Protein profiling of Haemonchus contortus found in sheep of Kashmir valley

Abstract

Economic losses due to helminth parasites in sheep throughout the world are considerable. Haemonchus contortus is a blood sucking intestinal helminth that lives in the abomasum of small ruminants worldwide. This parasite can be devastating to producers as it causes decreased production levels due to clinical signs such as anaemia, edema and death. For isolation of the proteins of the parasite, a well defined methodology was adopted. The abomasae of sheep in which this parasite resides were collected from abattoirs of various districts and were then carried to laboratory for screening. In case of collection sites falling in far areas, the organs were screened on spot. The parasites were collected in normal saline, washed and stored in 0.05 M PBS with pH of 7.4 at 0 °C. After refrigeration, frozen nematodes were thawed, homogenized and centrifuged at 1,000–15,000 rpm for 15 min. The supernatant was thus collected as a protein mixture and stored at −20 °C. Protein concentration of the samples was estimated by Lowry method. The samples were then analyzed through PAGE and then through SDS-PAGE. Protein estimation of the samples was estimated to be 4.2 mg/ml. The processed parasite samples were then subjected to PAGE and SDS-PAGE to determine the presence of the proteins. It showed high concentration of proteins in its whole protein profile. The proteins were seen as continuous bands intermixing with each other in PAGE analysis. The present study revealed two bands of molecular weights—55 and 33 kDa in PAGE analysis. The proteins when analyzed through SDS-PAGE were mostly found in the range of 25–70 kDa. The SDS-PAGE analysis showed four prominent bands. These bands were of the molecular weights of 66, 40, 33 and 26 kDa. The present work was a challenging one since only a single study was conducted in this region on this aspect and thus obviously was a big task to peep into the field where scanty input was available.