Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2

Fasciola hepatica Related Abstracts

2 The Differences between Direct Examination and ELISA Test during the Diagnosis of Fasciolosis in Jaundiced Slaughtered Sheep in Iraq

Authors: Azad A. Meerkhan, Alaa Hani Razak, Bayan M. S. Younis

Abstract:

The efficiency of enzyme-linked immunosorbent assay (ELISA) in sheep infected with Fasciola hepatica was studied. 232 jaundiced sheep among 5208 sheep slaughter in the Duhok abattoir (regardless of the age and gender) between the period of May. 2012 to Oct. 2012 were examined by direct examination (Searching of adult flukes in the bile duct) and by Enzyme-linked immunosorbent assay (ELISA) to detect the prevalence of fascioliasis in the studied population which showed a high observed infection ratio in Sep. 2012 (12.2%) with the high (ELISA) result of infection in May. 2012 (25.36%). Significant differences were found between the two ways in all of the months with the highest difference in May. 2012 and the net deference between the both ways was 6.91%.

Keywords: fascioliasis, Fasciola hepatica, layers, liver fluk, ELISA, direct examination

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1 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica

Authors: S. Samanta, J. Adeppa, O. K. Raina

Abstract:

Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: Fasciola hepatica, GST, fasciola gigantic, RT- PCR

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