• Journal of Sericultural and Entomological Science
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• v.13 no.1
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• pp.35-47
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• 1971

Current overseas research reveals that among the pathogens causing flacherie of silkworm, damage by infectious flacherie virus is the most serious, but little research in this fold has been reported in Korea. This experiment was undertaken to observe the occurrence of infectious flacherie virus by means of biological environmental conditions associated with occurrence of virus discase and interaction of the virus of flacherie and Bacillus spp. isolated from flacherie silkworm, and to determine ways to check infection by the virus during the rearing of silkworms. The results obtained are as follows: 1. The pathogen, infectious flacherie virus observed in Korea, is proved to be round in shape and 26-30m$\mu$ in diameter under observation with electron microscope, 2. The infectious flacherie virus-disease occurred apparently in conditions of nutritional disturbance such as shortage of diet or rearing in high temperature and humidity during the 3-4th instar. 3. The percentage of disease-occurrence was increased remarkably, and the latent period was shortened in the case of simple inoculation of virus suspension as compared with the suspension added with bacteria. 4. The application of calcium hydroxide in the silkworm-rearing bed is able to check infection of virus disease.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.27-31
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• 2003

Infectious flacherie of silkworm Bombyx mori is caused by B. mori infectious flacherie virus (BmIFV) and causes severe crop loss to sericulturists. In the present study, a colloidal textile dye-based dipstick immunoassay is developed for the detection of infectious flacherie in silkworms. Colloidal textile dye (blue D2R) with Aλ$_{max}$ at 620 nm was sensitised with 500 $\mu\textrm{g}$/ml of purified anti-BmIFV IgG. The dye-antibody reagent detects purified antigen up to 10 ng/ml and BmIFV infection in diseased larval extracts $(up to a dilution of {10^-5})$ and faecal matter extracts $(up to a dilution of {10^-2})$ by forming clear blue dot within 30 min. It was observed to be stable for three months period at $4^{\circ}C$. The efficacy of textile dye-based dipstick immunoassay was on pay with HRP-based dipstick immunoassay and fluorescent antibody test, and better than latex agglutination and ouchterlony tests in the detection of BmIFV The dye-based dipstick immunoassay method provides a simple, sensitive and less expensive test for the detection of BmIFV infection in silkworms.s.

• Journal of Sericultural and Entomological Science
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• v.20 no.2
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• pp.32-35
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• 1978

The infectious flacherie viruses are serious diseases in the silkworm, which affects the cocoon crops. However, there is only a few study on this diseases up-to-date in Korea and, in this experiment, the authors investigated the resistance of the silkworm varieties to flacherie and Ina-flacherie viruses by peroral infection. A cross, Hansaeng ＃1$\times$ Harisaeng ＃2, showed the highest resistance to the flacherie virus a Mudeung$\times$Geumho showed the lowest resistance among the examined varieties. It seemed that the varietal difference of resistance against Ina-flacherie virus was appeared and Jam117$\times$ Jam118, Gyeongchu$\times$Yeonil, Mudeung$\times$Geumho, Hansaeng＃1$\times$Hansaeng＃2 and Hansaeng＃3 $\times$ Hansaeng ＃4 showed non susceptibility by peroral infection. However it was shown that Jam115 $\times$Jam116 had the lowest resistance to Ina-flacherie virus.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.35-40
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• 2004

Bombyx mori densonucleosis virus type 1 (BmDNV1)- a non occluded virus causes flacherie disease in the susceptible stocks of the silkworm, Bombyx mori. However, some stocks are non-susceptible. Non-susceptibility to BmDNV1 in B. mori is a unique case where the virus infection is completely inhibited by a single gene of the host. A survey conducted by this institute in some parts of Karnataka state has revealed that, 43.05% of the total incidence of flacherie disease caused by non-occluded viruses, are due to the synergistic infection of B. mori densonucleosis and infectious flacherie virus. Earlier study indicated that rearing of BmDNV1 resistant silkworm stock is effective in protecting silkworm against BmIFV also. In the present study the response of 78 silkworm stocks which include 42 of non-diapausing and 36 of diapausing groups, to BmDNV1 is investigated. Newly ecdysed third instar larvae were inoculated per-os with 10% inoculum of BmDNV1 extracted from the mid-gut of infected silkworm. One non-diapausing and three diapausing silkworm stocks were found to be resistant to BmDNV1. Eleven silkworm stocks were found to possess moderate resistance whereas rest sixty three were found to be susceptible to BmDNV1. Genetic analysis has shown that the resistance to BmDNV1 is autosomally inherited and controlled by a major dominant or a major recessive gene in different silkworm stocks. These resistant stocks can be utilized as the resource material to develop BmDNV1 resistant commercial hybrids. The selection strategies, depending upon the mode of inheritance of resistance in the resource material chosen, are discussed.

• Journal of Sericultural and Entomological Science
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• v.32 no.1
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• pp.44-48
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• 1990

Monoclonal antibodies were prepared against Bombyx densonucleosis virus type-II(Yamanashi isolate). Four hybridoma clones, named C4, Fl, H2, M9 were only reacted with the DNV-II, but those were not reacted with Bombyx densonucleosis virus type-I(Ina isolate) and infectious flacherie virus(IFV) by double diffusion test in 0.8% agarose gel. C4, Fl and M9 of them were reacted with 53KDa polypeptide of DNV-II, and H2 was reacted with 46. 5KDa polypeptide of the virus.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.37-43
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• 1984

The prevalence of the infectious flacherie virus (FV) disease causes a severe damage to cocoon yield and various methods to control the disease have been studied. In this regard, guanidine hydrochloride (GH), one of the guanidine derivatives known as the most inhibitory agent against the replication of picorna virus, was applied to silkworms per os with mulberry leaves and the results were as follows. 1. The application of GH below 0.01% of the chemical concentration did not give any damage to silkworm larvae. 2. The transmission of the virus disease by introducing the FV infected larvae to the healthy larvae group was proportioned to the number of infected larvae. When l% of infected larvae was introduced to the rearing tray of healthy larvae, the pupation rate was 70.7%(79) and it was 38.4% (43) to 5% of infected larvae introduced, while the control of non-mixed with infected larvae gave 89.2% (100) of pupation rate. The cocoon yield from 10,000 larvae also showed the same tendency as the pupation rate. 3. The inhibitory effect of GH against the replication of FV showed ten times in treatment of 0,01% of the chemical agent compared to the non-treatment. 4. The successive application of GH after virus inoculation to silkworm larvae led to the most effective on the inhibition of the virus replication. 5. The immediate application of GH after the virus inoculation also gave the best effect on the inhibition of the virus replication in silkworm larvae. 6. The effect of GH on the inactivation of FV in vitro was not observed.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.95-100
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• 2006

Central Sericultural Research and Training Institute, Mysore have evolved several highly productive bivoltine hybrids which can produce international grade raw silk. Among them $CSR2{\times}CSR4,\;CSR2{\times}CSR5,\;CSR3{\times}CSR6,\;CSR17{\times}CSR16,\;CSR18{\times}CSR19$ and $CSR12{\times}CSR6$ are being popularized in the field. There is a minimum difference in their economic characters but they appear to differ in survival. Though they are productive under high input management conditions, they are very susceptible to different diseases under normal rearing practices. No systematic attempts have been made to test their susceptibility status / resistance. Thus the present study is a modest attempt to screen the above six productive bivoltine hybrids to two important pathogens viz., Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) and Bombyx mori Infectious Flacherie Virus (BmIFV) along with existing hybrid, $KA{\times}NB4D2$ to assess their susceptibility / resistance. The results shows that the productive hybrid $CSR2{\times}CSR4$ is the most resistant to BmNPV and it is suggested by its highest $LC_{50}$ value followed by $CSR12{\times}CSR6,\;KA{\times}NB4D2,\;CSR3{\times}CSR6,\;CSR17{\times}CSR16,\;CSR18{\times}CSR19,\;CSR2{\times}CSR5$. Based on the $LC_{50}$ value and $LT_{50}$ values for BmIFV, the hybrid $KA{\times}NB4D2$ was found to be the most resistant (1st position) one followed by $CSR3{\times}CSR6$ (2nd position) $CSR2{\times}CSR$ (3rd position) and $CSR12{\times}CSR6$ (4th position) $CSR17{\times}CSR16$, $CSR18{\times}CSR19$ (5th position) and $CSR2{\times}CSR5$ being the least. The response of 7 bivoltine hybrids to both the pathogens BmNPV and BmIFV indicates that, the hybrids $CSR2{\times}CSR4$, $CSR12{\times}CSR6$ and $KA{\times}NB4D2$ were found to be the most resistant when compared to others. Further, $KA{\times}NB4D2$ being less productive hybrid with a shell ratio of 20.08%, the other two hybrids $CSR2{\times}CSR4$ (Cocoon shell ratio, 21.44%) and $CSR12{\times}CSR6$ (cocoon shell ratio, 23.45%) can be considered to be most productive with superior quality cocoon and resistant to both BmNPV and BmIFV pathogens. The overall study indicated that the hybrid $CSR2{\times}CSR5$ is the most susceptible hybrid to both the pathogens.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.1
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• pp.8-12
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• 2009

A study has been made to know the effect of a phytoecdysteroid 'Sampoorna' on uniform maturation of silkworms during spinning and its effect on diseased silkworms infected by major silkworm disease viruses, Bombyx mori nuclear polyhedrosis virus (BmNPV) and Bombyx mori infectious flacherie virus (BmIFV). In the present investigation, the effect of the phytoecdysteroid "Sampoorna" on Grasserie disease caused by BmNPV have shown an average cocoon melting of 11.91% with a disease incidence of 5.83%. The values of 't' test for different treatments of BmNPV indicated low survival rate and cocoon traits were drastically reduced. Another major disease Flacherie caused by BmIFV has shown considerable levels of larval disease incidence (22-32%) and cocoon melting (3-7.67%) with an average melting of 12.95% and 20.24% disease incidence. There is a drastic reduction in survival rate, cocoon yield and other economic traits. The control batches were indicated negligible values for disease incidence and cocoon melting with Sapoorna application and without the inoculation of the two disease-causing viruses. The application of Sampoorna on already infected batches with major pathogens triggered high mortality and disease incidence and melting percentage was also significantly increased with reduced economic traits. Hence, it is suggested that application of Sampoorna in infected batches should be done only in the extreme conditions of rearing. Application of Sampoorna on healthy batches led to uniform maturation and improvement in productivity with the added advantage of better quality cocoons and labour saving.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.