• International Journal of Industrial Entomology and Biomaterials
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• v.21 no.1
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• pp.117-125
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• 2010

The tropical tasar silkworms, Antheraea mylitta (D) produce famous silk 'Kosa' in central part of India. Due to outdoor rearing it became susceptible to viral infection including cytoplasmic polyhedrosis virus (CPV). The common mode of entry of cytoplasmic polyhedrosis virus is per os and cause gresserie disease to the larvae. Histopathological studies elucidated the insect CPV virus produces infective polyhedral inclusion bodies (PIBs) in the midgut cell cytoplasm of virus infected fifth instar larvae. The PIBs multiply enormously in the cytoplasm without invading the nucleus. Ultrastructural studies confirmed the pathological effects of CPV on in midgut cell cytoplasm. The multiplication of polyhedral inclusion bodies took place into the vacuoles and form virogenic stromata in the cytoplasm of cells. However, the encapsulations of polyhedral inclusion bodies into the polyhedrin protein occurred and polyhedra were released into the lumen. At the late stage of infection, cells showed the regressed cytoplasmic organelles with large vacuoles and elongated mitochondria. Hence, the horizontal transmission of CPV causing the midgut cells disintegration in the tasar silkworm, Antheraea mylitta (D) confirmed during infection.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.352-355
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• 2004

Heliothis zea (HzAM1) insect cells were immobilized in microspheres by sodium-cellulosesulfate (NaCS) and polydiallyldimethylammoniumchloride (PDADMAC). The highest HzAMl cell density was $7.5{\times}10^7$ cells/mL in the microspheres. After infection of the immobilized cells by Heliothis zea single nuclear polyhedrosis virus (HzSNPV), the highest concentration of HzSNPV (polyhedral inclusion bodies: PIBs) produced was $2.87{\times}10^{10}$ PIBs/mL in the microspheres.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.395-400
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• 2004

Choristoneura fumiferana (Cf-2Cl) insect cells were cultured and immobilized by using cellulosesulfate (NaCS) and polydiallyldimethylammoniumchloride (PDADMAC). A concentration of CfMNPV (Choristoneura fumiferana multiple-mucleopolyhedrovirus) and a Cf-2Cl cell density in the microspheres have been achieved at the densities of $1.57\times10^{10}$ PIBs/ml and $7.5\times10^7$ cells/ml, respectively. Additionally, MTT-test was used to measure the viable cell density in the microspheres, and the confidence of MIT-test was investigated before and after baculovirus infection in the immobilized cell culture.

• Applied Microscopy
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• v.21 no.1
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• pp.21-26
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• 1991

Midgut cells of the dead Dendrolimus spectabilis larvae by infection of D. spectabilis nuclear polyhedrosis virus (DsNPV) were observed by transmission electron microscopy. DsNPV replicated in the nucleus of the infected midgut cells and the virogenic stroma of DsNPV appeared in the nucleus, from which nucleocapsids were formed. Also the formation of polyhedral inclusion bodies were observed in the nucleus.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.632-637
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• 2007

A cytoplasmic polyhedrosis virus (CPV) was isolated from the larvae of Thaumetopoea pityocampa and shown to cause an infection of midgut cells. This viral infection revealed several important diagnostic symptoms, including discoloration of the posterior midgut, reduced feeding, and extended development time of the larvae. The virus infection is lethal to Thaumetopoea pityocampa, and with the increasing doses kills the larvae within 4-5 days post infection. Electron microscopy studies showed typical cytoplasmic polyhedral inclusion bodies that are icosahedral, and ranged from 2.4 to $5.3{\mu}m$ in diameter. Electrophoretic analysis of the RNA genome showed that the virus has a genome composed of 10 equimolar RNA segments with the sizes of 3,907, 3,716, 3,628, 3,249, 2,726, 1,914, 1,815, 1,256, 1,058, and 899 bp, respectively. Based on morphology and nucleic acid analysis, this virus was named Thaumetopoea pityocampa cytoplasmic polyhedrosis virus (TpCPV), and belongs to the genus Cypovirus, family Reoviridae.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1485-1490
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• 2006

In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

• Korean journal of applied entomology
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• v.42 no.2
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• pp.159-163
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• 2003

This experiment was conducted to investigate pathogenicity of Spodoptera litura nucleopolyhedrovirus (SINPV) with different temperatures to mass production. In laboratory, LC$_{50}$ values of SINPV were 5.534$\times$10$^3$ PIBs/ml and 4.l0$\times$10$^2$PIBs/ml in 1st instar larvae at 2$0^{\circ}C$ and 28$^{\circ}C$, respectively, but those were increased at 32$^{\circ}C$. LC$_{50}$ at 3rd and 5th instar larvae were showed the lowest value at 28$^{\circ}C$. LT$_{50}$ values of SINPV in 1.0$\times$10$^{5.7}$ PIBs/ml were determined at various temperature conditions (20-32$^{\circ}C$). The result showed that treatments at 28$^{\circ}C$ and 32$^{\circ}C$ shortened LT$_{50}$ values, but treatments at 2$0^{\circ}C$ and 24$^{\circ}C$ was relatively longer. Also LT$_{50}$ values was shortened in high concentration and young larva. LT$_{50}$ values are dependent on the temperature, viral concentration and larval instar.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.239-255
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• 1997

Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.