Electrophoretic behaviour of proteins of Ostertagia ostertagi found in India sheep

Gurung Chowdhury, Gurung Kumar

Abstract

Economic losses due to helminth parasites in sheep throughout the world are considerable. Ostertagia ostertagi is a parasitic cattle nematode belonging to the superfamily of Strongyloidea and the family of Strongylidae. Ostertagia engenders immunity more slowly and is therefore the most important species in older cattle. These are able to induce structural, biochemical and immunological changes in the host like inappetence, diarrhoea, dull hair coat and weight loss, that is, ostertagiosis. At present, control is almost exclusively based on anthelmintics. 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 the laboratory for screening. The parasites were collected in normal saline, washed and stored in 0.05M PBS with pH of 7.4 at 0°C. After refrigeration, frozen nematodes were thawed, homogenized and centrifuged at 1000-15000 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 Polyacrylamide Gel Electrophoresis (PAGE) and then through Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Protein estimation of the samples was estimated to be 3.9 mg/ml. The processed parasite samples were then subjected to PAGE and SDS-PAGE to determine the presence of the proteins. The proteins were seen as continuous bands intermixing with each other in PAGE analysis. The present study revealed two bands of molecular weights - 67 and 20 kDa in PAGE analysis. The proteins when analyzed through SDS-PAGE were mostly found in the range of 20-70 kDa. The SDS-PAGE analysis showed seven prominent bands of molecular weights of 60, 50, 45, 40, 32, 28 and 20 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.