hemagglutinin-neuramidase (HN) to the cell-surface. Trafficking of viral glycoprotein to the surface of infected-cells results in syncytium formation in Newcastle disease virus (NDV)-infected baby hamster kidney (BHK) cells. Viral glycoprotein in the infected-cell is processed within the endoplasmic reticulum during routing into surface. The processing of viral glycoprotein like a N-linked oligosaccharide trimming by ${\alpha}$-glucosidase in cell is necessary for virus infection. Methanol extracts showed inhibitory activities ($IC_{50}$ $15{\mu}g/mL$) against ${\alpha}$-glucosidase. This suggested that E. sinica extracts inhibited the cell-surface expression of NDV-HN glycoprotein without significantly affecting HN glycoprotein synthesis in NDV-infected BHK cells.

• Food Science and Preservation
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• v.21 no.2
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• pp.170-174
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• 2014

Hepatitis A virus (HAV) infection leads to acute liver failure and death through the intake of contaminated food. The polymerase chain reaction (PCR) has been used to detect HAV in food samples. HAV detection takes a long time, however, due to the virus concentration step required before PCR assay. In this study, a rapid method of detecting the HAVs present in lettuce using immunomagnetic separation combined with quantum dots (IMS-QDs) assay was developed. The detection limit of IMS-QDs for HAV was 10 $TCID_{50}/mL$, similar to the result that was obtained using RT-PCR combined with PEG or IMS. The application of IMS-QDs assay completed the viral detection within one hour, but this was not possible using PEG combined with RT-PCR. In conclusion, IMS-QDs assay is a rapid and efficient method for detecting HAV at a low concentration in agricultural products.

• Food Science and Preservation
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• v.22 no.6
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• pp.901-907
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• 2015

Genetically modified (GM) pepper H15 containing the gene for cucumber mosaic virus (CMV) coat protein (CP) and its control line non-GM pepper P2377 were investigated for their allergic risk. Amino acid sequence of the inserted gene product CMV-CP was compared with those of known allergens. No known allergen had greater than 35% amino acid sequence homology over an 80 amino acid window or more than 8 consecutive identical amino acids. Protein patterns of GM and non-GM pepper extracts were evaluated by SDS-PAGE, which showed similar distribution of protein bands for both GM and non-GM pepper. Antigen-antibody reactions were compared between GM and its non-transgenic parental control. ELISA and immunoblot analysis of sera from allergic patients showed some IgE reactivity; however, no differences were observed between GM pepper H15 and P2377. We therefore conclude that CMV-CP is less likely to be an allergen; the protein composition and allergenicity of the GM pepper H15 is not different from that of P2377 and safe as a commercial host.

• Korean Journal of Poultry Science
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• v.49 no.2
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• pp.99-108
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• 2022

Avian leukosis virus (ALV) is a highly contagious retrovirus that causes tumors and has resulted in great economic loss worldwide owing to its high transmission rate. Various ALV viral subgroups exist, with infections occurring via specific host receptors. The susceptibility or resistance of avian species to the ALV-A and K subgroups is determined by the host receptor, the tumor virus locus A (tva) gene, while that to ALV-B depends on another host receptor, the tumor virus locus B (tvb) gene. The resistance alleles of tva and tvb have primarily been identified in China, but none have beendetected in Korea. We analyzed the frequencies of tva and tvb genotypes in White Leghorn (WL), Korean Ogye (KO), and Korean native chicken (KNC) breeds, and assessed the resistance to ALV subgroups. In WL, both tva and tvb had various genotypes, including susceptibility and resistance alleles, whereas in KO, tva and tvb resistance alleles were dominant. In KNC, tva susceptibility and resistance alleles were mixed, whereas tvb resistance alleles were dominant. In addition, we showed that there were differences in the splicing pattern of tva transcripts and the expression level of tvb transcripts within breeds. Finally, we confirmed that ALV resistance depended on KO and KNC genotypes by in vitro infection of chicken embryonic fibroblasts with ALV. These results highlight that some KO and KNC individuals are naturally resistant to ALV subgroups A, B, and K, and will facilitate the preservation of economically superior traits through selective breeding.

• Journal of Sericultural and Entomological Science
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• v.9
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• pp.67-87
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• 1969

1. Objectives and Importance. Many silkworms have been damaged by nuclear polyhedrosis virus diseases thoughout the country every year causing a decease in cocoon production by approximately ten per cent per year. The damage caused by the infections virus has occured in spite of complete disinfection. In this respect, it is well known it is impossible, at the present time, to protect the silkworm from these virus infections through chemical and physical control methods. Therefore, this author has attempted to solve this urgent problem from the view point of heredity and breeding, discovering the different resistances and heritabilities among 120 stains collected from throughout the country, and selecting the ones with highest resistance for the basic materials in the silkworm breeding. 2. Results of work 1) The strains with strong resistance to the nuclear polyhedrosis virus diseases are N$_4$, N$\sub$15/, N$\sub$48/, C$\sub$55/ and Em. the log ED$\sub$50/ values of them vary between 0.799 and 1.611. The susceptible strains are N$\sub$20/, C$\sub$62/, N$\sub$76/, N$\sub$79/ and C$\sub$108/, the log ED$\sub$50/ values of them vary between 5.159 and 7.258. (Reference Table 4) The Japanese strain with a log ED$\sub$50/, value of 3.770 is the strongest, followed by the Chinese strain with a log ED$\sub$50/ value of 3.564. The weakest is the European strain with a log ED$\sub$50/ value of 3.3381. The direction coefficient of the regression equation of the susceptibility varies between 0.1 and 0.6, the uniformness of the resistance of the preserved strains of this country is comparatively low. The hereditary henomena of the resistance of each strain and the conerete method of its application for silkworm breeding main the subjects for later studies. 2) The content of water and ash in silkworm has not been correlated with the capability for resistance to the virus diseases(Reference. Table 8). but it is very significantly correlated with mortality rate (in common reaning). In the case of the silkworms which have just completed the fourth moulting the content of water and ash is not related to the mortality rate. In the case of the silkworms which have just completed the third moulting, however, the water( +0.326) and ash (+0.362) registered a high significance. The ash content in the first ($\div$0.520) and second ($\div$0.386) moults is highly significant but water content in both cases is not significant (Reference Table 7). 3) The No. 205 strain proved to be the best in character among the various F$_1$ hybrids. No. 204 was very good in strength but a little lower in cocoon character than the control. No. 212 was a little low in coccon character and mortality was average, but the cocoon harvest was the best among all the varieties offered for (Reference Table 9). 4), In short, the above mentioned strains which are known to have strong resistance to the virus disease are expected to provide basic data for breeding strong varieties. It is proposed that continued research should be conducted on the characteristics of various strains for a satisfactory preservation of various characteristics.research should be conducted on the characteristics of various strains for a satisfactory preservation of various characteristics.

• Applied Biological Chemistry
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• v.38 no.5
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• pp.435-439
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• 1995

Some characteristics and pathogenicity of Hyphantria cunea nuclear polyhedrosis virus (HcNPV), a potential microbial pesticide was studied. H. cunea NPV replicated in the nucleus of S. frugiperda cells cultured in the TNMFH medium. In case of virus infected cell, prepolyhedra formation was observed at 24hrs post-infection. At 48 hrs post-infection, Most of the infected cell contained many mature polyhedra which were released into culture media 72 hrs post-infection, with the cells grown in suspension culture, pH of the culture medium increased during the virus replication: the pH of fresh medium was 6.35 and rose to 6.77 within 120 hrs. Polyhedra formed a band in linear density gradient of sucrose by centrifugation, which co-sedimented with $50{\sim}55%$ sucrose. The shape of the purified polyhedra was mostly tetragonal hexahedron and its size was about $2.5{\mu}m$. Electron microscopy and phase contrast microscopy showed that many bundled nucleocapsids were occluded in mature polyhedra at 48 hrs post infection. H. cunea larvae infected with NPV showed a higher motality in the second and third instar than in the fourth instar. Death rate of H. cunea larvae in the second and third instar fed with leaves coated with $1.5{\times}10^{9}{\sim}l.5{\times}10^{7}PIBs/ml$ reached more than 90%.