|
|
Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Original Research
5.
A comparative analysis on serum antibody levels of
sheep immunized with crude and thiol-purified
excretory/secretory antigen of Haemonchus
contortus -
S ArunKumar, S AbdulBasith and S Gomathinayagam
Vet World. 2012; 5(5): 279-284
doi:
10.5455/vetworld.201 2.279-284
Abstract
In vitro culture of live adult worms were made in
RPMI 1640 medium at a concentration of
approximately 50 worms / ml in a culture flask at
370 C for 24 hours and the culture supernatant was
used as antigen. The E/S antigen was purified by
thiol – sepharose affinity chromatography. On
western blot analysis, it was demonstrated that
the crude E/S antigen showed five reactive bands
at 24, 29, 46, 66 and 93 kDa and the Thiol –
purified antigen showed a single reactive band at
66 kDa. In immunization trial, sheep were
immunized with 500 µg of crude and thiol –
purified E/S antigen along with montanide as
adjuvant on day 0,30 and 60 intramuscularly.
Further, the assessment of serum antibody levels
in immunized sheep was made at weekly intervals by
enzyme linked immuno sorbent assay (ELISA). It was
observed that the mean absorbance values were
significantly (P≤0.01) higher up to 20 weeks post
immunization in Group-I (purified antigen) than
Group-II (crude antigen) compared to unimmunized
control group.
Key words: Haemonchus
contortus, Excretory / Secretory antigen, Thiol –
sepharose, antibody, ELISA.
|
|
