ANNUAL REPORT - Department of Biotechnology

and IISc, Bangalore, a recombinant baculovirus
expressing the nucleocapsid protein of peste des
petits virus(PPRV) was generated and used for
detection of PPRV-specific antibodies in infected
animals. Diagnostic methods were developed for
PPRV antigen and antibody detection. The lateral
flow method for qualitative detection of PPRV
antibody was found to be ideal for use as a fieldbased
kit. For quantitative estimation of PPRV
antibodies, single serum dilution ELISA using precoated
plates was developed using self-designed
modules. An antigen competition ELISA was also
developed as a 'lab-based' second tier test for PPRV
antigen detection. Some of these tests have been
validated at IVRI, Mukteswar and efforts are
underway to transfer the technology.
Phage display technique was successfully used as
an alternative to hybridoma to produce mono-specific
antibodies against recombinant gag antigen of
Bovine immunodeficiency Virus (BIV) at HSADL,
Bhopal. Five anti-gag recombinant antibodies (RAbs)
from two gag specific hybridoma clones were
developed from gag-immunized mouse. One of the
high reacting anti-gag RAb was successfully used in
competitive inhibition ELISA. RAb based ELISA
showed high sensitivity with reference to positive
serum in comparison to MAb based ELISA. Using
mRNA from mouse immunized with recombinant NP
antigen of Avian Influenza virus(AIV), five anti-NP
RAbs were generated. Preliminary results with one of
the anti-NP RAb has shown encouraging results for
developing a RAb based competitive ELISA for
detection of antibodies to nucleoprotein of AIV in
chicken sera.
Biological control of parasitic gastroenteritis by
nematode trapping fungi in livestock was studied at
Veterinary College, Anjora. Two promising fungal
isolates viz. Arthrobotrys oligospora and
Duddingtonia flagrans were studied for their ability as
bio-control agents using growth assay, predatory
activity, germination potential and ability to survive
passage through the guts of domestic livestock. A.
oligospora was better than D. flagrans in terms of
growth and predatory activity. However, only D.
flagrans produced profuse chlamydospores that
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germinated easily at 26 C and could survive passage
through the guts of both monogastric and polygastric
animals.
Efforts were made to develop diagnostics for
leptospira, at IVRI, Izatnagar and TANUVAS,
Chennai. An immunodominant surface outer
membrane lipoprotein, LipL41, was targeted as an
antigen for serodiagnosis using ELISA. The protein
gene from various serovars was amplified by PCR
using a set of designed primer and the gene from
Canicola serovar was cloned and expressed in E. coli
DH5 cells. The purified recombinant protein was
found to be seroreactive using western blotting and
ELISA. The 41 kDa protein and the 32 kDa
recombinant protein were used for serodiagnosis of
leptospirosis in various animal species. As
compared to the conventional microscopic
agglutination test, the recombinant proteins based
ELISAs were found to possess 100% sensitivity.
Molecular studies of pestivirus infections of small
ruminant was done at HSADL, Bhopal. Prevalence
rate of pestivirus antibodies was 26.1% in sheep and
29.1% in goats while it was 7% in sheep and 2.8% in
goats for border disease virus (BDV) antibodies
tested by ELISA. pestivirus antigen was detected in
11.8 % sheep and 8.7 % in goats providing
serological evidence of infection first time in Indian
small ruminants. A RT-PCR assay was standardized
for specific detection of border disease virus and a
nested PCR was developed for differentiation of
BDV, bovine viral diarrhea virus (BVDV) 1 and BVDV
2 using 5' UTR primers.
Animal Reproduction: Embryo Transfer (ET)
Technology and related areas :
Isolation, characterization and in vitro development
of buffalo preantral follicles for embryo development
was attempted at IVRI, Izatnagar. Mechanical
methods of isolation were found to be better as
compared to the enzymatic isolation as more number
of follicles of different sizes were recovered in short
period. Buffalo preantral follicles were cultured with
83 Research and Development