Released August 2007 - The Indian Society for Parasitology

26 Prasad et al.12060261 2 3 4 M170130100725540kDa1206032261 MkDa20012085706050403025201517111510Fig. 5: SDS-PAGE of adult somatic antigen of H. contortus.Lane 1-4. Adult somatic antigen of H. contortus (prominentbands are 15, 26, 60 and 120 kDa)Lane M. MarkerA1 2 3 4 5B1 2 3 4 5Fig. 6: SDS-PAGE of affinity purified bound fractions (pooledand concentrated of adult somatic antigen of H. contortus.Lane 1: Pooled fraction.Lane M. Markeranimals were treated with albendazole; coprologicalstthexamination was conducted from 1 week to 4 week.The animals were treated as having mono-specificinfection with H. contortus. Dot-ELISA wasperformed with mono-specific sera, because theanimals were infected with L 3 larval stage of H.contortus that was raised through coproculture in thelaboratory.Fig. 4: Dot-ELISA with the naturally infected sera andimmunoaffinity purified somatic antigen of H. contortus.A. (1-5) Purified antigen and naturally-infected sera.B. (6-10) Purified antigen and naturally-infected sera.CThe test was performed with both adult somaticantigen and immunoaffinity-purified somatic antigen,to detect antibodies in experimental sera duringstrdprepatency (1 week to 3 week post-infection) as wellas sera from sheep that were naturally-infected andhad confirmed infection of H. contortus. In dot-ELISA in which adult somatic antigen was used,rdantibodies could be detected only in 3 week sera andpositive control (by formation of solid dot) but not inst nd1 and 2 week sera, negative and uninfected control.rdC. Positive control, negative control, and control animal sera. However, a solid dot was formed with the 3 week seraand adult somatic antigen which indicated thepresence of sufficient antibodies against H. contortusinfection becomes patent by 27–28 days, and thatin the infected animal. When dot-ELISA waswhen the L 4 larvae and immature worms are present,performed with the same antigen and sera of naturallynoeggs are present in faeces. The L 4 larvae cause infected sheep, solid dot was formed with 50% of thesufficient damage, and before the infection becomes sera which indicated the presence of infection.patent, the animal may die suddenly. Other gastrointestinalnematode infections, except Dot-ELISA performed with immunoaffinity-purifiedoesophagostomosis, have less intensity of infection, somatic antigen gave different results in terms ofand the degree of pathogenicity is lower in earlier detection of infection. Solid dot formation tookst nd rdcomparison to H. contortus (Soulsby, 1982).place with 1 , 2 and 3 week sera as well as positivecontrol, and no dot formation was observed in case ofDuring the present study, care was taken that the negative controls. It indicated that immunoaffinityexperimentalsheep do not graze outside the shed purified antigen contained antigenic polypeptidespremises, and before the experiment started, all reactive to anti H. contortus antibodies in sheep as