ABSTRACT
An antigen capture enzyme-linked immunosorbent assay (ELISA) has been developed that could identify Haemonchus contortus antigens in dilute suspensions of worm homogenates and in faeces from sheep naturally infected with moderate burdens of the parasite. The ELISA is based on a polyclonal (rabbit or sheep) antibody coated solid-phase that captures parasite antigen from a suspension of worms or faeces. The captured antigen is detected by a mouse monoclonal antibody (MAb E2) that is reactive with a cuticle surface antigen of H. contortus. Binding of MAb E2 was shown to be specific for H. contortus and the ELISA was able to detect this antigen in faeces of some infected sheep. Characterisation of the antigen reacting with MAb E2 indicates it to be a collagenase and protease resistant surface protein of H. contortus that under reducing conditions in SDS-polyacrylamide gel electrophoresis exists as polypeptides of molecular weight 122, 56 and 49 kDa.
Subject(s)
Antigens, Helminth/analysis , Feces/parasitology , Haemonchiasis/veterinary , Haemonchus/isolation & purification , Sheep Diseases/diagnosis , Animals , Antibodies, Monoclonal/immunology , Antigens, Surface/analysis , Blotting, Western/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Fluorescent Antibody Technique/veterinary , Haemonchiasis/diagnosis , Haemonchus/immunology , Hybridomas , Immune Sera/immunology , Mice , Parasite Egg Count/veterinary , SheepABSTRACT
Quantification of a T6-lymphocyte antigen in peripheral blood mononuclear cells of sheep was used to select 15 from 48 one year old Merino ewes not previously exposed to Dermatophilus congolensis infection. These sheep were compared in response to challenge with D. congolensis zoospores and levels of T-6 lymphocyte antigen in peripheral blood mononuclear cells with 15 Merino ewes of similar age and strain from a different site that had been treated and recovered from chronic dermatophilosis. The T-6 lymphocyte antigen levels were significantly lower in the chronic dermatophilosis sheep and they developed significantly more severe lesions than the selected, previously unexposed sheep despite the former sheep having high serum antibody levels to D. congolensis. Measurement of the fleece characteristics, wax and suint concentration showed no differences between the groups that might have explained the considerable differences in their susceptibility to dermatophilosis.
