Dimorphisim In Nuclear Polyhedrosis Virus (Bmnpv) (Family:  Baculoviridae) Causing ‘grasserie’ Disease In Silkworm (Bombyx  Mori L.): Light And Electron Microscopy And Protein Profile

Satadal Chakrabarty; Suman Deb; A K  Saha; N Hazra; B Manna; B B Bindroo

doi:10.48165/

Authors

Satadal Chakrabarty Central Sericultural Research and Training Institute, Berhampore West Bengal, 742 101 (India)
Suman Deb Department of Zoology, The University of Burdwan, Burdwan, West Bengal, 713 104 (India)
A K Saha Central Sericultural Research and Training Institute, Berhampore West Bengal, 742 101 (India)
N Hazra Department of Zoology, The University of Burdwan, Burdwan, West Bengal, 713 104 (India)
B Manna Parasitology Research Unit, Department of Zoology, University of Calcutta, Kolkata, West Bengal - 700 019 (India)
B B Bindroo Central Sericultural Research and Training Institute, Berhampore West Bengal, 742 101 (India)

DOI:

https://doi.org/10.48165/

Keywords:

Bombyx mori, budded virus, Grasserie, polyhedra, occluded bodies, innate immune system

Abstract

The present study was aimed to microscopically assess dimorphism in nuclear polyhedrosis virus (BmNPV) responsible for ‘Grasserie’ disease in silkworm, Bombyx mori L. Tissue sections of infected larvae integument were found ruptured, had abundant lump cells and released milky fluid containing occluded bodies (OBs). Nuclei of columnar epithelial cells of integument were almost covered with BmNPV polyhedra. Scanning electron micrograph (SEM) revealed full lesions in infected integuments with very fragile hypo dermis. Transmission electron micrograph (TEM) showed that infecteddifferent parts of peripheral membrane contain lipid globules started to diffuse continuously to form a large structure to occupy maximum OBs. In advanced stage of infection, two viral phenotypes were noticed. 1st form occluded within polyhedra was responsible for primary infection in mid-gut cells. The 2nd form ‘budded virus (BV)’ never became occluded and was released into haemolymph and spread infection bycell to cell contact within insect body. The fully transformed lipid globules discharged the 1st form whereas BV was observed in integument, mid gut and Malpighian tissues. Interestingly, the infected mid gut cells were dilated in presence of OBs and gradually increased in number to form chain-like structure which broke up no sooner the number of OBs reached highest carrying capacity, releasing polyhedra in gut juice. Inner layer of infected Malpighian tissues were loose and fragmented with OBs in between the striations of Malphigian cells. BVs, the new finding from B. mori, are responsible for virulence of BmNPV in silkworm. 29 and 17 kDa proteins in multivoltine (Nistari) and bivoltine (NB4D2), respectively, were absent in the haemolymph of BmNPV infected counterpart. The total quantity of haemolymph protein was more in multivoltine samples than in bivoltine samples.

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References

Benz, G.T. 1963. Introduction: historical prospective. pp.1-35. In: The Biology of Baculoviruses, Vol.1. (Eds. R.R. Granados and B.A. Federici) CRC Press, Boca Raton, Florida, USA. , Bergold, G.H. 1958. Viruses of insects. pp. 413-456. In: Handbuch der Virus Forschung (Eds. C. Hallauer and K.F. Meyer), Springer-Verlag, Vienna.

Bhattacharya, J., Krishnan, N., Chandra, A.K., Prakash, O. and Sengupta, U. 1993. Use of simple slide agglutination test for the detection of nuclear polyhedrosis virus of silkworm, Bombyx mori L. Current Science, 8: 638-639.

Bradford, M. M.1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Annals of Biochemistry, 72 : 248-254. Brancalhao R.M.C., Torquato, E.F. and Fernandez, M.A.2009. Cytopathology of Bombyx mori

(Lepidoptera: Bombycidae) silk gland caused by multiple nucleopolyhedrovirus. Genetics and Molecular Research, 8: 162-172.

Chakrabarty S., Deb, S., Saha, A.K, Hazra, N., Manna, B. and Bindroo, B.B. 2011. Morphometrical, electron micrographical and innate protein profile study due to nuclear polyhedrosis virus infection in Bombyx mori L. In: Proceedings of 22nd All India Congress of Zoology & National Seminar on Recent Advances in Biological Sciences: Biodiversity and Human Welfare. Zoological Society of India, Bodh Gaya, Bihar, India (in press)

Cornalia, E. 1856. Monographia del bombice del gelso. Mem. I.R. 1st Lombardo di Scienzelet. Arti, Bernadoni di Gio, Milano. pp.348-351.

Funkoshi, M. and Aizawa, K. 1989. Anti-viral substance in the silkworm guts juice against a nuclear polyhedrosis virus of the silkworm. Journal of Invertebrate Pathology, 54: 151-155. Gamo, T. 1978. Low molecular weight lipoproteins in the haemolymph of the silkworm, Bombyx mori: Inheritance, isolation and some properties. Insect Biochemistry, 8: 457-470.

Gururaj, C.S., Sekarappa, B.M. and Sarangi, S.K. 1999a. Metabolic profile of the target tissues in silkworm, Bombyx mori L. infected with BmNPV. Indian Journal of Sericulture, 38: 89-94. Gururaj C.S., Sekarappa, B.M. and Sarangi, S.K. 1999b. Effect of BmNPV infection on the digestive enzyme activity in the silkworm. Indian Journal of Sericulture, 38: 102-106.

Haas-Stapleton, E.J., Washburn, J.O. and Volkman, L.E. 2005. Spodoptera frugiperda resistance to oral infection by Autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion derived virus binding in the mid gut. Journal of General Virology, 86: 1349-1355.

Herniou, E.A., Olszewski, J.A., Cory, J.S. and O’Reilly, D.R. 2003. The genome sequence and evolution of baculoviruses. Annual Review of Entomology, 48: 211-234.

Izumi, S., Kiguchi, K. and Tomino, S. 1984.Hormonal regulation of biosynthesis of major plasma proteins in Bombyx mori. Zoological Science, 1: 223-228.

Kawakami, K. 2001. Illustrated Working Process of New Bivoltine Silkworm Rearing Technology. JICA. PP.B.S.T. Project. Central Sericultural Research & Training Institute, Berhampore, Mysore, India, p. 11.

Khurad, A.M., Mahulikar, A, Rathod, M.K., Rai, M.M., Kanginakurdu, S. and Nagaraju, J. 2004. Vertical transmission of nuclear polyhedrovirus in the silkworm, Bombyx mori L. Journal of Invertebrate Pathology, 87: 8-15.

Kovacs-Simon, A., Titball, R.W., and Michell, S.L. 2011. Lipoproteins of bacterial pathogens. Infection and Immunity, 79: 548-561.

Krishnan, N., Chaudhuri, A., Sengupta, A.K., Chandra, A.K., Sen, S.K. and Saratchandra, B. 2001. NADP-dependent malate dehydrogenase activity and associated biometabolic changes in hemolymph and fat body tissues of silkworm Bombyx mori L. following Baculovirus infection, International Journal of Industrial Entomology, 2:149-153.

Satadal Chakrabarty et al.

Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

Nataraju, B., Datta, R.K., Baig, M., Balavenkatasubbaiah, M., Samson, M.V. and Sivaprasad, V. 1998. Studies on the prevalence of nuclear polyhedrosis of sericultural areas of Karnataka. Indian Journal of Sericulture, 37: 154-158.

Nataraju, B., Sivaprasad, V. and Datta, R.K. 2000. Studies on development of oral vaccine against NPV in silkworm, Bombyx mori L., Sericologia, 40: 421-427.

Patil, C.S.1995. Disinfection and maintenance of hygiene in Sericulture. Karnataka State Sericulture Research & Development Institute, Karnataka, India.

Rabindra, R.J. 1994. Investigation on the Polyhedral Virus Disease of Bombyx mori L. and their Management. Project Report, Tamil Nadu Agriculture University, Coimbatore, Tamil Nadu, India. Rahman, M.M. and Gopinathan, K.P. 2004. Systemic and in vitro infection process of Bombyx mori nucleopolyhedrovirus. Virus Research, 101: 109-118.

Reddy, J.T. and Locke, M. 1990. The size limited penetration of gold particles through insect basal lamina. Journal of Insect Physiology, 36: 397-407.

Sen, R., Patnaik, A.K., Maheshwari, M. and Dutta, R.K. 1999. Susceptibility status of the silkworm, Bombyx mori germplasm stock in India to BmNPV virus. Indian Journal of Sericulture, 36:51-54. Sheeba-Rajakumari D.V., Padmalata, C.S., Das, S.M. and Ranjitsingh, A.J.A. 2007. Efficacy of probiotic and neutraceutical feed supplements against flacherie disease in mulberry silkworm, Bombyx mori L. Indian Journal of Sericulture, 46: 179-182.

Steinhaus, E.A. 1949. Principles of Insect Pathology. McGraw Hill Book Co., Inc. London, UK. Torquato, E.F., de Miranda Neto, M.H., Brancalhão, R.M. and Franco, V.S. 2006. Nucleo polyhedrovirus: Scanning electron microscopy technique. Neotropical Entomology, 35: 787-790. Uchida, Y., Kawamoto, F., Himeno, M. and Hayashiya, K. 1984. A virus-inactivating protein isolated fromthe digestive juice of silkworm, Bombyx mori. Journal of Invertebrate Pathology, 43:182-189. Ujita, M., Kimura, A., Nishino, D., Yokoyama, E., Banno, Y., Fujii, H. and Hara, A. 2002. Specific binding of silkworm Bombyx mori 30 kDa lipoproteins to carbohydrates containing glucose. Bioscience Biotechnology and Biochemistry, 66: 2264-2266.

Ujita, M., Katsuno, Y., Kawachi, I., Ueno, Y., Banno, Y., Fujii, H. and Hara, A. 2005. Glucan-binding activity of silkworm 30 kDa apolipoprotein and its involvement in defense against fungal infection. Bioscience Biotechnology and Biochemistry, 69: 1178-1185.

Undeen, A.H. 1997. A handbook of biology and research technique. p. 14. In: Microsporidia (Protozoa) (Ed. J.D. Millar), Southern Cooperative Series, Bulletin No.387, Oklahama University, USA. Wang, X.Y., Cole, K.D. and Law, J.H. 1989. The nucleotide sequence of a microvitellogenin encoding gene from the tobacco hornworm, Manduca sexta. Gene, 80: 259-268.

Watanabe, H. and Kobayashi, M. 1969. Effect of virus infection on the protein synthesis in the silkworm, Bombyx mori L. infected with nuclear and cytoplasmic polyhedrosis virus, Japan. Journal of Application Entomology and Zoology, 15: 198-202.

Yao Hui-peng, W.U,, Xiao-feng and Gokulamma, K. 2006. Antiviral activity in the mulberry silkworm, Bombyx mori L. Journal of Zhejiang University SCIENCE A (Suppl.): 350-356.