Hudson Taylor J
Characterization Of structural Proteins Of Duck-Plague Virus - Mannuthy Department of Microbiology, College of Veterinary and Animal Sciences 1997
Two virulent strains of duck plague virus - DPV-I (IVRI)
and DPV-A (Alleppey isolate) and a vaccine strain - DPV-V (VBI
Palode) were investigated for the differences in clinical
manifestations in naturally and experimentally infected ducks,
morphological changes in developing duck/chicken embryo
(DDE/DCE) and cytopathic effects in duck embryo fibroblast/
chicken embryo fibroblast culture (DEFC/CEFC) and chicken
embryo fibroblast culture (CEFC).
Typical symptoms and lesions of duck plague were produced
by both the virulent strains. However, in DPV-A infection,
the level of mortality and severity of lesions like gizzard
muscle necrosis and haemorrhagic bands in the small intestine
were more pronounced. DPV-V did not produce any symptoms or
lesions on experimental inoculation into ducklings.
Embryonated duck eggs were used for passaging DPV-l and
isolating DPV-A, while embryonated chicken eggs were used for
propagating DPV-V. All the three strains produced mortality
of embryos with congestion on CAM and body of the embryo.
DPV-A produced more congestion on the extremities of the
embryos.
Duck embryo fibroblast cultures were used for culturing
the virulent strains while chicken embryo fibroblast cultures
were used for culturing the vaccine strain. All the three
strains produced characteristic CPE, with rounding and
clumping of cells, syncytium formation, vacuolation of
cytoplasm and formation of intranuclear inclusion bodies. The
time for the production of CPE decreased on successive
passage.
Titration of the three strains of DPV was done in
embryonated eggs (ELDso) and cell cultures (TCIDso). DPV-I,
DPV-A and DPV-V had an ELD50 of 105.27, 104.86 and 104 per ml
respectively. TCIDso of DPV-I and A in DEF culture were 105.75
per ml and 105.25 per ml respectively and that of DPV-V in CEFC
was 104.5 per ml. The tissue culture system gives the best
titre than the embryonating eggs for all the three DPV
strains.
On electron microscopy, the field isolate of DPV showed
particles ranging from 170-190 nm in diameter.
Protein analysis of the virulent strains viz. DPV-I and
DPV-A by SDS-PAGE revealed sixteen and fourteen proteins
respectively. Mild difference of two proteins (VP17 and VP22)
was noticed between the two strains. DPV-A lacked the 28KD
and 9KD protein bands.
The vaccine strain DPV-V on electrophoresis showed a
different pattern of protein bands from the virulent strains.
Eighteen proteins could be resolved in the vaccine strain.
The molecular weight of sixteen proteins of DPV-I ranged
from 9KD to 107KD while the fourteen proteins of DPV-A ranged
from l4KD to 107KD. The proteins of the vaccine strain also
ranged from 9 KD to 107 KD.
The possibility of the occurrence of strain variation as
indicated by the difference in the protein patterns of the DP
viruses under study is discussed.
636.089 6 / HUD/CH
Characterization Of structural Proteins Of Duck-Plague Virus - Mannuthy Department of Microbiology, College of Veterinary and Animal Sciences 1997
Two virulent strains of duck plague virus - DPV-I (IVRI)
and DPV-A (Alleppey isolate) and a vaccine strain - DPV-V (VBI
Palode) were investigated for the differences in clinical
manifestations in naturally and experimentally infected ducks,
morphological changes in developing duck/chicken embryo
(DDE/DCE) and cytopathic effects in duck embryo fibroblast/
chicken embryo fibroblast culture (DEFC/CEFC) and chicken
embryo fibroblast culture (CEFC).
Typical symptoms and lesions of duck plague were produced
by both the virulent strains. However, in DPV-A infection,
the level of mortality and severity of lesions like gizzard
muscle necrosis and haemorrhagic bands in the small intestine
were more pronounced. DPV-V did not produce any symptoms or
lesions on experimental inoculation into ducklings.
Embryonated duck eggs were used for passaging DPV-l and
isolating DPV-A, while embryonated chicken eggs were used for
propagating DPV-V. All the three strains produced mortality
of embryos with congestion on CAM and body of the embryo.
DPV-A produced more congestion on the extremities of the
embryos.
Duck embryo fibroblast cultures were used for culturing
the virulent strains while chicken embryo fibroblast cultures
were used for culturing the vaccine strain. All the three
strains produced characteristic CPE, with rounding and
clumping of cells, syncytium formation, vacuolation of
cytoplasm and formation of intranuclear inclusion bodies. The
time for the production of CPE decreased on successive
passage.
Titration of the three strains of DPV was done in
embryonated eggs (ELDso) and cell cultures (TCIDso). DPV-I,
DPV-A and DPV-V had an ELD50 of 105.27, 104.86 and 104 per ml
respectively. TCIDso of DPV-I and A in DEF culture were 105.75
per ml and 105.25 per ml respectively and that of DPV-V in CEFC
was 104.5 per ml. The tissue culture system gives the best
titre than the embryonating eggs for all the three DPV
strains.
On electron microscopy, the field isolate of DPV showed
particles ranging from 170-190 nm in diameter.
Protein analysis of the virulent strains viz. DPV-I and
DPV-A by SDS-PAGE revealed sixteen and fourteen proteins
respectively. Mild difference of two proteins (VP17 and VP22)
was noticed between the two strains. DPV-A lacked the 28KD
and 9KD protein bands.
The vaccine strain DPV-V on electrophoresis showed a
different pattern of protein bands from the virulent strains.
Eighteen proteins could be resolved in the vaccine strain.
The molecular weight of sixteen proteins of DPV-I ranged
from 9KD to 107KD while the fourteen proteins of DPV-A ranged
from l4KD to 107KD. The proteins of the vaccine strain also
ranged from 9 KD to 107 KD.
The possibility of the occurrence of strain variation as
indicated by the difference in the protein patterns of the DP
viruses under study is discussed.
636.089 6 / HUD/CH