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In vitro and in vivo studies on the protective potential of Withania somnifera against Infectious Bursal Disease Virus of poultry

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Title In vitro and in vivo studies on the protective potential of Withania somnifera against Infectious Bursal Disease Virus of poultry
 
Creator Ganguly, Bhaskar
 
Contributor Rastogi, S.K.
 
Subject in vitro, medicinal plants, withania somnifera, infectious bursal disease virus, poultry farming, poultry diseases, viral diseases
 
Description Thesis-PhD
Infectious Bursal Disease is a serious problem in poultry. Withania somnifera (WS), an important herb in traditional medicine, possesses prominent properties that may confer anti-IBDV activity. The present study aimed at determining the protective potential of Withania somnifera root powder against Infectious Bursal Disease virus of poultry in vitro and in vivo.
WS roots were extracted in acetone, water, aqueous methanol (1:1) and methanol: chloroform: water:: 12:5:3; the extraction recoveries were 0.162%, 11.64%, 16.82% and 14.39%, respectively. Methanol-chloroform-water (MCW) extract exhibited strongest absorbance and showed the highest content of phyto-chemical constituents, except tannins, as evident from qualitative and quantitative phyto-chemical tests, and high performance liquid chromatography. MCW extract also exhibited best anti-oxidant and reducing activity.
Based on the results of MTT assay, the three extracts were chosen to be used at a concentration of 300μg/mL, 240μg/mL and 160 μg/mLrespectively,in Chicken Embryo Fibroblast (CEF) cultures. MCW extract was found to exert greatest inhibitory activity against the virus in vitro, as determined by microtiter plate test (90.20% decrease in titer) and relative semi-quantitative PCR (60.20% decrease in viral load). Time-of-addition studies suggested that WS inhibited IBDV in vitro during early stages, involving neither virus binding nor viral internalization mechanisms, possibly through direct effects on the virion. Untreated virus-infected CEF underwent more apoptotic changes than those treated with MCW extract. MCW extract caused a rapid rise in nitric oxide (NO) levels that was inhibitory to the virus. However, other mechanisms for inhibition of the virus by the extract must also exist, as suggested by studies with aminoguanidine – a NO inhibitor. Strong reducing activity of MCW extract allowed its use for synthesizing silver nanoparticles that reduced the virus titer by 93.12%, likely through direct effects on the virion. Molecular docking analysis, undertaken to determine the ability of some maj direct interactions between WS extract and the virion.or phyto-constituents of WS to bind with viral proteins, also strengthened the possibility of direct interactions between WS extract and the virion.
In vivo studies were performed in White Leghorns to assess the protective effect of dietary WS supplementation, @ 1% of feed, against the virus. Continued dietary supplementation of layers with WS root powder @ 1% in feed ameliorated virus-induced hematological, biochemical and immunological alterations, and oxidative stress, resulting in lower viral persistence in the host.WS supplementation also provided protection against virus-induced lymphoid depletion and histopathological insult to the bursa and spleen.
The work concluded that Withania somnifera exerts protective activity against IBDV in vitro and in vivo.
 
Date 2016-06-09T12:06:37Z
2016-06-09T12:06:37Z
2015-01
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/67092
 
Language en
 
Format application/pdf
 
Publisher G.B. Pant University of Agriculture and Technology, Pantnagar-263145 (Uttarakhand)