Evaluation of immunogenic potential of 75kDa and 56kDa proteins of newcastle disease virus (NDV).

The R2B strain of virus of new castle disease virus (NDV) was propagated in 9-11 day old embryonated chicken eggs via allantoic cavity route and after seven serial passages virus was purified from allantoic fluid. Purified virus was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis which yielded six major polypeptides ranging from 38-200 kDa. Protein fractions, corresponding to 75 and 56kDa, resembling haemagglutinin-neuraminidase (HN) and fusion (F) proteins were used to ascertain their immunization potential. Immunization of viral proteins was compared with the whole virus vaccine. Among different group of birds, highest haemagglutination inhibition (HI) and enzyme linked immunosorbent assay (ELISA) titers were obtained in birds immunized with whole virus vaccine followed by viral proteins, 75 and 56kDa in combination which was comparable with birds immunized with 56kDa protein alone. Despite lower values of HI and ELISA titers elicited by viral subunits in immunized birds, when challenged with virulent NDV strain, protection accorded by viral proteins in combination (75 +56kDa) or 56kDa alone was comparable with whole virus vaccine.

Partial purification and cytotoxic effects of Salmonella proteinous moieties on chick embryo fibroblasts.

Salmonella enterica serovars, viz., S. Weltevreden, S. Typhimurium, S. Gallinarum and S. Bareilly were treated with cephotaxime to release of intracellular proteins. The cephotaxime extract (CE) was salt precipitated with ammonium sulphate (45-70%) and dialyzed, and denoted as precipitated dialyzed proteins (PDP). Further, both CE and PDP of Salmonella Weltevreden and PDP of rest of the serovars were subjected to gel filtration using Sephacryl S-200HR. Different fractions along with CE and PDP were studied for their cytotoxicity using chicken embryo fibroblast (CEF). All the CE and PDP exerted cytotoxic effects, characterized by rounding, detachment, shrinkage and clumping of cells with syncytia formation. Also, the fractions eluted in the 2nd and 3rd peaks through Sephacryl S-200HR column invariably had cytotoxic activity. It was concluded that in place of Vero cell line, CEF cells could also be used to test cytotoxicity.

Generation of anti-teliospores antibodies for immunolocalization and characterization of antigenic epitopes of teliospores of Karnal bunt (Tilletia indica) of wheat.

Polyclonal antibodies raised against intact teliospores of T. indica in New Zealand albino rabbits were used for the development of indirect immunofluorescence tests. Specificity of anti-teliospore antibodies was evaluated by cross reactivity studies on other bunt, smut and related pathogens. The characteristic reactivity pattern indicated that the antibodies reacted with Tilletia species only. Chemical modifications, heat and enzyme treatments followed by indirect immunofluorescence tests were employed to delineate the molecular nature of the surface antigens. There was partial or no loss in immunoreactivity by methanol, periodate, heat or trypsin treatments. Extensive periodate treatment altered the fluorescence pattern due to changes in configuration of carbohydrate antigen present in episporium. Sequential treatment of periodate and trypsin showed diminished fluorescence due to access of proteolytic enzyme into inner site of episporium thereby cleaving peptide epitope(s) after reorientation of carbohydrate moietiesby periodate treatment. It indicated glycoprotein nature or peptide nature of epitopes on the teliospore surface.

Anthocyanin, bisabolol and phenylammonialyase activity in cell cultures of Populus deltoides.

Phenolics, anthocyanin and alpha-bisabolol production from poplar (Populus delotides) in tissue culture was determined. A number of phenolic acids were identified by HPLC. PAL activity in response to phytohormones, cells growth and anthocyanin production showed a positive correlation. A component, alpha-bisabolol, was identified using gas chromatography and UV spectroscopy. In vitro production of said metabolites was influenced by phytohormones.