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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



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 g 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.