Identification and Characterization of Antigenic Proteins of Excretory/Secretory and Somatic Products of Fasciola gigantica Collected from Ruminants

Abstract

Objectives Fascioliasis is foodborne trematodes disease of ruminants caused by Fasciola gigantica and F. hepatica causing huge economic losses in term of reduced growth, fertility, meat and milk yield. The early diagnostic approaches and anthelmintic resistance are major obstacles in the control of fascioliasis. To improve the diagnosis and find alternative therapeutic measures, the current study was designed for the identification and characterization of somatic and ES products of adult worms collected from bile ducts of cattle and buffaloes. The immunogenicity of identified polypeptides in adult worm extracts were tested by Western Blot and indirect ELISA method. Materials and Methods Adult F. gigantica were collected from the bile ducts slaughtered of cattle and buffaloes. These flukes were processed to obtain somatic and ES products. Somatic proteins were obtained by homogenizing and centrifugation and ES products were obtained by incubation in PBS in which flukes were cultured. Protein concentration was quantified by Bradford method. The somatic proteins and ES products were then separated on SDS PAGE and electro-transferred to a nitrocellulose membrane to detect their immunogenicity. By Western blotting and Indirect ELISA immunogencity of different antigens and their reactivity against sera of F. gigantica infected animals. Chisqaure analysis was performed to find association of different risk factors with infection. Results The overall prevalence of fasciolosis was 4.7% in cattle and buffaloes. The disease was found to be associated with host locality (χ2=42.247p=0.000), age of host (χ2=22.828 p=0.000) and grazing management of the host (χ2 =22.828 p=0.000). Somatic proteins separated by SDS were ranged of 15 to 146 kDa. Most frequent protein bands were ranged 16,27,44,45,78,80,90,110, and 130 kDa. Molecular weights of ES products were in range of 93,100,130,145,147, and 148 kDa. Western Blot of somatic products detected a polypeptide in these extracts is a range of size 130 to 250 kDa. Conclusion The immunogenic proteins identified were 38 to 72 kDa and 95 to 250 kDa and a band range of 130 to 250 kDa by Western blotting and indirect ELISA which can be used for early diagnosis. Immunogenic proteins in the somatic extracts must be tested in future for the development of vaccine. The antigenic proteins in the ES products should also be identified. Conclusion The immunogenic proteins identified were 38 to 72 kDa and 95 to 250 kDa and a band range of 130 to 250 kDa by Western blotting and indirect ELISA which can be used for early diagnosis. Immunogenic proteins in the somatic extracts must be tested in future for the development of vaccine. The antigenic proteins in the ES products should also be identified.